Cell subsets responding to purified hepatocytes and evidence of indirect recognition of hepatocyte major histocompatibility complex class I antigen. I. The role of L3T4+ T cells in the development of allospecific cytotoxicity in hepatocyte-sponge matrix allografts.
Cell subsets responding to purified hepatocytes and evidence of indirect recognition of hepatocyte major histocompatibility complex class I antigen. I. The role of L3T4+ T cells in the development of allospecific cytotoxicity in hepatocyte-sponge matrix allografts.
复制标题
对纯化肝细胞作出反应的细胞亚群以及间接识别肝细胞主要组织相容性复合物 I 类抗原的证据。
作者：
G. Bumgardner;Sally C. Chen;P. Almond;Fritz H. Bach;N. Ascher;A. J. Matas
本研究的目的是确定L3T4+、Ly2- T细胞在肝细胞-海绵基质同种异体移植（HC-SMA）中体内纯化的同种异体MHC I+、II类肝细胞对同种异体细胞溶解T细胞的反应中所起的作用。在之前的研究中，我们已经证明99%纯小鼠肝细胞在体外的混合淋巴细胞-肝细胞培养（MLHC）和体内的HC-SMA中刺激同种异体细胞溶解T细胞的发育。此外，在混合淋巴细胞-肝细胞培养中，反应性脾细胞的L3T4+， Ly2- T细胞的消耗抑制了对纯化肝细胞的同种异体ctl的发育。在这里，我们使用抗L3T4单克隆抗体，测试了体内L3T4+， Ly2- T细胞免疫缺失对HC-SMA中allo- ctl后续发展的影响。对照组和处理组分别于移植后第4天和第12天收获海绵细胞，并通过FACS和免疫荧光标记进行表型分析。同样对来自同一动物的脾细胞进行分析，以评估免疫清除的完整性。移植后第12天评估同种异体细胞毒性。我们发现抗L3T4单抗免疫治疗可有效清除脾脏中的L3T4+、Ly2- T细胞；然而，对照组和处理组从海绵中分离出的L3T4+细胞数量相似。此外，抗l3t4单抗的局部和全身免疫治疗完全消除了HC-SMA中肝细胞应答的同种异体ctl的发展。我们从这些和先前的数据得出结论，在体外和体内，MLHC中的宿主或应答者L3T4+、Ly2- T细胞和应答者辅助细胞或HC-SMA中的宿主巨噬细胞可能参与肝细胞I类抗原的“间接”识别。
The purpose of this study was to determine the role of L3T4+, Ly2- T cells in the development of allospecific cytolytic T cells in response to purified allogeneic MHC class I+, class II- hepatocytes in vivo in hepatocyte-sponge matrix allografts (HC-SMA). In previous studies we have shown that 99% pure murine hepatocytes stimulate the development of allospecific cytolytic T cells in vitro in mixed lymphocyte-hepatocyte culture (MLHC) and in vivo in HC-SMA. Furthermore, depletion of L3T4+, Ly2- T cells from responder splenocytes inhibits the development of allo-CTLs in response to purified hepatocytes in mixed lymphocyte-hepatocyte culture. Here, using an anti-L3T4 monoclonal antibody, we tested the effect of in vivo immunodepletion of L3T4+, Ly2- T cells on the subsequent development of allo-CTLs in HC-SMA. Sponge cells were harvested on day 4 and day 12 after grafting from control and treated groups and phenotypically analyzed by FACS and immunofluorescent labelling. Splenocytes from the same animals were similarly analyzed to assess for completeness of immunodepletion. Allospecific cytotoxicity was assessed on day 12 after grafting. We found that immunotherapy with anti-L3T4 mAb was effective in depleting L3T4+, Ly2- T cells from the spleen; however, a similar number of L3T4+ cells was isolated from the sponge between control and treated groups. Furthermore, the development of allo-CTLs in response to hepatocytes in HC-SMA was completely abrogated by both local and systemic immunotherapy with anti-L3T4 mAb. We conclude from these and previous data that host or responder L3T4+, Ly2- T cells and responder accessory cells in MLHC or host macrophages in HC-SMA may participate in "indirect" recognition of hepatocyte class I antigen both in vitro and in vivo.