Імуногістохімічна характеристика субпопуляцій лімфоцитів селезінки кролів

Abstract

Представлені результати імуногістохімічної характеристики субпопуляцій лімФоцитів 6ілої пульпи селезінки статевозрілих кролів каліфорнійської породи віком 6-8-ми місяців. З’ясовано розміщення, кількісний вміст субпопуляцій лімфоцитів CD4+, CD8+, CD19+, CD20+, вираховано імунорегуляторний індекс. Встановлено, що лімфоцити CD4+, CD8+ більше зосереджувалися в мантійній зоні, ніж у світлому центрі лімфоїдного вузлика. Імунорегуляторний індекс періартеріальних лімфоїдних піхв становив 1,34, білої пульпи 0,92. Серед популяцій В-лімфоцитів незначно переважали СД 19+. Найбільша їх кількість знаходилась в мантійній зоні, найменша в періартеріальних лімфоїдних піхвах. В маргінальній зоні наявні всі субпопуляції.

Представлены результаты иммуногистохимической характеристики субпопуляций лимфоцитов белой пульпы селезёнки половозрелых кроликов калифорнийской породы возрастом 6-8-ми месяцев. Выяснено размещение, количественное содержание субпопуляций лимфоцитов CD4+, CD8+, CD19+, CD20+, рассчитан иммунорегуляторный индекс. Установлено, что лимфоциты Cd4+, Cd8+ больше сосредоточивались в мантийний зоне, чем в светлом центре лимфоидного фолликула. Иммунорегуляторный индекс периартериальных лимфоидных муфт составлял 1,34, белой пульпы 0,92. Cреди популяций В-лимфоцитов незначительно преобладали ОД 19+. Наибольшее их количество находилось в мантийний зоне, наименьшее в периартериальных лимфоидных муфтах. В маргинальной зоне имеющиются все субпопуляции.

The modern achievements of histo - and cytoimmunohistochemistry with the monoclonal antibodies and enzyme-linked immunosorbent assay are allowed detecting on the surface of lymphocytes and other immunocompetent cells the specific receptors that can serve as markers of various subpopulations of cells. Such immunohistochemical studies should be carried out under pathological conditions, primarily, disorders of the immune system, study the effects on populations of different factors. This is especially important to implement for productive of farm animals with the aim of obtaining the safe food. Object and methods. For research were conducted selections of mature spleen сalifornia breed rabbits (6-8 mons) of both sexes (ratio female: male was 1:1). For histological studies pieces of material recorded in the 10-12% refrigerated neutral formalin solution, with subsequent filling in paraffin. Paraffin sections were made at sledge microtome MC-2, with a thickness less than 5 microns. To identify and study subpopulations of lymphocytes by light microscopy are used murine monoclonal antibodies from Danish company DAKO, expressing antigenic markers CD4+, CD8+, CD19+, CD20+. The placement, content (absolute and relative quantity) and proportion of populations were determined. The immunoregulatory index was determined as the ratio of CD4+ cells to CD8+ cells. Research results. The subpopulations of cells with CD4+, CD8+, CD19+, CD20+ in the splenic pulp was placed solitary and diffuses, often formed of a chain or cluster. The number of CD4+ in lymphoid sheaths near the vessels made up of 20,25 % of the total number of populations of the white pulp, and the rest (79,75 %) were located in pieces per unit area. The bright center of the lymphoid nodule number was 45,29±13,34 pieces per unit area, in mantle zone - 34,14± 9,96 pieces per unit area which amounted to 24,61 % and 75,39 %, respectively. The cells with CD8+ in lymphoid sheaths near the vessels were 8,6±4,18 pieces per unit area (13,99 % of the total number of population in the white pulp), in lymphoid nodule - 52,86±19,02 pieces per unit area. In the mantle zone of lymphoid nodule this population was predominant to compared to the bright center of lymphoid nodule in 1,8 times (21,00± 8,94 and 37,80±14,88 pieces per unit area respectively). The immunoregulatory index of the lymphoid sheaths near the vessels was > 1 (1,34), <1 in the white pulp (0,92), the lowest immunoregulatory index in the light center (0,53), in the mantle zone this index was equal to 0,90. The light center number of CD19+ made up 32,64 % of the total population of the lymphoid nodule in the white pulp, their number is totaled 18,33±2,36 pieces per unit area. In the light center number of CD 20+ was increased by approximately 2 % and accounted for 34,75 % of the total population of the lymphoid nodule in white pulp, their number have consisted of 19,85±3,59 pieces per unit area. Such a small difference is because CD 19+cells are at a very early stage of development; the CD 20+cells are at the stages of differentiation of b-cells. Quantitative analysis of subpopulations of lymphocytes of the splenic white pulp of rabbits is indicated that among the studied T-cells is dominated the CD8+. This is due to the results which obtained in the study of mice. The CD8+ cells can additionally expressive also in dendritic cells. A significant content of B-cells is explained to the feature of the primary immune response and the presence of considerable amount of soluble antigens which the most appealing these populations.