Імуногістохімічне дослідження селезінки свині

Abstract

Представлені результати розміщення, кількісного вмісту субпопуляцій лімфоцитів СD4+, СD8+, СD19+, СD20+ селезінки статевозрілих свиней віком 8-10 місяців, вирахувано імунорегуляторний індекс. Встановлено, що лімфоцити СD4+, СD8+ більше зосереджувалися в періартеріальних лімфоїдних піхвах та періартеріальних зонах лімфоїдного вузлика. Імунорегуляторний індекс пульпи селезінки дорівнював 1,71. Серед популяцій В-лімфоцитів незначно переважали СD 19+. Найбільше лімфоцитів СD19+, СD20+ локалізувалось в мантійній, маргінальній зонах та світлому центрі лімфоїдних вузликів; Представлены результаты размещения, количественного содержания субпопуляций лимфоци- тов СD4+, СD8+, СD19+, СD20+ пульпы селезёнки половозрелых свиней возрастом 8-10 месяцев, рассчитан иммунорегуляторный индекс. Установлено, что лимфоциты СD4+, СD8+ больше сосредоточивались в периартериальных лимфоидных влагалищах и периартериальных зонах лимфоидного фолликула. Иммунорегуляторный индекс пульпы селезёнки был равен 1,71. Среди популяций В-лимфоцитов незначительно преобладали СD19+. Больше всего лимфоцитов СD19+, СD20+ локализовалось в мантийной, маргинальной зонах и светлом центре лимфоидных фолликулов; Immunohistochemical studies should be carried out at the pathological states of the organism, first of all, with violations in the work of the immune system, the study of the impact on the population of various factors. It is especially important to implement it for productive farm animals in order to obtain safe food products. Object and methods. For research was selection of mature spleens of pig (8-10 months) of both sexes (ratio female: male was 1:1). For histological studies, pieces of material are recorded in a 10-12% refrigerated neutral formalin solution, with subsequent filling in paraffin. The paraffin sections were made at the sledge microtome MC-2, with a thickness of fewer than 5 microns. For the detection and study of lymphocyte subpopulations, light microscopy used mouse monoclonal antibodies from the Danish firm DAKO, markers of CD4 +, CD8 +, CD19 +, and CD20 +. The placement and content (absolute and relative quantity) and the quantitative population ratio were determined. Research results. Subpopulations of lymphocytes with CD4+, CD8+, CD19+, and CD20+ were isolated and diffuse, often forming chains or clusters. The lymphocytes with markers CD4+, CD8+, CD19+, CD20+ have a rarefied location in the red pulp of the spleen. CD4+ lymphocytes were unevenly distributed in the pulp: the overwhelming majority was localized in lymphoid nodule (66,97 %), in lymphoid sheats near the vessels of white splenic pulp was 30,73 % of the total number of pulp populations, the smallest remainder (2,30 %) was located in red pulp. In the structure of lymphoid nodule, these lymphocytes were also distributed unevenly: 39,95 % were localized in the zone near the vessels, 6,52 % – in mantle zone of lymphoid nodule (or 4,24±1,28 pieces per unit area), in the light centre of lymphoid nodule their number was equal to 20,43±5,12 pieces per unit area, in the marginal zone 14,63±3,07 pieces per unit area). The CD8+ lymphocytes were predominantly concentrated in the lymphoid nodule of white pulp (60,28 %), the remainder – in lymphoid sheats near the vessels, to wit 38,1 % or 22,28±4,17 pieces per unit area and insignificant in red pulp – 1,62 %, which was only 0,95±0,78 pieces per unit area. The morphometric studies have shown that in the mantle zone of the lymphoid nodule of this population was significantly less in 5,95 times the light center of the lymphoid nodule (2,39±0,85 pieces per unit area and 10,86±2,83 pieces per unit area, respectively). The amount of lymphocytes CD8+ in the in a zone near the vessels of the lymphoid nodule was 14,21±2,38 pieces per unit area, which was 40,3 % of the total number of lymphoid nodule white pulp spleen. Among the populations of B-lymphocytes, the spleen of the pig’s spleen was slightly superior to CD19 + -lymphocytes compared with lymphocytes with markers of СD20+ (75,14±7,13 and 71,95±7,36 pieces per unit area, respectively). The largest number of these lymphocytes were found in the lymphoid nodule (77,93 %), and the lowest – in red pulp (8,6 %). In lymphoid sheats near the vessels of the white pulp, their share was 13,47 %. In lymphoid nodule, lymphocytes CD19+ were dominated in the marginal zone (44,96 %). In the light center of a lymphoid nodule, the number of lymphocytes CD 19+ was amounted to 29,92 % of the total number of this lymphoid nodule in white pulp lymphocytes and was 17,52±3,41 pieces per unit area. In the mantle zone and zone near the vessels of lymphoid nodule lymphocytes with markers CD19+ were occupied 14,38 % and 10,74 % of their total lymphoid nodule number. Immunohistochemical studies have established the distribution of lymphocytes with markers of СD20+ as follows: 76,96 % was localized in the lymphoid nodule of white pulp, 13,0 % in a zone near the vessels of white pulp and 7,23 % in red pulp.