IMMUNOHISTOCHEMICAL INVESTIGATION OF THE PIG’S SPLEEN

Dunaіevska О. F.

BIOLOGY

Scentific article

Immunohistochemical studies should be carried out at the pathological states of the organism, frst of all, with violatons in the work of the immune system, the study of the impact on the populaton of various factors. It is especially important to implement it for productve farm animals in order to obtain safe food products. Object and methods. For research was selecton of mature spleens of pig (8-10 months) of both sexes (rato female: male was 1:1). For histological studies, pieces of material are recorded in a 10-12% refrigerated neutral formalin soluton, with subsequent flling in parafn. The parafn sectons were made at the sledge microtome MC-2, with a thickness of fewer than 5 microns. For the detecton and study of lymphocyte subpopulatons, light microscopy used mouse monoclonal antbodies from the Danish frm DAKO, markers of CD4 +, CD8 +, CD19 +, and CD20 +. The placement and content (absolute and relatve quantty) and the quanttatve populaton rato were determined. Research results. Subpopulatons of lymphocytes with CD4+, CD8+, CD19+, and CD20+ were isolated and diffuse, ofen forming chains or clusters. The lymphocytes with markers CD4+, CD8+, CD19+, CD20+ have a rarefed locaton 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 populatons, 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 insignifcant 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 populaton was signifcantly less in 5,95 tmes the light center of the lymphoid nodule (2,39±0,85 pieces per unit area and 10,86±2,83 pieces per unit area, respectvely). 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 populatons 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, respectvely). 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 distributon 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.

spleen, immunohistochemistry, CD-lymphocytes, morphology, morphometry, pig, pulp, lymphoid follicles

• Kvaratsheliya AG, Klochkova SV, Nikityuk DB, Alekseeva NT. Morfologicheskaya harakteristika timusa i selezyonki pri vozdeystvii faktorovrazlichnogo proishozhdeniya. Zhurnal anatomii i gistopatologii. 2016;5(3):77-83. [in Russiаn].
• Panіkar II. Morfogenez organіv іmunnoyi sistemi svіyskoyi svinі na rannіh etapah postnatalnogo perіodu ontogenezu [avtoreferat]. K.: NYBiP;2015. 42 s. [in Ukrainian].
• Brinza L, Djebali S, Tomkowiak M, Maflle J, Loiseau C, Schicklin S, et al. Immune signatures of protectve spleen memory CD8 T cells. Scientfc Reports. 2016. DOI: 10.1038/srep37651
• Dunaіеvska OF. Imunogіstohіmіchna harakteristka subpopulyatsіy lіmfotsitіv selezіnki krolіv. Vіsnik problem biologіyi і meditsini.2017;3;2(138):60-3. [in Ukrainian].
• Dunaіеvska OF. Imunogistohimichna harakteristka subpopulyatsiy limfotsitv selezinki golubiv ta kurey. Ukrainian Journal of Ecology.2018;8(1):282-8. Dostupno: DOI: htp://dx.doi.org/ 10.15421/2018_213 [in Ukrainian].
• Xiaoxu D, Gao S, Li J, Wu L, Zhang Y, Li W, et al. Acute arsenic exposure induces inflammatory responses and CD4+Tcell subpopulatons differentaton in spleen and thymus with the involvement of MAPK, NF-kB, and Nrf2 / Nrf2. Molecular Immunology. 2017;81:160-72. Availablefrom: htp://dx.doi.org/10.1016/j.molimm. 2016.12.005 0161-5890
• Benkisser-Petersen M, Buchner M, Dörffel A, Dühren-von-Minden M, Claus R, Kläsener K, et al. Spleen Tyrosine Kinase Is Involved in theCD38 Signal Transducton Pathway in Chronic Lymphocytc Leukemia. PLOS ONE. 2016 December 30. DOI: 10.1371/journal.pone.0169159
• Fedorovskaya NS, Dyakonov DA. Immunomorfologicheskaya harakteristka selezyonki pri tsitopeniyah immunnogo genezisa. Kirov: Avers;2013. 101 s. [in Russiаn].
• Seo YJ, Jothikumar P, Suthar MS, Zhu C, Grakoui A. Local Cellular and Cytokine Cues in the Spleen Regulate In Situ T Cell Receptor Affnity, Functon, and Fate of CD8+ T Cells. Immunity. 2016 [cited 10 Dec 2016];45:988-98. Available from: htp://dx.doi.org/10.1016/j.immuni.2016.10.024
• Goralskiy LP, Homich VT, Kononskiy OI. Osnovi gіstologichnoyi tehniki і morfofunktsіonalnі metodi doslіdzhen u normі ta pri patologіyi.Zhitomir: Polissya; 2012. 288 s. [in Ukrainian].
• Dunaіеvska OF. Morfometrichni osoblivosti selezinki sviney. Visnik Lvіvskogo un-tu. Ser.: biologichna. 2016;72:202-9. [in Ukrainian]

«Bulletin of problems biology and medicine» Issue 1 Part 2 (143), 2018 year, 52-55 pages, index UDK 636.92:591.441

10.29254/2077-4214-2018-1-2-143-52-55