Cote P J, Gerin J L
Department of Microbiology and Immunology, Georgetown University Medical Center, Rockville, MD 20852, USA.
Hepatology. 1995 Sep;22(3):687-99.
Cellular immune responses to hepatitis B virus (HBV) play an important role in the resolution of acute infection. They also influence the course of chronic infection and disease but are inadequate to completely clear the infection. Woodchuck hepatitis virus (WHV) infection of the woodchuck can provide a model to study these processes. Lymphocyte responses of woodchucks were assessed by in vitro proliferation and/or interleukin (IL)-2 assays using mitogen (Concanavalin A [ConA]), cytokine (IL-2), superantigen (Staphylococcus aureus enterotoxin B [SEB]), major histocompatibility complex (MHC) allo-antigen (mixed lymphocyte reaction [MLR]), and viral antigens (woodchuck hepatitis virus core antigen [WHcAg] and woodchuck hepatitis virus surface antigen [WHsAg]). ConA-stimulated woodchuck lymphocytes underwent cell division based on cell counting experiments and produced IL-2 as detected using an IL-2-dependent murine cell line but failed to incorporate sufficient tritiated thymidine; however, they did incorporate sufficient tritiated adenosine and deoxyadenosine to permit development of a meaningful proliferation assay. The IL-2 assay was sensitive and specific for detection of woodchuck IL-2 induced by mitogen, superantigen, and MLR, as shown by quantitative titration analysis and anti-body neutralization of ConA-supernatant activity. Cyclosporin A and FK506 specifically inhibited ConA- and SEB-induced IL-2 production by woodchuck lymphocytes. Positive two-way MLRs were detected by IL-2 production and proliferation assay between woodchucks from different geographic regions, thus indicating divergence among MHC molecules; however, occasional negative MLR reactions among indigenous pairs of woodchucks indicated that some woodchucks were mutually immunocompatible to some degree. The radioadenosine proliferation assay was sensitive for detecting peripheral blood lymphocyte responses to WHcAg and WHsAg in adult woodchucks with recently resolved acute infections. The above systems should facilitate the design of adoptive therapy and liver transplantation experiments in the woodchuck, and also enable modeling of immune responses that promote and maintain chronic hepadnavirus infection.
对乙型肝炎病毒（HBV）的细胞免疫反应在急性感染的消退过程中发挥着重要作用。它们也会影响慢性感染和疾病的进程，但不足以完全清除感染。土拨鼠感染土拨鼠肝炎病毒（WHV）可为研究这些过程提供一个模型。通过体外增殖和/或白细胞介素（IL）-2检测来评估土拨鼠的淋巴细胞反应，检测使用了丝裂原（刀豆蛋白A [ConA]）、细胞因子（IL-2）、超抗原（金黄色葡萄球菌肠毒素B [SEB]）、主要组织相容性复合体（MHC）同种抗原（混合淋巴细胞反应 [MLR]）以及病毒抗原（土拨鼠肝炎病毒核心抗原 [WHcAg] 和土拨鼠肝炎病毒表面抗原 [WHsAg]）。根据细胞计数实验，ConA刺激的土拨鼠淋巴细胞发生了细胞分裂，并产生了IL-2，这是使用依赖IL-2的小鼠细胞系检测到的，但未能掺入足够的氚标记胸腺嘧啶；然而，它们确实掺入了足够的氚标记腺苷和脱氧腺苷，从而能够开展有意义的增殖检测。如通过定量滴定分析以及ConA上清液活性的抗体中和所显示的，IL-2检测对于检测由丝裂原、超抗原和MLR诱导的土拨鼠IL-2具有敏感性和特异性。环孢素A和FK506特异性抑制土拨鼠淋巴细胞由ConA和SEB诱导的IL-2产生。通过来自不同地理区域的土拨鼠之间的IL-2产生和增殖检测，检测到了阳性双向MLR，这表明MHC分子之间存在差异；然而，在本地土拨鼠对之间偶尔出现的阴性MLR反应表明，一些土拨鼠在一定程度上相互免疫相容。放射性腺苷增殖检测对于检测近期急性感染已消退的成年土拨鼠外周血淋巴细胞对WHcAg和WHsAg的反应具有敏感性。上述系统应有助于设计土拨鼠的过继性治疗和肝移植实验，还能够对促进和维持慢性嗜肝DNA病毒感染的免疫反应进行建模。
