Antigenic properties of a water soluble fraction of sheep erythrocytes

Séman, Michel; Mazié, J. C.; Bussard, A

doi:10.1002/eji.1830020421

Cited by 19 publications

(7 citation statements)

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“…However, homologous PFC were inhibited to an extent of only 60-70 % [6]. Inhibition features were better in mice immunized with the soluble antigen when 85 % of homologous PFC were inhibited.…”

Section: Discussionmentioning

confidence: 89%

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Bispecific cells among IgM and IgG producers during the early phase of primary and secondary responses

et al. 1975

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Simultaneous immunization of mice with sheep (SRBC) and horse (HRBC) erythrocytes regularly resulted in the appearance of hemolytic plaque-forming cells (PFC) specific for each type of erythrocyte and also of PFC lysing both types of erythrocytes. After primary stimulation the highest number of bispecific cells (42/10(6) cells) was found among PFC as revealed by the direct procedure (IgM producers). Among PFC enhanced with anti-mouse Fab serum (IgG producers), bispecific cells were less numerous (8/10(6) cells). In preparations enhanced by anti-mouse-gammaFc serum which reveals IgG producers without inhibiting IgM antibody, the number of bispecific PFC equalled the sum of bispecific cells revealed by direct and anti-Fab enhanced procedures. The number of direct bispecific PFC during primary and secondary response was approximately the same, whereas the number of IgG-producing, bispecific PFC increased considerably during the secondary response. Another difference was the time limitation of the appearance of bispecific cells: after primary immunization direct bispecific PFC were detected only on days 3, 4 and 5, but enhanced bispecific PFC were present from day 4 up to day 12. However, during the secondary reaction, bispecific PFC were detected by all three procedures only between days 3 and 6. Studies on the cross-reactivity between SRC and HRBC gave negative results at the humoral level, even when the mice were primed with a minimal amount of both erythrocytes and then two months later, boosted with one of them. Studies at the cellular level showed that after immunization with one antigen, only 0.4 to 0.7 direct or enhanced PFC/10(6) cells could simultaneously lyse both erythrocyte types. Thus, a hundred times more bispecific PFC were constantly found after double immunization of the animals. Moreover, sudden disappearance of all bispecific PFC on the 7th day after secondary stimulation makes it unlikely that all bispecific PFC are simply cross-reacting cells.

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Section: Discussionmentioning

confidence: 89%

“…Before use they were washed three times in phosphate buffered 0.05 M 0.9 % saline, pH 7 (PBS). Soluble SRBC and HRBC antigens were prepared according t o the method of Seman et al [6]. In short, washed erythrocytes were lysed by the addition of distilled water (1.…”

Section: Antigensmentioning

confidence: 99%

Bispecific cells among IgM and IgG producers during the early phase of primary and secondary responses

et al. 1975

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“…Influenza virus strain A/PR/8 and water-soluble antigen of sheep erythrocytes obtained as described elsewhere [5] were used as T cell-dependent antigens. Polyvinylpyrrolidone with a molecular weight of 350 kD and dinitrophenyl-Ficoll served as TI-2 antigens.…”

Section: Methodsmentioning

confidence: 99%

Role of various subpopulations of B lymphocytes in production of antigen-induced nonspecific immunoglobulins

et al. 1999

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Congeneric CBA and CBA/N mice (with and without Lyb5+ B cells, respectively) respond to T cell-dependent antigens by the appearance of antibody-producing cells and accumulation of cells synthesizing nonspecific immunoglobulins. Type 2 cell-independent antigens induce similar changes only in CBA mice having mature B lymphocytes. The data suggest that B lymphocytes are responsible for the appearance of antibody-producing cells and polyclonal activation. Key Words: antibodies; nonspecific immunoglobulins; polyclonal activation; B cell subpopulationContact with antigens stimulates the synthesis of specific antibodies and sharply enhances production of nonspecific immunoglobulins [3]. Induction of the immune response usually requires the presence of T helper cells. Some antigens induce the immune response without T helpers, so-called T cell-independent (TI) antigens. TI can be divided into 2 types: TI-I and TI-2 antigens. TI-1 antigens are polyclonal activators stimulating production of antibodies by mature (Lyb5+) and immature (Lyb5-) B lymphocytes, while TI-2 antigens possess no mitogenic properties and stimulate the production of antibodies only by mature Lyb5+ B lymphocytes [4,6]. The mechanisms of the effect of TI-2 remain unclear. It is unknown whether nonspecific immunoglobulins are synthesized only by mature B lymphocytes or they are also produced by immature B cells. The factors contributing to polyclonal activation are still unstudied.Here we studied the role of Lyb5+ and Lyb5-B lymphocytes in polyclonal activation induced by TI-2 antigens. Laboratory of Biosynthesis of Immunoglobulins, Institute of Viral Preparations, Russian Academy of Medical Sciences, Moscow MATERIALS AND METHODSExperiments were performed on congeneric CBA and CBA/N mice (with and without Lyb5+ B cells, respectively) obtained from Stolbovaya nursery and gifted by B. V. Nikonenko (Institute of Tuberculosis).Influenza virus strain A/PR/8 and water-soluble antigen of sheep erythrocytes obtained as described elsewhere [5] were used as T cell-dependent antigens. Polyvinylpyrrolidone with a molecular weight of 350 kD and dinitrophenyl-Ficoll served as TI-2 antigens.In in vivo experiments, the mice were intravenously injected with 1-2 ~tg polyvinylpyrrolidone, 5 pg influenza virus, or 500-700 ~tg sheep erythrocytes antigen. Antibody-producing (APC) and immunoglobulinsynthesizing (IGSC) cells of splenocyte suspensions were counted on day 4 using ELISPOT [1].During in vitro induction of the immune response, splenocytes from nonimmunized animals were incubated in 96-well plates with 5 pg/ml influenza virus and 10 ng/ml polyvinylpyrrolidone or 30 ng/ml dinitrophenyl-Ficoll in RPMI-1640 medium containing 10% fetal serum and other additives for 4 days in a CO 2 incubator at 37~ The number of APC and IGSC

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“…В качестве ТЗ АГ использовали водораство-римый антиген бараньих эритроцитов (ВРА-БЭ) [11], ТН-1 АГ липополисахарид Escherichia coli cеротип 0111: В4 (LPS) (Sigma), ТН-2 АГ -α(1→3) декстран L. mesenteroides (Декс) и поли-винилпирролидон 350 кДа (ПВП) (Sigma). ТЗ АГ вносили в культуры клеток в дозе 25 мкг/мл, ТН-1 АГ -1 нг/мл и ТН-2 АГ -10 нг/мл.…”

Section: материалы и методыunclassified

INFLUENCE OF Breg AND IL-10 UPON HUMORAL IMMUNE RESPONSE

Gavrilova¹,

Снегирева²,

Sidorova³

2016

Med. immunol.

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Адрес для переписки:Гаврилова Марина Викторовна ФГБНУ «Научно-исследовательский институт вакцин и сывороток им. И.И. Мечникова» 115533, Россия, Москва, Нагатинская наб., 18, кв. 90. Тел.: 8 (499) 618-49-93; (495) 674-08-42. E-mail: gavrilovamv@gmail.com Address for correspondence: Gavrilova Marina V. Vaccines and Sera 115533, Russian Federation, Moscow, Nagatinskaya emb., 18, apt 90. Phone: 7 (499) 618-49-93; (495) 674-08-42. E-mail: gavrilovamv@gmail.com Образец цитирования: М.В. Гаврилова, Н.А. Снегирева, Е.В. Сидорова «Влияние Breg и IL-10 на гуморальный иммунный ответ» // Медицинская иммунология, 2016. Т. 18, № 4. С. 331-338. doi: 10.15789/1563-0625-2016-4-331-338 © Гаврилова М.В. и соавт., 2016 For citation: M.V. Gavrilova, N.A. Snegireva, E.V. Sidorova "Influence of Breg and IL-10 upon humoral immune response", Medical Immunology (Russia)/Meditsinskaya Immunologiya, 2016, Vol. 18, no. 4, pp. 331-338. doi: 10.15789/1563-0625-2016 Резюме. Breg угнетают патологические процессы при воспалительных и аутоиммунных заболева-ниях. Регуляторные функции Breg обусловлены в основном секрецией IL-10. Роль Breg в гумораль-ном иммунном ответе не изучалась. В данной работе было исследовано влияние IL-10 и Breg при иммунном ответе В1-и В2-клеток мыши на Т-зависимые и Т-независимые антигены в модельной системе in vitro. В качестве Т-зависимого антигена использовали водорастворимый антиген бараньих эритроцитов, Т-независимого антигена 1 типа -LPS, Т-независимых антигенов 2 типа поливинил-пирролидон и α(1→3) декстран. В1-и В2-лимфоциты были выделены из клеток перитонеальной по-лости и селезенки мышей линии СВА соответственно. Клетки культивировались в среде RPMI 1640 с 10% ЭТС и всеми необходимыми добавками с антигенами, IL-10 и без них. Число антитело-и им-муноглобулин-образующих клеток определяли методом ELISPOT. I.I. Mechnikov Research Institute forПод действием водорастворимого антигена бараньих эритроцитов число антитело-и иммуногло-булин-образующих клеток в культурах В1-и В2-лимфоцитов возрастало. Добавление IL-10 приво-дило к снижению числа антителопродуцентов в среднем на 27%. IL-10 снижал также число индуци-рованных LPS антитело-и иммуноглобулин-образующих клеток в культуре В2-лимфоцитов на 75%. Под действием IL-10 в культуре В1-клеток, активированных Т-независимыми антигенами 2 типа, число антителопродуцентов уменьшалось в среднем на 50%.Для изучения функциональной активности Breg выделяли из клеток перитонеальной полости и селезенки мышей линии СВА. Количество Breg после активации В-клеток LPS, иономицином и форболовым эфиром возрастало в 20 раз (с 4 до 96%). Основным изотипом иммуноглобулинов, секретируемых Breg, являлся IgM. При чистоте Вreg 96% иммуноглобулины секретировали ~12% этих клеток. Это означает, что некоторая часть Breg одновременно синтезирует иммуноглобулины и IL-10.Получение высокоочищенных Breg и использование трансвеллов позволило исследовать бес-контактное влияние Breg на иммунный ответ в культуре спленоцитов xid-мышей линии CBA/N. Было установлено, что как специфический,...

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Antigenic properties of a water soluble fraction of sheep erythrocytes

Cited by 19 publications

References 5 publications

Bispecific cells among IgM and IgG producers during the early phase of primary and secondary responses

Bispecific cells among IgM and IgG producers during the early phase of primary and secondary responses

Role of various subpopulations of B lymphocytes in production of antigen-induced nonspecific immunoglobulins

INFLUENCE OF Breg AND IL-10 UPON HUMORAL IMMUNE RESPONSE

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