Dimier-Poisson I H, Bout D T, Quéré P
Equipe associé INRA Im munologie Parasitaire et vaccinologie UFR des Sciences Pharmaceutiques, 37200 Tours, France.
Avian Dis. 2004 Sep;48(3):617-24. doi: 10.1637/7180-031604R.
A reproducible and original method for the preparation of chicken intestine epithelial cells from 18-day-old embryos for long-term culture was obtained by using a mechanical isolation procedure, as opposed to previous isolation methods using relatively high concentrations of trypsin, collagenase, or EDTA. Chicken intestine epithelial cells typically expressed keratin and chicken E-cadherin, in contrast to chicken embryo fibroblasts, and they increased cell surface MHC II after activation with crude IFN-gamma containing supernatants, obtained from chicken spleen cells stimulated with concanavalin A or transformed by reticuloendotheliosis virus. Eimeria tenella was shown to be able to develop until the schizont stage after 46 hr of culture in these chicken intestinal epithelial cells, but it was not able to develop further. However, activation with IFN-gamma containing supernatants resulted in strong inhibition of parasite replication, as shown by incorporation of [3H]uracil. Thus, chicken enterocytes, which are the specific target of Eimeria development in vivo, could be considered as potential local effector cells involved in the protective response against this parasite.
通过机械分离程序，获得了一种可重复且新颖的方法，用于从18日龄胚胎制备鸡肠上皮细胞以进行长期培养，这与先前使用相对高浓度胰蛋白酶、胶原酶或乙二胺四乙酸（EDTA）的分离方法不同。与鸡胚成纤维细胞相比，鸡肠上皮细胞通常表达角蛋白和鸡E-钙黏蛋白，并且在用含有粗制γ干扰素的上清液激活后，它们的细胞表面主要组织相容性复合体II（MHC II）增加，这些上清液是从用刀豆球蛋白A刺激或经网状内皮组织增殖病病毒转化的鸡脾细胞中获得的。结果表明，柔嫩艾美耳球虫在这些鸡肠上皮细胞中培养46小时后能够发育到裂殖体阶段，但无法进一步发育。然而，用含有γ干扰素的上清液激活导致寄生虫复制受到强烈抑制，这通过[3H]尿嘧啶掺入得以证明。因此，鸡肠上皮细胞作为柔嫩艾美耳球虫在体内发育的特定靶细胞，可被视为参与针对这种寄生虫的保护性反应的潜在局部效应细胞。
