Louis J A, Moedder E, MacDonald H R, Engers H D
J Immunol. 1981 May;126(5):1661-6.
The proliferative response of nylon wool purified, primed lymph node cells to L. tropica parasites in vitro was found to be restored by the addition of either syngeneic or allogeneic adherent spleen cells as a putative source of macrophages. These results suggested a lack of H-2 restriction in Leishmania-specific T cell responses. However, when T cell blasts generated in vitro in response to the parasite were separated on Percoll density gradients and subsequently maintained for 4 days in the presence of TCGF, their response to L. tropica was found to be strictly dependent on the presence of syngeneic spleen cells. Further studies using congenic recombinant mice demonstrated that proliferation of a parasite-specific blasts required the presence of spleen cells compatible with the responding cells in the I-A region of the MHC. This requirement for I-A compatible adherent cells in the spleen cell populations was further confirmed by a lack of proliferative responses in the presence of spleen cells treated with monoclonal anti-la antibodies and complement. Leishmania-immune F1 blasts responding to the parasite in the context of either parental la-bearing accessory cell could be obtained by positive selection from a F1 hybrid responding cell population. Using flow microfluorometry, the T cell phenotype of the L. tropica-specific blasts was determined to be Thy-1+, lyt-1+, and Lyt-2-.
研究发现,通过添加同基因或异基因贴壁脾细胞作为假定的巨噬细胞来源,尼龙毛纯化的、经致敏的淋巴结细胞在体外对热带利什曼原虫寄生虫的增殖反应得以恢复。这些结果表明利什曼原虫特异性T细胞反应缺乏H-2限制。然而,当体外针对寄生虫产生的T细胞母细胞在Percoll密度梯度上分离,随后在TCGF存在下维持4天时,发现它们对热带利什曼原虫的反应严格依赖于同基因脾细胞的存在。使用同源重组小鼠的进一步研究表明,寄生虫特异性母细胞的增殖需要在MHC的I-A区域存在与反应细胞相容的脾细胞。在用单克隆抗Ia抗体和补体处理的脾细胞存在下缺乏增殖反应,进一步证实了脾细胞群体中对I-A相容贴壁细胞的这一需求。在亲本携带Ia的辅助细胞的背景下对寄生虫作出反应的利什曼原虫免疫F1母细胞可以通过从F1杂交反应细胞群体中进行阳性选择获得。使用流式细胞荧光术,确定热带利什曼原虫特异性母细胞的T细胞表型为Thy-1+、lyt-1+和Lyt-2-。