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Dissertationen zum Thema „Immunoglobulins“
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1
Das, Mrinmoy. „The regulatory effects of circulating normal immunoglobulins on autophagy and Th17 response“. Thesis, Paris 6, 2017. http://www.theses.fr/2017PA066153/document.
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Les immunoglobulines circulantes jouent un rôle critique dans l’homéostasie immune en modulant les fonctions des cellules du système immunitaire. Au cours de ma thèse, j’ai exploré les effets régulateurs des immunoglobulines G thérapeutiques (IVIG) et des immunoglobulines A monomériques circulantes (mIgA) sur l’autophagie et les réponses Th17 respectivement. Les IVIg sont une préparation thérapeutique d’IgG normales poolées. Elles ont utilisées comme agent anti-inflammatoire dans le traitement de maladies auto-immunes et inflammatoires variées. Cependant, les mécanismes ne sont pas complètement élucidés et plusieurs mécanismes mutuels et non exclusifs ont été proposés. L’autophagie est un important processus biologique impliquant la dégradation lysosomale des composants cellulaires endommagés et des protéines mal repliées. Il y a plusieurs preuves montrant l’implication de l’autophagie dans les maladies auto-immunes et auto-inflammatoires incluant la découverte de polymorphismes dans des gènes liés à l’autophagie. J’ai montré que l’induction de l’autophagie par les IVIG représente un nouveau mécanisme d’action permettant leur effet thérapeutique dans les maladies auto-immunes et inflammatoires. Les Th17 représentent une cible attractive pour traiter plusieurs maladies inflammatoires et auto-immunes. Malgré le fait qu’elles sont le deuxième anticorps le plus abondant dans la circulation, la function immunorégulatrice des IgA n’est relativement pas explorée. J’ai montré que les IgA monomériques (mIgA) inhibent la différentiation et l’amplification des cellules Th17 humaines et la production de leur cytokine effectrice IL-17ACirculating immunoglobulins play a critical role in the immune homeostasis by modulating the functions of immune cells. In my thesis, I investigated the regulatory effects of therapeutic immunoglobulin G (IVIG) and circulating monomeric immunoglobulin A (mIgA) on autophagy and human Th17 response respectively. IVIG is a therapeutic preparation of pooled normal IgG. It is used as an anti-inflammatory agent in the treatment of a wide variety of autoimmune and inflammatory diseases. However, the mechanisms are not yet fully elucidated and several mutually non-exclusive mechanisms have been proposed. Autophagy is an important biological process involving lysosomal degradation of damaged cellular components and misfolded proteins. There are several evidences that support the involvement of autophagy in autoimmune and auto- inflammatory disorders including the discovery of polymorphisms in autophagy-related genes. I show that induction of autophagy by IVIG represents a novel mechanism of action in achieving therapeutic effect in autoimmune and inflammatory diseases. Th17 cells represent an attractive target to treat several inflammatory and autoimmune diseases. Despite being second most abundant antibody in the circulation, the immunoregulatory function of IgA is relatively unexplored. I have shown that monomeric IgA (mIgA) inhibits differentiation and amplification of human Th17 cells and the production of their effector cytokine IL-17A
2
Almroth, Gabriel. „Immunoglobulins, immunoglobulin subclass-distributions and serologic markers in some renal and systemic disorders /“. Linköping : Univ, 2000. http://www.bibl.liu.se/liupubl/disp/disp2000/med646s.pdf.
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3
Rogers, Kenneth Alton. „Immunoglobulins and Immunoglobulin Fc Receptors in Nonhuman Primates Commonly Used in Biomedical Research“. Digital Archive @ GSU, 2006. http://digitalarchive.gsu.edu/biology_diss/6.
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Antibodies neutralize and eliminate pathogens, malignancies, and toxins by acting either alone or in association with Fc receptors which, once engaged, activate the elimination mechanisms of phagocytic cells. Based on structural differences, antibodies are divided into functionally distinct classes (IgM, IgD, IgG, IgE and IgA). Structure-function relationships within these classes are not well characterized. In addition, animal models for the assessment of potential therapeutic strategies for the modulation of the interaction between antibodies and Fc receptors are not established. Nonhuman primates are widely used to model human diseases and, represent excellent in vivo systems for this assessment. Therefore, we have studied nonhuman primate IgD as well as IgG and IgA specific Fc receptors in rhesus macaques, cynomolgus macaques, baboons and sooty mangabeys. IgD genes had not been identified in nonhuman primates nor the IgD receptors characterized in any species. We characterized IgD genes of the four monkey species, as well as chimpanzees and dogs. In contrast to other antibody classes, the IgD hinge regions are highly conserved between human and nonhuman primates, thus indicating a role in Fc receptor binding. In humans, Fc receptors CD16a (natural killer cells) and CD16b (neutrophils) bind IgG1 and IgG3, and CD89 (myeloid cells) binds IgA. To assess ligand binding and glycosylation properties of nonhuman primate CD16a, CD16b, and CD89, we sequenced, cloned, and generated recombinant molecules in a mammalian expression system. Our results verify the presence of CD16a, but not CD16b in nonhuman primates. CD16a is expressed on monocytes and a subpopulation of lymphocytes. In sooty mangabeys, CD16 is also expressed on neutrophils. Recombinant sooty mangabey/baboon CD16a binds to human IgG1 and IgG2, but not IgG3 and IgG4. Monkey CD89 has the same peripheral blood leukocyte expression profiles as humans, and binds human and recombinant macaque IgA. Blocking of N-glycans inhibited expression of CD89, but only marginally CD16a expression. Although extensive similarities of antibody/Fc receptor interactions exist between human and nonhuman primates, several differences must be considered when evaluating therapeutic strategies. However, these differences can be exploited to further characterize the structure-function relationships existing within antibody molecules and respective receptors.
4
Teng, Su Fern. „Immunoglobulins binding ligands“. Thesis, University of Cambridge, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.627345.
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5
Ding, Cheng. „Siglec-G is a negative regulator of NF-[kappa]B activation and has pivotal roles in B-1 cell development and resistance to sepsis /“. Columbus, Ohio : Ohio State University, 2008. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1226876722.
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6
Smith, David G. E. „Activities of anti-lipopolysaccharide immunoglobulins“. Thesis, University of Edinburgh, 1989. http://hdl.handle.net/1842/19300.
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7
Barua, Utpal. „Allergen specific immunoglobulins during pregnancy“. Thesis, Sheffield Hallam University, 1993. http://shura.shu.ac.uk/19325/.
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In this study the serum concentration of IgE and IgG4 (total and allergen specific taking Timothy grass pollen as the model allergen) have been investigated prospectively during and after pregnancy in healthy women and women suffering from allergic rhinitis. The results show that the total serum IgG concentration remained unchanged in both groups during pregnancy. There was no significant difference in the serum concentration of IgG 4 between pregnant allergic women and non-pregnant allergic women. Levels of IgG4 were approximately twice as high (p< 0.01) in non-allergic pregnant women compared to the non-allergic nonpregnant control group. Total IgG4 concentrations were similar in allergic and non-allergic women during pregnancy; however, in the non-pregnant state allergic women had significantly (p = 0.017) higher levels of IgG4 than non-allergic women. The results show that both during pregnancy and in the non-pregnant state there was a highly significantly (p< 0.001) greater serum concentration of total IgE in allergic than non-allergic subjects. Although the level of IgE was significantly (p=0.004) lower in pregnancy, the differences are relatively small and seem unlikely to be of great physiological significance. Allergic symptomatology did not correspond to IgE levels during pregnancy. In both the pregnant and non-pregnant women the serum concentration of antigen-specific IgE was highly significantly greater in the allergic than the non-allergic subjects. However, in allergic women, the concentration of antigen-specific IgE was very much lower during pregnancy, at about 6% of the non-pregnant level (p < 0.001). The concentration of antigen-specific IgG4 was also reduced in pregnancy in allergy sufferers, being about half of the level found in the non-pregnant individuals (p < 0.001). There appeared to be an increase in spontaneous first trimester abortion in women who suffered symptoms of allergy. From the case histories of all 418 pregnancies at the Langold Health Centre ante-natal clinic attending between September 1976 and December 1990, 192 were to allergy sufferers and 226 were to normal women. The abortion rate was 16.7% in the allergic group and 5.3% in the normal pregnant women (p < 0.001).
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Jeffrey, Philip D. „The structure and specificity of immunoglobulins“. Thesis, University of Oxford, 1989. http://ora.ox.ac.uk/objects/uuid:79bcae45-289d-4401-9916-d719bc2751a4.
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An investigation into the structures of the antigen-binding fragments (Fab) of mouse monoclonal immunoglobulins by X-ray crystallography, is presented. The family of immunoglobulins studied, Gloops 1-5, possess the ability to bind to both the peptide antigen and the parent protein: Hen Egg-white Lysozyme. The Gloop1 and Gloop4 Fabs were generated by proteolytic cleavage of the antibody, and crystallisation trials yielded crystals of uncomplexed Fabs of both species. Attempts to grow Fab:antigen complex crystals proved unsuccessful. Partial purification of the heterogeneous Gloop4 Fab was found to be essential for the success of the crystallisations. Data were collected on crystal forms of Gloop1, Gloop2 and Gloop4 on an area detector. The structures of three Gloop2 crystal forms were solved by the molecular replacement technique, using models consisting of Fab domain pairs. Two of the crystal forms were refined by molecular dynamics methods at maximum resolutions of 3.3Å and 2.8Å, and the third by rigid body methods alone at 3.5Å resolution. Analysis of the Gloop2 structures between the crystal forms and with other Fabs indicated no atypical features in the domains. The elbow bend of the Gloop2 Fabs differed by up to 7°. The relative association of variable and constant domain pairs was also seen to vary between the Fabs. The changes in domain pairing caused significant differences in the relative disposition of the complementarity determining regions (CDRs), although there was no evidence for conformational change within individual CDRs. The Gloop2 combining site is dominated by a groove of approximate dimensions 12Åx9Åx7Å, containing many aromatic side-chains and a pair of glutamic acid residues in analogous positions to a those found in a FabrHen egg-white Lysozyme complex.
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Prinsloo, Earl Adin Gerard. „An investigation of the binding capacities of recombinant domain mutants of the human Polymeric Immunoglobulin Receptor (pIgR)“. Thesis, Nelson Mandela Metropolitan University, 2006. http://hdl.handle.net/10948/403.
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The membrane bound glycoprotein, polymeric immunoglobulin receptor (pIgR) is the primary transport molecule of the polymeric immunoglobulins, dimeric IgA and pentameric IgM, across epithelial cells. This process, known as transcytosis, is essential in order to establish immunity at mucosal surfaces. Typically, pIgR binds to the polymeric immunoglobulin at the basolateral surface of the epithelial cell, via five homologous immunoglobulin-like domains of the ectodomain. Binding is covalent to IgA and non-covalent to IgM; the IgM binding varying among species. The pIgR-bound complex is released at the apical surface of the cell after cleavage of pIgR at Arg585, thereafter referred to as secretory component (SC). SC confers protective and immunologic functions to the polymeric immunoglobulin. Free SC, i.e. not complexed with polymeric immunoglobulins, is also known to be released into mucosal secretions; and binds to pathogenic bacteria and bacterial products. It is known that domain I of the ectodomain is the primary domain in the interaction with polymeric immunoglobulins, while domain V is involved in a covalent linkage with IgA. However, little is known of domains II-IV and their role in immunoglobulin binding, particularly to IgM. This study aimed to characterize the binding of recombinant human pIgR domain mutants to polymeric IgM using immunological, biophysical and cell based techniques; thereby allowing greater insight into the contribution of each of the five domains. The unique domain structure allowed for selective amplification of single and multiple domain mutants from cloned human PIGR ectodomain cDNA. Mutants were cloned and expressed in Esherichia coli BL21 (DE3) as inclusion bodies. Recombinant mutant proteins were refolded in vitro by equilibrium gradient dialysis and purified to homogeneity. Equilibrium binding data show significant contributions to specific binding as a factor of domain presence. Binding kinetics determined by biophysical surface plasmon resonance measurements show the interplay between association and dissociation rates as defined by individual domains. In vitro competitive binding studies using the human intestinal carcinoma, HT29, known to constitutively express pIgR, show that the constructed recombinant domain mutants outcompete native pIgR. The level of competition is shown to be dependant on the domains downstream of domain I. The data also confirm the biological activity of the first in vitro refolded recombinant human SC.
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李瑞山 und Shui-shan Lee. „Anti-neutrophil cytoplasmic antibody: a clinical & experimental study“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1993. http://hub.hku.hk/bib/B31981513.
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11
Lee, Shui-shan. „Anti-neutrophil cytoplasmic antibody : a clinical & experimental study /“. [Hong Kong : University of Hong Kong], 1993. http://sunzi.lib.hku.hk/hkuto/record.jsp?B13762679.
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12
Hirano, Ayumi. „T dependent B cell help in cattle : immunoregulatory function of interleukin-4 and CD40-CD40L interactions /“. free to MU campus, to others for purchase, 1997. http://wwwlib.umi.com/cr/mo/fullcit?p9841150.
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13
Rodríguez, Lobato Luis Gerardo. „Factores pronósticos en pacientes con amiloidosis sistémica por cadenas ligeras“. Doctoral thesis, Universitat de Barcelona, 2018. http://hdl.handle.net/10803/663901.
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La amiloidosis de cadenas ligeras (AL) es una neoplasia rara, que se caracteriza por la presencia de células plasmáticas (CP) clonales en la médula ósea (MO), las cuales producen una inmunoglobulina (Ig) de cadena ligera monoclonal frecuentemente de isotipo lambda. Las Ig de cadena ligera amiloidogénicas se agregan y se depositan en el tejido como fibrillas de amiloide. La presentación clínica es muy heterogénea y el diagnóstico requiere una alto índice de sospecha. El pronóstico depende principalmente del órgano afecto, así como del tamaño y biología de la CP clonal. El objetivo del tratamiento es suprimir la producción de la proteína precursora de la fibrilla generada por la clona de CP, y minimizar la toxicidad relacionada al tratamiento. Alcanzar una adecuada respuesta hematológica se asocia a una prolongación de la supervivencia; sin embargo, para muchos pacientes aún es una enfermedad con un pronóstico ominoso. Existen otros factores de riesgo potenciales aún no estudiados, como la aparición de bandas oligoclonales (BOC), la presencia de inmunoparesia (IP) al diagnóstico y el porcentaje de infiltración de la MO por CP, aún no completamente bien caracterizados en esta patología.
Los objetivos de la presente tesis doctoral fueron: Determinar la incidencia, historia natural e impacto pronóstico de las BOC tras el tratamiento de pacientes con amiloidosis AL. Investigar el impacto pronóstico en términos de supervivencia y respuesta a tratamiento de la IP al dianóstico. Analizar la correlación entre el porcentaje de infiltración de la MO por CP y características clínicas de los pacientes, así como los resultados clínicos en términos de supervivencia.
En el primer trabajo publicado, analizamos el papel de las BOC, definidas como la presencia de una banda monoclonal detectada por inmunofijación de proteínas séricas y/o urinarias, distinta a la proteína M original al diagnóstico, ya sea en la cadena pesada y/o ligera. Descubrimos por primera vez que la incidencia de BOC, después de la terapia de primera línea, es incluso mayor que en pacientes con mieloma múltiple. Este fenómeno humoral oligoclonal fue más frecuente en los pacientes que lograron una respuesta completa. Aquellos pacientes que mantuvieron las BOC durante más de un año tuvieron un mejor pronóstico, incuso en aquellos con características de mal pronóstico (estratificación de riesgo de la Clínica Mayo del 2004), reflejando la importancia de una respuesta inmune humoral más robusta en amiloidosis AL.
En el segundo trabajo publicado, analizamos el papel de la IP definida como la supresión de todas las Ig no involucradas por debajo del límite inferior de la normalidad para nuestro laboratorio. Demostramos que la presencia de IP en el momento del diagnóstico, tiene un impacto negativo en la supervivencia, especialmente en etapas tempranas de la enfermedad, y podría ser un indicador pronóstico discriminatorio adicional. Además, demostramos que la recuperación de la IP un año después del inicio del tratamiento podría ser un marcador independiente de supervivencia a largo plazo.
En el tercer y último trabajo publicado, analizamos la correlación entre la infiltración de CPMO, las características clínicas y los resultados de la amiloidosis AL. En este trabajo, demostramos que una mayor infiltración de CPMO (>10%) se asoció con un mayor daño sistémico orgánico, en particular, afectación cardíaca, una mayor mortalidad temprana y un peor pronóstico, y rara vez, se relacionó con el desarrollo de las características del mieloma múltiple. Curiosamente, los pacientes con menor infiltración de CPMO se asoció con una mayor prevalencia de daño renal.Systemic immunoglobulin light chain amyloidosis (AL) is a rare plasma cell neoplasm, characterized by a clonal population of bone marrow (BM) plasma cells (PC) that produces a monoclonal immunoglobulin (Ig) light-chain, more frequently of the lambda isotype. The amyloidogenic Ig light-chains aggregate and deposit in tissue as amyloid fibrils with a predominant β-pleated sheet structure. The clinical presentation is heterogeneous, and its diagnosis requires a high index of suspicion. Prognosis depends on the organ involvement as well as the size and biology of the plasma cell clone. The treatment goal is to suppress the production of the fibril precursor protein by the underlying plasma cell clone, while minimizing the treatment-related toxicity. In the first publication, we studied the oligoclonal bands (OB), defined as the presence of a serum and/or urine immunofixation electrophoresis (IFE) monoclonal spike that was different from the original M-protein either in heavy and/or light chains or with a different IFE migration pattern. We uncovered for first-time that the incidence of OB after fist-line therapy is even higher than in patients with multiple myeloma. This oligoclonal humoral phenomenon was more frequent in patients who achieved a complete response, and those patients with OB for more than one year had a significantly better outcome, even in those with poor prognostic features, thereby reflecting the relevance of a more robust humoral immune response. The second biomarker was immunoparesis (IP) defined as the suppression of all uninvolved the immunoglobulins below the lower limit of normality. We showed that the presence of IP at diagnosis could have a negative impact on survival, especially in early stages of the disease, and could be an additional powerful discriminatory prognostic indicator. Also, we showed that the recovery of IP one-year after treatment onset could be an independent long-term marker of survival. In the third and last publication, we analyze the correlation between the BMPC infiltration, clinical features and outcomes in AL amyloidosis. We showed that a higher BMPC infiltration (>10%) was associated with increased systemic organ damage (cardiac involvemen), a higher early mortality, and worst prognosis, being rarely related to the development of myeloma features.
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Didriksen, Nancy A. (Nancy Andrews). „Psychological Stress: Effect on Humoral Immune Functioning as Measured by Immunoglobulin Levels“. Thesis, North Texas State University, 1986. https://digital.library.unt.edu/ark:/67531/metadc331278/.
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The purpose of the present study was to determine if psychological stress, defined as academic examination stress, would systematically produce changes in immune parameters (immunoglobulin concentration) and psychological functioning. It was hypothesized that as examination stress occurred there would be an effect on immunological function consistent with heightened psychological activity/stress. Subjects were 23 master's and doctoral students in psychology who volunteered for the research project. All subjects were administered a series of psychological tests to measure stress, personality factors, emotional states, and anxiety levels. All tests were administered and.blood samples drawn over a period of 15 months across two lowstress and two high-stress periods. Immunological tests included white blood cell (WBC) differential count and radial immunodiffusion (RID) for the determination of concentration of different immunoglobulin classes (IgA, IgG, IgM) in serum. Data were treated to a one-way analysis of variance (ANOVA) with repeated measures, t /test for correlated samples correlational matrix between variables across assessments and discriminant function analysis. Results showed (1) increased immunoglobulin levels during periods of stress; (2) immunoglobulin G most consistently related to stress and probably most indicative of the stressed condition and biological resistance to stress; (3) anxiety related to external events; (4) increase in anxiety under stress; and (5) anxiety inversely correlated with emotional stability and coping skills while positively related to tension, increased number of somatic complaints, and obsessive-compulsive trends. Firm support was provided for the hypothesis that as stress occurred, there would be consistent changes in immunological functioning associated with heightened psychological activity/stress. It was concluded that a response pattern to stress was adaptive along both psychological- and immunological dimensions and that the concept of bodymind interaction was the most realistic approach to understanding the total response patterns.
15
Laurencikiene, Jurga. „Regulation of germline transcription in the immunoglobulin heavy chain locus /“. Stockholm, 2004. http://diss.kib.ki.se/2004/91-7349-989-7/.
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16
Mohamed, Alahlafi Abdelaziz H. „The significance of immune reactants deposited in the basement membrane zone of the skin in patients with lupus erythematosus“. Thesis, University of Oxford, 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.275408.
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17
Head, Jared G. „Structure and function of the #beta#-sheet proteins, titin and CD2“. Thesis, University of Bristol, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.265387.
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Costello, Mark Eugene. „Growth and productivity of hybridoma cell lines in vitro“. Thesis, Manchester Metropolitan University, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.280629.
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Davies, Julian. „Antibody VH domains as small recognition units“. Thesis, University of Cambridge, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.263483.
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20
Kwong, Wai-ip Eric. „Development of an in-house anti-Ro52 ELISA and a study of the clinical applications of anti-Ro52 in rheumatic diseases“. Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B42925009.
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21
Ng, Kin-man. „Application of p63 4A4 antibody in detection of cervical cancer and precursors in cytology samples“. Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B42905357.
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22
Engström, Gunilla Norhagen. „Mucosal conditions in immunodeficient individuals with special emphasis on salivary immunoglobulins“. Stockholm : Department of Clinical Immunology, and the Department of Periodontology, Karolinska Institutet, 1993. http://catalog.hathitrust.org/api/volumes/oclc/28436070.html.
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Low, Douglas Graham. „NMR investigation of molecular contacts in a hapten-antibody complex“. Thesis, University of St Andrews, 1996. http://hdl.handle.net/10023/13986.
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An antibody Fv fragment was cloned from hybridoma cells of a monoclonal antibody against the steroid derivative estrone-3-glucuronide (E3G). The detection of this molecule is important in a method for predicting fertility. The variable heavy and light chain genes were isolated and amplified and placed into a modified pUC 19 vector to allow expression of active protein. The immunochemistry of the fragment was determined by ELISA, demonstrating that the fragment did bind E3G, that this binding was specific, the antibody fragment competed against the parent monoclonal antibody, and that the fragment could inhibit binding of the monoclonal antibody to an E3G-alkaline phosphotase conjugate. The expression of the fragment in E. coli was optimised in various media with levels of 4 mg.L-1 achieved. The structure of E3G was probed in solution and in complex with the Fv fragment, utilising E3G with a fully labelled glucuronic acid moiety: estrone-[U-13C]-glucuronide. Firstly in the in the solution structure studies by giving 13C and 1H assignments via a 13C-13C COSY and a 1H-13C HSQC, and secondly in the bound state by allowing the use of isotope editing techniques. In solution the glycan was found to exist in multiple conformations, with particularly large fluctuations about the glycosidic linkage. The antibody selects a conformation from the free solution which does not correspond to either of the two minimum energy conformations of the free glycan. The glucuronic acid moiety undergoes a stacking interaction with an aromatic ring in the binding site, and both NOEs and ring current shifts are in good agreement with the predicted bound state conformation. The predicted bound state conformation is also in good agreement with preliminary X-ray data. A homology modelled protein structure of the Fv was built and compared with the X-ray crystal structure.
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Stephens, Lauren Ellis. „Interaction of immunoglobulins with primate Fc[gamma]RIIIa“. Thesis, University of Cambridge, 2014. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.708276.
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Al-Lazikani, Bissan. „Canonical structures of immunoglobulins and T cell receptors“. Thesis, University of Cambridge, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.624098.
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Krick, Kari Elizabeth. „Exercise and Immunodeficiency Affect Immunoglobulins in Endurance Horses“. Thesis, Virginia Tech, 2002. http://hdl.handle.net/10919/43971.
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Two studies were conducted on endurance horses predominantly of Arabian breeding participating in an 80 km ride dedicated to research in April 2001 (Trial 1) and April 2002 (Trial 2). Objectives were to determine effects of endurance exercise, antioxidant supplementation, and a feed rich in fiber and fat vs. a high fat sweet feed on immunoglobulin A and G concentrations as well as identify selective IgA deficiency in endurance horses of Arabian breeding. There were no effects of distance in Trial 1 on IgA (P = 0.73) or IgG (P = 0.18) concentrations. In Trial 2, IgA concentrations increased (P = 0.05) and IgG concentrations increased (P = 0.006) after the start of the race. There were no effects of antioxidant supplementation on IgA (P = 0.16), IgG (P = 0.16), and IgM (P = 0.70) concentrations. There were no diet effects on IgA (P = 0.80), IgG (P = 0.59), and IgM (P = 0.54) concentrations. There were horses in both trials that were deficient in IgA only. Concentrations of IgG and IgM were within normal ranges, and there were no differences in training, performance and transportation variables, IgG concentrations, antioxidant supplementation, and feed supplementation compared to the horses with normal IgA concentrations. The concentration of IgM was higher in IgA deficient horses in Trial 1 (P = 0.035) and Trial 2 (P = 0.017). Horses with deficient IgA tended to be associated with health problems commonly found in humans and dogs affected with selective IgA deficiency.Master of Science
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Velez, Maria-Gabriela. „Development and function of allelically included B cells /“. Connect to full text via ProQuest. Limited to UCD Anschutz Medical Campus, 2008.
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Thesis (Ph.D. in Immunology) -- University of Colorado Denver, 2008.Typescript. Includes bibliographical references (leaves 153-162). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;
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Arasteh, Nikoo. „Passive immunization of rainbow trout with chicken immunoglobulins (IgY)“. Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0011/NQ56496.pdf.
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Galens, Kevin. „Knowledge based structure modeling of the third hypervariable region of antibodies /“. Link to online version, 2006. https://ritdml.rit.edu/dspace/handle/1850/1877.
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Van, Zyl Dwain George. „Production of recombinant human CD21 and CD23 : towards a better understanding of their interaction“. Thesis, Nelson Mandela Metropolitan University, 2013. http://hdl.handle.net/10948/d10211135.
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The prevalence of allergic diseases has dramatically increased over the last three decades. Presently, it is estimated that 20-30 per cent of the developed world suffers from allergic diseases. The majority of allergic diseases are rooted in the activities of IgE; an immunoglobulin which exerts its effector functions by interacting with a network of proteins. This network includes its low affinity receptor CD23. Cross linking of membrane IgE and CD21 by soluble CD23 results in an increase in IgE synthesis. This marks the interaction between CD23 and CD21 as an attractive therapeutic target. However, details regarding this interaction are inadequate for rational drug design. To obtain a deeper understanding of the CD23-CD21 interaction recombinant human CD21 (SCR1-2 and SCR5-8) and CD23 (16 kD and 25 kDa) were produced. The cloning, expression and purification of recombinant proteins comprised a significant portion of this study. Recombinant CD23 was expressed as inclusion bodies, refolded by rapid dilution and purified by size exclusion chromatography. Conversely, recombinant CD21 was expressed as soluble MBP-fusions and purified with an amylose affinity resin. The interaction between recombinant CD23 and CD21 was analysed by flow cytometry and ELISA experiments. Flow cytometry showed that 16 kDa and 25 kDa CD23 interacted with SCR5-8 to the same extent. Semi-quantitative ELISA experiments showed that both SCR1-2 and SCR5-8 were able to interact with 16 kDa and 25 kDa CD23. This suggests that the binding sites of SCR1-2 and SCR5-8 occur on 16 kDa CD23. Furthermore, since proteins were expressed in E. coli it suggests that the CD23-CD21 interaction does not require glycosylation. Furthermore, considering what is known about the SCR1-2-CD23 interaction from previous NMR studies; i.e. that the C-terminal tail (residues residues 289-298) of CD23 is responsible for binding SCR1-2, indicates that SCR5-8 binds somewhere within the lectin domain of CD23. This indicates that the CD23-CD21 interaction involves C-terminal tail-SCR1-2 and lectin domain-SCR5-8 interactions.
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梁子明 und Tze-ming Leung. „Immunological studies of a phosphorylcholine-specific antibody using gene transfer techniques“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1994. http://hub.hku.hk/bib/B31233740.
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Kwong, Wai-ip Eric, und 鄺偉業. „Development of an in-house anti-Ro52 ELISA and a study of the clinicalapplications of anti-Ro52 in rheumatic diseases“. Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2009. http://hub.hku.hk/bib/B42925009.
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Upton, I. „The immune response of the pig to infections with Steptococcus suis type II“. Thesis, University of Cambridge, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.377260.
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34
Bidgood, Susanna Ruth. „Antibody mediated intracellular immunity“. Thesis, University of Cambridge, 2013. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.648288.
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Wright, Deborah Emma. „A study into the effect of antibody on murine gammaherpesvirus 68 replication“. Thesis, University of Cambridge, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.609834.
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Leung, Tze-ming. „Immunological studies of a phosphorylcholine-specific antibody using gene transfer techniques /“. [Hong Kong] : University of Hong Kong, 1994. http://sunzi.lib.hku.hk/hkuto/record.jsp?B13762680.
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Engström, Per-Erik. „Mucosal and systemic antibody networks with special emphasis on IgA subclasses“. Stockholm : Department of Clinical Immunology and the Department of Periodontology, Karolinska Institute, 1993. http://catalog.hathitrust.org/api/volumes/oclc/28436118.html.
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Dooley, Helen. „Characterisation of single domain antibody fragments from the nurse shark Ginglymostoma cirratum, using phage display“. Thesis, University of Aberdeen, 2001. http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=219953.
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Navarro, López Alba. „Estudi de l'estat mutacional dels gens de les immunoglobulines i heterogeneïtat clínico biològica del linfoma de les cèl•lules del mantell“. Doctoral thesis, Universitat de Barcelona, 2012. http://hdl.handle.net/10803/91069.
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El limfoma de cèl•lules del mantell (LCM) és una neoplàsia heterogènia en relació a les característiques clíniques, biològiques i moleculars que presenten els pacients. La morfologia, l’immunofenotip i el genotip no són suficients per explicar aquesta gran diversitat. Encara que es considera una neoplàsia agressiva alguns pacients segueixen un curs clínic indolent sense la necessitat de tractament en el moment del diagnòstic i presenten una supervivència llarga.
Els resultats d’aquesta tesi demostren que els LCM amb elevada càrrega mutacional en els gens de les immunoglobulines (IGHV)(<97% d’identitat amb la línia germinal) utilitzen preferencialment certes famílies de gens IGHV, suggerint la presència de processos de selecció antigènica com a mecanisme de patogènesi d’aquesta neoplàsia i donant importància a la caracterització del receptor de les cèl•lules B (BCR).
Els LCM amb gens IGHV mutats (M-LCM) presenten una menor complexitat genòmica, són preferencialment negatius per a l’expressió del factor de transcripció SOX11 i presenten afectació ganglionar amb més freqüència que els casos sense mutacions (U-LCM). Per altra banda, el perfil d’expressió gènica entre casos amb i sense mutacions en els gens IGHV, també mostra expressió diferencial de programes cel•lulars relacionats amb poblacions de cèl•lules B normals incidint amb la ontogènia d’ambdós grups. El perfil d’expressió de microRNAs també permet diferenciar entre dos grups de pacients amb diferent supervivència, reforçant la idea de la presència de dos subgrups que pertanyen a una mateixa entitat.
La identificació i la correcte identificació d’aquests pacients de limfoma de cèl•lules del mantell són rellevants per al posterior maneig i tractament dels mateixos.Mantle cell lymphoma is a malignant and heterogeneous disease regarding the clinical, biological and molecular characteristics presented by patients, all of them with the translocation t(11;14) and CCND1 overexpression. Although is considered an aggressive neoplasm some patients follow and indolent clinical course without treatment at the time of diagnosis and long survival. The results of this studies demonstrate that LCM with high mutational load in the immunoglobulins genes (IGHV) (<97% of identity with the germ line) use preferentially specific IGHV gene families, suggesting the presence of antigenic selection process as a mechanism of pathogenesis of this neoplasm and giving importance to the characterization of the B cell receptor. The LCM cases with IGHV mutated genes (M-LCM) present less genomic complexity are preferentially negative for the expression of the transcriptor factor SOX11 and also present lymphadenopathies more frequently compared with the IGHV unmutated cases (U-LCM). Furthermore, the gene expression profile between cases with and without IGHV mutations also shows differential expression of cellular programs related to normal B cell populations maybe related with the ontogeny for both groups. The expression profile of microRNAs also differentiates between two groups of patients with different survival and other molecular characteristics, reinforcing the idea of the presence of two subgroups that belong to the same entity. The correct identification and classification of these patients are relevant for subsequent handling and treatment of the patients.
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Fernández, Codina Andreu. „Malaltia relacionada amb la IgG4“. Doctoral thesis, Universitat Autònoma de Barcelona, 2019. http://hdl.handle.net/10803/667368.
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La malaltia relacionada amb la IgG4 (IgG4-RD) és una patologia fibroinflamatòria
autoimmune sistèmica, caracteritzada pel desenvolupament de masses
fibròtiques que poden portar a l’alteració de l’arquitectura dels teixits i a la
disfunció orgànica. En aquesta tesi doctoral s’ha estudiat per primera vegada
aquesta malaltia a Espanya, creant un registre nacional, basat en els criteris de
consens internacional en patologia en IgG4-RD. El primer estudi ha mostrat que
els pacients amb IgG4-RD han estat predominantment homes d’edat mitjana. La
malaltia va afectar múltiples òrgans en un 47% dels casos. Les zones més
freqüentment afectades van ser el retroperitoneu, l’òrbita, les glàndules salivals i
el pàncrees. Després de completar un primer tractament, el 39% dels pacients
van presentar una recidiva. El segon estudi va mostrar que només un 55% dels
participants tenien valors en sèrum de IgG4 >135 mg/dL. Els tractaments més
utilitzats (constituint un 73% dels realitzats) van ser glucocorticoides (GC), GC
amb cirurgia, GC amb azatioprina , i cirurgia sola. Tots els tractaments utilitzats
van tenir un elevat grau de resposta però amb freqüents recidives. L’IgG4
responder index (IgG4-RI) modificat va correspondre’s amb l’activitat de la
malaltia. Cap pacient va desenvolupar càncer en el primer any des del diagnòstic
de la malaltia. En conclusió, les característiques de la IgG4-RD a Espanya
s’ajusten a les descrites a la literatura. L’elevació sèrica de IgG4 va ser inferior
que en poblacions asiàtiques. El tractament amb GC va ser el més utilitzat, però
els fàrmacs antireumàtics modificadors de la malaltia podrien ser d’utilitat.
L’IgG4-RI modificat va ser útil per valorar la resposta. La incidència de neoplàsies
va ser escassa.IgG4-Related disease (IgG4-RD) is a rare systemic autoimmune fibroinflammatory condition, characterized by the development of fibrotic masses and organ dysfunction. In this doctoral thesis, IgG4-RD was studied in Spanish patients for the first time creating a nationwide registry, based on the international consensus criteria on IgG4-RD pathology. The first study showed that patients with IgG4-RD were predominantly middle-aged men. The disease involved multiple organs in 47% of the cases. The most frequently involved zones were retroperitoneum, orbit, salivary glands and pancreas. After completing the first treatment, 39% of the patients had flares. The second study found that only 55% of the participants had serum IgG4 levels over 135 mg/dL. The most commonly used treatments (73% of all) were glucocorticoids (GC), GC with surgery, GC with azathioprine, and surgery alone. They all had a high response rates, but relapses were frequent. The modified IgG4-responder index (IgG4-RI) corresponded to the disease’s activity. No patients developed cancer in the first year after the disease’s diagnosis. In conclusion, IgG4-RD characteristics in Spain were similar to the ones described in the literature. Serum IgG4 elevation was lower than in Asian populations. Treatment with GC was the most frequently used, but disease modifying antirheumatic drugs could be useful. Modified IgG4-RI was useful to monitor the outcomes. The incidence of malignancy was low.
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Benadda, Samira. „Le rôle du système endosomal dans la fonction des récepteurs activateurs aux fragments Fc des immnunoglobulines : exemples du RFcyI“. Thesis, Université de Paris (2019-....), 2019. http://www.theses.fr/2019UNIP7075.
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Les Récepteurs aux fragments Fcs des immunoglobulines (RFcs) sont des récepteurs majeurs du système immunitaire, présents à la surface des cellules immunes et faisant l’interface entre l’immunité innée et l’immunité adaptative. Ils reconnaissent la partie Fc des immunoglobulines des complexes immuns (IC) constitués d’un antigène et d’une immunoglobuline spécifique déclenchant des voies de signalisation qui déterminent la production de médiateurs anti- ou pro-inflammatoires ainsi que d’autres réactions permettant l’élimination de l’infection. Après la liaison de la partie Fc des ICs, le récepteur et les ICs sont internalisés. Il est connu que cette internalisation permet la présentation de l’antigène contenu dans les ICs et l’élimination de pathogènes internalisés, mais le rôle de l’internalisation dans la transduction du signal via les RFcs n’avait pas été bien étudié auparavant. Nous avons étudié le rôle du système endosomal dans la fonction des RFcs et montré que le RFcγI, récepteur de haute affinité aux IgG, continue à signaler via des plates-formes de signalisation endosomale, ce qui serait un mécanisme essentiel pour l'activation complète des fonctions clés des RFcs. Les endosomes caractérisés par l’aminopeptidase insulinodépendante IRAP constituerait une plateforme de signalisation pour le RFcγI. De manière similaire, nous avons mis en évidence que la sous unité CD3ζ du TCR, récepteur au lymphocyte T, forme un pool intracellulaire dans les endosomes contenant IRAP et la syntaxine STX6. Nos résultats montrent que le TCR continue de signaler après l'endocytose et que cette signalisation intracellulaire est particulièrement importante pour l’activation de la cellule T par des complexes peptide-CMH de faible affinitéImmunoglobulin Fc receptors (FcR) are cell surface immune receptors at the interface between innate and adaptive immunity. They recognize the Fc part of the immunoglobulin of the immune complexes consisting of an antigen and a specific immunoglobulin. They trigger signaling pathways that determine the production of anti- or pro-inflammatory mediators and other reactions allowing the elimination of infection. After the binding of ICs, the receptor and the ICs are internalized. It is known that this internalization allows the presentation of the antigen contained in the ICs and the elimination of internalized pathogens, but the role of internalization in signal transduction via RFCs has not been well studied before.We investigated the role of the endosomal system in FcR function and we showed that the FcγR1, a high affinity IgG receptor, continues to signal via endosomal signaling platfrorms, which would be essential for the complete activation of the receptor functions. The endosomes characterized by insulin-dependent aminopeptidase IRAP constituted a signaling platform for RFcγI. Similarly, we have demonstrated that the CD3ζ subunit of the TCR, the lymphocyte T receptor, form an intracellular pool in the endosomes containing IRAP and the syntaxine STX6. Our results demonstrate that the TCR continues to signal after endocytosis and that this intracellular signaling is particularly important for T cell activation by low affinity peptide-MHC complexes
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Mohammed, Mansoor S. „Uptake of human immunoglobulins into the egg yolk of hens“. Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape8/PQDD_0004/NQ43267.pdf.
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Maury, Michael [Verfasser]. „Aggregation and structural changes in spray dried immunoglobulins / Michael Maury“. Aachen : Shaker, 2005. http://d-nb.info/1186577290/34.
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Sedano, Rodolfo Canseco. „Effect of immunoglobulins on early bovine embryo development in vitro“. Thesis, Virginia Tech, 1985. http://hdl.handle.net/10919/41575.
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Bovine morulae (day 6: n=257) were obtained to evaluate
the [effect of immunoglobulins (Ig) on early bovine embryo
development in vitro. Fifty-four cows superovulations were
conducted in 36 cows with follicle stimulating hormone.
Embryos were collected by non-surgical procedures and
morphologically evaluated and randomly assigned to culture.
Embryos were cultured in Ham's F-10 containing 10%
(6.4 mg/ml) steer serum (SS), 1% (.64 mg/ml) bovine gamma .
globulins (GG), 1% (.64 mg/ml) bovine IgG, 1% (.64 mg/ml)
bovine 1gM, 10% SS plus 1% GG, 10% SS plus 1% 1gG, or 10% SS
plus 1% 1gM. Embryos were cultured to the hatched blastocyst stage or degeneration and evaluated at 12 h intervals.Master of Science
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Ariño, Rodríguez Helena. „Neurological syndromes associated with antibodies against the glutamic acid decarboxylase (GAD)“. Doctoral thesis, Universitat de Barcelona, 2017. http://hdl.handle.net/10803/666797.
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This thesis focuses on antibodies against the glutamic acid decarboxylase (GAD-ab) in neurological disorders. GAD-ab were first identified in 1988 in the serum and CSF from one patient with stiff-person syndrome (SPS) and diabetes mellitus (T1DM). Since then, GAD-ab have become excellent biomarkers in several autoimmune conditions affecting the endocrine system, the CNS or both simultaneously. High serum GAD-ab levels have expanded the neurological spectrum and have been described in patients with cerebellar ataxia (CA), epilepsy and limbic encephalitis (LE) of autoimmune origin. Even though all the syndromes share the same autoimmune biomarker, it may well be that GAD-ab have a different role in each syndrome. Whether all GAD-associated syndromes share the same pathogenic mechanisms, or what renders certain brain regions vulnerable to autoantibody attack is not clear. In recent years, other antibodies have been found in patients with neurological syndromes attributed to GAD-ab, such as antibodies directed against the alpha subunit of the glycine receptor (GlyRα1) in patients with syndromes of the stiff-person spectrum, or antibodies against GABA receptors (GABAA in patients with severe epilepsy, and GABAB in patients with limbic encephalitis, GAD-ab and an unknown lung cancer), yielding the possibility of an alternative immunological response coexisting with GAD autoimmunity that might be more relevant in certain neurological conditions.
In this thesis we explored the immunological determinants linked to the different neurological phenotypes with state-of-the-art techniques, and investigated the prognostic value of GAD-ab in neurological disorders. After studying the largest cohort of patients (121) with neurological syndromes and GAD-ab, we found that: 1) the presence of additional antibodies against antigens of the inhibitory synapse or a different reactivity against particular GAD isoforms or sites of GAD65 do not explain the diversity of the clinical phenotype in non-paraneoplastic neurological syndromes associated with GAD-ab; 2) the immunological response against GAD is different in serum and CSF, indicating a process of antigen-driven intrathecal maturation in patients with non-paraneoplastic syndromes; 3) patients with cerebellar ataxia and GAD-ab may respond to immunotherapy, and maintain good functional status at long-term. Early initiation of treatment likely offers a greater chance of improvement; 4) neurological syndromes with paraneoplastic criteria in the context of GAD autoimmunity have a different clinical presentation and humoral immunity profile. Patients presenting neurological syndromes not typically associated with GAD-ab should be screened for an underlying cancer; 5) among patients with stiff-person spectrum disorders, the immunological classification is an independent predictor of outcome. Those patients with GAD-ab have worse prognosis than antibody-negative patients and patients with GlyR-ab.
Our results confirmed that autoimmunity regarding the humoral response is similar among different neurological syndromes, and that GAD-ab is still the most important biomarker in these diseases. From a clinical perspective we contributed to fill some clinical gaps, like the value of GAD-ab in SPS, the management of patients with GAD-associated CA, and he clues to suspect paraneoplastic neurological syndromes.Esta tesis se centra en síndromes neurológicos asociados a anticuerpos contra el enzima descarboxilasa del ácido glutámico (anti-GAD), descritos asociados a multitud de síndromes neurológicos y enfermedades endocrinológicas. El rol de estos anticuerpos en cada una de estas entidades no está perfectamente establecido. Los resultados de esta tesis contribuyen a clarificar qué valor tiene el hallazgo de anticuerpos anti-GAD en pacientes con determinados síndromes neurológicos y a caracterizar el perfil de algunos de los grupos grupos clínicos asociados a anti-GAD más desconocidos, como son pacientes con ataxia cerebelosa (AC) o síndromes paraneoplásicos. Algunas de las conclusiones más relevantes, recogidas en las 4 publicaciones principales que conforman esta tesis, son: 1) la presencia adicional de anticuerpos contra antígenos de la sinapsis inhibitoria, una reactividad diferente contra alguna de las isoformas de GAD o contra algún epítopo particular de GAD65, no determinan la diversidad fenotípica en síndromes neurológicos asociados a anti-GAD ; 2) la respuesta inmune contra GAD difiere en suero y líquido cefalorraquídeo; 3) los pacientes con AC y anti-GAD pueden responder a la inmunoterapia y mantener un buen estado funcional a largo plazo, pero es importante la instauración del tratamiento de forma precoz; 4) los síndromes neurológicos paraneoplásicos asociados a anti-GAD tienen una presentación clínica y un perfil inmunológico diferente; 5) En pacientes con síndromes del espectro de la persona rígida, la clasificación inmunológica es un factor pronóstico independiente. Aquellos pacientes con anticuerpos anti-GAD tienen un peor pronóstico.
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Eddahri, Fouad. „Caractéristation des lymphocytes T auxiliaires impliqués dans la régulation de la réponse humorale“. Doctoral thesis, Universite Libre de Bruxelles, 2007. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/210685.
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Barden, Timothy John. „The synthesis of novel O-alkyl analogues of the energy-repartitioning [beta]-agonist clenbuterol and their physiological and immunological characterisations“. Thesis, [Campbelltown, N.S.W. : The Author], 1995. http://handle.uws.edu.au:8081/1959.7/29572.
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It was proposed that some O-alkyl analogues of the beta-adrenergic agonist clenbuterol would be effective structural and functional congeners of clenbuterol which may then be used for the production of clenbuterol-specific idiotypic antibodies. These antibodies could possibly then be used to generate anti-idiotypic antibodies that mimic the energy-repartitioning effects of clenbuterol. Therefore, the aim of this work was to synthesise and characterise these compounds, evaluate their physiological effects, characterise the specificity of antibodies produced in response to protein conjugates of two of the novel compounds, and then use this data to determine the utility of these compounds for the generation of anti-idiotype antibodies which mimic clenbuterol. The target compounds were synthesised in five steps from 3,5-dichloro-4-hydroxyacetophenone in overall yields of 5-28%. A synthetic scheme similar to that which has led to clenbuterol was used to form the phenylethanolamine backbone, with modifications to include the O-alkyl moiety via a modified Williamson ether synthesis, and elimination of a synthetic chlorination step. Overall, 15 new compounds were synthesised, which were characterised and their structure confirmed from proton and carbon-13 NMR, IR and mass spectral data. The two haptenic analogues were then conjugated to carrier proteins using carbodiimide-based chemistries. In conclusion, the results indicated that the O-alkyl analogues, although structurally similar, were ineffective functional mimics of clenbuterol. Therefore, the anti-clenbuterol antobodies produced from the novel O-alkyl analogues would appear to be unsuitable for production of anti-idiotypic antibodies that mimic the energy-repartitioning effects of clenbuterol since the antibodies were unable to distinguish between the compound which demonstrated energy-repartitioning effects (clenbuterol) and those that did not (O-alkyl clenbuterol analogues).
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Ramsland, Paul Allen. „Variable region gene expression and structural motifs of human polyreactive immunoglobulins“. University of Technology, Sydney.|bFaculty of Science, 1997. http://hdl.handle.net/2100/372.
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Polyreactive immunoglobulins (Ig) exhibit a capacity to recognise multiple, structurally dissimilar antigens through a single combining site. This characteristic differentiates these Igs from monoreactive Igs which bind to a single antigen, usually with high specificity and affinity. Chronic B lymphocytic leukaemia (B CLL) is a malignancy identified by the incessant accumulation, in the peripheral circulation, of B lymphocytes of a mature and resting morphology. B CLL malignant cells generally express both surface IgM and the pan T cell antigen CD5. Moreover, the IgM on the surface of these CD5 positive B CLL cells is frequently polyreactive. This thesis examines the structural diversity found in the combining sites of B CLL derived Igs in an attempt to elucidate the structural basis of polyreactive antigen binding displayed by a significant proportion of human Igs. The genes encoding the variable (V) domains of five B CLL derived IgM antibodies (Bel, Tre, Yar, Hod and Jak) were cloned and sequenced (Chapter Two). When the light chain V domain genes were aligned with the closest germline VL and JL coding DNA sequences it was determined that there was either a complete absence of somatic mutation (Tre, Yar and Jak) or a minimal number of mutations (Bel and Hod) present in the rearranged VL domain genes. A remarkable fidelity in the splicing of VL to JL genes was noted suggesting that the diversity, normally introduced through variability of splicing VL to JL, is reduced in Igs expressed by B CLL cells. Furthermore, the markedly reduced primary structural diversity was highlighted when two of the VL domain genes (Yar and Hod) were found to be different in sequence by only four nucleotides and two amino acids. The heavy chain V domain genes of the same five Igs were sequenced in another study (Brock, 1995), however, it was interesting to analyse the sequences of the VH domain genes and compare them with the VL domain genes. The naive or gerrnline nature of the B CLL antibodies was reflected in the VH genes by either an absence or a low frequency of mutations within these sequences compared with germline immunoglobulin gene sequences. No obvious conserved motif, which could be related to polyreactivity, was observed when the primary protein sequence was analysed for distribution of identical or similar amino acids. Thus, homology modelling was used to construct three-dimensional models of the Fv (VL-VH) portions of the five B CLL IgM molecules to examine the structures of the combining sites of these Igs (Chapter Three). Framework regions were constructed using X-ray coordinates taken from highly hon~ologous human variable domain structures. Complementarity determining regions (CDR) were predicted by grafting loops, taken from known Ig structures, onto the Fv framework models. The CDR templates were selected, where possible, to be of the same length and of high residue identity or similarity. If a single template CDR was not appropriate to model a particular CDR the loop was built from loop sterns of known conformation, followed by chain closure with a p-turn. Template models were refined using standard molecular mechanics simulations. The binding sites were either relatively flat or contained a deep cavity at the VL-VH domain interface. Further differences in topology were the result of some CDR loops protruding into the solvent. Examination of the electrostatic molecular surface did not reveal a common structural feature within the binding sites of the five polyreactive Fv. While two of the binding cavities were positively charged the other three structures displayed either negatively charged or predominantly hydrophobic combining sites. These findings suggested that a diversity of structural mechanisms are involved in polyreactive antigen binding. Rcsidues within CDRs which have aromatic side-chains and are partially exposed to solvent were distributed across large regions of the combining sites. It is possible that these aromatic residues are responsible for the conserved binding to mouse Igs observed (Chapter Two) for the B CLL derived polyreactive IgM molecules. Two Fv molecules (Be1 and Tre) were cloned as dicistronic constructs, into the bacterial expression vector pFLAG. The expression of the Fvs was fully characterised and unfortunately the VL and VH of Be1 and Tre Igs did not associate in an appropriate manner to yield large quantities of purified Fv (Chapter Four). Expression of correctly folded and stabilised fragments of human polyreactive immunoglobulins would enable the structural basis for the polyreactive binding phenomenon to be fully explored using protein crystallography.
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McCannel, Anne Marie. „Separation of immunoglobulins from egg yolk using metal chelate interaction chromatography and ion exchange chromatography“. Thesis, University of British Columbia, 1988. http://hdl.handle.net/2429/27987.
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The immune response of chickens immunized with β-N-acetylglucosaminidase was monitored in the egg yolks of the birds using an enzyme-linked immunosorbent assay. Significantly higher levels of specific antibodies were detected in the yolks of the birds immunized with the enzyme when compared with the yolks of a control bird collected over the same period significant differences also were found in the response within the immunized group of birds, indicating individual variability to the injections.
Immunoglobulins were isolated from egg yolk after a preliminary purification using alginate to precipitate lipoproteins. A ten millilitre DEAE-Sephacel ion exchange chromatography column resulted in a final product containing 16 mg of IgG with a purity of 60% when 50 mL of an egg yolk supernatant was applied. Specific antibody activity toward the antigens β-lactoglobulin and E. coli lipopolysaccharide was higher in both cases in the isolated immunoglobulin fractions which contained lower purity (40%). Lower antibody activity was observed in the 60% purified fractions. Separation of specific antibodies from non-specific antibodies appeared to occur, possibly due to the given characteristics of the specific antibodies, or due to the differences exhibited by chicken IgG subpopulations.
Using metal chelate interaction chromatography, a 10 mL copper-loaded column was able to yield 104 mg of IgG with a purity of 75% when 200 mL egg yolk supernatant was applied. Again, the heterogeneous nature of chicken IgG was illustrated.
A comparison of the two chromatographic techniques indicated the advantages of metal chelate interaction chromatography over ion exchange chromatography under the conditions examined.
Applications of the chicken IgG isolated by metal chelate interaction chromatography to an enzyme-linked immunosorbent assay for the detection of β-N-acetylglucosaminidase was attempted. Linear relationships were obtained when standard solutions of the enzyme were used in the assay. The results indicate that MCIC-isolated chicken immunoglobulins have excellent potential for use in analytical tests.Land and Food Systems, Faculty of
Graduate
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Andersson, Tove. „Transcription factors regulating the immunoglobulin heavy chain locus /“. Stockholm, 1999. http://diss.kib.ki.se/1999/91-628-3559-9/.
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