Splíchal I, Reháková Z, Sinkora M, Sinkora J, Trebichavský I, Laude H, Charley B
Division of Immunology and Gnotobiology, Academy of Sciences of the Czech Republic, Nový Hrádek (Czech Republic).
Res Immunol. 1997 May;148(4):247-56. doi: 10.1016/s0923-2494(97)80866-8.
Non-infectious UV-inactivated transmissible gastroenteritis virus (TGEV) was previously shown to induce interferon alpha (IFN alpha) secretion following in vitro incubation with blood mononuclear cells. In this study, pig foetuses at different stages of gestation were injected in utero with (a) partially UV-inactivated wild TGEV or (b) fully UV-inactivated wild or dm49-4 mutant TGEV coronavirus. Nucleated cells from foetal liver, bone marrow, spleen and blood were isolated 10 or 20 h after injection and assayed ex vivo for IFN alpha secretion by ELISPOT and ELISA techniques. The administration of TGEV induced IFN alpha-secreting cells in foetal lymphohaematopoietic organs at mid-gestation. In contrast, IFN alpha was not detected in control sham-operated foetuses. A specific point mutation in the amino acid sequence of the viral membrane glycoprotein M of TGEV mutant dm49-4 was associated with lower or absent IFN alpha in utero inducibility by mutant virus as compared with wild virus. Flow cytometry analysis did not show differences in leukocyte surface marker expression between control and TGEV- or between dm49-4 and wild virus-treated foetus cells, with the exception of a reduction in percentages of polymorphonuclear cells in TGEV-treated lymphohaematopoietic tissues, which is probably due to IFN alpha secretion. The present data provided in vivo evidence of IFN alpha secretion at the cell level in foetal lymphohaematopoietic organs. Such IFN alpha-secreting cells in lymphohaematopoietic tissues may be the source of IFN alpha detected during foetal infections.
先前的研究表明,经紫外线灭活的非感染性传染性胃肠炎病毒(TGEV)与血液单核细胞进行体外孵育后可诱导α干扰素(IFNα)分泌。在本研究中,对处于不同妊娠阶段的猪胎儿进行子宫内注射:(a)部分紫外线灭活的野生TGEV,或(b)完全紫外线灭活的野生或dm49 - 4突变TGEV冠状病毒。注射后10或20小时,分离胎儿肝脏、骨髓、脾脏和血液中的有核细胞,并通过ELISPOT和ELISA技术在体外检测IFNα分泌。在妊娠中期,TGEV的给药诱导胎儿淋巴造血器官中分泌IFNα的细胞。相比之下,在假手术对照胎儿中未检测到IFNα。与野生病毒相比,TGEV突变体dm49 - 4的病毒膜糖蛋白M氨基酸序列中的特定点突变与突变病毒在子宫内诱导IFNα的能力降低或缺失有关。流式细胞术分析显示,对照与TGEV处理组或dm49 - 4与野生病毒处理组的胎儿细胞之间白细胞表面标志物表达没有差异,但TGEV处理的淋巴造血组织中多形核细胞百分比降低,这可能是由于IFNα分泌所致。本研究数据提供了胎儿淋巴造血器官细胞水平上IFNα分泌的体内证据。淋巴造血组织中这些分泌IFNα的细胞可能是胎儿感染期间检测到的IFNα的来源。