Kreuzer S, Rieger J, Strucken E M, Thaben N, Hünigen H, Nöckler K, Janczyk P, Plendl J, Brockmann Gudrun A
Humboldt-Universität zu Berlin, Breeding Biology and Molecular Genetics, Invalidenstr. 42, D-10115 Berlin, Germany.
Freie Universität Berlin, Department of Veterinary Medicine, Institute of Veterinary Anatomy, Koserstraße 20, 14195 Berlin, Germany.
Vet Immunol Immunopathol. 2014 Apr 15;158(3-4):143-55. doi: 10.1016/j.vetimm.2014.01.001. Epub 2014 Jan 8.
The aim of the present study was to test the effect of Enterococcus faecium NCIMB 10415 (E. faecium) on CD4+ T helper immune cell subpopulations and CD25+ cells in ileal lymphatic tissue after challenge with Salmonella (S.) Typhimurium DT 104. German Landrace piglets treated with E. faecium (n=16) as a feed additive and untreated controls (n=16) were challenged with S. Typhimurium 10 days after weaning. The expression of lineage specific T helper cell subtype master transcription factors on mRNA level was measured in the whole tissue of the gut associated lymphoid tissues (ileocecal mesenteric lymph node, ileum with Peyer's patches and papilla ilealis) and in magnetically sorted T helper cells from blood and ileocecal mesenteric lymph nodes at two and 28 days post infection. CD25 protein expression of T helper cells was studied by flow cytometry in ileal Peyer's patches, lymph nodes and blood. Distribution and morphology of CD25+ cells was demonstrated in situ by immunohistochemistry in paraffin embedded specimens of the ileum and the ileocecal mesenteric lymph nodes. The data provide evidence for a higher T helper 2 cell driven immune response in the control group compared to the E. faecium treated group (P<0.05) in CD4+ magnetically sorted lymphocytes from the ileocecal mesenteric lymph nodes at two and 28 days post infection. We did not observe differences for CD25+ cells in immunohistochemistry and flow cytometry between E. faecium fed pigs and the control group, but provided a detailed description of the occurrence and morphology of these cells in the gut associate lymphoid tissues of piglets. In conclusion we suggest that (i) prolonged feeding with E. faecium can result in changes of the T helper cell response leading to a stronger infection with S. Typhimurium and (ii) that it is important to examine purified immune cells to be able to detect effects on T helper cell subpopulations.
本研究的目的是检测屎肠球菌NCIMB 10415（屎肠球菌）对经鼠伤寒沙门氏菌DT 104攻击后回肠淋巴组织中CD4⁺辅助性T免疫细胞亚群和CD25⁺细胞的影响。断奶10天后，将作为饲料添加剂用屎肠球菌处理的德国长白仔猪（n = 16）和未处理的对照仔猪（n = 16）用鼠伤寒沙门氏菌进行攻击。在感染后第2天和第28天，在肠道相关淋巴组织（回盲肠系膜淋巴结、带有派尔集合淋巴结和回肠乳头的回肠）的全组织以及从血液和回盲肠系膜淋巴结中磁性分选的辅助性T细胞中，测量谱系特异性辅助性T细胞亚群主转录因子在mRNA水平上的表达。通过流式细胞术研究回肠派尔集合淋巴结、淋巴结和血液中辅助性T细胞的CD25蛋白表达。通过免疫组织化学在回肠和回盲肠系膜淋巴结的石蜡包埋标本中原位显示CD25⁺细胞的分布和形态。数据表明，在感染后第2天和第28天，来自回盲肠系膜淋巴结的CD4⁺磁性分选淋巴细胞中，与屎肠球菌处理组相比，对照组中辅助性T2细胞驱动的免疫反应更强（P<0.05）。我们未观察到喂食屎肠球菌的猪和对照组在免疫组织化学和流式细胞术中CD25⁺细胞存在差异，但详细描述了这些细胞在仔猪肠道相关淋巴组织中的出现情况和形态。总之，我们认为（i）长期喂食屎肠球菌可导致辅助性T细胞反应发生变化，从而导致鼠伤寒沙门氏菌感染增强，以及（ii）检查纯化的免疫细胞对于能够检测对辅助性T细胞亚群的影响很重要。
