Shinoda M, Hudson J L, Strömberg I, Hoffer B J, Moorhead J W, Olson L
Department of Neuroscience, Karolinska Institute, Stockholm, Sweden.
Exp Neurol. 1996 Oct;141(2):173-80. doi: 10.1006/exnr.1996.0151.
Microglia express cytokines, major histocompatibility (MHC) loci, and several other immunologically important constituents. The aim of this study was to detect immunological responses of microglial cells following allogeneic dopaminergic transplantation using active and adoptive immunizations. Adult inbred Fisher 344 (F344 RT1) rats were unilaterally dopamine (DA) depleted in striatum by injection of 6-hydroxydopamine. The degree of degeneration was assessed by recording the rotational response to apomorphine. Fetal ventral mesencephalic tissue containing DA neuroblasts from Wistar-Furth (WF, RT1u) rat donors (9-12 mm CRL) were later implanted in striatum on the lesioned side. Lymph nodes and spleen cells were collected aseptically, resuspended, and diluted for isovolumetric injections. Animals selected for active immunization were injected intraperitoneally with varying amounts of WF lymphocytes. Animals selected for adoptive immunization (transferred immunity) were intraperitoneally injected with 10(8) F344 lymphocytes prepared from animals actively immunized 3 weeks previously. Monoclonal antibodies against CD4 (OX38), CD8 (OX8), CD11b (OX42), MHC class I (OX18), monomorphic MHC class II (OX-6), and ED1 and polyclonal antibodies against tyrosine hydroxylase (TH) were used for immunohistochemistry. We found that the degree of ED1-positive cell proliferation was well correlated to the immunization patterns. Groups that were actively immunized with or without prior adoptive immunization had a larger amount of reactive microglial proliferation. ED1 immunohistochemistry revealed patterns of immunolabeling of engrafted areas: 8-12 weeks after grafting in nonimmunized and adoptively immunized groups reactive microglial proliferation occurred only at the graft periphery. Active and adoptive + active immunization led to ED1-IR within the grafts themselves. At early stages nonimmunized groups had an ED1 pattern which was partially inside the grafts. At early time points nonimmunized groups contained ameboid microglial cells within the grafts which disappeared at later stages and were absent in the immunized groups. ED1-positive ameboid microglial cells within the grafts may be of graft origin and constitute a part of a continued normal development of the fetal tissue.
小胶质细胞表达细胞因子、主要组织相容性（MHC）位点以及其他几种免疫重要成分。本研究的目的是通过主动免疫和过继免疫来检测同种异体多巴胺能移植后小胶质细胞的免疫反应。成年近交系Fisher 344（F344 RT1）大鼠通过注射6-羟基多巴胺使纹状体单侧多巴胺（DA）耗竭。通过记录对阿扑吗啡的旋转反应来评估退变程度。随后将来自Wistar-Furth（WF，RT1u）大鼠供体（9 - 12 mm头臀长）的含有DA神经母细胞的胎儿腹侧中脑组织植入损伤侧的纹状体。无菌收集淋巴结和脾细胞，重悬并稀释以进行等体积注射。选择用于主动免疫的动物腹腔注射不同量的WF淋巴细胞。选择用于过继免疫（转移免疫）的动物腹腔注射10⁸个从3周前主动免疫的动物制备的F344淋巴细胞。抗CD4（OX38）、抗CD8（OX8）、抗CD11b（OX42）、MHC I类（OX18）、单态MHC II类（OX - 6）的单克隆抗体以及抗酪氨酸羟化酶（TH）的多克隆抗体用于免疫组织化学。我们发现ED1阳性细胞增殖程度与免疫模式密切相关。主动免疫组无论有无预先过继免疫都有更多的反应性小胶质细胞增殖。ED1免疫组织化学揭示了植入区域的免疫标记模式：在未免疫和过继免疫组中，移植后8 - 12周反应性小胶质细胞增殖仅发生在移植周边。主动免疫和过继 + 主动免疫导致移植体内出现ED1免疫反应性。在早期，未免疫组的ED1模式部分位于移植体内。在早期时间点，未免疫组在移植体内含有阿米巴样小胶质细胞，这些细胞在后期消失，且在免疫组中不存在。移植体内的ED1阳性阿米巴样小胶质细胞可能起源于移植组织，并且是胎儿组织持续正常发育的一部分。
