Вопросы вирусологии. 2020; 65: 35-40
Характеристика В1-клеток в процессе экспериментального лейкомогенеза
https://doi.org/10.36233/0507-4088-2020-65-1-35-40Аннотация
Введение. Вирус лейкоза крупного рогатого скота (КРС) вызывает значительную поликлональную экспансию CD5+ , IgM+ B-лимфоцитов, известных как персистирующий лимфоцитоз приблизительно у 30% заражённого КРС. Однако пока не ясно, что происходит с данной субпопуляцией В-клеток в ранний период инфицирования животных.
Цель исследования – количественная характеристика IgM+ и CD5+ В-клеток в процессе иммунного ответа на заражение вирусом лейкоза крупного рогатого скота (BLV), которая может дать важную информацию о механизмах прайминга лимфоцитов при инфицировании BLV.
Материал и методы. В эксперименте использовали BLV-отрицательных телят чёрно-пёстрой породы в возрасте 8 мес (n = 11). Животным опытной группы (n = 8) внутривенно вводили кровь BLV-положительной коровы. Телятам контрольной группы (n = 3) вводили физиологический раствор. Исследования проводили до и после заражения на 5, 7, 14, 21, 28 и 65-е сутки иммунного ответа. Количество В-лимфоцитов в крови определяли методом иммунопероксидазного окрашивания на основе моноклональных антител к IgM и CD5.
Результаты. В результате проведённых исследований установлено, что уровень CD5+ В-клеток повышается на 14-е сутки первичного иммунного ответа, характеризующегося поликлональной пролиферацией CD5+ В-клеток, которые являются первичной мишенью для BLV. Данные исследований подтверждают, что в лимфоцитах экспериментально заражённого КРС поверхностная агрегация молекул IgM и CD5 на В-лимфоцитах отсутствует.
Обсуждение. Известно, что именно от субпопуляции В1-клеток зависит волнообразный характер синтеза IgM, который был показан в предыдущих исследованиях. После 7-х суток иммунного ответа показатели IgM+ и CD5+-клеток не коррелируют, что показывает их функциональное различие. Возможно, увеличение числа CD5+-клеток связано не с В-клетками, а с дифференцирующимися под влиянием вируса Т-лимфоцитами.
Выводы. Субпопуляция В1-клеток является первичной мишенью вируса лейкоза КРС. 65-е сутки иммунного ответа характеризуются экспансией IgM+ В-клеток, снижением числа CD5+-клеток и равномерным распределением рецепторов по периметру клеток.
Список литературы
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4. Brym P., Ruść A., Kamiński S. Evaluation of reference genes for qRT-PCR gene expression studies in whole blood samples from healthy and leukemia-virus infected cattle. Vet. Immunol. Immunopathol. 2013; 153(3-4): 302-7. http://doi.org/10.1016/j.vetimm.2013.03.004
5. Mayr B., Vogel I., Graninger W., Schlerka G., Wöckl F., Schleger W. Circulating immune cells and immune complexes in peripheral blood of healthy and of bovine leukemia virus-infected cows and lymphosarcomatous calves. Vet. Immunol. Immunopathol. 1982; 3(5): 475-84. http://doi.org/10.1016/0165-2427(82)90013-7
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16. Сantor C.H., Pritchard S.M., Dequiedt F., Willems I., Kettmann R., Davis W.C. CD5 is dissociated from the B-cell receptor in B cells from BLV-infected, PL cattle: consequences to B-cell receptor-mediated apoptosis. J. Virol. 2001; 75(4): 1689-96. http://doi.org/10.1128/JVI.75.4.1689-1696.2001
17. Gillet N., Florins A., Boxus M., Burteau C., Nigro A., Vandermeers F., et al. Mechanisms of leukemogenesis induced by BLV: prospects for novel anti-retroviral therapies in human. Retrovirology. 2007; 4: 18. http://doi.org/10.1186/1742-4690-4-18
18. Suzuki S., Konnai S., Okagawa T., Ikebuchi R., Nishimori A., Kohara J., et al. Increased expression of the regulatory T cellassociated marker CTLA-4 in bovine leukemia virus infection. Vet. Immunol. Immunopathol. 2015; 163(3-4): 115-24. http://doi.org/10.1016/j.vetimm.2014.10.006
19. Hamilton V.T., Stone D.M., Cantor G.H. Translocation of the B cell receptor to lipid rafts is inhibited in B cells from BLV-infected, persistent lymphocytosis cattle. Virology. 2003; 315(1): 135-47. http://doi.org/10.1016/s0042-6822(03)00522-1
Problems of Virology. 2020; 65: 35-40
Characterization of B1-cells during experimental leukomogenesis
https://doi.org/10.36233/0507-4088-2020-65-1-35-40Abstract
Background. Bovine leukemia causes a significant polyclonal expansion of CD5+ , IgM+ B lymphocytes, known as persistent lymphocytosis (PL), in approximately 30% of infected cattle. However, it is not yet clear what happens to this subpopulation of B cells in the early period of infection of animals.
Purpose. Quantitative characterization of IgM+ and CD5+ B cells during the immune response, which can provide important information on the mechanisms of lymphocyte priming in BLV infection.
Material and methods. The experiment used BLV-negative calves of black-motley breed at the age of 8 months (n = 11). Animals (n = 8) were intravenously injected with blood of a BLV-positive cow. Control calves (n = 3) were injected with saline. Studies were performed before and after infection on days 5, 7, 14, 21, 28 and 65 of the immune response. The determination of the number of B-lymphocytes in the blood was carried out by the method of immunoperoxidase staining based on monoclonal antibodies to IgM, CD5.
Results. As a result of the studies, it was found that the level of CD5+ B cells increases on the 14th day of the primary immune response, characterized by polyclonal proliferation of CD5+ B cells, which are the primary target for BLV. Our research data confirm that in the lymphocytes of experimentally infected cattle, surface aggregation of IgM and CD5 molecules on B-lymphocytes is absent.
Discussion. It is known that the wave-like nature of IgM synthesis, which was shown in previous studies, depends on a subpopulation of B1 cells. After 7 days of the immune response, IgM+ and CD5+ cells do not correlate, which shows their functional difference. The increase in CD5+ cells is probably not associated with B cells, but with T cells differentiating under the influence of the virus.
Conclusions. A subset of B1 cells is the primary target of cattle leukemia virus. The 65th day of the immune response is characterized by the expansion of IgM+ B cells, a decrease in the number of CD5+ cells and a uniform distribution of receptors around the perimeter of the cells.
References
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2. Frie M.C., Sporer K.R., Wallace J.C., Maes R.K., Sordillo L.M., Bartlett P.C., et al. Reduced humoral immunity and atypical cell-mediated immunity in response to vaccination in cows naturally infected with bovine leukemia virus. Vet. Immunol. Immunopathol. 2016; 182: 125-35. http://doi.org/10.1016/j.vetimm.2016.10.013
3. Florins A., Boxus M., Vandermeers F., Verlaeten O., Bouzar A.B., Defoiche J., et al. Emphasis on cell turnover in two hosts infected by bovine leukemia virus: A rationale for host susceptibility to disease. Vet. Immunol. Immunopathol. 2008; 125(1-2): 1-7. http://doi.org/10.1016/j.vetimm.2008.04.007
4. Brym P., Ruść A., Kamiński S. Evaluation of reference genes for qRT-PCR gene expression studies in whole blood samples from healthy and leukemia-virus infected cattle. Vet. Immunol. Immunopathol. 2013; 153(3-4): 302-7. http://doi.org/10.1016/j.vetimm.2013.03.004
5. Mayr B., Vogel I., Graninger W., Schlerka G., Wöckl F., Schleger W. Circulating immune cells and immune complexes in peripheral blood of healthy and of bovine leukemia virus-infected cows and lymphosarcomatous calves. Vet. Immunol. Immunopathol. 1982; 3(5): 475-84. http://doi.org/10.1016/0165-2427(82)90013-7
6. Murakami K., Sentsui H., Inoshima Y., Inumaru S. Increase in gammadelta T cells in the blood of cattle persistently infected with bovine leukemia virus following administration of recombinant bovine IFN-gamma. Vet. Immunol. Immunopathol. 2004; 101(1-2): 61-71. http://doi.org/10.1016/j.vetimm.2004.04.016
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9. Murakami K., Inumaru S., Yokoyama T., Okada K., Sentsui H. Expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor on B-1a cell from persistent lymphocytosis (PL) cows and lymphoma cell induced by bovine leukemia virus. Vet. Immunol. Immunopathol. 1999; 68(1): 49-59. http://doi.org/10.1016/s0165-2427(99)00011-2
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13. Mirsky M.L., Olmstead C.A., Da Y., Lewin H.A. The prevalence of proviral bovine leukemia virus in peripheral blood mononuclear cells at two subclinical stages infection. J. Virol. 1996; 70(4): 2178-83.
14. Juliarena M.A., Barrios C.N., Ceriani C.M., Esteban E.N. Hot topic: Bovine leukemia virus (BLV)-infected cows with low proviral load are not a source of infection for BLV-free cattle. J. Dairy Sci. 2016; 99(6): 4586-9. http://doi.org/10.3168/jds.2015-10480
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16. Santor C.H., Pritchard S.M., Dequiedt F., Willems I., Kettmann R., Davis W.C. CD5 is dissociated from the B-cell receptor in B cells from BLV-infected, PL cattle: consequences to B-cell receptor-mediated apoptosis. J. Virol. 2001; 75(4): 1689-96. http://doi.org/10.1128/JVI.75.4.1689-1696.2001
17. Gillet N., Florins A., Boxus M., Burteau C., Nigro A., Vandermeers F., et al. Mechanisms of leukemogenesis induced by BLV: prospects for novel anti-retroviral therapies in human. Retrovirology. 2007; 4: 18. http://doi.org/10.1186/1742-4690-4-18
18. Suzuki S., Konnai S., Okagawa T., Ikebuchi R., Nishimori A., Kohara J., et al. Increased expression of the regulatory T cellassociated marker CTLA-4 in bovine leukemia virus infection. Vet. Immunol. Immunopathol. 2015; 163(3-4): 115-24. http://doi.org/10.1016/j.vetimm.2014.10.006
19. Hamilton V.T., Stone D.M., Cantor G.H. Translocation of the B cell receptor to lipid rafts is inhibited in B cells from BLV-infected, persistent lymphocytosis cattle. Virology. 2003; 315(1): 135-47. http://doi.org/10.1016/s0042-6822(03)00522-1
