Dissertations / Theses: 'Antibody treatment'

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Alsughayyir, Jawaher. "CD49d-specific Single Domain Antibodies for the Treatment of Multiple Sclerosis." Thesis, Université d'Ottawa / University of Ottawa, 2012. http://hdl.handle.net/10393/23528.

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Multiple sclerosis is a neurodegenerative disorder affecting the central nervous system (CNS). Currently, the disease is incurable and immunomodulating drugs are the only option to control the disease. CD49d is an adhesion receptor expressed on most immune cells. Antibodies that bind to CD49d and block immune cells from trafficking toward the CNS are being pursued as one class of therapeutics. In this work, by combining recombinant antibody and phage display technologies we isolated 10 anti-CD49d single domain antibodies from a synthetic antibody light chain variable domain (VL) phage display library. Isolated VLs (~ 12 kDa) were expressed in Escherichia coli, purified and analysed for biophysical characteristics. The majority were expressed in good yields and were non-aggregating. All 10 VLs bound recombinant CD49d by ELISA, and 7 bound to CD49d-expressing cells in flow cytometry experiments. To empower the VLs for better therapeutic efficacy (thru increasing avidity and half-life), three of the lead VLs were re-engineered as fusions to fragment crystallisable (Fc) of human immunoglobulin gamma (IgG). The engineered hFc-VL fragments (~ 70 – 90 kDa) retained their specificity for CD49d by flow cytometry. With (i) being less immunogenic due to their human nature, (ii) their efficient access to cryptic epitopes (iii) having half-lives comparable to IgGs’ and (iv) being more cost effective compared to IgGs, these novel antibody fragments (monovalent VLs and bivalent hFc-VLs) provide a promising therapeutic platform against multiple sclerosis.

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Chen, Chao, and 陳超. "Identification of a novel cancer therapeutic antibody against human epidermal growth factor receptor 2 (Her2) and antibody engineering for development of cancer therapeutics." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2013. http://hdl.handle.net/10722/196461.

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Cancer is one of the leading causes of death worldwide. Monoclonal antibodies (mAbs) have been proved effective for cancer therapy. MAbs possess advantages over chemical drugs and small molecular drugs in cancer treatment, such as high specificity, low toxicity, effector function, long half-life in circulation system and less side effects. There are eight FDA-approved anti-cancer antibody drugs now, and many more are under development. Antibodies have two functional domains, the Fab region that is responsible for antigen recognition, and the Fc region that couples the antibody to immune effector pathways. Fab-mediated interference with cancer cell signalling may lead to growth inhibition and direct cell death, while Fc-mediated effector function through interactions with Fc-gamma receptors (FcrRs) expressed in immune cells or through complement cascades may lead to target cell cytotoxicity. Antibody engineering to increase the binding affinity and effector function may improve antibody in vivo efficacy. Anti-Her2 mAb herceptin (trastuzumab) is effective in treatment of Her2-overexpressing breast cancer patients. However, only 25–30% of patients with Her2-overexpressing tumors respond to single agent trastuzumab, and drug resistance develops even in responding patients. Accumulating evidence showed that cross-talk between Her2 and the insulin-like growth factor receptor type I (IGF-IR), including receptor heterodimerization and transactivation, and elevated IGF-IR signalling have been associated with trastuzumab resistance. Therefore, we hypothesized that dual specific antibodies co-targeting both IGF-IR and Her2 may prevent or delay the emergence of resistance to mono-specific antibodies. Mouse monoclonal antibody, M590 showed very good binding activity to IGF-IR. By engineering the CH3 domain of human Fc in pDR12 plasmid, we developed a “knobs-into-holes” hybrid IgG expression system, and successfully produced M590-Herceptin bi-specific IgG, which showed high binding avidity for both antigens and preserved antibody-dependent cell-mediated cytotoxicity (ADCC), a main route of immune protections conferred by therapeutic antibodies in vivo. M590-Herceptin dual specific antibody inhibited breast cancer and ovarian cancer cell proliferation in vitro, and inhibited cancer growth in a SKOV-3 Her2- and IGF-IR-overexpressing ovarian cancer xenograft mouse model. M590-Herceptin hybrid showed better anti-cancer activity compared with M590 and Herceptin alone, or in combination. Meantime, I also constructed a phage display antibody Fab library using the mRNA of rabbits immunized by membrane proteins of SKOV-3 cells, and isolated a novel anti-Her2 mAb, designated as 1C6. Results from in vitro assays showed that 1C6 had anti-cancer activity which was comparable to that of herceptin. M590-1C6 hybrid IgG was also constructed, and the results from in vitro assays and mouse study showed that M590-1C6 hybrid IgG also possess better inhibitory activity of Her2 positive tumours compared with m590 or 1C6 alone. In summary, this study indicates that bi-specific antibodies co-targeting two elevated cancer receptors are more effective than mono-specific antibodies for cancer therapy.
published_or_final_version
Microbiology
Doctoral
Doctor of Philosophy

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Schroeder, Krista Marie. "Disparities in Monoclonal Antibody Treatment of Elderly Metastatic Colorectal Cancer Patients." ScholarWorks, 2015. https://scholarworks.waldenu.edu/dissertations/1421.

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Multiple research studies have demonstrated racial, socioeconomic status (SES), and neighborhood disparities in first-line treatment of colorectal cancer patients, including those with metastatic colorectal cancer. However, disparities in adjunct monoclonal antibody treatment disparities have not been explored. The purpose of this study was to assess racial, SES, and neighborhood disparities in adjunct monoclonal antibody treatment of elderly metastatic colorectal cancer patients. The research was rooted in 3 theories: the fundamental cause theory, the diffusion of innovations theory, and theory of health disparities and medical technology. Data from the SEER-Medicare database and logistic regression were used to assess the relationship between the variables of interest and adjunct monoclonal antibody therapy. In this study, race (p = 0.070), SES (p = 0.881), and neighborhood characteristics (p = 0.309) did not significantly predict who would receive monoclonal antibody therapy. The results demonstrated a potential improvement in historically documented colorectal cancer treatment disparities. Specifically, historical treatment disparities may not be relevant to newer therapies prescribed to patients with severe disease. The difference could be related to improved access to care or a change in treatment paradigm due to the severity of metastatic colorectal cancer. Future studies aimed at understanding the causes of this social change (i.e., reduced treatment disparities) are warranted. Understanding the root cause of the reduced treatment disparities observed in this study could be used to reduce treatment disparities in other cancer populations.

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Odili, Joy Ifeyinewa. "Development of specific antibody fragments for the detection and treatment of melanoma." Thesis, University College London (University of London), 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.430128.

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Alberts, Justin Charles John. "Bispecific antibody mediated targeting cytotoxic lymphocytes for the treatment of colorectal carcinoma." Thesis, University College London (University of London), 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.248396.

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Douglass, Angela. "The use of an antibody in the diagnosis and treatment of liver fibrosis." Thesis, University of Aberdeen, 2009. http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=131533.

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This thesis examines the use of an antibody and single chain antibody to target myofibroblasts, for the diagnosis and treatment of liver fibrosis. Furthermore the effects of myofibroblasts depletion on inflammation, fibrosis and regeneration are investigated in animal models of chronic and acute liver disease. Liver myofibroblasts have become a focus as a potential therapeutic target as removal or prevention of their activation attenuates liver fibrosis severity. One of the major issues surrounding fibrosis treatment is developing a therapeutic that targets myofibroblasts without causing additional damage to the liver. Using an antibody may provide a solution due to their high specificity for their target antigen. Several antibodies are currently used in the clinic to treat a range of diseases. However no treatment is currently indicated for fibrosis, with transplantation being the only option at the later stages of disease. Our lab has developed a single chain antibody fragment (scab) capable of targeting activated myofibroblasts. Here we examine the use of this antibody as a drug delivery method in vitro and in vivo. This thesis also describes the generation of a human monoclonal IgG to target myofibroblasts and recruit the natural immune effector functions as a safer alternative to toxic drug conjugates. The presence of myofibroblasts is indicative for fibrosis and as such we have used myofibroblasts as a potential imaging target. Using C1-3 conjugated to a fluorophore, we investigated the possibility of grading fibrosis severity, using histological and imaging techniques.

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DI, CINTIO FEDERICA. "Nanoparticles anti-GPC1 for glioblastoma multiforme treatment." Doctoral thesis, Università degli Studi di Trieste, 2022. http://hdl.handle.net/11368/3015204.

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Glioblastoma multiforme (GBM) the most aggressive (WHO grade IV) diffuse glioma, is also by far the most frequent one. After standard treatment, the 2-year overall survival of GBM patients is approximately only 25%. Although numerous experimental drugs have been tested in clinical trials, GBM patients have not yet profited of curative treatments. To overcome the big limitations regarding GBM treatment, we address the challenge of developing a drug delivery system based on highly biocompatible chitosan nanobubbles (NBs) conjugated with an anti-glypican1 (GPC1) antibody loaded with docetaxel as chemotherapeutic agent. This drug delivery approach has been proposed to counter major challenges as overcoming the BBB, allowing the therapeutic release exclusively to tumor cells, and minimizing the possible side effects in cancer patients. The GPC1 proteoglycan has been chosen as useful target for drug delivering with NBs, therefore GPC1 expression was characterized in-vitro, being found expressed in GBM cell lines (e.g., T98G, U87-MG) but not found expressed in non-GBM cell line. Consistently, we assessed the localization of GPC1 protein expression on the cell surface and in the cytoplasm of GBM cell lines whereas it was lacked in the negative control cells. Of note, in primary tumor sections of these 10 GBM cases, GPC1 was found overexpressed whereas in normal tissues was found not expressed. To obtain a specific anti-GPC1 antibody recognizing the last 70 amino acid of GPC1 protein and therefore the cell-surface form of GPC1, mouse immunization has been performed. Hybridomas have produced three different anti-GPC1 specific clones (A, B, C). By using the B and C clones, GPC1 expression was detected in GBM cells at levels comparable to the levels obtained by using the commercially available antibody by the B and C clones. On the contrary, the A clone was not capable to recognize GPC1. Therefore, we purified the B and C clones to obtain specific anti-GPC1 monoclonal Abs. Moreover, C and B appeared to be more efficient than the a-GPC1c for detection of GPC1 expression levels. According to the results of antibody testing in GBM cell lines and negative control cell lines, the B clone was chosen to be conjugated to the NBs to develop the active drug delivery strategy. To select the drug to be loaded in the NBs, the killing capability of temozolomide (TMZ), paclitaxel (PTX) and docetaxel (DTX) was evaluated in GBM cells. DTX have the highest killing capability compared to PTX and TMZ. Therefore, we used DTX for the NBs loading encapsulation. The in-vitro characterization of NBs showed the average diameter of about 350 nm and a positive charge and spherical morphology. In-vitro analysis of the treatment of NBs in GBM cells, showed the localization of NBs conjugated with B antibody in cell cytoplasm around the nucleus. In contrast, a lower mean fluorescence intensity was observed for the cells treated with unconjugated NBs. For the in-vitro cytotoxic effect of NBs, NB loaded with DTX, NBs loaded with DTX and conjugated with B antibody, showed a killing capability correlated with the concentration in each evaluated point, with cell viable levels comparable to those of free DTX for some concentrations. Blank NBs, NB conjugated with Cy 5.5, and NB conjugated with B antibody were not toxic at all tested concentrations. In-vivo and ex-vivo test of the biodistribution of anti-GPC1 NBs in xenograft GBM mouse models, showed that the presence of the conjugation with the B antibody seems to be allow a major accumulation of the injected NBs in the tumor as well as a higher retention time at least until the last time point of 96 h of treatment. In conclusion, the proposed active drug delivery approach using anti-GPC1 conjugated NBs loaded with DTX could be useful for the treatment of GBM.
Glioblastoma multiforme (GBM) the most aggressive (WHO grade IV) diffuse glioma, is also by far the most frequent one. After standard treatment, the 2-year overall survival of GBM patients is approximately only 25%. Although numerous experimental drugs have been tested in clinical trials, GBM patients have not yet profited of curative treatments. To overcome the big limitations regarding GBM treatment, we address the challenge of developing a drug delivery system based on highly biocompatible chitosan nanobubbles (NBs) conjugated with an anti-glypican1 (GPC1) antibody loaded with docetaxel as chemotherapeutic agent. This drug delivery approach has been proposed to counter major challenges as overcoming the BBB, allowing the therapeutic release exclusively to tumor cells, and minimizing the possible side effects in cancer patients. The GPC1 proteoglycan has been chosen as useful target for drug delivering with NBs, therefore GPC1 expression was characterized in-vitro, being found expressed in GBM cell lines (e.g., T98G, U87-MG) but not found expressed in non-GBM cell line. Consistently, we assessed the localization of GPC1 protein expression on the cell surface and in the cytoplasm of GBM cell lines whereas it was lacked in the negative control cells. Of note, in primary tumor sections of these 10 GBM cases, GPC1 was found overexpressed whereas in normal tissues was found not expressed. To obtain a specific anti-GPC1 antibody recognizing the last 70 amino acid of GPC1 protein and therefore the cell-surface form of GPC1, mouse immunization has been performed. Hybridomas have produced three different anti-GPC1 specific clones (A, B, C). By using the B and C clones, GPC1 expression was detected in GBM cells at levels comparable to the levels obtained by using the commercially available antibody by the B and C clones. On the contrary, the A clone was not capable to recognize GPC1. Therefore, we purified the B and C clones to obtain specific anti-GPC1 monoclonal Abs. Moreover, C and B appeared to be more efficient than the a-GPC1c for detection of GPC1 expression levels. According to the results of antibody testing in GBM cell lines and negative control cell lines, the B clone was chosen to be conjugated to the NBs to develop the active drug delivery strategy. To select the drug to be loaded in the NBs, the killing capability of temozolomide (TMZ), paclitaxel (PTX) and docetaxel (DTX) was evaluated in GBM cells. DTX have the highest killing capability compared to PTX and TMZ. Therefore, we used DTX for the NBs loading encapsulation. The in-vitro characterization of NBs showed the average diameter of about 350 nm and a positive charge and spherical morphology. In-vitro analysis of the treatment of NBs in GBM cells, showed the localization of NBs conjugated with B antibody in cell cytoplasm around the nucleus. In contrast, a lower mean fluorescence intensity was observed for the cells treated with unconjugated NBs. For the in-vitro cytotoxic effect of NBs, NB loaded with DTX, NBs loaded with DTX and conjugated with B antibody, showed a killing capability correlated with the concentration in each evaluated point, with cell viable levels comparable to those of free DTX for some concentrations. Blank NBs, NB conjugated with Cy 5.5, and NB conjugated with B antibody were not toxic at all tested concentrations. In-vivo and ex-vivo test of the biodistribution of anti-GPC1 NBs in xenograft GBM mouse models, showed that the presence of the conjugation with the B antibody seems to be allow a major accumulation of the injected NBs in the tumor as well as a higher retention time at least until the last time point of 96 h of treatment. In conclusion, the proposed active drug delivery approach using anti-GPC1 conjugated NBs loaded with DTX could be useful for the treatment of GBM.

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Arrowood, Michael James. "Cryptosporidium: Oocyst production and hybridoma generation for examining colostrum and monoclonal antibody roles in cryptosporidial infections." Diss., The University of Arizona, 1988. http://hdl.handle.net/10150/184335.

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Techniques for the large-scale isolation of Cryptosporidium oocysts and sporozoites, obtained from the feces of experimentally infected Holstein calves, were developed employing discontinuous sucrose gradients and isopycnic Percoll gradients. The three step oocyst recovery method utilized two sequential discontinuous sucrose gradients followed by one Percoll gradient. Recovered oocysts were essentially free of debris and bacteria and represented 34% of the original oocyst suspension. Sporozoites were recovered from excystation mixtures on a single Percoll gradient. Sixty-three percent of the original sporozoites were recovered with 2.2% contamination by intact oocysts and virtually no oocyst walls. Eight anti-oocyst hybridomas were derived from oocyst-immunized mice: five from BALB/c mice and three from RBF/Dn mice. The monoclonal antibody (Mab) OW3 reacted specifically with C. parvum oocysts in immunofluorescent assays (IFA) and was shown to be superior to conventional stains for detecting oocysts in fecal smears from infected individuals. Sixteen anti-sporozoite hybridomas were derived from sporozoite-immunized BALB/c mice. The Mabs appeared to react with cell surface and cytoplasmic antigens by IFA. Two anti-sporozoite Mabs (C8C5, C6B6) reacted with a 20 kDa sporozoite antigen in western blots while the Mab C4A1 reacted with multiple antigens in western blots. These three Mabs (C8C5, C6B6, C4A1) were examined for potential modulation of cryptosporidial infections in vivo by oral Mab administration to oocyst-inoculated neonatal mice. The role for colostrum and breast milk in controlling cryptosporidial infections was examined by immunizing mouse dams and experimentally infecting their neonatal offspring. Colostrum and Mab-treated neonatal mice were sacrificed four days post infection. No difference in infection rates was observed among the treatment groups. Suckling mice treated daily with orally administered mixtures of Mabs (purified or ascitic fluid) showed significantly reduced parasite loads compared to control mice at four days post infection. In vitro cultivation of C. parvum was successful through asexual stages in human fetal lung, bovine turbinate and murine L929 cells. Parasite numbers that developed in the cell cultures varied from infection run to infection run.

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Lopez-Oliva, Santa Cruz Isabel. "Rheumatoid arthritis and periodontitis : antibody response, oral microbiome, cytokine profile and effect of periodontal treatment." Thesis, University of Birmingham, 2018. http://etheses.bham.ac.uk//id/eprint/8058/.

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Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease that affects about 1% of the world population. This common disease is characterized by chronic inflammation of the synovium that leads to destruction of cartilage and bone in the join, and the cause of this exacerbated inflammatory reaction remains unknown. Periodontitis (PD) is also a chronic inflammatory disease characterized by destruction of bone and other connective tissue that shares notable similarities with RA. Over the last 20 years, numerous studies have found an epidemiological connection between RA and periodontitis. However the biological mechanisms that explain the interrelations between the two conditions are not known. The aim of this thesis was to investigate the role of periodontitis in RA and the effect of periodontal therapy on immunological and microbiological parameters. To do that, different biological samples were collected from two pilot studies, comparing RA and periodontitis patients to the appropriate controls and from a selected group of randomized RAPD patients before and after periodontal therapy. The antibody response and subgingival microbiome of patients with RA and periodontitis were compared to the appropriate controls (no RA no PD, RA no PD, no RA PD). The effect of periodontal therapy on these parameters and on the cytokine changes in gingival crevicular fluid was also investigated. The findings from this thesis lend further credence to the link between RA and the oral microbiome, with RA patients having a disrupted and more anaerobic microflora and an exacerbated immunological reaction against periodontal bacteria and citrullinated proteins.

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Locker, Kathryn CS. "Molecular mechanisms underlying treatment of acute type 1 diabetes with an anti-TLR4/MD2 antibody." University of Cincinnati / OhioLINK, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1601993060493285.

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Wetzel, Hanna N. "Preclinical Development of the Anti-cocaine Monoclonal Antibody h2E2 for the Treatment of Cocaine Addiction." University of Cincinnati / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1529333855259163.

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Leung, Chieh-wing Jervis, and 梁倢榮. "Development of a real-time PCR-based method for the measurement of neutralizing antibody to interferon-beta in multiple sclerosis patients." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2014. http://hdl.handle.net/10722/206488.

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Background Multiple sclerosis (MS) is a chronic inflammatory demyelinating disorder of the central nervous system (CNS). In Hong Kong, the prevalence rates of MS is 4.8/100,000. First line disease modifying agent (DMA) type 1 interferon β (IFN-β) is the most commonly use therapy for relapsing and remitting MS(RRMS). Depending on the administration type and route of IFN-β, up to 80% of patients develop harmless binding antibody (BAb),which binds to IFN molecules but not necessary interfere its bioactivity. When IFN-β therapy continues, maturation of BAb response can lead to the formation of high affinity neutralizing antibody (NAb). About 45% of MS patients develop NAb against IFN-β in one year of IFN-β treatment. NAb shows a loss of IFN-β clinical effect by increasing MRI activity and disease progression. As the clinical effect of NAb is lagging behind the initial appearance of NAb in the body, it is suggested to develop a NAb assay to predict treatment failure and advice switching therapy for patients when NAb is present. Aim The aim of this study was: I. To develop and evaluate a qPCR-based method for the measurement of NAb to IFN-β in MS patient. II. To establish the normal reference range of NAb in Chinese population. III. To seek the possibility of using anti-IFN-β BAb assay and in vivo MxA gene expression assay as a screening test for NAb IV. To compare the performance between MxA induction qPCR, ELISA, WB assay and luciferase IFN-β reporter gene assay Materials and methods23Chinese RRMS patients who treated with IFN-β-1a therapy for a minimum of12 months were recruited in this study. Serum and PBMC were collected12 hours after the IFN-β-1a injection. MxA, IFNAR1 and IFNAR2 mRNA from PBMC were tested byin vivo MxA gene expression assay. NAb containing serum was tested by anti-IFN-β BAb assay, IFN-β reporter gene assay, in vitro MxA induction WB, ELISA and qPCR assay. In addition, blood samples from 3 Chinese volunteers without any known autoimmune disease history were collected to evaluate the baseline of NAb titer and MxA expression. Result The experimental condition of MxA induction qPCR assay was optimized by using 2.5×105A549 cells plating density, 10% FCS concentration,5 hours IFN-β stimulation time and GAPDH normalization. Assay accuracy was validated by reference anti-IFN-β antibody. Starting from 2.5 TRU, linear relationship could be observed (r2= 0.9873). The lower limit of quantification (LLOQ) was 0.02 LU/mL, the upper limit of quantification (ULOQ) was 16LU/mL and the limit of detection (LOD) was 0.002 LU/mL. The reproducibility of the assay was measured, the intra-and inter-assay imprecision(%CV)for high value were 5.95% and 7.17% respectively, while the intra-and inter-assay impression were8.31% and 15.95%respectively.Results of the qPCR-based method were concurring with that of luciferase IFN-β reporter gene assay. The upper limit of the NAb reference range in Chinese population was 40.3 TRU (n=3, 95% CI = 31.7-48.8). The performance observed in MxA induction ELISA assay swas unsatisfactory. The correlation of anti-IFN-β BAb assay and in vivoMxA gene expression assay results with NAb status indicated both tests were sensitive enough for NAb screening. Conclusion A normal range of NAb titer in Chinese population was established in this study. Anti-IFN-β BAb assay and in vivo MxA gene expression assay were proved suitable for NAb screening. The performance of the developed MxA induction qPCR assay was superior to MxA induction ELISA, WB assay and comparable to luciferase IFN-β reporter gene assay. By using MxA induction qPCR assay, actual efficacy of IFN-β therapy could be measured and monitored. Any treatment failure could be predicted earlier.
published_or_final_version
Pathology
Master
Master of Medical Sciences

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vanzolini, tania. "Development of new biological drugs for the treatment of fungal infections." Doctoral thesis, Urbino, 2021. http://hdl.handle.net/11576/2692691.

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Lichvar, Alicia B. "Proteasome Inhibitor Treatment of Antibody Mediated Rejection and Mixed Acute Rejection: Defining Factors that Predict Long-Term Outcomes." University of Cincinnati / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1490698721411028.

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Nonomura, Yumi. "Peripheral blood Th9 cells are a possible pharmacodynamic biomarker of nivolumab treatment efficacy in metastatic melanoma patients." 京都大学 (Kyoto University), 2017. http://hdl.handle.net/2433/225479.

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Sartor, Chiara <1988&gt. "Research of predictive biomarkers to anti-CD22 antibody-drug conjugate treatment in B-cell acute lymphoblastic leukemia." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2022. http://amsdottorato.unibo.it/10252/1/PhD%20thesis_Chiara%20Sartor_XXXIV%20ciclo.pdf.

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Background: The treatment of B-cell acute lymphoblastic leukemia (B-ALL) has been enriched by novel agents targeting surface markers CD19 and CD22. Inotuzumab ozogamicin (INO) is a CD22-calicheamicin conjugated monoclonal antibody approved in the setting of relapse/refractory (R/R) B-ALL able to induce a high rate of deep responses, not durable over time. Aims: This study aims to identify predictive biomarkers to INO treatment in B- ALL by flow cytometric analysis of CD22 expression and gene expression profile. Materials and methods: Firstly, the impact on patient outcome in 30 R/R B-ALL patients of baseline CD22 expression in terms of CD22 blast percentage and CD22 fluorescent intensity (CD22-FI) was explored. Secondly, baseline gene expression profile of 18 R/R B-ALL patient samples was analyzed. For statistical analysis of differentially expressed genes (DEGs) patients were divided in non-responders (NR), defined as either INO-refractory or with duration of response (DoR) < 3 months, and responders (R). Gene expression results were analyzed with Ingenuity pathway analysis (IPA). Results: In our patient set higher CD22-FI, defined as higher quartiles (Q2-Q4), correlated with better patient outcome in terms of CR rate, OS and DoR, compared to lower CD22-FI (Q1). CD22 blast percentage was less able to discriminate patients’ outcome, although a trend for better outcome in patients with CD22 ≥ 90% could be appreciated. Concerning gene expression profile, 32 genes with corrected p value <0.05 and absolute FC ≥2 were differentially expressed in NR as compared to R. IPA upstream regulator and regulator effect analysis individuated the inhibition of tumor suppressor HIPK2 as causal upstream condition of the downregulation of 6 DEGs. Conclusions: CD22-FI integrates CD22-percentage on leukemic blasts for a more comprehensive target pre-treatment evaluation. Moreover, a unique pattern of gene expression signature based on HIPK2 downregulation was identified, providing important insights in mechanisms of resistance to INO.

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Montanini, M. "ANTI-CD25 MONOCLONAL ANTIBODY (MAB):AN IMMUNOMODULATING DRUG CANDIDATE FOR THE TREATMENT OF T-CELL MEDIATED DISEASES." Doctoral thesis, Università degli Studi di Milano, 2014. http://hdl.handle.net/2434/245741.

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INTRODUCTION AND BACKGROUND Anti-CD25 is a fully human IgG1 kappa monoclonal antibody (mAb) against the alpha subunit of the interleukin 2 receptor (IL2Rα, also known as CD25 or TAC antigen) of potential clinical interest in autoimmune diseases, including multiple sclerosis, uveitis, type 1 diabetes and psoriasis. Anti-CD25 acts mainly by inhibiting the proliferation of T cells and by consequence T cell clonal expansion and cytokine production. IL2 is a potent immunomodulator whose major function is the activation of various cells of the immune system, i.e. T cells (including CD4+ CD25+ regulatory T cells), B cells, NK cells and macrophages, which express CD25 upon antigen stimulation. Anti-CD25 is presented with a potential therapeutic application in T cell mediated diseases, including organ transplant rejection and autoimmune disease, such as multiple sclerosis (MS). The key to immunotherapeutic success with an anti-CD25 is to elicit the correct balance of effector and regulatory T cells. Nevertheless, the effects of an anti-CD25 antibody on the balance between pro-inflammatory T cells versus anti-inflammatory regulatory T cells are still unclear. The potential advantages of Anti-CD25 are the high affinity for human CD25 and the fact that it is a fully humanized mAb, potentially less immunogenic, leading to longer duration of therapeutic effect.It is difficult to foresee which subpopulation of CD25 cells will be the most inhibited by blocking the IL-2Rα as a consequence of in vivo administration of Anti-CD25 mAb. Such aspects were investigated in non-human primates. The ICH S6 guidance providing general principles for designing scientifically acceptable preclinical safety evaluation programs, to support clinical development and marketing authorization, was followed. The purpose of the thesis was to assess the potential adverse effects resulting from the repeated administration of Anti-CD25 mAb in Cynomolgus monkeys. Selection of the relevant animal species for preclinical safety studies was based first on interspecies amino-acid sequence homology of CD25 extracellular domain. The capacity of Anti-CD25 to bind monkey (Rhesus, Cynomolgus and marmoset), minipig, mouse, rat and rabbit CD25 receptor expressed on CD3+ T cells was evaluated using resting or activated Peripheral Blood Mononuclear Cells (PBMCs). The results obtained from Anti-CD25 cross-reactivity evaluation to PBMCs showed that Cynomolgus monkey was the only species where binding to CD25-bearing cells was shown at potentially relevant in vivo concentrations. Therefore the Cynomolgus monkey was considered the relevant species to be used in the preclinical development program of Anti-CD25. EXPERIMENTAL DESIGN The thesis will show data from a study where Anti-CD25 mAb was given by intravenous route (i.v.) at doses of 5, 25, and 125 mg/kg or subcutaneously (s.c.) at a dose of 75 mg/kg to 22 males and 22 females (4 animals/sex for groups 1, 2, and 3 and 5 animals/sex for groups 4 and 5) once a week for 4 consecutive weeks (total of 5 doses) as shown in the table below: Group Doses Volume of administr. Concentr. In vehicle No. of males No. of females Group (mg /kg/ week) (ml/Kg) (mg/ml) identification [admin.Route] --------------------------------------------------------------------------------------------------------------------------------------------------------------- 1 0 (vehicle) [IV] 2 0 4 4 white 2 5 [IV] 2 2.5 4 4 yellow 3 25 [IV] 2 12.5 4 4 green 4 125 [IV] 2 62.5 5 5 red 5 75 [SC] 2 37.5 5 5 blue During the study, general clinical observations, clinical pathology, CD25 expression, CD25 saturation and down-modulation on lymphocyte subsets, NK cell activity, cytokine release markers, toxicokinetics and host anti-drug Ab (ADA) were evaluated. One week after the last treatment 3 monkeys/sex/group were sacrificed for pathology investigations, while the remaining animals were subjected to a recovery period and then sacrificed. RESULTS General examinations: No animals died during the study. No clinical signs (general and local) or behavioral changes were observed in any monkey. No treatment-related hematological changes were seen in any animal treated at the various dosages by either intravenous or subcutaneous route at the end of the treatment or at any test point of the recovery period. No treatment-related changes and no systemic changes were noted at either sacrifice. Toxicokinetics evaluations demonstrated that serum levels increased between the first and the last treatment at all doses by both administration routes. A good compound bioavailability was detected when administered by subcutaneous route at 75 mg/kg. No gender difference was found. Male animals of all the treated groups (from group 2 to group 5) were found positive for the presence of binding anti-drug antibodies to Anti-CD25 whereas females were negative. Antibodies in males were revealed starting from week 6 of the recovery period with a mean titer of >100. They were detected till the last week of recovery. Leukocyte and immunophenotyping analysis showed no effects at any dose and by either administration route. Special examinations: 1. PD markers leukocyte and lymphocyte subsets; CD25 expression, saturation and down-modulation on lymphocyte subsets Immunophenotyping analysis and CD25 expression/saturation/down-modulation were performed on peripheral blood collected in EDTA and analyzed by flow cytometry.CD25 expression in healthy Cynomolgus monkeys was detected only on CD4+ T-cells and this expression was revealed in about 4% of this lymphocyte subpopulation. Anti-CD25 mAb administered at dosages of 5, 25 and 125 mg/kg by iv route, and 75 mg/kg by sc route was able to completely bind and saturate IL-2Rα expressed on CD4+ T-cells, as detected 3 and 6 hours after the treatment, confirming data obtained in previous experiments. CD25 saturation and CD25 down-modulation was observed in all dosed animals until the end of treatment (Day 28). CD25 desaturation was observed during recovery in a dose-dependent manner, and this effect was more evident in female animals. 2. Other biomarkers Natural killer (NK) activity was measured by the ability of effector cells (PBMCs) to kill the target cells (human K562 cell line). Heparinized blood samples were obtained from all animals at the following time point: pre-dose, at the end of treatment (on week 5) and at three timepoints during the recovery period (in weeks 13, 25 and 28). No toxicologically relevant variations were observed in NK activity, in any animal at any time point. CRS (cytokine release syndrome) MARKERS: TNFα, IL-6, IFNγ, IL-1β, IL-2, IL-4, IL-10, IL-8, MCP-1 Blood samples (plasma) were collected from animals at the following time points: day 0 (predose), day1 (2h, 6h, 24h) and day 29 (2h, 6h, 24h). Simultaneous detection of several cytokines/chemokines was performed with Luminex100 system. IL-1β, IL-2 and IFN-γ as well as TNF-α, IL-4 and IL-10 did not show any significant change over time. Nor was any effect seen on IL-8. MCP-1 showed a time effect but this was not statistically relevant. The modulation was similar both in the control group and in treated groups. IL-6 level showed a rise at 2h and 6h after administration of the two higher i.v. doses of Anti-CD25 mAb. This was observed both after the 1st and the 5th administration, with the female animals showing a slightly higher response. The increase in IL-6 was also clearly seen over time. 3. Functional test T-cell proliferation assay and Treg cells number evaluation The functional tests were set to evaluate the antiproliferative and immunomodulatory effect of Anti-CD25 mAb .T cell proliferation assay and FoxP3 staining were performed at the following time point: pre-dose, 24h after the last administration and at 3 recovery time-points (8 , 18, 25 weeks after the last treatment). There were no effects on the ex-vivo T-cell proliferation assay. Anti 25 mAb significantly decreased the percentage of T regulatory cells 24h after the last administration of 125 mg/kg i.v. and 75 mg/kg s.c. CONCLUSION Neither toxicologically relevant changes in clinical observations on hematology, blood chemistry and coagulation parameters nor in histology examinations were found. Toxicokinetics demonstrated the compound accumulated in serum upon prolonged dosing by both administration routes. A good s.c. bioavailability was shown. Anti-CD25 mAb was shown to bind and saturate CD25 (IL-2Rα) expressed on blood CD4 T-cells. This effect was observed at all doses and by both administration routes since the first dosing and was maintained through the dosing period. CD25 desaturation occurred dose-dependently during the recovery period. A trend towards down-modulation was observed at all doses. There was no elevation in the CRS (cytokine release) markers measured, except for IL-6 which showed transient increases at the two higher i.v. doses. This effect could indicate a possible CRS (cytokine release syndrome) in vivo. Regulatory T cells percentage was significantly decreased at the highest i.v. or s.c. doses. This effect seems to correlate with the CD25 expression down-modulation. In conclusion, Anti-CD25 mAb did not result in any adverse toxicological effect either on clinical observations or clinical and morphological pathology, when administered by intravenous or subcutaneous route. However, transient increases in serum IL-6 and reduction in the percentage of blood regulatory T-cells were observed. The results presented in this thesis support further investigations on the potential therapeutic value of the Anti-CD25 mAb evaluated in human diseases where autoimmunity may play a pathogenetic role.

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MATRICARDI, SARA. "Epileptic phenotypes, treatment options and long-term outcomes of autoimmune epilepsies." Doctoral thesis, Università Politecnica delle Marche, 2021. http://hdl.handle.net/11566/288143.

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Obiettivi Le crisi epilettiche possono essere sintomo di esordio o prominente di encefaliti autoimmuni. Le crisi sono generalmente farmacoresistenti gli antiepilettici, ma possono beneficiare di trattamenti immunomodulanti. Nonostante le crescenti conoscenze scientifiche in questo ambito, le crisi epilettiche ad eziologia autoimmune sono spesso sotto-diagnosticate e non vi sono linee guida standardizzate per la diagnosi e la gestione terapeutica. Lo scopo dello studio è di analizzare e descrivere i fenotipi epilettici ad eziologia autoimmune, le opzioni terapeutiche, nonché i possibili esiti a lungo termine. Metodi Studio di coorte osservazionale retrospettivo, condotto in un periodo di 10 anni (dal 2010 al 2020). Si tratta di uno studio nazionale multicentrico, che ha coinvolto 34 centri per l’epilessia. Sono stati arruolati pazienti con crisi di nuova insorgenza ad eziologia autoimmune, definita in base alla rilevazione di anticorpi antineuronali specifici, o sulla base della presentazione clinica e degli esami laboratoristici e strumentali. Risultati Sono stati arruolati 263 pazienti (138 di sesso femminile, con età di 55 anni, range 4-86) seguiti per un periodo di tempo di 30 mesi (range 12-20). L’età all’esordio era di 48 anni (range 2-82). Anticorpi antineuronali sono stati identificati nel 63.50% dei casi (79.65% dei quali aveva anticorpi diretti controllo antigeni della superficie neuronale). Nessun tipo di crisi è risultato essere correlato alla positività anticorpale, ad eccezione delle crisi distoniche facio-brachiali, patognomoniche della encefalite da anticorpi anti-LGI1 (p<0.001). La maggior parte dei pazienti positivi presentava più tipi di crisi (p=0.01); e un coinvolgimento prevalente delle strutture temporali (p=0.02). All’analisi multivariata però queste caratteristiche non sono risultate essere fattori predittivi indipendenti la rilevazione anticorpale. Interessante è risultata essere la maggiore prevalenza di episodi di stato epilettico nei pazienti negativi (p<0.001), correlata anche una prognosi meno favorevole. Durante la fase acuta, l’87.73% dei pazienti presentava crisi farmacoresistenti agli antiepilettici, senza che vi fosse una prevalenza significativa della positività anticorpale. Oltre alle crisi, la fase acuta era caratterizzata da altri sintomi associati nel 93% dei casi; soprattutto disturbi cognitivi, psichiatrici e del sonno, prevalenti nei pazienti con anticorpi negativi (p< 0.001). La maggior parte dei pazienti (88.60%) ha ricevuto un trattamento immunomodulante, e il 61.80% ha presentato una risposta favorevole. Fattori indipendenti predittivi di una risposta favorevole alla terapia sono risultati essere l’inizio precoce della terapia immunomodulante (entro 3 mesi; p<0.001) e la presenza di anticorpi diretti contro antigeni della superficie neuronale (p=0.01). Gli esiti a lungo termine erano rappresentati dalla persistenza di crisi oltre la fase acuta dell’encefalite nel 43.73% dei casi, con prevalenza nei pazienti negativi (p<0.001), e con associati anche deficit neuropsicologici e sintomi psichiatrici nell’81.73% dei casi. Conclusioni Il crescente riconoscimento di forme di epilessia a genesi autoimmune e l’ampio spettro di encefaliti autoimmuni con crisi epilettiche quali sintomo predominante ha determinato un crescente interesse clinico e scientifico verso un nuovo ambito di difficile diagnosi e gestione terapeutica. Il confronto tra i vari sottogruppi in base alla determinazione anticorpale e alla risposta terapeutica permette una migliore definizione e caratterizzazione di questi pazienti. Un’eziologia autoimmune delle crisi epilettiche rappresenta una opportunità terapeutica, e il trattamento precoce a livello patogenetico può ridurre il rischio di sviluppare irreversibili esiti a lungo termine. Questo studio presenta un livello di evidenza di Classe IV per raccomandazioni nella gestione complessiva di questi pazienti.
Background and aims Epileptic seizures may be a presenting or prominent symptom of brain dysfunction in autoimmune encephalitis. They are usually resistant to symptomatic therapy with antiseizure medications (ASMs) but may benefit from immunomodulatory treatments. Despite the increasing knowledge in this field and progress in research interest, autoimmune epilepsy is still an under-recognized condition without standardized diagnostic and management guidelines. This study aims to analyze the epileptic phenotypes of seizures of autoimmune etiology, assessing their clinical presentation, seizure semiology, and associated paraclinical findings. Treatment options, management, and overall outcomes at the long-term were also provided. Methods An observational cohort study was retrospectively performed over 10 years period (from 2010 to 2020). This is a nationwide study, carried out in 34 Italian epilepsy centers being part of the network of high expertise centers of the Italian League Against Epilepsy [LICE]. Patients with new-onset seizures of an autoimmune etiology were enrolled. This latter was defined by the detection of antineuronal antibodies or suspected on the basis of clinical presentation and paraclinical findings. Results Overall, 263 patients (138 females; median age 55 years, range 4-86) were enrolled and followed-up for a median time of 30 months (range 12-120). The median age at seizure onset was 48 years (range 2-82). Antineuronal antibodies were detected in 63.50% of cases (79.65% of them had antibodies targeting neuronal cell-surfaceantigens). No specific seizure semiology was found to be related to antibody-positivity, except for facio-brachial dystonic seizures, which are pathognomonic of LGI1 encephalitis (p< 0.001). Most antibody-positive patients had multiple seizure types (p=0.01); and a prevalent involvement of the temporal regions (p=0.02), but when performing the multivariate analysis, these features were not confirmed to be independent predictors for antibody detection. Of interest, a higher prevalence of episodes of statusepilepticus was found in the antibody-negative patients (OR 0.23, 95% CI 0.12-0.45; p<0.001), which also usually leads to a less favorable prognosis in these cases.During the acute phase, 87.73% presented seizures drug-resistant to most common ASMs, without any significant prevalence in antibody detection. Besides seizures, the acute phase was also marked by associated symptoms in 93%; mostly cognitive impairment, psychiatric symptoms, and sleep disorders were prevalent in antibody-positive subgroup (OR 4.70, 95% CI 2.33-9.47; OR 3.43, 95% CI 1.81-6.50; OR 5.41, 95% CI 2.16-13.52, respectively; p< 0.001). Most patients (88.60%) were treated with immunotherapy, and 61.80% of them were considered responders. Independent predictors of a favorable outcome were confirmed to be early immunotherapy (within 3 months from the onset; OR 12.08, 95% CI 5.50-26.50; p<0.001) and the detection of antineuronal surface antibodies (OR 2.38; 95% CI 1.15-4.92; p=0.01). Long-term outcomes were marked by persisting seizures beyond the acute phase of the encephalitis in 43.73%, with a prevalence in antibody-negative patients (p<0.001), associated with neuropsychological deficits and psychiatric disorders in81.73% of them. Conclusions The increasing recognition of an autoimmune basis of epilepsies and the broad spectrum of autoimmune encephalitis with predominant epileptic seizures has raised interest in scientific and clinical epileptology, opening a new field with challenging issues in diagnosis and treatment. The comparison between subgroups of antibody findings and outcomes may improve the operative definition and characterization. An autoimmune etiology represents a rare chance in seizures management, and an early treatment at the pathogenic level may reduce the risk of irreversible sequelae at the long-term. This study provides Class IV evidence for management recommendations.

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Hänel, Mathias, Friedrich Fiedler, and Christoph Thorns. "Anti-CD20 Monoclonal Antibody (Rituximab) and Cidofovir as Successful Treatment of an EBV-Associated Lymphoma with CNS Involvement." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-135105.

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Background: Epstein-Barr virus(EBV)-associated posttransplant lymphoproliferative disease (PTLD) is a serious complication after allogeneic hematopoietic stem cell transplantation (HSCT). Especially in cases with involvement of the central nervous system (CNS) treatment is difficult because the efficacy of most chemotherapeutic agents as well as EBV-specific cytotoxic donor T cells in liquor is uncertain. In the last years the anti-CD20 monoclonal antibody Rituximab was intensively investigated in the treatment of EBV-PTLD. However, only 8 patients with B-cell lymphoma and CNS involvement treated with Rituximab were reported. Case Report: A 24-year-old female patient with acute T-lymphoblastic leukemia in second complete remission had received allogeneic, unrelated, T-cell depleted HSCT. 10 months later an EBV-associated PTLD was diagnosed. Beside peripheral lymphomas and B symptoms the patient showed neurological symptoms. Examination of the cerebrospinal fluid (CSF) revealed a meningeosis lymphoblastica caused by the EBV lymphoma. Treatment with Rituximab and the antiviral drug Cidofovir led to complete remission with regression of the peripheral lymphomas and disappearance of the neurological symptoms. In addition, the PCR control on EBV DNA became negative in the plasma as well as in CSF. Conclusion: The combination of Rituximab and Cidofovir appears as an interesting alternative treatment in patients with EBV-associated PTLD and CNS involvement
Hintergrund: Die Epstein-Barr-Virus(EBV)-assoziierte Posttransplantations-lymphoproliferative Disease (PTLD) ist eine gefürchtete Komplikation nach allogener hämatopoetischer Stammzelltransplantation (HSCT). Insbesondere bei Befall des zentralen Nervensystems (ZNS) ist die Behandlung auf Grund der unsicheren Liquorwirksamkeit der meisten Chemotherapeutika als auch von EBV-spezifischen zytotoxischen T-Spenderzellen schwierig. Der monoklonale Anti-CD20-Antikörper Rituximab wurde in den letzten Jahren bei Patienten mit EBV-PTLD intensiv untersucht. Allerdings wurde bislang lediglich von 8 Patienten mit ZNS-Befall eines B-Zell-Lymphoms berichtet, bei denen eine Therapie mit Rituximab erfolgte. Kasuistik: Eine 24-jährige Patientin hatte wegen einer akuten T-lymphoblastischen Leukämie in zweiter kompletter Remission eine allogen-unverwandte, T-Zelldepletierte HSCT erhalten. 10 Monate später wurde eine EBV-assoziierte PTLD diagnostiziert. Neben peripheren Lymphomen und B-Symptomen zeigte die Patientin neurologische Symptome. Die Liquoruntersuchung erbrachte den Befund einer Meningeosis lymphoblastica im Rahmen des EBV-Lymphoms. Die Behandlung mit Rituximab und dem Virustatikum Cidofovir führte zu einer kompletten Remission mit Rückbildung der peripheren Lymphome und Verschwinden der neurologischen Symptomatik. Außerdem wurde die PCR-Kontrolle auf EBV-DNA sowohl im Plasma als auch im Liquor negativ. Schlussfolgerung: Die Kombination von Rituximab und Cidofovir erscheint als eine interessante Therapiealternative für Patienten mit EBV-assoziierter PTLD und ZNS-Befall
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich

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20

Wareham, Carol. "Exploring the efficacy of anti-GD2 and anti-4-1BB monoclonal antibody therapy for the treatment of neuroblastoma." Thesis, University of Southampton, 2014. https://eprints.soton.ac.uk/376894/.

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Hänel, Mathias, Friedrich Fiedler, and Christoph Thorns. "Anti-CD20 Monoclonal Antibody (Rituximab) and Cidofovir as Successful Treatment of an EBV-Associated Lymphoma with CNS Involvement." Karger, 2001. https://tud.qucosa.de/id/qucosa%3A27618.

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Background: Epstein-Barr virus(EBV)-associated posttransplant lymphoproliferative disease (PTLD) is a serious complication after allogeneic hematopoietic stem cell transplantation (HSCT). Especially in cases with involvement of the central nervous system (CNS) treatment is difficult because the efficacy of most chemotherapeutic agents as well as EBV-specific cytotoxic donor T cells in liquor is uncertain. In the last years the anti-CD20 monoclonal antibody Rituximab was intensively investigated in the treatment of EBV-PTLD. However, only 8 patients with B-cell lymphoma and CNS involvement treated with Rituximab were reported. Case Report: A 24-year-old female patient with acute T-lymphoblastic leukemia in second complete remission had received allogeneic, unrelated, T-cell depleted HSCT. 10 months later an EBV-associated PTLD was diagnosed. Beside peripheral lymphomas and B symptoms the patient showed neurological symptoms. Examination of the cerebrospinal fluid (CSF) revealed a meningeosis lymphoblastica caused by the EBV lymphoma. Treatment with Rituximab and the antiviral drug Cidofovir led to complete remission with regression of the peripheral lymphomas and disappearance of the neurological symptoms. In addition, the PCR control on EBV DNA became negative in the plasma as well as in CSF. Conclusion: The combination of Rituximab and Cidofovir appears as an interesting alternative treatment in patients with EBV-associated PTLD and CNS involvement.
Hintergrund: Die Epstein-Barr-Virus(EBV)-assoziierte Posttransplantations-lymphoproliferative Disease (PTLD) ist eine gefürchtete Komplikation nach allogener hämatopoetischer Stammzelltransplantation (HSCT). Insbesondere bei Befall des zentralen Nervensystems (ZNS) ist die Behandlung auf Grund der unsicheren Liquorwirksamkeit der meisten Chemotherapeutika als auch von EBV-spezifischen zytotoxischen T-Spenderzellen schwierig. Der monoklonale Anti-CD20-Antikörper Rituximab wurde in den letzten Jahren bei Patienten mit EBV-PTLD intensiv untersucht. Allerdings wurde bislang lediglich von 8 Patienten mit ZNS-Befall eines B-Zell-Lymphoms berichtet, bei denen eine Therapie mit Rituximab erfolgte. Kasuistik: Eine 24-jährige Patientin hatte wegen einer akuten T-lymphoblastischen Leukämie in zweiter kompletter Remission eine allogen-unverwandte, T-Zelldepletierte HSCT erhalten. 10 Monate später wurde eine EBV-assoziierte PTLD diagnostiziert. Neben peripheren Lymphomen und B-Symptomen zeigte die Patientin neurologische Symptome. Die Liquoruntersuchung erbrachte den Befund einer Meningeosis lymphoblastica im Rahmen des EBV-Lymphoms. Die Behandlung mit Rituximab und dem Virustatikum Cidofovir führte zu einer kompletten Remission mit Rückbildung der peripheren Lymphome und Verschwinden der neurologischen Symptomatik. Außerdem wurde die PCR-Kontrolle auf EBV-DNA sowohl im Plasma als auch im Liquor negativ. Schlussfolgerung: Die Kombination von Rituximab und Cidofovir erscheint als eine interessante Therapiealternative für Patienten mit EBV-assoziierter PTLD und ZNS-Befall.
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

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Kao, Daniel Joseph. "Development of a synthetic peptide vaccine and antibody therapeutic for the prevention and treatment of Pseudomonas Aeruginosa infection /." Connect to full text via ProQuest. Limited to UCD Anschutz Medical Campus, 2007.

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Thesis (Ph.D. in Pharmacology) -- University of Colorado Denver, 2007.
Typescript. Includes bibliographical references (leaves 203-212; 260-261). Free to UCD affiliates. Online version available via ProQuest Digital Dissertations;

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Rahnama, Samira. "The development of a novel treatment for equine laminitis." Thesis, Queensland University of Technology, 2020. https://eprints.qut.edu.au/199907/1/Samira_Rahnama_Thesis.pdf.

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Each year hundreds of horses are destroyed due to a crippling hoof disease known as laminitis. This project set out to discover how high concentrations of insulin can cause damage in the hoof and to determine if the disease can be prevented using a therapeutic antibody that blocks receptors for an insulin-like growth factor. Therapeutic antibodies have been developed to treat illness in humans but have never been used in horses. The results showed that the antibody can limit the damage caused by insulin and that it may become a useful treatment in the future.

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Hu, Eileen Yifan. "Developing Methods and Targeted Therapeutics to Address Complications of Ibrutinib Treatment in Chronic Lymphocytic Leukemia." The Ohio State University, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=osu1587494624201361.

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McBride, Harry Michael. "The recruitment of ribosomal inactivating protein or T cells by antibody derivatives in the treatment of B cell lymphoma." Thesis, University of Southampton, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.295851.

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Barbarino, Verena [Verfasser], Christian [Gutachter] Pallasch, and Marcus [Gutachter] Krüger. "Macrophage-mediated antibody dependent effector function in aggressive B cell lymphoma treatment / Verena Barbarino ; Gutachter: Christian Pallasch, Marcus Krüger." Köln : Universitäts- und Stadtbibliothek Köln, 2021. http://nbn-resolving.de/urn:nbn:de:hbz:38-531485.

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Jayaram, Rohith. "A Local, Sustained Delivery System for Zoledronic Acid and RANKL-Inhibitory Antibody as a Potential Treatment for Metastatic Bone Disease." UKnowledge, 2015. http://uknowledge.uky.edu/cbme_etds/34.

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Cancerous solid tumors can migrate and lead to metastatic bone disease. Drugs prescribed to reduce bone resorption from metastasis, such as zoledronic acid and the RANKL-inhibitory antibody Denosumab, cause side effects such as osteonecrosis of the jaw when delivered systemically. This project used two biocompatible materials, acrylic bone cement (PMMA) and poly(lactic-co-glycolic acid) (PLGA), to incorporate and sustain release of anti-resorptive agents. Results showed similar mechanical properties for acrylic bone cements loaded up to 6.6% drug by weight. Results showed sustained zoledronic acid release for 8 weeks from both systems, with PMMA releasing up to 22% of loaded drug and PLGA films releasing over 95%. The antibody release rate was lower, with the majority of antibody still inside the PLGA films after 8 weeks. In vitro bioactivity remained above 50% for zoledronic acid eluted from both materials at early, middle, and late time points. This study sheds light on the behavior of these biocompatible polymers at high drug weight percent loadings compared to previous studies. PLGA demonstrated superior release kinetics but inferior bioactivity of eluted drug. By incorporating anti-resorptive drugs into locally implantable materials, this work could lead to a treatment offering improved quality of life for cancer patients.

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Oden, Felix [Verfasser]. "Generation of an antibody targeting B cell maturation antigen for the treatment of multiple myeloma and autoimmune diseases / Felix Oden." Berlin : Freie Universität Berlin, 2014. http://d-nb.info/1063934494/34.

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Ishida, Yoshihiro. "Killer immunoglobulin-like receptor genotype did not correlate with response to anti-PD-1 antibody treatment in a Japanese cohort." Kyoto University, 2020. http://hdl.handle.net/2433/253207.

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Akgün, Katja, Imke Metz, Hagen H. Kitzler, Wolfgang Brück, and Tjalf Ziemssen. "Rescue therapy with alemtuzumab in B cell/antibody-mediated multiple sclerosis." Sage, 2018. https://tud.qucosa.de/id/qucosa%3A35543.

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Alemtuzumab exerts its clinical efficacy by its specific pattern of depletion and repopulation of different immune cell subsets. Recently, single cases of multiple sclerosis patients who developed severe exacerbation after the first alemtuzumab application, accompanied by re-appearance of peripheral B cells, were reported. Here we present a case with underlying B cell-driven multiple sclerosis that impressively improves after alemtuzumab, although peripheral B cell repopulation took place. Our detailed clinical, histopathological, imaging and immunological data suggest that alemtuzumab can act as an effective rescue treatment in highly active B cell-driven and antibody/complement-mediated multiple sclerosis type II patients.

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Haack, Stephanie [Verfasser], and Niklas [Gutachter] Beyersdorf. "A novel mouse model for systemic cytokine release upon treatment with a superagonistic anti-CD28 antibody / Stephanie Haack ; Gutachter: Niklas Beyersdorf." Würzburg : Universität Würzburg, 2021. http://d-nb.info/123439152X/34.

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Johansson, Jeannette. "Antibodies for better or worse or Antibody variability in an egg-laying mammal and a novel strategy in the treatment of allergies." Doctoral thesis, Uppsala University, Department of Cell and Molecular Biology, 2002. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-2533.

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Antibodies are a central part of the immune defense system, and a large variability in their specificity is needed in order to be able to react against all possible foreign substances we may encounter during our lives. In this thesis, results are presented from investigations into how an egg-laying mammal, the Australian duck-billed platypus (Ornithorhynchus anatinus) creates antibody variability. Our results show that despite the lack of many V gene families the antibody repertoire in the platypus seems to be well developed. A long and highly variable complementarity-determining region (CDR) 3 compensates for the limited germline diversity. Interestingly, the presence of additional cysteine residues in the CDRs may form stabilizing disulfide bridges in the antigen binding loops and thereby increasing the affinity of the antibody-antigen interaction.

Although the immune system is necessary for survival, it must be strictly controlled since it may otherwise over-react and cause more harm than benefits. Allergies and autoimmune diseases are examples of such over-reactions by the immune system. Allergies are increasing in the western world and have become one of the main medical issues of the 21^st century. IgE is the central mediator in atopic allergies such as hay fever, eczema and asthma; it is therefore a prime target in the development of allergen-independent preventative treatments. Here we present results from several studies of a novel vaccine strategy aimed at reducing the levels of IgE antibodies. The vaccine results in the induction of anti-IgE antibodies, and the skin reactivity upon allergen challenge was significantly reduced in vaccinated animals. Our results suggest that active immunization against IgE has the potential to become a therapeutic method for humans. In addition, an evaluation of possible adjuvants that could be used as immune stimulators and thus help break self-tolerance at the time of vaccination is presented.

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Marckel, Jordan A. "The in-vivo Preclinical Development of a Humanized Anti-cocaine Monoclonal Antibody and its Fab Fragment for the Treatment of Cocaine Abuse." University of Cincinnati / OhioLINK, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1613745458699203.

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Nicolay, Uwe. "Health-related quality of life, treatment satisfaction and clinical aspects of patients with primary antibody deficiency receiving subcutaneous IgG self-infusions at home /." Stockholm, 2006. http://diss.kib.ki.se/2006/91-7140-826-6/.

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Santos, Filipe Miguel Barão dos. "Canine lymphoma immunophenotype prevalence and predisposition and evaluation of SN38 to be used in an antibody-drug conjugate for B-cell lymphoma treatment." Master's thesis, Universidade de Lisboa, Faculdade de Medicina Veterinária, 2021. http://hdl.handle.net/10400.5/21061.

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Dissertação de Mestrado Integrado em Medicina Veterinária
Lymphoma is one of the most diagnosed neoplasias in dogs and one of the most recurring in clinical practice. This disease features various immunophenotypes, clinical and anatomopathological presentations, being the B-cell lymphoma the most reported in literature. In this species, lymphoma is similar to non-Hodgkin human lymphoma regarding therapy response, immunophenotype and anatomopathological presentation. Therefore, the dog is a very good spontaneous model for comparative studies. In this way, a statistical study was conducted in order to investigate the frequency for each immunophenotype and a possible breed predisposition for them, in comparison with previous studies. The sample included 165 records of canine lymphoma cases from 36 breeds kept by the Laboratory of Pathological Anatomy of the Faculty of Veterinary Medicine in the last 7 years with an immunohistochemistry diagnosis. Hitherto, conventional therapy comprises chemotherapy, corticotherapy and radiotherapy. However, these approaches have their limitations. This arises the need for novel therapies. Amongst those, immunotherapy with antibody-drug conjugates (ADCs) stands out since they allow for a more specific and directed therapy with less side effects. Given the promising potential of these strategies, the aim of this project consisted in contributing to the development of an ADC through the cloning of previously selected single-domain antibodies (sdAbs) in a pET21a vector for large-scale production and the validation of SN38 as a cytotoxic drug to link to these antibodies. The final aim was the ADC therapeutic application in canine B-cell lymphoma. Making use of descriptive statistics and Fisher’s exact tests, the obtained results confirmed a large majority of B-cell lymphoma cases (72.7%) but, contrarily to previous studies, an association with lymphoma immunophenotype was not found in the majority of the studied breeds. These results are not very statistically reliable, due to the small sample population. The cell viability assays performed with SN38 in two B-cell lymphoma lines, a canine lymphoma (CLBL-1) and a human lymphoma cell line (Raji), disclosed IC50 ranging between 4 to 300 nM, promising results which reinforce the importance in investing in more studies in order to obtain more data that can validate this compound for canine lymphoma therapy.
ABSTRACT - Predisposição e prevalência de imunofenótipos do linfoma canino e avaliação do SN38 para utilização num imunoconjugado para tratamento do linfoma de células B - O linfoma é uma das neoplasias mais diagnosticada em cães e uma das mais recorrentes na prática clínica. Esta doença apresenta vários imunofenótipos e apresentações clínicas e anatomopatológicas, sendo o linfoma de células B o mais registado na literatura. Nesta espécie, o linfoma assemelha-se ao linfoma não-Hodgkin no Homem, nomeadamente na resposta à terapia, imunofenótipo e apresentação anatomopatológica. Assim, o cão apresenta-se como um excelente modelo espontâneo de estudos comparativos com o Homem. Desta forma, foi efectuado um estudo estatístico de forma a averiguar a frequência de cada imunofenótipo e uma possível predisposição de raça para os mesmos em comparação com estudos anteriores. A amostra incluiu 165 registos de casos de linfoma canino de 36 raças mantidos pelo Laboratório de Anatomia Patológica da Faculdade de Medicina Veterinária nos últimos 7 anos com diagnóstico imunohistoquímico. Até à data, a terapêutica convencional para o linfoma resume-se à terapêutica com anti-neoplásicos, à corticoterapia e à radioterapia. Contudo, estas abordagens têm limitações. Atendendo a estas adversidades, surge a necessidade de novas terapias. Destas, destaca-se a imunoterapia com anticorpos conjugados com fármacos citotóxicos (ADCs) que permitem uma terapia mais específica e direccionada, com menos efeitos adversos. Dado o potencial promissor destas estratégias, este trabalho teve como objectivo contribuir para o desenvolvimento de um ADC através da clonagem de anticorpos de domínio único (sdAbs), previamente seleccionados, num vector pET21a para produção em larga escala e validação do SN38 como composto citotóxico para ligação a estes anticorpos, tendo em vista a sua aplicação terapêutica em linfoma canino de células B. Com recurso a estatística descritiva e testes exactos de Fisher, os resultados obtidos confirmaram uma larga maioria de casos de linfoma de células B (72,7%), mas, contrariamente a estudos prévios, não se encontrou uma associação com imunofenótipo na maioria das raças estudadas, resultados estes estatisticamente pouco significativos devido à pequena amostra do estudo. Os ensaios de viabilidade celular realizados com o SN38 em duas linhas celulares de linfoma de células B, uma de linfoma canino (CLBL-1) e outra de linfoma humano (Raji), revelaram valores de IC50 na ordem dos 4 a 300 nM, resultados promissores que reforçam a importância de investir em mais estudos de forma a obter mais dados que possibilitem validar este composto na terapêutica do linfoma canino.
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Caudana, Pamela. "Translating IL-2/anti-IL-2 antibody complexes treatment for human tumor immunotherapy IL-2/anti-IL-2 antibody complex combined with CTLA-4- but not PD1- blockade rescues anti-tumor NK function by modulating intra-tumoral regulatory T cells." Thesis, Sorbonne Paris Cité, 2018. http://www.theses.fr/2018USPCB146.

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L'IL-2 à forte dose a été la première immunothérapie administrée pour le traitement de cancers humains, atteignant dans les mélanomes métastatiques des réponses objectives similaires à celles obtenues avec les traitements avec des Ac bloquants les points de contrôle immunitaires (checkpoint) (autour de 15% de répondeurs). Toutefois, sa toxicité est importante et son mécanisme d'action chez les patients répondant reste mal connu. Sa faible efficacité est due en partie à l'action de l'IL-2 sur les cellules T regulatrices (Tregs). Par conséquent, différentes approches ont été envisagées pour amener à nouveau l'IL-2 en clinique. L'une de ces stratégies est basée sur les complexes IL-2/Ac anti-IL-2 (IL-2Cx). Selon la spécificité de l'anticorps utilisé, ce complexe permet de diriger l'action de l'IL-2 sur les Tregs ou les CD8+/NK. Les IL-2Cx stimulant les Tregs ont déjà été utilisé avec succès dans des modèles précliniques de maladies autoimmunes telles que le diabète, l'EAE, etc. D'un autre côté, les IL-2Cx stimulant les cellules immunes effectrices ont montré leur efficacité dans des modèles précliniques de mélanomes, leucémies et carcinomes du rein. Jusqu'à maintenant, les résultats précliniques publiés ont été obtenus avec des IL-2Cx utilisant des Ac commerciaux anti-IL-2 murine ou anti-IL-2 humaine (hIL-2), mais il n'existe pas d'IL-2Cx formés avec des Ac anti-hIL-2 de grade clinique. Dans ce contexte, mon travail de thèse avait pour objectif : i) de générer des Ac humains anti-hIL-2 qui sous forme de complexes seraient capables d'activer préférentiellement les cellules NK et les effecteurs T CD8+, pour le traitement du cancer ; et ii) l'évaluation préclinique de traitements combinés avec les IL-2Cx et les traitements bloquant les checkpoints pour la thérapie anti-cancer. Pour générer les Ac humains anti-hIL-2, nous avons utilisé 2 stratégies : i) phage display contre l'hIL-2 basée sur une librairie pre-immune d'Ac de lama; et ii) l'isolation d'auto-anticorps anti-hIL-2 produits par les cellules B de patients atteints de diabète de type 1, décrit auparavant dans notre équipe. A l'aide de ces 2 approches techniques, nous avons généré avec succès un grand nombre d'Ac anti-hIL-2, mais nous avons rencontré de difficultés pour isoler les Ac qui auraient l'activité souhaitée sous forme d'IL-2Cx. Pour résoudre cette difficulté technique, en collaboration avec une équipe de l'Institut Pasteur de Paris, nous avons produit de nouvelles variantes de l'hIL-2 pour simplifier la sélection des Ac anti-hIL-2. En parallèle, nous avons évalué l'efficacité antitumorale des IL-2Cx combinés au blocage des checkpoint CTLA4 ou PD1. Dans un modèle de carcinome du poumon spontané inductible, nous avons montré que la combinaison IL-2Cx/anti-PD1 contrôle de façon durable l'évolution de la tumeur et induit des réponses T spécifiques de la tumeur. Par ailleurs, dans le modèle syngénique de la tumeur B16-OVA, naturellement résistante au blocage des checkpoints, la combinaison des IL-2Cx avec le blocage de la voie CTLA4 ou PD1 permet de contrôler la résistance. Concernant les mécanismes d'action de ces traitements, les 2 combinaisons induisent la « re-activation » des cellules T CD8+ intratumorales et augmentent l'amplitude de la réponse T spécifique de la tumeur. Toutefois, seule la combinaison IL-2Cx/anti-CTLA4 diminue les Treg intratumoraux, conduisant à une augmentation du rapport NK/Treg. De plus, cette combinaison est strictement dépendante des cellules NK pour son effet thérapeutique in vivo, en plus de l'implication des cellules T CD8. Par conséquent, nos résultats suggèrent que les thérapies basées sur l'utilisation de l'hIL-2 devraient être à nouveau prise en considération. De plus, notre observation que les combinaisons des IL-2Cx avec le blocage de CTLA4 ou de PD1 passent par des mécanismes cellulaires distincts, ouvre la voie pour la conception rationnelle de thérapies combinatoires contre le cancer
High-dose IL-2 was the first immunotherapy ever assessed for the treatment of human cancer, achieving similar objective responses in metastatic melanoma as with a-checkpoint mAbs treatment (15%). However, less than 10% of eligible patients receive this potentially curative treatment, likely due to its associated toxicity and need of hospitalization, as well as concern on the expansion of Treg cells. Consequently, different approaches have been applied to bring back IL-2 to the clinics, trying to bypass Treg activation and to improve IL-2 pharmacodynamics. One of these strategies is the use of IL-2/anti-IL-2 mAb complexes (IL-2Cx). In mouse studies it has been shown that when IL-2 is complexed with an anti-IL-2 antibody (Ab), IL-2 pharmacodynamics is improved, toxicity is lowered and depending on the Ab, the IL-2Cx can direct IL-2 action to effector or to regulatory immune cells. In the past years, Treg-stimulating IL-2Cx have been successfully used in preclinical models of autoimmune diseases like EAE, Arthritis, renal diseases and allergy. On the other hand, effector cell-stimulating IL-2Cx have shown to delay tumor growth in preclinical models of melanoma, leukemia and renal cell carcinoma. Heretofore, all published preclinical data have been obtained using IL-2Cx formed with commercial rat anti-mouse IL-2 or rat-anti human IL-2 Abs, but yet, there are no available IL-2Cx formed with clinical-grade anti-human (h) IL-2 mAbs. In this context, my thesis work was dedicated to: i) the generation of human anti-human IL-2 Abs that when complexed with hIL-2 preferentially activate NK and effector CD8+ T cell, for cancer treatment; and ii) the pre-clinical evaluation of the efficacy of combinatory treatments using IL-2Cx with immunecheckpoint (CKP) blockade for cancer therapy. To generate human anti- hIL-2 Abs we used two strategies: i) phage display against hIL-2 using a pre-immune lama library; and ii) isolation of anti-hIL-2 auto-antibodies from B cells obtained from Type 1 diabetic patients, previously described by our team. Using both techniques, we succeeded in generating large amounts of anti-hIL-2 Abs, but we encountered many difficulties to isolate those with the desired IL-2Cx activity. To solve this technical issue, in collaboration with a team at Institut Pasteur in Paris, we designed new hIL-2 variants allowing a simplified screening of the IL-2 Abs (currently under patenting process). In parallel, we evaluated the anti-tumoral efficacy of effector cell-stimulating IL-2Cx combined with blockade of CTLA-4 and PD1 pathway, with a translational perspective. First, in an inducible spontaneous lung adenocarcinoma model we showed that the IL-2Cx/anti-PD1 combo durably controlled tumor growth and induced tumor-specific T cell responses. In the B16-OVA syngeneic model, naturally resistant to anti-checkpoint inhibition, combination of IL-2Cx with PD1 or CTLA-4 pathway blockade reversed resistance. Mechanistically, both combos worked by re-invigorating intratumoral CD8+ T cells and increasing the breath of tumor-specific T cell responses. However, only the IL-2Cx/anti-CTLA-4 combo diminished intratumoral Treg cells, leading to an increase in the NK/Treg cell ratio and is non-redundantly dependent on NK cells. No treatment toxicity was observed when IL-2 was used in the form of IL-2Cx. Overall, our results suggest that IL-2-based therapies should be reconsidered for the treatment of cancer. Moreover, our observation that the combinations of IL-2Cx with PD1 or CTLA-4 pathway blockade act by different cellular mechanisms, paves the way for the rational design of combinatorial anti-tumoral therapies

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Buteyn, Nathaniel J. "Role of Innate Immunity Activators in the Treatment of Acute Myeloid Leukemia." The Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1574343556916953.

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Anzicek, Nika. "Studies towards a second-generation synthesis of the aplyronines." Thesis, University of Cambridge, 2017. https://www.repository.cam.ac.uk/handle/1810/267831.

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The aplyronines are a family of 24-membered macrolides of polyketide origin, isolated from the Japanese sea hare Aplysia kurodai. They exhibit an exceptional biological activity profile, acting through an actin and tubulin dual-targeting mechanism, with subnanomolar growth inhibitory potency against a diverse range of cancer cell lines. These characteristics render the aplyronines ideal payloads for antibody-drug conjugates but their prohibitively low natural abundance calls for an efficient total synthesis to overcome the supply issue. This dissertation describes the efforts towards developing a second-generation Paterson synthesis of the macrocyclic core of the aplyronines, focused on improving the scalability and selectivity of key transformations. Chapter 1 details the isolation, biological background and previous synthetic efforts towards the aplyronines to illustrate their therapeutic potential and the challenges associated with material sourcing by chemical synthesis. Chapter 2 presents the existing body of work on the aplyronine project within the Paterson group, highlighting the lessons learned over the past two decades and shortcomings to be addressed. Chapter 3 discusses a revised protecting group strategy towards the C1-C27 macrocyclic alcohol 159 with fewer manipulation steps. A refined reaction sequence featuring titanium aldol methodology and an enzymatic desymmetrisation process delivered multigram stocks of the C15-C27 aldehyde 161 upon scale- up, testifying to the robustness of the devised route. Synthesis of the C1-C14 northern fragment 253 closely followed the existing boron aldol approach with optimisation of the C11-C12 alkylation step, geared towards enhancing the regioselectivity. Chapter 4 describes the coupling of the two major fragments using an Horner-Wadsworth-Emmons reaction to assemble the C1-C27 backbone of the cyclic aplyronine core and suitably adjusted endgame steps to enable a one-step oxidative unmasking of the macrolactonisation sites. The first-generation intermediate 159 was accessed via site-specific Yamaguchi esterification and orthogonal deprotection of the C27 allyl carbonate. Discussion in Chapter 5 includes the appendage of the C28-C34 side chain 118, prepared by the known sequence, and suggestions for the future direction of the second-generation route with the outlook of linker appendage for the purposes of antibody-drug conjugate development.

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Dias, Joana Nunes Ribeiro. "Clinical and immunological characterization of naturally occurring canine lymphoma : development and application of engineered recombinant antibodies for diagnosis and treatment." Doctoral thesis, Universidade de Lisboa, Faculdade de Medicina Veterinária, 2019. http://hdl.handle.net/10400.5/17735.

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Tese de Doutoramento em Ciências Veterinárias. Especialidade de Sanidade Animal
Non-Hodgkin lymphoma (NHL) is one of the most common causes of cancer-related death worldwide. Although the outcome of NHL patients has improved with current therapies, the rate of mortality is still high. A plethora of new drugs is entering clinical development for NHL treatment; however, the approval of new treatments remains low due in part to the paucity of clinically relevant models for validation. Canine lymphoma (cNHL) shares remarkable similarities with its human counterpart, making the dog an excellent animal model to explore novel therapeutic options. Therefore, driven by the great success achieved by immunotherapies in human NHL, comparative research has focused on the development of similar immunotherapeutic approaches for dogs. However, the successful use of this animal model remains challenging, still lacking the characterization of the canine immune system, of common tumor epitopes, the development of canine-specific/cross-reactive agents and the establishment of preclinical models. Within this context, we aimed to develop novel antibody-based therapies for cNHL, while contributing for the characterization and validation of the cNHL model for translational immune-oncology research. For that purpose, a cNHL biobank was successfully constructed. The clinical cytokine patterns in patients with cNHL were investigated, confirming a local and systemic dysregulation in cytokine response. Furthermore, a positive correlation between intratumoral immune response and a favorable response to chemotherapy indicates that the modulation of the immune response might contribute to improve patient outcomes. With that in mind, a through characterization of canine CD20 expression was conducted and contributed for the validation of this receptor as a potential immunotherapeutic target for cNHL. This motivated the development and identification of a panel of single domain antibodies (sdAbs) with high binding activity and specificity to canine and human CD20. In addition, to develop a novel drug delivery system for cNHL treatment, we described novel methodologies to identify potential targets, while selecting highly specific sdAbs against NHL. This work allowed to select a promising pool of sdAbs that specifically target NHL tumor receptors for the development of a novel antibody drug conjugate (ADC). Furthermore, we conducted a thorough investigation of a novel ADC payload – panobinostat - a potent histone deacetylase (HDAC) inhibitor with strong in vitro and in vivo antitumor properties in cNHL. Finally, we established a new bioluminescent murine model for monitoring tumor progression and treatment response in preclinical studies. In summary, the work presented herein allowed the establishment of a solid platform for the acceleration of the translational research of novel immunotherapeutic approaches for comparative oncology.
RESUMO - Caracterização clínica e imunológica do linfoma canino como modelo animal : desenvolvimento de anticorpos recombinantes para tratamento e diagnóstico - O linfoma não-Hodgkin (LNH) é uma das principais causas de morte por neoplasia em todo o mundo, representando 90% de todos os linfomas. O LNH abrange um grupo heterogéneo de tumores, caracterizado pela proliferação de linfócitos malignos, 85-90% dos quais de linfócitos B. Os anticorpos direccionados para o recetor CD20, combinados com quimioterapias convencionais revolucionaram o tratamento do linfoma de células B, melhorando o tempo de remissão e aumentando a taxa de sobrevivência. No entanto, independentemente da terapêutica utilizada, a taxa de mortalidade mantém-se elevada. Uma grande diversidade de novos fármacos encontra-se em fase de desenvolvimento clínico para o tratamento do LNH; no entanto, a aprovação de novos tratamentos permanece baixa devido, em parte, à escassez de modelos clinicamente relevantes para validação. O cLNH e o LNH humano (hLNH) partilham muitas características histopatológicas, moleculares, genéticas e clínicas, desta forma o cão é considerado um excelente modelo animal para explorar novas opções terapêuticas. Consequentemente, a investigação em oncologia comparativa, motivada pelo grande sucesso alcançado pelas imunoterapias no tratamento do hLNH, tem-se focado no desenvolvimento de abordagens imunoterapêuticas semelhantes para cães. Contudo, o sucesso do cLNH como modelo animal tem vindo a revelar-se desafiante, na medida que a validação do seu uso para o desenvolvimento de imunoterapias ainda carece da caracterização do sistema imune canino, das células imunitárias e das moléculas efectoras, da avaliação dos epítopos tumorais comuns, do desenvolvimento de agentes imunoterapêuticos específicos para a espécie canina com potencial reacção cruzada com a espécie humana e do estabelecimento de modelos pré-clínicos para oncologia veterinária. Neste contexto, o presente trabalho teve como principal objectivo validar o cLNH como modelo animal para o desenvolvimento de novas estratégias imunoterapêuticas para LNH, visando estabelecer uma linha de investigação inovadora para o desenvolvimento de anticorpos recombinantes para o tratamento do cLNH. Neste sentido, desenvolveu-se uma estratégia multidisciplinar constituída por etapas complementares de modo a construir uma sólida plataforma para o desenvolvimento de ensaios clínicos de imunoterapias em oncologia comparativa...
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Setoyama, Takeshi. "Development of novel bioassay for the measurement of bioactive insulin-like growth factors in blood samples and treatment strategy targeting the bioactive insulin-like growth factors for non-islet cell tumor hypoglycemia." Kyoto University, 2016. http://hdl.handle.net/2433/204576.

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Lind, Anne-Li. "Biomarkers for Better Understanding of the Pathophysiology and Treatment of Chronic Pain : Investigations of Human Biofluids." Doctoral thesis, Uppsala universitet, Anestesiologi och intensivvård, 2017. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-326180.

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Chronic pain affects 20 % of the global population, causes suffering, is difficult to treat, and constitutes a large economic burden for society. So far, the characterization of molecular mechanisms of chronic pain-like behaviors in animal models has not translated into effective treatments. In this thesis, consisting of five studies, pain patient biofluids were analyzed with modern proteomic methods to identify biomarker candidates that can be used to improve our understanding of the pathophysiology chronic pain and lead to more effective treatments. Paper I is a proof of concept study, where a multiplex solid phase-proximity ligation assay (SP-PLA) was applied to cerebrospinal fluid (CSF) for the first time. CSF reference protein levels and four biomarker candidates for ALS were presented. The investigated proteins were not altered by spinal cord stimulation (SCS) treatment for neuropathic pain. In Paper II, patient CSF was explored by dimethyl and label-free mass spectrometric (MS) proteomic methods. Twelve proteins, known for their roles in neuroprotection, nociceptive signaling, immune regulation, and synaptic plasticity, were identified to be associated with SCS treatment of neuropathic pain. In Paper III, proximity extension assay (PEA) was used to analyze levels of 92 proteins in serum from patients one year after painful disc herniation. Patients with residual pain had significantly higher serum levels of 41 inflammatory proteins. In Paper IV, levels of 55 proteins were analyzed by a 100-plex antibody suspension bead array (ASBA) in CSF samples from two neuropathic pain patient cohorts, one cohort of fibromyalgia patients and two control cohorts. CSF protein profiles consisting of levels of apolipoprotein C1, ectonucleotide pyrophosphatase/phosphodiesterase family member 2, angiotensinogen, prostaglandin-H2 D-isomerase, neurexin-1, superoxide dismutases 1 and 3 were found to be associated with neuropathic pain and fibromyalgia. In Paper V, higher CSF levels of five chemokines and LAPTGF-beta-1were detected in two patient cohorts with neuropathic pain compared with healthy controls. In conclusion, we demonstrate that combining MS proteomic and multiplex antibody-based methods for analysis of patient biofluid samples is a viable approach for discovery of biomarker candidates for the pathophysiology and treatment of chronic pain. Several biomarker candidates possibly reflecting systemic inflammation, lipid metabolism, and neuroinflammation in different pain conditions were identified for further investigation.
Uppsala Berzelii Technology Centre for Neurodiagnostics

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Ferrière, Stephen. "Etude du mode d'action d’agents contournant l'action du facteur VIII dans l’hémophilie A Antibodies in the Treatment of Haemophilia A-A Biochemical Perspective A single-domaine antibody that blocks factor VIIa activity in the absence but not presence of tissue factor A hemophilia A mouse model for the in vivo assessment of emicizumab function." Thesis, université Paris-Saclay, 2020. http://www.theses.fr/2020UPASQ017.

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L’hémophilie A est une maladie hémorragique due à un déficit en facteur VIII de la coagulation (FVIII). Le traitement est une thérapie dite de substitution utilisant des concentrés de FVIII. Toutefois, lors d’apparition d’inhibiteurs dirigés contre le FVIII, la stratégie thérapeutique consiste à utiliser des agents contournant l’action du FVIII tel que le FVIIa recombinant (rFVIIa) ou d’un anticorps bispécifique mimant l’activité du FVIII (emicizumab). Le but de cette thèse a consisté à étudier les modes d’action de ces deux thérapies de contournement à l’aide d’outils biologiques innovants développés spécialement. En ce qui concerne le rFVIIa, bien qu’il soit efficace cliniquement, son mode d’action était encore sujet à discussion au début de ce travail. Dans une première partie de cette thèse, nous avons étudié le mode d’action du rFVIIa au moyen d’un anticorps de type « nanobody » que nous avons généré et caractérisé. Nous montrons dans un premier temps que cet anticorps appelé KB-FVIIa-004, possède une activité inhibitrice du FVIIa uniquement lorsque le FVIIa est libre et non lié à son cofacteur, le facteur tissulaire (FT). Cette caractéristique originale nous a permis de montrer in vivo que l’activité du rFVIIa est indépendante en grande majorité de son cofacteur FT. Dans une seconde partie de ce travail, nous avons étudié le mode d’action de l’emicizumab in vivo. En effet, la mesure de son efficacité et l’équivalence en activité FVIII sont compliquées à mesurer et manque de fiabilité dans les tests in vitro. Nous avons développé un modèle murin innovant semi-humanisé en utilisant des souris hémophilie A dans lesquelles nous avons injectés du Facteur IX et du Facteur X humain. Nous avons montré une activité « FVIII-like » de l’anticorps (pour une dose ≥1,5 mg/kg) correspondante à une équivalence de 4,5 U FVIII/kg (soit 9,0 U/dL). De manière intéressante, l’association d’une faible dose de FVIII (5 U/kg) avec l’Emicizumab nous a permis de montrer un arrêt complet du saignement dans notre modèle murin, suggérant un effet additif entre le FVIII et l’emicizumab
Hemophilia A is a hemorrhagic disorder linked to the functional deficiency of coagulation factor VIII (FVIII). Clinical management of hemophilia A mostly relies on substitution therapy using FVIII concentrates. However, since patients may develop inhibitory antibodies against FVIII, alternative treatment strategies are required that bypass FVIII function, such as the use of recombinant Factor VIIa (rFVIIa) or a bispecific antibody that mimics FVIII activity, emicizumab. The aim of this thesis was to study the mode of action of both FVIII-bypassing therapeutics using novel, innovative tools specifically designed for this purpose. Although rFVIIa is clinically efficient, its mode of action was still unclear at the start of my studies. In a first part of this thesis, we have studied the mode of action of rFVIIa using a novel single-domain antibody (nanobody) that we generated and characterized. We first established that this nanobody, designated KB-FVIIa-004, inhibits FVIIa activity selectively in the absence but not presence of its cofactor, tissue factor (TF). This unique characteristic allowed us to show the the in vivo activity of rFVIIa is predominantly independent of its cofactor TF. In a second part of the study, we have examined the in vivo mode of action of the FVIII-mimetic emicizumab. Indeed, to measure its efficacy and equivalence to FVIII using in vitro assays is complicated and will generate inaccurate data. We therefore developed an innovative semi-humanized mouse model for hemophilia A, in which infused human factors IX and X. We were able to establish a FVIII-like activity of emicizumab (for a dose ≥1,5 mg/kg) corresponding to FVII dose of 4,5 U/kg (i.e. 9,0 U/dL). Interestingly, combining a low dose of FVIII (5 U/kg) with emicizumab resulted in a complete correction of the bleeding in our mouse model, suggesting an additive effect between FVIII and emicizumab

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Chakraborti, Srinjoy. "Therapeutic Antibody Against Neisseria gonorrhoeae Lipooligosaccharide, a Phase-variable Virulence Factor." eScholarship@UMMS, 2017. https://escholarship.umassmed.edu/gsbs_diss/905.

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Neisseria gonorrhoeae (Ng) which causes gonorrhea has become multidrug-resistant, necessitating the development of novel therapeutics and vaccines. mAb 2C7 which targets an epitope within an important virulence factor, the lipooligosaccharide (LOS), is a candidate therapeutic mAb. Ninety-four percent of clinical isolates express the 2C7-epitope which is also a vaccine target. Ng expresses multiple LOS(s) due to phase-variation (pv) of LOS glycosyltransferase (lgt) genes. mAb 2C7 reactivity requires a lactose extension from the LOS core Heptose (Hep) II (i.e. lgtG ‘ON’ [G+]). Pv results in HepI with: two (2-), three (3-), four (4-), or five (5-) hexoses (Hex). How HepI glycans impact Ng infectivity and mAb 2C7 function are unknown and form the bases of this dissertation. Using isogenic mutants, I demonstrate that HepI LOS glycans modulate mAb 2C7 binding. mAb 2C7 causes complement (C’)-dependent bacteriolysis of three (2-Hex/G+, 4-Hex/G+, and 5-Hex/G+) of the HepI mutants in vitro. The 3-Hex/G+ mutant (resistant to C’-dependent bacteriolysis) is killed by neutrophils in the presence of mAb and C’. In mice, 2- and 3-Hex/G+ infections are significantly shorter than 4- and 5-Hex/G+ infections. A chimeric mAb 2C7 that hyperactivates C’, attenuates only 4- and 5-Hex/G+ infections. This study enhances understanding of the role of HepI LOS pv in gonococcal infections and shows that longer HepI glycans are necessary for prolonged infections in vivo. This is the first study that predicts in vitro efficacy of mAb 2C7 against all four targetable HepI glycans thereby strengthening the rationale for development of 2C7-epitope based vaccines and therapeutics.

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Chakraborti, Srinjoy. "Therapeutic Antibody Against Neisseria gonorrhoeae Lipooligosaccharide, a Phase-variable Virulence Factor." eScholarship@UMMS, 2005. http://escholarship.umassmed.edu/gsbs_diss/905.

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Neisseria gonorrhoeae (Ng) which causes gonorrhea has become multidrug-resistant, necessitating the development of novel therapeutics and vaccines. mAb 2C7 which targets an epitope within an important virulence factor, the lipooligosaccharide (LOS), is a candidate therapeutic mAb. Ninety-four percent of clinical isolates express the 2C7-epitope which is also a vaccine target. Ng expresses multiple LOS(s) due to phase-variation (pv) of LOS glycosyltransferase (lgt) genes. mAb 2C7 reactivity requires a lactose extension from the LOS core Heptose (Hep) II (i.e. lgtG ‘ON’ [G+]). Pv results in HepI with: two (2-), three (3-), four (4-), or five (5-) hexoses (Hex). How HepI glycans impact Ng infectivity and mAb 2C7 function are unknown and form the bases of this dissertation. Using isogenic mutants, I demonstrate that HepI LOS glycans modulate mAb 2C7 binding. mAb 2C7 causes complement (C’)-dependent bacteriolysis of three (2-Hex/G+, 4-Hex/G+, and 5-Hex/G+) of the HepI mutants in vitro. The 3-Hex/G+ mutant (resistant to C’-dependent bacteriolysis) is killed by neutrophils in the presence of mAb and C’. In mice, 2- and 3-Hex/G+ infections are significantly shorter than 4- and 5-Hex/G+ infections. A chimeric mAb 2C7 that hyperactivates C’, attenuates only 4- and 5-Hex/G+ infections. This study enhances understanding of the role of HepI LOS pv in gonococcal infections and shows that longer HepI glycans are necessary for prolonged infections in vivo. This is the first study that predicts in vitro efficacy of mAb 2C7 against all four targetable HepI glycans thereby strengthening the rationale for development of 2C7-epitope based vaccines and therapeutics.

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Maho, Maud. "Evaluation des effets des traitements par Rituximab versus corticothérapie seule sur la réponse auto-réactive des patients atteints de pemphigus. First-line Rituximab combined with short-term Prednisone versus Prednisone alone for the treatment of Pemphigus (RITUX 3) : a prospective, multicentre, parallel-group, open-label randomised trial Risk factors for short-term relapse in patients with pemphigus treated by Rituximab as first-line therapy Rituximab and corticosteroid effect on Desmoglein specific B cells and T follicular helper cells in patients with Pemphigus Modifications or the transcriptomic profile of autoreactive B cells from pemphigus patients after treatment with Rituximab or standard corticosteroid regimen Long-term increase of Kcnn4 potassium channel surface expression on B cells in pemphigus patients after Rituximab treatment Rituximab is an effective treatment in patients with Pemphigus Vulgaris and demonstrates a steroid-sparing effect Modifications of the BAFF/BAFF-Receptor axis in patients with pemphigus treated with rituximab versus standard corticosteroids regimen. CD11C+ B cells are mainly memory cells prone to differentiate into antibody-secreting cells." Thesis, Normandie, 2019. http://www.theses.fr/2019NORMR132.

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Le pemphigus est une maladie auto-immune spécifique de la peau et des muqueuses provoqué par des auto-anticorps (Ac) spécifiques des desmogléines (Dsg) 1 ou 3. Ces Ac pathogéniques inhibent l'adhésion cellulaire des kératinocytes. Le pemphigus se déclenche par la conjonction d’événements rares impliquant l’émergence puis la coopération de lymphocytes B (LB) et de lymphocytes LT auto-réactifs dans un contexte génétique et environnemental particulier. Jusqu’à présent, la première ligne de traitement du pemphigus était constituée de fortes doses de corticoïdes, qui sont de puissants immunosupresseurs systèmiques. Le Rituximab (RTX), un Ac monoclonal chimérique anti-CD20, constitue une thérapeutique innovante aboutissant à l’élimination des LB. L’étude clinique RITUX 3 a été conçue pour évaluer l’efficacité et l’innocuité du traitement utilisant le RTX associé à une courte corticothérapie dans le traitement de première intention du pemphigus par rapport au traitement de référence par la corticothérapie standard (CS). Dans un premier temps, notre analyse clinico-biologique des patients après 24 mois a démontré que l’utilisation du RTX associé à de la prednisone à court terme en traitement de première intention chez les patients atteints de pemphigus foliacé et vulgaire modéré à sévère est à la fois plus efficace et mieux toléré que le traitement de référence par la prednisone seule (89% de patients versus 34%). Cette efficacité a été confortée à plus long terme après la reconstitution du répertoire lymphocytaire B avec un risque de rechute de 2% à 36 mois. La présence d’une forme sévère de pemphigus au diagnostic (PDAI ≥ 45) et d’un taux d’Ac anti-Dsg à 3 mois supérieur aux valeurs seuils (anti-Dsg1 ≥ 20 ou anti-Dsg3 ≥ 120) sont associés à un risque de rechute précoce de 50%. Ces deux facteurs prédictifs permettent d'identifier un sous-groupe de patients présentant un risque élevé de rechute nécessitant une perfusion d'entretien de RTX au 6ème mois. Dans un deuxième temps, nous avons étudié l’impact des traitements par RTX et par CS chez les patients atteints de pemphigus afin de mieux appréhender la réponse auto-immune. La caractérisation phénotypique des LB auto-réactifs et l’analyse de la fréquence des LB capables de sécréter des immunoglobulines (Ig)G anti-Dsg par une approche ELISPOT a permis d’établir que l’efficacité du traitement par RTX dans le pemphigus semble liée à l’élimination des LB mémoires CD27+IgG+ spécifiques des Dsg. Des LB auto-réactifs Dsg restent détectables après RTX suite à la reconstitution lymphocytaire B, mais ces LB ont un phénotype naïf et non commuté (IgM) et ne secrètent plus d’IgG. En revanche, la persistance des LB auto-réactifs capables de sécréter des IgG anti-Dsg après traitement par CS est certainement à l’origine des rechutes fréquentes. L’analyse de l’expression génique ciblée à l’échelle unicellulaire a démontré qu’initialement, les LB spécifiques des Dsg ont un profil pro-inflammatoire avec l’expression de trois gènes codant pour les interleukines (IL)-1β, IL-12p35 et IL-23p19 et pour le gène de l’IRF5 (Interferon regulatory factor 5) par rapport aux LB non auto-réactifs. Le RTX et la CS ont des effets différents sur l'expression de ces gènes mais les deux réduisent l’expression génique d’IL-1β qui semble jouer un rôle important dans la physiopathologie du pemphigus. Parallèlement, l’analyse transcriptomique puis protéique des LB isolés des patients en rémission complète ou incomplète 6 ans après l’étude RITUX 1 a mis en évidence une augmentation d'expression de KCNN4 (Potassium calcium-activated channel subfamily N member 4) à la surface des LB chez les patients atteints de pemphigus en rémission complète pouvant influencer la maturation des LB
Pemphigus is an autoimmune disease of the skin and mucous membranes caused by autoantibodies (Ab) specific to desmoglein (Dsg) 1 or 3. These pathogenic Ab inhibit cell adhesion of keratinocytes. The development of pemphigus is associated with the conjunction of many uncommon events involving the emergence and then the cooperation of auto-reactive B cells and T cells link to genetic and environmental factors. Until now, the first line of treatment consisted of high doses of corticosteroids. Rituximab (RTX), an anti-CD20 chimeric monoclonal antibody, is an innovative therapy that results in B cells depletion. The RITUX 3 clinical trial was designed to evaluate the efficacy and safety of RTX combined with a short-course glucocorticoid therapy as a first-line treatment of pemphigus versus the standard treatment with standard corticosteroids (CS). As a first step, our clinico-biological analysis of patients after 24 months has shown that the use of RTX combined with short-term prednisone as a first-line treatment in patients with moderate to severe pemphigus is both more effective and better tolerated than the reference treatment with prednisone alone. Respectively, 89% of patients versus 34% in each group and both pemphigus foliaceus and pemphigus vulgaris patients responded. This efficacy was confirmed in the longer term after reconstitution of the B lymphocyte repertoire with a risk of relapse of only 2% at 36 months. The presence of a severe form of pemphigus at diagnosis (PDAI ≥ 45) and an anti-Dsg Ab level at 3 months above threshold values (anti-DSG1 ≥ 20 or anti-DSG3 ≥ 120) are associated with 50% risk of early relapse. These two predictive factors make it possible to identify a subgroup of patients at high risk of relapse requiring a maintenance infusion of RTX at the 6th month. In a second step, we studied the impact of RTX and CS treatments in patients with pemphigus in order to better understand the autoimmune response. The phenotypic characterization of auto-reactive B cells and the analysis of the frequency of B cells able of secreting anti-Dsg immunoglobulin (Ig) G by an ELISPOT approach demonstrated that the efficacy of RTX treatment in pemphigus seems related to the elimination of IgG-switched Dsg memory B-cells. Dsg specific B cells remain detectable after RTX when B cells return, but these B cells have a naïve and non-switched (IgM) phenotype and no longer secrete IgG. On the other hand, the persistence of self-reactive Dsg B cells capable of secreting IgG anti-Dsg after treatment with CS is certainly at the origin of the frequency of relapses. The unicellular targeted gene expression analysis demonstrated that initially, Dsg-specific B cells have a pro-inflammatory profile with the overexpression of three genes encoding Interleukin (IL) -1β, IL-12p35 and IL-23p19 and for the IRF5 gene (Interferon regulatory factor 5) compared to non-self-reactive B cells. RTX and CS have different effects on the expression of these genes, but both reduce the gene expression of IL-1β, which seems to play an important role in the pathophysiology of pemphigus

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Gustafsson, Gabriel. "Alpha-Synuclein Oligomers : Cellular Mechanisms and Aspects of Antibody Treatment." Doctoral thesis, 2017. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-326320.

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In Parkinson’s disease (PD) and dementia with Lewy bodies (DLB), aggregated α-synuclein deposit inside cells within the brain. Smaller soluble α-synuclein aggregates, oligomers, are present both intra- and extracellularly. The α-synuclein oligomers are known to be particularly harmful, although the underlying neurotoxic mechanisms are not fully understood. The aim of this thesis was to investigate the pathogenic roles of α-synuclein oligomers and the possibility to target such species with antibody treatment. Passive immunotherapy with α-synuclein antibodies can lead to reduced pathology and ameliorated symptoms in transgenic mice. However, it remains unknown whether the antibodies are taken up by cells or whether they act extracellularly. In Paper I, we assessed cellular internalization of various α-synuclein monoclonal antibodies. The oligomer selective mAb47 displayed the highest uptake, which was promoted by the extracellular presence of α-synuclein. Alpha-synuclein aggregates can be found in both neurons and glial cells, but the pathogenic role of glial deposits has only been sparsely investigated. In Paper II, co-cultures of neurons and glia were exposed to α-synuclein oligomers. The astrocytes in the cultures rapidly accumulated oligomers, which were only partially degraded by lysosomes. The sustained intracellular α-synuclein deposits were associated with mitochondrial stress reactions in the astrocytes. In Paper III, we sought to explore whether the astrocytic pathology induced by α-synuclein oligomers could be ameliorated by antibody treatment. Pre-incubation of oligomers with mAb47 promoted α-synuclein clearance, reduced astrocytic accumulation and rescued cells from mitochondrial stress. We could demonstrate that binding of the antibody to its antigen in the extracellular space was crucial for these effects to occur. The progressive pathology in PD is believed to be driven by cell-to-cell spreading of α-synuclein aggregates, potentially via exosomes and other extracellular vesicles (EVs). In Paper IV, we found that either fusing α-synuclein to a non-physiological protein tag or introducing the PD-causing A53T mutation directed α-synuclein towards EV secretion. Also, EV-associated α-synuclein was particularly prone to induce toxicity in recipient cells. In conclusion, this thesis sheds new light on the cellular dysfunction related to α-synuclein pathology and on how the underlying pathogenic processes may be targeted by antibody treatment.

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Shih-Jen, Liu, and 劉士任. "Treatment of B-cell Lymphoma with Chimeric Antibody-Cytokine Fusion Proteins." Thesis, 1998. http://ndltd.ncl.edu.tw/handle/20302514994859455580.

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博士
國防醫學院
生命科學研究所
86
Monoclonal antibodies have been applied to the treatment of cancer in a variety of ways. In this study, we have sought to combine targeting and immuno-potentiation activities by generating a fusion protein that possesses both tumor reactivity and cytokines function. Murine monoclonal antibody S5A8, which recognizes an idiotypic epitope expressed on a murine B cell lymphoma 38C13, was engineered to make anti-Id-cytokine fusion proteins. The heavy- and light-chain variable genes of S5A8 were cloned by PCR and ligated to the corresponding human constant genes in different Ab-expression vectors to produce chimeric S5A8 antibodies (chS5A8) and antibodies with cytokines attached to the carboxyl terminal of the heavy chain to produce chS5A8- IL-2, chS5A8-IL-4, and chS5A8-GM-CSF. The cloned VL and VH genes of S5A8 were ligated with a designed linker with the IL-2 gene conjugated 3'' end to the VH gene to make the single- chain Ab-IL-2 fusion protein (scFvS5A8-IL-2). All engineered proteins retained their specificity to recognize tumor idiotypic antigen, however, the binding affinity varied. The chS5A8, chS5A8- IL-2 , chS5A8-IL-4, and chS5A8-GM-CSF had similar binding affinity as compared with mouse S5A8, but the binding affinity of scFvS5A8-IL-2 was about 12-fold lower than the F(ab) fragment of S5A8. The chS5A8-IL-2, scFvS5A8-IL-2, and chS5A8-GM-CSF maintained full biological activity compared with recombinant IL-2 and GM-CSF, but chS5A8-IL-4 was 5-fold lower than recombinant IL-4. In pharmacokinetics analysis, the cytokine fusion proteins were cleared from the circulation rapidly than the parent antibody S5A8 or chS5A8. In vitro cytotoxicity revealed that chS5A-IL-2 and scFvS5A8-IL-2 could enhance lymphokine-activated-killer (LAK) cell-mediated tumor cell lysis. Furthermore, we showed that chS5A8-IL-2 was proficient in inhibiting tumor growth in vivo more effectively than a combination therapy with anti-Id Abs and IL-2, whereas scFvS5A8-IL-2 did not show any therapeutic effect. These results indicate that the anti-Id-IL-2 fusion protein represents a potent reagent for the treatment for B-cell lymphoma. The effector mechanisms involved in this tumor eradication are not dependent on T cells, since the therapeutic effect of chS5A8-IL2 was not altered in T cell-deficient SCID mice, even in syngeneic mice with depleted CD4+ or CD8+ T cell subsets. In contrast, NK cells are essential for the observed antitumor effect, since therapy with chS5A8-IL-2 is unable to induce tumor eradication in NK- depleted mice or NK function-deficient beige mice. Our data demonstrate that an immunotherapeutic approach using cytokine target by antibodies to tumor sites has a potent effect against B-cell lymphoma.

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WANG, XING-MIN, and 王興民. "Treatment of cancer with prodrugs activated by antibody targeted enzyme conjugates." Thesis, 1992. http://ndltd.ncl.edu.tw/handle/30176710188631192529.

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Yen, Ta-Li, and 閻大立. "Establishment of anti-TMPRSS2 antibody for diagnostic and treatment in prostate cancer." Thesis, 2019. http://ndltd.ncl.edu.tw/handle/ef5wxm.

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碩士
國立臺灣大學
生物化學暨分子生物學研究所
107
Androgen signaling and pericellular proteolysis play important roles in prostate cancer (PCa) progression and metastasis. Among them, type II transmembrane serine proteases II (TMPRSS2) receives an attention because we found that TMPRSS2 could promote PCa cells invasion, tumor growth and metastasis through activation of its proteolytic cascade and extracellular matrix degradation. TMPRSS2 has been also reported to be shed out from cells after its activation. Thus, TMPRSS2 may serve as a biomarker for PCa diagnosis/prognosis and a therapeutic target in PCa. In this thesis, I aimed to generate anti-TMPRSS2 antibodies with a potential of diagnosis, prognosis or therapeutic usages. I used a mammalian secretory expression system to express recombinant TMPRSS2 proteins in CHO cells and purify those proteins from the conditioned media. I successfully used the purified rTMPRSS2 to be antigens for polyclonal antibody generation from a guinea pig and a rabbit. Guinea pig anti-TM2 antibody could recognize the protease domain of TMPRSS2. Rabbit anti-TM2 polyclonal antibody showed a great specificity against TMPRSS2 in cell lysates and could also recognize the shed TMPRSS2 in the conditioned media of PCa cells and PCa patients’ urine samples. The establishment of neutralizing monoclonal antibodies against TMPRSS2 are still undergoing. Thus, the results together indicate that the anti-TM2 antibodies are successfully generated from the animals of guinea pig and rabbit with a potential of clinical application.

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Alghuneim, Arwa. "Evaluating the regulation of signaling pathways downstream of CD44 antibody treatment in AML." Thesis, 2019. http://hdl.handle.net/10754/656305.

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Acute myeloid leukemia (AML) is a subset of leukemia that is characterized by the clonal expansion of cytogenetically and molecularly abnormal myeloid blasts. These blasts are highly proliferative accumulating in bone marrow and blood which leads to severe infections, anemia, and bone marrow failure. The poor prognosis of AML patients caused by the low tolerance to intensive chemotherapy has encouraged the pursuit of alternative therapeutic approaches. Differentiation therapy which involves the use of agents that can release the differentiation block in these leukemic blasts has emerged as a promising therapeutic approach. The use of All-trans retinoic acid (ATRA) represents a successful example of such an approach, nonetheless its efficacy is restricted to one subtype of AML. Efforts have been focused on finding differentiation agents which are effective for the other more common AML subtypes. Anti-CD44 targeted antibodies that activate the CD44 cell surface antigen are a promising candidate. Previous studies have shown that anti-CD44 treatment has been able to release the differentiation block in AML1 through AML5 subtypes. The exact mechanism by which anti-CD44 treatment is able to induce its effects has not been fully elucidated. Recent studies highlight the role that epigenetic mechanisms play during haematopoiesis and leukemogenesis and therefore, in this work we investigated the epigenetic mechanisms associated with anti-CD44 induced differentiation. Using AML cell lines from different subtypes, we demonstrated that anti-CD44-induced differentiation results in an extensive change of histone modification levels. We found that inhibiting enzymes responsible for the H3K9ac, H3K4me, H3K9me, and H3K27me modifications, attenuated the anti-proliferative and differentiation promoting effects of antic-CD44 treatment. Taken together, these data highlight the promising potential of using anti-CD44 as a therapeutic agent across multiple subtypes in AML

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Dissertations / Theses on the topic 'Antibody treatment'

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