Khatri Mahesh, Sharma Jagdev M
Department of Veterinary and Biomedical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN 55108, USA.
Vet Immunol Immunopathol. 2009 Feb 15;127(3-4):316-24. doi: 10.1016/j.vetimm.2008.10.327. Epub 2008 Nov 5.
We exposed chicken embryos at embryonation day 18 (ED18) to a classical virulent infectious bursal disease virus (IBDV; cIBDV) and an attenuated strain of IBDV (aIBDV) and examined the response of embryonic lymphoid cells to these viruses. Embryos responded much more vigorously to cIBDV than to aIBDV. Following cIBDV exposure, embryonic thymus and bursa showed cellular destruction, enhanced rate of apoptosis and presence of viral proteins detectable by immunohistochemistry. At ED21, thymocytes from cIBDV-exposed embryos were severely deficient (P<0.05) in responding to stimulation in vitro with mitogens containing mouse anti-chicken CD28 mAb, PMA and ionomycin. Because purified CD3(+) T cells were also refractory to the mitogens, the mitogenic inhibition of embryonic thymocytes was not attributed to the presence of non-T cell suppressors. Cell suspensions prepared from embryonic thymus and spleen had upregulated gene expression of IFN-gamma and IL-6 cytokines and of chemokine IL-8. In sharp contrast to cIBDV, embryos exposed to aIBDV had minimal detectable changes in the thymus and bursa, although the rate of apoptosis was enhanced in the thymus. Viral antigen was not detectable in the bursa until after hatch. Thymocytes from these embryos responded vigorously to the mitogens, similar to the response of thymocytes from unexposed control embryos. In addition, aIBDV induced a modest gene upregulation of IFN-gamma, IL-6 and IL-8 in thymus and spleen. Relatively modest response of the embryo to aIBDV is significant because in ovo vaccination with aIBDV-type viruses and several other non-pathogenic viruses result in protective immunity that is well pronounced at hatch.
我们在胚胎发育第18天(ED18)将鸡胚暴露于经典强毒传染性法氏囊病病毒(IBDV;cIBDV)和IBDV减毒株(aIBDV),并检测胚胎淋巴细胞对这些病毒的反应。胚胎对cIBDV的反应比对aIBDV的反应强烈得多。暴露于cIBDV后,胚胎胸腺和法氏囊出现细胞破坏、凋亡率增加以及免疫组化可检测到病毒蛋白。在ED21时,来自暴露于cIBDV的胚胎的胸腺细胞在用含小鼠抗鸡CD28单克隆抗体、佛波酯和离子霉素的丝裂原进行体外刺激时反应严重不足(P<0.05)。由于纯化的CD3(+) T细胞对丝裂原也不敏感,胚胎胸腺细胞的丝裂原抑制作用并非归因于非T细胞抑制因子的存在。从胚胎胸腺和脾脏制备的细胞悬液中,IFN-γ、IL-6细胞因子以及趋化因子IL-8的基因表达上调。与cIBDV形成鲜明对比的是,暴露于aIBDV的胚胎胸腺和法氏囊中可检测到的变化极小,尽管胸腺中的凋亡率有所增加。直到孵化后,法氏囊中才检测到病毒抗原。这些胚胎的胸腺细胞对丝裂原反应强烈,类似于未暴露的对照胚胎的胸腺细胞的反应。此外,aIBDV诱导胸腺和脾脏中IFN-γ、IL-6和IL-8的基因适度上调。胚胎对aIBDV的相对适度反应很重要,因为用aIBDV型病毒和其他几种非致病性病毒进行卵内接种可产生在孵化时就很明显的保护性免疫。