Journal articles: 'Murine mode'

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Itoh, Masahiro, Hiroshi Moriyama, Akiko Yano, Xiu-Qin Li, and Yoshiki Takeuchi. "Mode of migration of normal lymphocytes inside murine testis." Anatomical Record 251, no. 2 (June 1998): 152–60. http://dx.doi.org/10.1002/(sici)1097-0185(199806)251:2<152::aid-ar2>3.0.co;2-0.

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Arakawa, Tsutomu, Yasunori Kurosawa, Michael Storms, Toshiaki Maruyama, C. J. Okumura, and Yoshiko Kita. "Capto MMC mixed-mode chromatography of murine and rabbit antibodies." Protein Expression and Purification 127 (November 2016): 105–10. http://dx.doi.org/10.1016/j.pep.2016.07.010.

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Fabre, Pierre-Henri, Yuli S. Fitriana, Gono Semiadi, Marie Pagès, Ken Aplin, Nanang Supriatna, and Kristofer M. Helgen. "New record of Melomys burtoni (Mammalia, Rodentia, Murinae) from Halmahera (North Moluccas, Indonesia): a review of Moluccan Melomys." Mammalia 82, no. 3 (April 25, 2018): 218–47. http://dx.doi.org/10.1515/mammalia-2016-0137.

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AbstractMosaic-tailed rodents of the genusMelomysbelong to the Australasian old endemic murine radiation and exhibit a rat-like morphology with arboreal or scansorial specializations. Here we report a new population ofMelomys burtonifrom the island of Halmahera (in the North Moluccas, Indonesia). Our molecular phylogenetic results highlight close relationships and recent evolutionary divergences amongM. burtonifrom Halmahera and the Australo-Papuan taxaM. burtoniandM. lutillusand other Moluccan taxa, includingM. paveli. Multivariate as well as geometric morphometric analyses of cranial, and dental features support the recognition ofM. burtonifrom Halmahera as a slightly distinctive insular population, preventing us from elevating it as a new taxa. This population is recorded from lowland secondary forest and forest edge habitats in south-central Halmahera. As with other Moluccan endemic murines, colonization by an Australo-Papuan ancestor and subsequent isolation is the probable mode of diversification forM. burtoniin Halmahera. The discovery ofMelomysin Halmahera fills a previously puzzling gap in knowledge of the murine fauna of the Moluccas and the biogeography of the Wallacean region.

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Kolpakov, Sergei, Elena Kolpakova, Elena Zlatnik, Evgeniya Nepomnyashchaya, Inna Novikova, Oksana Shulgina, Alexandr Sagakyants, and Yuri Sidorenko. "Evaluation of antitumor activity in strains of a new rotavirus group in Reoviridae family on a model of transplantable murine melanoma." Problems in oncology 66, no. 6 (December 30, 2020): 712–17. http://dx.doi.org/10.37469/0507-3758-2020-66-6-712-717.

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The purpose was to study antitumor effects of the group K rotaviruses strains No. 228 and No. 100 in the experiment on a model of transplantable murine melanoma. Material and methods. The study included 65 С57Black/6 mice with transplantable B16/F10 melanoma and two strains of the Reoviridae family members characterized as rotaviruses, not belonging to the known groups, with the working title “group K rotaviruses (RVK)”. Animals received RVK in “vaccination” (before tumor transplantation) and “treatment” (after tumor formation) modes. Live and inactivated strains were used. RVK were administered intramuscularly as 0.3 ml of virus-containing culture fluid with at least 5x109 viral particles per 1 ml, with a total of 4 injections. Life span of mice and morphological characteristics of tumors were evaluated. Results. Injections of both strains increased the survival of tumor-bearing mice by 1.7-1.9 times in 4 of 8 experimental groups, compared to controls. The modes of RVK administration showed some differences: the survival was longer in mice with the “vaccination” mode compared to the “treatment” mode. Morphological changes in tumors were similar after application of both modes and inclued dystrophic changes of tumor cells, formation of extensive necrosis areas, and leukocyte infiltration. Discussion. Live and inactivated RVK had unidirectional effects implying its association with immunomodulatory action rather than with a direct lytic effect on the tumor. Conclusions. Both studied strains of rotaviruses in the group K had antitumor effect in the model of transplantable В16/F10 murine melanoma in the «vaccination» mode.

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Prabhakara, Ranjani, Janette M. Harro, Jeff G. Leid, Megan Harris, and Mark E. Shirtliff. "Murine Immune Response to a ChronicStaphylococcus aureusBiofilm Infection." Infection and Immunity 79, no. 4 (January 31, 2011): 1789–96. http://dx.doi.org/10.1128/iai.01386-10.

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ABSTRACTStaphylococcus aureushas reemerged as an important human pathogen in recent decades. Although many infections caused by this microbial species persist through a biofilm mode of growth, little is known about how the host's adaptive immune system responds to these biofilm infections. In this study,S. aureuscells adhered to pins in culture and were subsequently inserted into the tibiae of C57BL/6 mice, with an infecting dose of 2 × 105CFU. This model was utilized to determine local cytokine levels, antibody (Ab) function, and T cell populations at multiple time points throughout infection. Like human hosts,S. aureusimplant infection was chronic and remained localized in 100% of C57BL/6 mice at a consistent level of approximately 107CFU/gram bone tissue after day 7. This infection persisted locally for >49 days and was recalcitrant to clearance by the host immune response and antimicrobial therapy. Local inflammatory cytokines of the Th1 (interleukin-2 [IL-2], IL-12 p70, tumor necrosis factor alpha [TNF-α], and IL-1β) and Th17 (IL-6 and IL-17) responses were upregulated throughout the infection, except IL-12 p70, which dwindled late in the infection. In addition, Th1 Ab subtypes against a biofilm antigen (SA0486) were upregulated early in the infection, while Th2 Abs and anti-inflammatory regulatory T cells (Tregs) were not upregulated until later. These results indicate that early Th1 and Th17 inflammatory responses and downregulated Th2 and Treg responses occur during the development of a chronic biofilm implant infection. This unrestrained inflammatory response may cause tissue damage, thereby enablingS. aureusto attach and thrive in a biofilm mode of growth.

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Gareau, Daniel S., Glenn Merlino, Christopher Corless, Molly Kulesz-Martin, and Steven L. Jacques. "Noninvasive Imaging of Melanoma with Reflectance Mode Confocal Scanning Laser Microscopy in a Murine Model." Journal of Investigative Dermatology 127, no. 9 (September 2007): 2184–90. http://dx.doi.org/10.1038/sj.jid.5700829.

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Poole, Aaron, Phyllis Gamble, Esther Tamayo, Igor Patrikeev, Jingna Wei, Kathleen Vinvent, Gayle Olson, George Saade, Alison Stuebe, and Egle Bytautiene. "41: Effect of lactation on maternal postpartum cardiometabolic status–a murine mode." American Journal of Obstetrics and Gynecology 210, no. 1 (January 2014): S27. http://dx.doi.org/10.1016/j.ajog.2013.10.074.

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Li, Pei-Lin, Rodger E. Tiedemann, S. Louise Moffat, and John D. Fraser. "The Superantigen Streptococcal Pyrogenic Exotoxin C (SPE-C) Exhibits a Novel Mode of Action." Journal of Experimental Medicine 186, no. 3 (August 4, 1997): 375–83. http://dx.doi.org/10.1084/jem.186.3.375.

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Recombinant streptococcal pyrogenic exotoxin C (SPE-C) is a potent superantigen that stimulates Vβ2-bearing human T cells, but is inactive in mice. SPE-C binds with high affinity to both human HLA-DR and murine I-E molecules, but not to murine I-A molecules in a zinc-dependent fashion. Competition binding studies with other recombinant toxins revealed that SPE-C lacks the generic low affinity major histocompatibility complex (MHC) class II α-chain binding site common to all other bacterial superantigens. Despite this, SPE-C cross-links MHC class II to induce homotypic aggregation of class II–bearing B cells. Nondenaturing sodium dodecyl sulfate electrophoresis and size exclusion chromatography revealed that both wild-type and recombinant SPE-C exist in a stable dimer at neutral or alkaline pH. These data support a recent crystal structure of SPE-C and reveal yet another mechanism by which bacterial superantigens ligate and cross-link MHC class II.

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Stypmann, Jörg, Markus A. Engelen, Clemens Troatz, Markus Rothenburger, Lars Eckardt, and Klaus Tiemann. "Echocardiographic assessment of global left ventricular function in mice." Laboratory Animals 43, no. 2 (April 2009): 127–37. http://dx.doi.org/10.1258/la.2007.06001e.

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Doppler-echocardiographic assessment of cardiovascular structure and function in murine models has developed into one of the most commonly used non-invasive techniques during the last decades. Recent technical improvements even expanded the possibilities. In this review, we summarize the current options to assess global left ventricular (LV) function in mice using echocardiographic techniques. In detail, standard techniques as structural and functional assessment of the cardiovascular phenotype using one-dimensional M-mode echocardiography, two-dimensional B-mode echocardiography and spectral Doppler signals from mitral inflow respective aortal outflow are presented. Further pros and contras of recently implemented techniques as three-dimensional echocardiography and strain and strain rate measurements are discussed. Deduced measures of LV function as the myocardial performance index according to Tei, estimation of the mean velocity of circumferential fibre shortening, LV wall stress and different algorithms to estimate the LV mass are described in detail. Last but not least, specific features and limitations of murine echocardiography are presented. Future perspectives in respect to new examination techniques like targeted molecular imaging with advanced ultrasound contrast bubbles or improvement of equipment like new generation matrix transducers for murine echocardiography are discussed.

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Verbeke, Frederick, Nathan Debunne, Yorick Janssens, Bart De Spiegeleer, and Evelien Wynendaele. "A bioanalytical screening method for Enterococcus faecalis RNPP-type quorum sensing peptides in murine feces." Bioanalysis 14, no. 3 (February 2022): 151–67. http://dx.doi.org/10.4155/bio-2021-0225.

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Background: Bacteria coordinate their behavior as a group via communication with their peers, known as ‘quorum sensing’. Enterococcus faecalis employs quorum sensing via RNPP-peptides which were not yet reported to be present in mammalian biofluids. Results: Solid phase extraction of murine feces was performed, followed by ultra high performance liquid chromatography (UHPLC–MS/MS) in multiple reaction monitoring (MRM) mode (in total <90 min/sample) for the nine known RNPP peptides. Limits of detection ranged between 0.045 and 52 nM. Adequate identification criteria allowed detection of RNPP quorum sensing peptides in 2/20 wild-type murine feces samples (i.e., cAM373 and cOB1). Conclusion: A fit-for-purpose UHPLC–MS/MS method detected these RNPP peptides in wild-type murine feces samples.

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de Harven, E., and H. Christensen. "High-resolution immunogold labeling of cell surfaces: The hypothetical homodimeric nature of the cd5 receptor." Proceedings, annual meeting, Electron Microscopy Society of America 46 (1988): 380–81. http://dx.doi.org/10.1017/s0424820100103966.

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Molecules exposed on cell surfaces and labeled with colloidal gold markers can be optimally demonstrated using the backscattered electron imaging (BEI) mode of the scanning electron microscope (SEM). Steric hindrance, however, limits labeling efficiency, making it necessary to use gold markers of small size for labeling at the molecular level. Using a JEOL 840 SEM equipped with a lanthanum hexaboride (LaB6) cathode, 13 nm gold particles were demonstrated. This, however, seems to represent the limit of the resolution of this type of instrument in the BEI mode. Fortunately, it has been demonstrated by Walther and Muller that 5 nm gold particles can be seen in the BEI mode, using field emission SEM.We have confirmed this observation, using the JEOL 890 field emission SEM and a solid state backscattered electron detector. Human peripheral blood lymphocytes prefixed with 0.1% glutaraldehyde, incubated with the murine monoclonal antibody LEU-1 (CD5), were labeled with a goat anti-murine IgG adsorbed on 5 nm gold particles (GAM-G5, Janssen Pharmaceutica, Beerse, Belgium) according to previously described procedures.

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Nakamura, Morihiko, Hiroyuki Ogawa, and Tokugoro Tsunematsu. "Mode of action of monoclonal-nonspecific suppressor factor (MNSF) produced by murine hybridoma." Cellular Immunology 116, no. 1 (October 1988): 230–39. http://dx.doi.org/10.1016/0008-8749(88)90223-7.

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Pallavicini, M. G., L. J. Summers, F. J. Giroud, P. N. Dean, and I. W. Gray. "Multivariate analysis and list mode processing of murine hemopoietic subpopulations for cytokinetic studies." Cytometry 6, no. 6 (November 1985): 539–49. http://dx.doi.org/10.1002/cyto.990060608.

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Gareau, Daniel S., James Lagowski, Vincent M. Rossi, John A. Viator, Glenn Merlino, Molly Kulesz-Martin, and Steven L. Jacques. "Imaging Melanoma in a Murine Model Using Reflectance-Mode Confocal Scanning Laser Microscopy and Polarized Light Imaging." Journal of Investigative Dermatology Symposium Proceedings 10, no. 2 (November 2005): 164–69. http://dx.doi.org/10.1111/j.1087-0024.2005.200408.x.

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Nair, Sreenath, Abigail Davis, Olivia Campagne, John D. Schuetz, and Clinton F. Stewart. "Development and Validation of a Sensitive and Specific LC-MS/MS Method for IWR-1-Endo, a Wnt Signaling Inhibitor: Application to a Cerebral Microdialysis Study." Molecules 27, no. 17 (August 25, 2022): 5448. http://dx.doi.org/10.3390/molecules27175448.

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IWR-1-endo, a small molecule that potently inhibits the Wnt/β-catenin signaling pathway by stabilizing the AXIN2 destruction complex, can inhibit drug efflux at the blood–brain barrier. To conduct murine cerebral microdialysis research, validated, sensitive, and reliable liquid chromatography–tandem mass spectrometry (LC-MS/MS) methods were used to determine IWR-1-endo concentration in the murine plasma and brain microdialysate. IWR-1-endo and the internal standard (ISTD) dabrafenib were extracted from murine plasma and microdialysate samples by a simple solid-phase extraction protocol performed on an Oasis HLB µElution plate. Chromatographic separation was executed on a Kinetex C18 (100A, 50 × 2.1 mm, 4 µm particle size) column with a binary gradient of water and acetonitrile, each having 0.1% formic acid, pumped at a flow rate of 0.6 mL/min. Detection by mass spectrometry was conducted in the positive selected reaction monitoring ion mode by monitoring mass transitions 410.40 > 344.10 (IWR-1-endo) and 520.40 > 307.20 (ISTD). The validated curve range of IWR-1-endo was 5–1000 ng/mL for the murine plasma method (r2 ≥ 0.99) and 0.5–500 ng/mL for the microdialysate method (r2 ≥ 0.99). The lower limit of quantification (LLOQ) was 5 ng/mL and 0.5 ng/mL for the murine plasma and microdialysate sample analysis method, respectively. Negligible matrix effects were observed in murine plasma and microdialysate samples. IWR-1-endo was extremely unstable in murine plasma. To improve the stability of IWR-1-endo, pH adjustments of 1.5 were introduced to murine plasma and microdialysate samples before sample storage and processing. With pH adjustment of 1.5 to the murine plasma and microdialysate samples, IWR-1-endo was stable across several tested conditions such as benchtop, autosampler, freeze–thaw, and long term at −80 °C. The LC-MS/MS methods were successfully applied to a murine pharmacokinetic and cerebral microdialysis study to characterize the unbound IWR-1-endo exposure in brain extracellular fluid and plasma.

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Lorenz, Robert R., Olivia Dan, Marc Nelson, Michael A. Fritz, and Marshall Strome. "Immunosuppressive Effect of Irradiation in the Murine Laryngeal Transplantation Model: A Controlled Trial." Annals of Otology, Rhinology & Laryngology 112, no. 8 (August 2003): 712–15. http://dx.doi.org/10.1177/000348940311200811.

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Since the first successful human laryngeal transplantation in 1998, research continues toward developing less-morbid immunosuppressive protocols. Although irradiation of donor organs is known to decrease acute rejection, the most advantageous method of radiation delivery is still unknown. Using a rat laryngeal transplant model, we sought to determine the most beneficial timing and delivery method of irradiation. A prospective study was undertaken including 16 treatment arms of 10 to 30 animals each (189 transplantations). The animals received 800 cGy before transplantation to the donor larynx in vivo, the donor larynx in vitro, or the recipient animal's neck. The transplantation occurred at 24 hours, 5 days, or 10 days after irradiation. The transplanted larynges were harvested 15 days after transplantation and histologically scored for rejection. Irradiation of allogeneic transplantations demonstrated a strongly protective effect from rejection as compared to no irradiation (p < .001), regardless of the method of radiation delivery or the amount of time between irradiation and transplantation (p = .78). Irradiation of donor larynges between allogeneic rats has a protective effect, reducing the degree of acute rejection when irradiation is used as the single mode of immunosuppression, and is unrelated to either the timing or the mode of delivery of the irradiation.

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de Harven, Etienne, Davide Soligo, Roy McGroarty, Hilary Christensen, Richard Leung, and Cameron Ackerley. "Quantification of gold labeled cell surface antigens by SEM: Current progress and remaining problems." Proceedings, annual meeting, Electron Microscopy Society of America 48, no. 3 (August 12, 1990): 34–35. http://dx.doi.org/10.1017/s042482010015770x.

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Taking advantage of the high elemental contrast of particles of colloidal gold observed in the backscattered electron imaging(BEI) mode of the SEM (1,2), the human T lymphocyte was chosen as a model system to study the potential value of immunogold labeling for the quantification of cell surface expressed molecules. The CD3 antigen which is expressed on all human T lymphocytes and is readily identified by the LEU-4 murine monoclonal antibody (Becton Dickinson, Mountain View, CA) followed by a gold conjugated goat anti-mouse Ig polyclonal antibody was chosen as a model target antigen. When quantified by non-EM methods, using radio-iodinated probes or FACS analysis, approximately 30,000 to 50,000 copies of this antigen per cell are enumerated.The following observations were made while attempting to quantify the same molecule by SEM after specific immunogold labeling:Imaging in the SE vs BE mode: The numbers of gold markers counted in the secondary electron (SE) imaging mode are considerably lower than those counted on the same cells in the backscattered electron (BE) imaging mode.

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Dovey, Oliver M., Jonathan L. Cooper, Annalisa Mupo, Carolyn S. Grove, Claire Lynn, Nathalie Conte, Robert M. Andrews, et al. "Molecular synergy underlies the co-occurrence patterns and phenotype of NPM1-mutant acute myeloid leukemia." Blood 130, no. 17 (October 26, 2017): 1911–22. http://dx.doi.org/10.1182/blood-2017-01-760595.

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Key Points Npm1c and Nras-G12D co-mutation in mice leads to AML with a longer latency and a more mature phenotype than the Npm1c/Flt3-ITD combination. Mutant Flt3 or Nras allele amplification is the dominant mode of progression in both Npm1c/Flt3-ITD and Npm1c/Nras-G12D murine AML.

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Robert Kane, J., Junfei Zhao, Takashi Tsujiuchi, Brice Laffleur, Aayushi Mahajan, Ganesh Rao, Angeliki Mela, et al. "IMMU-42. CD8+ T-CELLS MEDIATE IMMUNOEDITING, AND INFLUENCE GENOTYPE, TUMOR ONCOGENIC PATHWAYS AND MICROENVIRONMENT DURING PROGRESSION OF MURINE GLIOMAS." Neuro-Oncology 21, Supplement_6 (November 2019): vi128. http://dx.doi.org/10.1093/neuonc/noz175.534.

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Abstract Cancer immunoediting shapes tumor progression by the immunological selection of tumor cell variants that can evade immune recognition. Given the immune evasive cellular diversity of glioblastoma, we hypothesized that CD8+ T-cells mediate immunoediting in this tumor. We evaluated tumor progression in the absence of CD8+ T-cells by depleting this immune cell population in a transgenic murine glioma model. Tumors generated in the absence of CD8+ T-cells developed poorly in recipients with intact immunity, implying a more immunogenic profile. These tumors demonstrated increased chromosomal instability, gene fusions, MAPK signaling, and macrophage infiltration. These observations were stochastic, suggesting variability in the mode of tumor evolution in the absence of this immune effector. MAPK activation was correlated with macrophage recruitment in two transgenic murine models and the human disease. Our results indicate that CD8+ T-cells mediate a strong immunoediting selection in glioblastoma that protect against the hallmarks of cancer and drive immune evasion.

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Lozano-Aponte, Jorge, Thomas Scior, Francisco Noé Mendoza Ambrosio, Minerva González-Melchor, and Christian Alexander. "Exploring electrostatic patterns of human, murine, equine and canine TLR4/MD-2 receptors." Innate Immunity 26, no. 5 (December 25, 2019): 364–80. http://dx.doi.org/10.1177/1753425919894628.

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Electrostatic interactions between phosphate anions and Toll-like receptor 4 / Myeloid differentiation factor-2 (TLR4/MD-2) protein complexes of human, murine, equine and canine species were computed. Such knowledge can provide mechanistic information about recognising LPS-like ligands, since anionic phosphate groups belong to the structural features of LPS with their diphosphorylated diglucosamine backbone. Sequence composition analyses, electrostatic interaction potentials and docked energies as well as molecular dynamics studies evaluated the phosphate interactions within the triangular LPS binding site (wedge). According to electrostatic analyses, human, horse and dog wedges possess phosphate-binding sites with indistinct positive and negative charge distributions, but the murine wedge shows a unique strong negative net charge at the site where antagonists bind in other species (Pan). Docking of a phosphate mono-anion (probe) confirmed its repulsion at this Pan site, but the Pag site of the murine wedge attracted the probe. It is occupied by phosphate groups of agonists in other species (Pag). Molecular dynamics trajectories show a variable degree of random walk across the wedges, that is, not following electrostatic preferences (neither Pag nor Pan). In summary, two opposing electrostatic patterns exist –murine versus human, equine and canine species – all of which reflect the potential dual activity mode of under-acylated ligands such as lipid IVA.

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Hoa, Tran Thu, Le Hong Duc, Rachele Isticato, Loredana Baccigalupi, Ezio Ricca, Pham Hung Van, and Simon M. Cutting. "Fate and Dissemination of Bacillus subtilis Spores in a Murine Model." Applied and Environmental Microbiology 67, no. 9 (September 1, 2001): 3819–23. http://dx.doi.org/10.1128/aem.67.9.3819-3823.2001.

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ABSTRACT Bacterial spores are being consumed as probiotics, although little is known about their efficacy or mode of action. As a first step in characterizing spore probiotics, we have studied the persistence and dissemination of Bacillus subtilis spores given orally to mice. Our results have shown that spores do not appear to disseminate across the mucosal surfaces. However, we found that the number of spores excreted in the feces of mice was, in some experiments, larger than the original inoculum. This was an intriguing result and might be explained by germination of a proportion of the spore inoculum in the intestinal tract, followed by limited rounds of cell growth and then sporulation again. This result raises the interesting question of whether it is the spore or the germinated spore that contributes to the probiotic effect of bacterial spores.

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Flynn, Mark C., Thomas R. Scott, Thomas C. Pritchard, and Carlos R. Plata-Salamán. "Mode of action of OB protein (leptin) on feeding." American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 275, no. 1 (July 1, 1998): R174—R179. http://dx.doi.org/10.1152/ajpregu.1998.275.1.r174.

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OB protein (leptin) decreases food intake in a variety of species. Here we investigated the effects of the intracerebroventricular administration of recombinant murine OB protein on food consumption and meal parameters in Wistar rats maintained ad libitum. The intracerebroventricular administration of OB protein (0.56–3.5 μg/rat) decreased feeding in a dose-dependent manner. Computer analysis of meal parameters demonstrated that OB protein (3.5 μg/rat, n = 10) decreased nighttime meal size by 42%, whereas meal frequency and meal duration were unaffected. Derived analyses for the nighttime also showed that OB protein decreased the feeding rate (meal size/meal duration) by 30%, whereas the satiety ratio (intermeal intervals/meal size) increased by 100%. A similar profile was observed during the daytime and total daily periods. The intracerebroventricular administration of heat-inactivated OB protein (3.5 μg/rat, n = 10) had no effect on any meal parameter. The results show that OB protein administered intracerebroventricularly inhibits feeding through a specific reduction of meal size.

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Valkov, Nedyalka, Avash Das, Nathan R. Tucker, Guoping Li, Ane M. Salvador, Mark D. Chaffin, Getulio Pereira De Oliveira Junior, et al. "SnRNA sequencing defines signaling by RBC-derived extracellular vesicles in the murine heart." Life Science Alliance 4, no. 12 (October 18, 2021): e202101048. http://dx.doi.org/10.26508/lsa.202101048.

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Extracellular vesicles (EVs) mediate intercellular signaling by transferring their cargo to recipient cells, but the functional consequences of signaling are not fully appreciated. RBC-derived EVs are abundant in circulation and have been implicated in regulating immune responses. Here, we use a transgenic mouse model for fluorescence-based mapping of RBC-EV recipient cells to assess the role of this intercellular signaling mechanism in heart disease. Using fluorescent-based mapping, we detected an increase in RBC-EV–targeted cardiomyocytes in a murine model of ischemic heart failure. Single cell nuclear RNA sequencing of the heart revealed a complex landscape of cardiac cells targeted by RBC-EVs, with enrichment of genes implicated in cell proliferation and stress signaling pathways compared with non-targeted cells. Correspondingly, cardiomyocytes targeted by RBC-EVs more frequently express cellular markers of DNA synthesis, suggesting the functional significance of EV-mediated signaling. In conclusion, our mouse model for mapping of EV-recipient cells reveals a complex cellular network of RBC-EV–mediated intercellular communication in ischemic heart failure and suggests a functional role for this mode of intercellular signaling.

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HUNT, G., and A. J. THODY. "AGOUTI PROTEIN CAN ACT INDEPENDENTLY OF MELANOCYTE-STIMULATING HORMONE TO INHIBIT MELANOGENESIS." Journal of Endocrinology 147, no. 2 (November 1995): R1—R4. http://dx.doi.org/10.1677/joe.0.147r001.

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Abstract In animals, the coat-darkening effects of α-melanocyte stimulating hormone (α-MSH) are opposed by agouti protein. Although agouti protein has been shown to be a competitive antagonist of the melanocyte-associated MC-1 melanocortin receptor, the possibility that agouti protein can affect melanogenesis independently of its ability to antagonise melanocortin activity cannot be excluded. This study demonstrates that murine agouti protein causes both a time- and concentration-dependent suppression of melanogenesis in B16 F1 murine melanoma cells. In addition, human agouti protein decreases melanogenesis in cultured human epidermal melanocytes. However, agouti protein has little effect on the ability of α-MSH to stimulate melanogenesis. These observations raise fundamental questions about the mode of action of agouti protein in regulating melanogenesis.

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Weicker, Sean, Wanda Cromlish, Sonia Lamontagne, Jacques-Yves Gauthier, Sylvie Desmarais, Renata Oballa, Marc Ouellet, et al. "Cathepsin S in a Murine Model of Allergic Asthma (B108)." Journal of Immunology 178, no. 1_Supplement (April 1, 2007): LB22—LB23. http://dx.doi.org/10.4049/jimmunol.178.supp.b108.

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Abstract Cathepsin S (Cat S), expressed predominately on antigen presenting cells, has been proposed as a therapeutic target for asthma. We used genetic and pharmacological tools to investigate the role of Cat S in murine models of allergic asthma. Mice null for Cat S were protected from OVA-induced pulmonary inflammation, but exhibited no protection from OVA-induced airway hyper-reactivity. To determine the role of Cat S during the challenge phase, we identified a potent and selective Cat S inhibitor, Compound A (Cpd A, IC50 mCat S = 0.6 nM, ≥470 fold selective vs mCat B, K, L), which inhibited antigen presentation in a mouse cell-based assay (IC50 = 44 nM). The prodrug of Cpd A, Cpd B, gave excellent plasma levels of Cpd A when dosed in mice by gavage, or in food. In vivo competition in mice with an irreversible pan-selective cysteine cathepsin probe showed that Cpd B gave selective inhibition of lung and spleen Cat S at 1 mpk, but lost selectivity at 50 mpk. Cpd B was dosed in mice (10, 100 mpk in food) over 4 days of the challenge period in the murine ovalbumin model. Both doses had no effect on bronchoalveolar lavage infiltrating cells, despite showing high levels of Cat S inhibition. Thus, Cat S inhibition in a therapeutic mode does not attenuate airway inflammation in antigen sensitised mice suggesting that anti-Cat S therapy would not be an effective asthma treatment.

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Terenzi, Fulvia, Srabani Pal, and Ganes C. Sen. "Induction and mode of action of the viral stress-inducible murine proteins, P56 and P54." Virology 340, no. 1 (September 2005): 116–24. http://dx.doi.org/10.1016/j.virol.2005.06.011.

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Wang, Hui, Ke Xue, Zhuwen Duan, Yuyun Yang, Zixu He, Chuanchen Wu, Wei Zhang, Wen Zhang, Ping Li, and Bo Tang. "A photoacoustic and fluorescence dual-mode probe for LTA4H imaging reveals inflammation site in murine." Sensors and Actuators B: Chemical 286 (May 2019): 243–49. http://dx.doi.org/10.1016/j.snb.2019.01.154.

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Babu, Dinesh, Stefaan J. Soenen, Koen Raemdonck, Georges Leclercq, Ole De Backer, Roberto Motterlini, and Romain A. Lefebvre. "TNF-α/Cycloheximide-Induced Oxidative Stress and Apoptosis in Murine Intestinal Epithelial MODE-K Cells." Current Pharmaceutical Design 18, no. 28 (August 8, 2012): 4414–25. http://dx.doi.org/10.2174/138161212802481291.

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Berg-Johansen, Britta, Ellen C. Liebenberg, Alfred Li, Brandon R. Macias, Alan R. Hargens, and Jeffrey C. Lotz. "Spaceflight-induced bone loss alters failure mode and reduces bending strength in murine spinal segments." Journal of Orthopaedic Research 34, no. 1 (August 31, 2015): 48–57. http://dx.doi.org/10.1002/jor.23029.

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Jadus, M. R., and R. Parkman. "The selective growth of murine newborn-derived suppressor cells and their probable mode of action." Journal of Immunology 136, no. 3 (February 1, 1986): 783–92. http://dx.doi.org/10.4049/jimmunol.136.3.783.

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Abstract Naturally occurring suppressor cells residing in the spleens of newborn mice of less than 5 days old are known to suppress various lymphocyte activities. A population of these suppressor cells can be maintained and expanded in the supernatants derived from Wehi-3 cells. These suppressor cells, designated as Wehi-3-expanded neonatal splenocytes (WENS), can suppress mixed lymphocyte reactions (MLR) and T and B cell mitogen responses without any genetic restrictions. The WENS bear the Ly-5, J11d, and class I molecules. WENS suppression is not mediated through an interleukin 1 or interleukin 2 absorptive mechanism. To achieve maximum suppression of MLR, WENS must be present for at least 24 hr. WENS inhibited the proliferation of Wehi-164 cells but not other tumor cells. The inhibition of Wehi-164 growth was due to the action of natural cytotoxic cells, because WENS lysed Wehi-164 cells but not the natural killer target cell YAC-1. Maximum lysis of Wehi-164 by WENS required 18 to 24 hr. Five WENS cell lines were cultured for more than 6 mo; three of the cell lines lost their capacity to lyse Wehi-164 targets (natural cytotoxicity) and simultaneously lost their natural suppressor activity. The two WENS lines that retained natural cytotoxicity also retained natural suppressor activity. Thus, natural suppressor cells may manifest their suppression through a natural cytotoxicity mechanism.

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Kuznetsov, Y. G., A. Low, H. Fan, and A. McPherson. "Atomic Force Microscopy Investigation of Isolated Virions of Murine Leukemia Virus." Journal of Virology 79, no. 3 (February 1, 2005): 1970–74. http://dx.doi.org/10.1128/jvi.79.3.1970-1974.2005.

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ABSTRACT Virions of mouse leukemia virus spread on glass substrates were visualized by atomic force microscopy. The size distribution mode was 145 nm, significantly larger than that for human immunodeficiency virus particles. The distribution of particle sizes is broad, indicating that no two particles are likely identical in content or surface features. Virions possess knoblike protrusions, which may represent vestiges of budding from cell membranes. Particles which split open allowed imaging of intact cores with diameters of 65 nm. They also permitted estimation of viral shell thickness (35 to 40 nm) and showed the presence of a distinct trough between the shell and the core surface.

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Siri, F. M., L. A. Jelicks, L. A. Leinwand, and J. M. Gardin. "Gated magnetic resonance imaging of normal and hypertrophied murine hearts." American Journal of Physiology-Heart and Circulatory Physiology 272, no. 5 (May 1, 1997): H2394—H2402. http://dx.doi.org/10.1152/ajpheart.1997.272.5.h2394.

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Transgenic murine models are being used increasingly to explore the molecular basis of heart disease. Until recently, there were no means for noninvasive assessment of changes in mass and function of the murine heart because of its very small size and high heart rate. Transthoracic echocardiography has now been utilized to obtain noninvasive estimates of murine left ventricular (LV) wall thicknesses, internal dimension, and mass. However, this approach is based on one-dimensional (M-mode) measurements of the LV at its midwall that take no account of variations in LV chamber and wall dimensions along other minor axes and at other anatomic levels. Thus asymmetries in LV geometry, which can affect LV mass estimates, may be undetected. In this study, gated (diastolic) magnetic resonance imaging (MRI) was utilized to obtain two-dimensional images of the LV at four anatomic levels in intact, anesthetized mice. In 17 normal CD-1 mice (body mass, 18-47 g; gravimetric LV mass, 51-135 mg), LV mass estimates produced from the MRI data correlated well (r = 0.87) with LV mass determined gravimetrically. In addition, this approach identified changes in LV mass and wall thickness-to-chamber diameter ratio in a group of seven aortic-constricted mice (body mass, 32-39 g; gravimetric LV mass, 119-198 mg) with compensated and decompensated LV hypertrophy. These findings suggest that utility of MRI for serial, noninvasive assessment of experimentally induced alterations in mass and geometry of the murine heart.

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An, Dong Sung, Robert P. Wersto, Brian A. Agricola, Mark E. Metzger, Stephanie Lu, Rafael G. Amado, Irvin S. Y. Chen, and Robert E. Donahue. "Marking and Gene Expression by a Lentivirus Vector in Transplanted Human and Nonhuman Primate CD34+Cells." Journal of Virology 74, no. 3 (February 1, 2000): 1286–95. http://dx.doi.org/10.1128/jvi.74.3.1286-1295.2000.

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ABSTRACT Recently, gene delivery vectors based on human immunodeficiency virus (HIV) have been developed as an alternative mode of gene delivery. These vectors have a number of advantages, particularly in regard to the ability to infect cells which are not actively dividing. However, the use of vectors based on human immunodeficiency virus raises a number of issues, not the least of which is safety; therefore, further characterization of marking and gene expression in different hematopoietic lineages in primate animal model systems is desirable. We use two animal model systems for gene therapy to test the efficiency of transduction and marking, as well as the safety of these vectors. The first utilizes the rhesus animal model for cytokine-mobilized autologous peripheral blood CD34+ cell transplantation. The second uses the SCID-human (SCID-hu) thymus/liver chimeric graft animal model useful specifically for human T-lymphoid progenitor cell reconstitution. In the rhesus macaques, detectable levels of vector were observed in granulocytes, lymphocytes, monocytes, and, in one animal with the highest levels of marking, erythrocytes and platelets. In transplanted SCID-hu mice, we directly compared marking and gene expression of the lentivirus vector and a murine leukemia virus-derived vector in thymocytes. Marking was observed at comparable levels, but the lentivirus vector bearing an internal cytomegalovirus promoter expressed less efficiently than did the murine retroviral vector expressed from its own long terminal repeats. In assays for infectious HIV type 1 (HIV-1), no replication-competent HIV-1 was detected in either animal model system. Thus, these results indicate that while lentivirus vectors have no apparent deleterious effects and may have advantages over murine retroviral vectors, further study of the requirements for optimal use are warranted.

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Luongo, Diomira, Lucia Treppiccione, Francesco Maurano, Mauro Rossi, and Paolo Bergamo. "The murine enterocyte cell line Mode-K is a novel and reliable in vitro model for studies on gluten toxicity." Food and Chemical Toxicology 140 (June 2020): 111331. http://dx.doi.org/10.1016/j.fct.2020.111331.

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Signor, Luca, Theo Paris, Caroline Mas, Adrien Picard, Georges Lutfalla, Elisabetta Boeri Erba, and Laure Yatime. "Divalent cations influence the dimerization mode of murine S100A9 protein by modulating its disulfide bond pattern." Journal of Structural Biology 213, no. 1 (March 2021): 107689. http://dx.doi.org/10.1016/j.jsb.2020.107689.

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Nakamura, Jiro. "Calcium ionophore, A23187 alters the mode of cAMP formation in wild-type S49 murine lymphoma cells." Biochimica et Biophysica Acta (BBA) - Molecular Cell Research 1313, no. 1 (August 1996): 6–10. http://dx.doi.org/10.1016/0167-4889(96)00045-6.

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Wu, Mengfan, Dany Y. Matar, Zhen Yu, Ziyu Chen, Samuel Knoedler, Brian Ng, Oliver A. Darwish, et al. "Continuous NPWT Regulates Fibrosis in Murine Diabetic Wound Healing." Pharmaceutics 14, no. 10 (October 6, 2022): 2125. http://dx.doi.org/10.3390/pharmaceutics14102125.

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Scarring is associated with significant morbidity. The mechanical signaling factor yes-associated protein (YAP) has been linked to Engrailed-1 (En1)-lineage positive fibroblasts (EPFs), a pro-scarring fibroblast lineage, establishing a connection between mechanotransduction and fibrosis. In this study, we investigate the impact of micromechanical forces exerted through negative pressure wound therapy (NPWT) on the pathophysiology of fibrosis. Full-thickness excisional dorsal skin wounds were created on diabetic (db/db) mice which were treated with occlusive covering (control) or NPWT (continuous, −125 mmHg, 7 days; NPWT). Analysis was performed on tissue harvested 10 days after wounding. NPWT was associated with increased YAP (p = 0.04) but decreased En1 (p = 0.0001) and CD26 (p < 0.0001). The pro-fibrotic factors Vimentin (p = 0.04), α-SMA (p = 0.04) and HSP47 (p = 0.0008) were decreased with NPWT. Fibronectin was higher (p = 0.01) and collagen deposition lower in the NPWT group (p = 0.02). NPWT increased cellular proliferation (p = 0.002) and decreased apoptosis (p = 0.03). Western blotting demonstrated increased YAP (p = 0.02) and RhoA (p = 0.03) and decreased Caspase-3 (p = 0.03) with NPWT. NPWT uncouples YAP from EPF activation, through downregulation of Caspace-3, a pro-apoptotic factor linked to keloid formation. Mechanotransduction decreases multiple pro-fibrotic factors. Through this multifactorial process, NPWT significantly decreases fibrosis and offers promising potential as a mode to improve scar appearance.

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Neagoe, Raluca A. I., Elizabeth E. Gardiner, David Stegner, Bernhard Nieswandt, Steve P. Watson, and Natalie S. Poulter. "Rac Inhibition Causes Impaired GPVI Signalling in Human Platelets through GPVI Shedding and Reduction in PLCγ2 Phosphorylation." International Journal of Molecular Sciences 23, no. 7 (March 29, 2022): 3746. http://dx.doi.org/10.3390/ijms23073746.

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Rac1 is a small Rho GTPase that is activated in platelets upon stimulation with various ligands, including collagen and thrombin, which are ligands for the glycoprotein VI (GPVI) receptor and the protease-activated receptors, respectively. Rac1-deficient murine platelets have impaired lamellipodia formation, aggregation, and reduced PLCγ2 activation, but not phosphorylation. The objective of our study is to investigate the role of Rac1 in GPVI-dependent human platelet activation and downstream signalling. Therefore, we used human platelets stimulated using GPVI agonists (collagen and collagen-related peptide) in the presence of the Rac1-specific inhibitor EHT1864 and analysed platelet activation, aggregation, spreading, protein phosphorylation, and GPVI clustering and shedding. We observed that in human platelets, the inhibition of Rac1 by EHT1864 had no significant effect on GPVI clustering on collagen fibres but decreased the ability of platelets to spread or aggregate in response to GPVI agonists. Additionally, in contrast to what was observed in murine Rac1-deficient platelets, EHT1864 enhanced GPVI shedding in platelets and reduced the phosphorylation levels of PLCγ2 following GPVI activation. In conclusion, Rac1 activity is required for both human and murine platelet activation in response to GPVI-ligands, but Rac1’s mode of action differs between the two species.

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Mbow, M. Lamine, Robert D. Gilmore, and Richard G. Titus. "An OspC-Specific Monoclonal Antibody Passively Protects Mice from Tick-Transmitted Infection by Borrelia burgdorferi B31." Infection and Immunity 67, no. 10 (October 1, 1999): 5470–72. http://dx.doi.org/10.1128/iai.67.10.5470-5472.1999.

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ABSTRACT A murine monoclonal antibody directed against Borrelia burgdorferi B31 outer surface protein C (OspC) antigen was generated by a method whereby borreliae were inoculated into the mouse via the natural transmission mode of tick feeding. Passive immunization with this antibody resulted in protection of C3H/HeJ and outbred mice from a tick-transmitted challenge infection. Immunofluorescence staining of borrelia cells indicated surface exposure of the OspC epitope reactive with the monoclonal antibody.

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Battaglia, Manuela, Angela Stabilini, and Maria-Grazia Roncarolo. "Rapamycin selectively expands CD4+CD25+FoxP3+ regulatory T cells." Blood 105, no. 12 (June 15, 2005): 4743–48. http://dx.doi.org/10.1182/blood-2004-10-3932.

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Abstract Rapamycin is an immunosuppressive compound that is currently used to prevent acute graft rejection in humans. In addition, rapamycin has been shown to allow operational tolerance in murine models. However, a direct effect of rapamycin on T regulatory (Tr) cells, which play a key role in induction and maintenance of peripheral tolerance, has not been demonstrated so far. Here, we provide new evidence that rapamycin selectively expands the murine naturally occurring CD4+CD25+FoxP3+ Tr cells in vitro. These expanded Tr cells suppress proliferation of syngeneic T cells in vitro and prevent allograft rejection in vivo. Interestingly, rapamycin does not block activation-induced cell death and proliferation of CD4+ T cells in vitro. Based on this new mode of action, rapamycin can be used to expand CD4+CD25+FoxP3+ Tr cells for ex vivo cellular therapy in T-cell-mediated diseases. (Blood. 2005;105:4743-4748)

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Matsui, Takeshi, Nanako Kadono-Maekubo, Yoshiro Suzuki, Yuki Furuichi, Keiichiro Shiraga, Hiroyuki Sasaki, Azusa Ishida, et al. "A unique mode of keratinocyte death requires intracellular acidification." Proceedings of the National Academy of Sciences 118, no. 17 (April 23, 2021): e2020722118. http://dx.doi.org/10.1073/pnas.2020722118.

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The stratum corneum (SC), the outermost epidermal layer, consists of nonviable anuclear keratinocytes, called corneocytes, which function as a protective barrier. The exact modes of cell death executed by keratinocytes of the upper stratum granulosum (SG1 cells) remain largely unknown. Here, using intravital imaging combined with intracellular Ca2+- and pH-responsive fluorescent probes, we aimed to dissect the SG1 death process in vivo. We found that SG1 cell death was preceded by prolonged (∼60 min) Ca2+ elevation and rapid induction of intracellular acidification. Once such intracellular ionic changes were initiated, they became sustained, irreversibly committing the SG1 cells to corneocyte conversion. Time-lapse imaging of isolated murine SG1 cells revealed that intracellular acidification was essential for the degradation of keratohyalin granules and nuclear DNA, phenomena specific to SC corneocyte formation. Furthermore, intravital imaging showed that the number of SG1 cells exhibiting Ca2+ elevation and the timing of intracellular acidification were both tightly regulated by the transient receptor potential cation channel V3. The functional activity of this protein was confirmed in isolated SG1 cells using whole-cell patch-clamp analysis. These findings provide a theoretical framework for improved understanding of the unique molecular mechanisms underlying keratinocyte-specific death mode, namely corneoptosis.

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Milenic, D. E., B. Detrick, J. C. Reynolds, and D. Colcher. "Characterization of Primate Antibody Responses to Administered Murine Monoclonal Immunoglobulin." International Journal of Biological Markers 5, no. 4 (October 1990): 177–87. http://dx.doi.org/10.1177/172460089000500403.

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Murine monoclonal antibodies (MAb) are currently being assessed for their utility as tools in cancer management. Anti-murine immunoglobulin responses have been observed in many patients receiving monoclonal antibody treatment. In this study, we evaluated the response of primates to the administration of a monoclonal antibody. MAb B6.2, an antibody generated against a human breast tumor metastasis, was used as a prototype MAb. Baboons were inoculated with MAb B6.2 whole IgG, Fab', or F(ab')2 fragments. Blood samples were drawn at periodic intervals post-inoculation and the sera collected. Anti-murine immunoglobulin responses were detected using a solid-phase radioimmunoassay. The specificity of the antibody response was analyzed to determine if the response was directed against the species of origin of the MAb (species specificity), against the class of the MAb (isotype specificity), or against the hypervariable region of the MAb (idiotype specificity). We found that primates develop a humoral immune response against all three forms of the monoclonal antibody [IgG, Fab', and F(ab')2]. Furthermore, this antibody response demonstrated a high degree of specificity for the antigen binding site suggesting an idiotypic specificity. Using a competitive radioimmunoassay, the antibody response was found to interfere with antigen binding of MAb B6.2. These studies suggest that monoclonal antibody treatment can generate an anti-idiotypic response which may alter the efficacy of this mode of treatment.

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Weissman, A. M., P. Ross, E. T. Luong, P. Garcia-Morales, M. L. Jelachich, W. E. Biddison, R. D. Klausner, and L. E. Samelson. "Tyrosine phosphorylation of the human T cell antigen receptor zeta-chain: activation via CD3 but not CD2." Journal of Immunology 141, no. 10 (November 15, 1988): 3532–36. http://dx.doi.org/10.4049/jimmunol.141.10.3532.

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Abstract TCR stimulation by Ag or anti-receptor antibodies in murine T cells results in the activation of two independent protein kinases, protein kinase C (PKC) and a protein tyrosine kinase. Similarly, stimulation of murine Thy-1 or Ly-6 with mAb also results in activation of both of these kinase pathways. Tyrosine phosphorylation in all cases occurs on the TCR zeta-chain. It is known that Ag and anti-receptor antibodies activate PKC in human T cells. In this study we demonstrate that mitogen or anti-CD3 antibodies activate tyrosine phosphorylation of the human TCR-zeta-chain. PMA, which activates PKC, does not result in zeta-chain tyrosine phosphorylation. Stimulation of human T cells by antibodies that bind the CD2 molecule is an alternate mode of inducing T cell proliferation. These antibodies surprisingly do not induce tyrosine phosphorylation of the zeta-chain. Thus, different methods of cellular activation can result in distinguishable patterns of receptor-mediated biochemical signaling events.

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Taguchi, Fumihiro, and Shutoku Matsuyama. "Soluble Receptor Potentiates Receptor-Independent Infection by Murine Coronavirus." Journal of Virology 76, no. 3 (February 1, 2002): 950–58. http://dx.doi.org/10.1128/jvi.76.3.950-958.2002.

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ABSTRACT Mouse hepatitis virus (MHV) infection spreads from MHV-infected DBT cells, which express the MHV receptor CEACAM1 (MHVR), to BHK cells, which are devoid of the receptor, by intercellular membrane fusion (MHVR-independent fusion). This mode of infection is a property of wild-type (wt) JHMV cl-2 virus but is not seen in cultures infected with the mutant virus JHMV srr7. In this study, we show that soluble MHVR (soMHVR) potentiates MHVR-independent fusion in JHMV srr7-infected cultures. Thus, in the presence of soMHVR, JHMV srr7-infected DBT cells overlaid onto BHK cells induce BHK cell syncytia and the spread of JHMV srr7 infection. This does not occur in the absence of soMHVR. soMHVR also enhanced wt virus MHVR-independent fusion. These effects were dependent on the concentration of soMHVR in the culture and were specifically blocked by the anti-MHVR monoclonal antibody CC1. Together with these observations, direct binding of soMHVR to the virus spike (S) glycoprotein as revealed by coimmunoprecipitation demonstrated that the effect is mediated by the binding of soMHVR to the S protein. Furthermore, fusion of BHK cells expressing the JHMV srr7 S protein was also induced by soMHVR. These results indicated that the binding of soMHVR to the S protein expressed on the DBT cell surface potentiates the fusion of MHV-infected DBT cells with nonpermissive BHK cells. We conclude that the binding of soMHVR to the S protein converts the S protein to a fusion-active form competent to mediate cell-cell fusion, in a fashion similar to the fusion of virus and cell membranes.

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Nelles, Megan Elizabeth, Alain Labbe, Jagdeep Walia, Lintao Jia, Caren Furlonger, Takahiro Nonaka, Jeffrey A. Medin, and Christopher J. Paige. "Interleukin-12 initiates an immune response that leads to tumour rejection (B151)." Journal of Immunology 178, no. 1_Supplement (April 1, 2007): LB31—LB32. http://dx.doi.org/10.4049/jimmunol.178.supp.b151.

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Abstract Immunotherapies are developed based on the notion that cancer cells can be targeted and eliminated by an appropriately stimulated immune system. Integral to both the innate and acquired immune systems, cytokines are ideal candidates for use in immunotherapy. We have developed two systems of interleukin-12 (IL-12) therapy, in a murine model, that elicit protective immune responses against the acute lymphoblastic leukemia (ALL) cell line, 70Z/3. The first system involves direct injection of IL-12, and the second is a tumour cell-mediated approach. Initially we found that direct delivery of low doses of IL-12 is sufficient to elicit a long-term protective immune response against an established tumour burden, mediated by both CD4+ and CD8+ T cells. Based on this knowledge, we created a lentiviral vector expressing murine IL-12, and transduced 70Z/3 cells to obtain IL-12 secreting tumor cells. We demonstrated that anti-tumour immunity in the second model is long lasting but is primarily dependent on the CD4+ T cell subset alone. Our results highlight that the mode of IL-12 delivery has a distinct impact on the immune response that is initiated and can lead to tumour clearance by way of disparate mechanisms. Furthermore, we demonstrate some of the underlying conditions within the tumour microenvironment that may have resulted in these differential outcomes.

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Garland, Russell, Christopher Kirkham, Michelle Yap, Louise Brackenbury, Tommaso Iannitti, Robert Nunan, and S. Jenkinson. "6 Pre-clinical pharmacodynamic biomarker assays of immune modulation can translate to inform exploratory endpoints of target engagement in first-in-human clinical trial stages of drug discovery." Journal for ImmunoTherapy of Cancer 8, Suppl 3 (November 2020): A6. http://dx.doi.org/10.1136/jitc-2020-sitc2020.0006.

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BackgroundLack of efficacy is a common cause of failure in Phase I and Phase II clinical trials. Pharmacodynamic (PD) biomarker assays can demonstrate target engagement and proof of mechanism; both key components to improve trial success. Biomarkers established at the pre-clinical phase can serve as exploratory endpoints in early phase clinical trials, to confirm the mode of action of the therapeutic. We show examples of human in vitro assays and murine T cell adoptive transfer models, which can be used to establish potential PD biomarkers for inclusion in the clinical phases.MethodsHuman peripheral blood mononuclear cell (PBMC) were incubated with SEB in the presence of Pembrolizumab or Ipilimumab. IL-2 and IFNgamma levels were quantified by Luminex. To identify biomarkers of checkpoint inhibition, mice transferred with a defined population of ovalbumin (OVA)-specific T cells were challenged with OVA antigen or EG7 tumour. Activation and proliferation of antigen-specific T cells was determined and Nanostring gene expression analysis performed. Flow cytometry staining panels for human immune markers including CD4, CD14, CD25 and FOXP3 were established pre-clinically. As part of the assay validation process for a clinical trial, whole blood SEB activation was performed in normal donors, with Luminex analysis of IL-2, IL-17, IFNgamma and TNFalpha.ResultsImmune checkpoint inhibitors resulted in increased IL-2 and IFNgamma secretion in human PBMC stimulated with SEB. In the murine PD model, anti-PD-L1 caused upregulation of CD25, IFNgamma and granzyme B by antigen-specific CD8 T cells. Gene expression analysis of murine tumours elucidated changes in response to a vaccine. Flow cytometry panel staining determined the frequencies of human Treg and monocytes, which are common targets of immune-modulating therapies. Fit-for-purpose validation was performed for a human SEB activation assay resulting in robust changes in cytokine production.ConclusionsThe experiments here show the flow of experiments that can be performed to identify a PD biomarker for use in first in man trials; the pre-clinical human PBMC SEB screening assay provides a simple assay demonstrating that a therapy can enhance T cell function and would be translatable to the clinic. The murine PD model provides a platform to screen for biomarkers of T cell function and monitor gene expression modulation. Biomarkers identified in the murine setting provide a good starting point for exploratory assessment in early phase clinical trials, where inclusion of exploratory PD biomarker endpoints in can confirm proof of mechanism and improve study success rates.Ethics ApprovalHuman tissues used in this study were collected with ethical approval from UK Research Ethics Committee South West, Bristol (UK), approval number 15/SW/0029.

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Greulich, Franziska, Kirsten Adele Bielefeld, Ronny Scheundel, Aikaterini Mechtidou, Benjamin Strickland, and Nina Henriette Uhlenhaut. "Enhancer RNA Expression in Response to Glucocorticoid Treatment in Murine Macrophages." Cells 11, no. 1 (December 23, 2021): 28. http://dx.doi.org/10.3390/cells11010028.

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Glucocorticoids are potent anti-inflammatory drugs; however, their molecular mode of action remains complex and elusive. They bind to the glucocorticoid receptor (GR), a nuclear receptor that controls gene expression in almost all tissues in a cell type-specific manner. While GR’s transcriptional targets mediate beneficial reactions in immune cells, they also harbor the potential of adverse metabolic effects in other cell types such as hepatocytes. Here, we have profiled nascent transcription upon glucocorticoid stimulation in LPS-activated primary murine macrophages using 4sU-seq. We compared our results to publicly available nascent transcriptomics data from murine liver and bioinformatically identified non-coding RNAs transcribed from intergenic GR binding sites in a tissue-specific fashion. These tissue-specific enhancer RNAs (eRNAs) correlate with target gene expression, reflecting cell type-specific glucocorticoid responses. We further associate GR-mediated eRNA expression with changes in H3K27 acetylation and BRD4 recruitment in inflammatory macrophages upon glucocorticoid treatment. In summary, we propose a common mechanism by which GR-bound enhancers regulate target gene expression by changes in histone acetylation, BRD4 recruitment and eRNA expression. We argue that local eRNAs are potential therapeutic targets downstream of GR signaling which may modulate glucocorticoid response in a cell type-specific way.

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Smethurst, Peter A., Lotta Joutsi-Korhonen, Marie N. O'Connor, Erica Wilson, Nicola S. Jennings, Stephen F. Garner, Yanjun Zhang, et al. "Identification of the primary collagen-binding surface on human glycoprotein VI by site-directed mutagenesis and by a blocking phage antibody." Blood 103, no. 3 (February 1, 2004): 903–11. http://dx.doi.org/10.1182/blood-2003-01-0308.

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Abstract Glycoprotein (GP) VI is the major receptor responsible for platelet activation by collagen, but the collagen-binding surface of GPVI is unknown. To address this issue we expressed, from insect cells, the immunoglobulin (Ig)–like ectodomains (residues 1-185) of human and murine GPVI, called hD1D2 and mD1D2, respectively. Both proteins bound specifically to collagen-related peptide (CRP), a GPVI-specific ligand, but hD1D2 bound CRP more strongly than did mD1D2. Molecular modeling and sequence comparison identified key differences between hD1D2 and mD1D2. Ten mutant hD1D2s were expressed, of which 4 had human residues replaced by their murine counterpart, and 6 had replacements by alanine. CRP binding studies with these mutants demonstrated that the exchange of lysine at position 59 for the corresponding murine glutamate substantially reduced binding to CRP. The position of lysine59 on the apical surface of GPVI suggests a mode of CRP binding analogous to that used by the related killer cell Ig-like receptors to bind HLA. This surface was confirmed as critical for collagen binding by epitope mapping of an inhibitory phage antibody against GPVI. This anti-GPVI, clone 10B12, gave dose-dependent inhibition of the hD1D2-collagen interaction. Clone 10B12 inhibited activation of platelets by CRP and collagen in aggregometry and thrombus formation by the latter in whole blood perfusion. Antibody 10B12 showed significantly reduced binding to the hD1D2-E59, and, on that basis, the GPVI:10B12 interface was modeled.

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Sifaoui, Ines, Rubén L. Rodríguez-Expósito, María Reyes-Batlle, Robert Sutak, José E. Piñero, and Jacob Lorenzo-Morales. "Amoebicidal Effect of COVID Box Molecules against Acanthamoeba: A Study of Cell Death." Pharmaceuticals 17, no. 6 (June 20, 2024): 808. http://dx.doi.org/10.3390/ph17060808.

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Acanthamoeba spp. can cause a sight threatening disease. At present, the current treatments used to treat Acanthamoeba spp. Infections, such as biguanide-based antimicrobials, remain inefficacious, with the appearance of resistant forms and high cytotoxicity to host cells. In this study, an initial screening was conducted against Acanthamoeba castellanii Neff and murine macrophages J774A.1 using alamarBlue™. Among the 160 compounds included in the cited box, 90% exhibited an inhibition of the parasite above 80%, while only 18.75% of the compounds inhibited the parasite with a lethality towards murine macrophage lower than 20%. Based on the amoebicidal activity, the cytotoxicity assay, and availability, Terconazole was chosen for the elucidation of the action mode in two clinical strains, Acanthamoeba culbertsoni and Acanthamoeba castellanii L10. A fluorescence image-based system and proteomic techniques were used to investigate the effect of the present azole on the cytoskeleton network and various programmed cell death features, including chromatin condensation and mitochondria dysfunction. Taking all the results together, we can suggest that Terconazole can induce programmed cell death (PCD) via the inhibition of sterol biosynthesis inhibition.

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Chen, Zirong, Peng Peng, Xiaolin Zhang, Barbara Mania-Farnell, Guifa Xi, and Feng Wan. "Advanced Pediatric Diffuse Pontine Glioma Murine Models Pave the Way towards Precision Medicine." Cancers 13, no. 5 (March 5, 2021): 1114. http://dx.doi.org/10.3390/cancers13051114.

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Diffuse intrinsic pontine gliomas (DIPGs) account for ~15% of pediatric brain tumors, which invariably present with poor survival regardless of treatment mode. Several seminal studies have revealed that 80% of DIPGs harbor H3K27M mutation coded by HIST1H3B, HIST1H3C and H3F3A genes. The H3K27M mutation has broad effects on gene expression and is considered a tumor driver. Determination of the effects of H3K27M on posttranslational histone modifications and gene regulations in DIPG is critical for identifying effective therapeutic targets. Advanced animal models play critical roles in translating these cutting-edge findings into clinical trial development. Here, we review current molecular research progress associated with DIPG. We also summarize DIPG animal models, highlighting novel genomic engineered mouse models (GEMMs) and innovative humanized DIPG mouse models. These models will pave the way towards personalized precision medicine for the treatment of DIPGs.

Journal articles on the topic 'Murine mode'

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