A Research on the Induction of Apoptosis in Lymphocytes by Malaria Infection

(1)伯氏疟原虫诱导小鼠淋巴细胞凋亡我们从三组小鼠中分离和收获外周血淋巴细胞，1)感染伯氏疟原虫nk65毒株的小鼠20只，表现为中等水平的寄生虫血症（D-4），感染同一菌株的小鼠20只，表现为高水平的寄生虫血症（D-5），对照小鼠10只，未感染。各组淋巴细胞分别与微珠偶联抗mla L3T4 （CD4）辅助T细胞和抗mla ly2 （CD8a）抑制T细胞反应，并通过磁性细胞分离器进行分选。从每个标记的淋巴细胞组中提取dna并在琼脂糖凝胶中电泳，发现高度寄生小鼠CD8a阳性T细胞的dna片段化。(2)恶性疟原虫（Plasmodium falciparum， P.f.）诱导人淋巴细胞凋亡我们分离并收获了6例恶性疟原虫感染患者的外周血淋巴细胞。这些淋巴细胞与抗fas单克隆抗体（小鼠）反应，然后与fitc偶联的抗小鼠lgs反应。荧光显微镜显示约1%的外周血淋巴细胞荧光阳性，提示疟疾感染诱导了细胞死亡的异常程序。(3)凋亡淋巴细胞电镜观察与抗fas单克隆抗体（小鼠）反应的外周淋巴细胞与微珠偶联抗小鼠lgs（大鼠）孵育，磁细胞分离机处理。电子显微图显示单核细胞细胞核断裂或染色质分布异常，有微珠附着。

(1) Plasmodium bergbei (P.b.) induces apoptosis in mouse lymphocytesWe isolated and harvested peripheral lymphocytes from three groups of mice, 1) 20 mice infected with virulent strain of P.b.NK65 showing moderate level of parasitemia (D-4), 20 mice infected with the same strain of P.b.showing high parasitemia (D-5), and 10 control mice with no infection. Lymphocytes from each group were reacted with microbeads-conjugated anti-MLA L3T4 (CD4) helper T-cells and anti-MLA ly2 (CD8a) suppressor T cells respectively, and subjected to sorting through the Magnetic Cell Separator. Extraction of the DNAs from the each labeled group of lymphocytes and electrophoresis of the DNAs in agarose gels revealed the fragmentation of DNAs from CD8a positive T cells of highly parasitized mice.(2) Plasmodium falciparum (P.f.) induces apoptosis in human lymphocytesWe isolated and harvested peripheral lymphocytes from 6 patients contracting P.f.infection. Those lymphocytes were reacted with anti-Fas monoclonal antibody (mouse) and then with FITC-conjugated anti mouse lgs. Fluorescent microscopy revealed that about 1% of the peripheral lymphocytes showed positive fluorescence, suggesting the induction of an abnormal program of cell death by malaria infection.(3) Electron micrography of apoptotic lymphocytesPeripheral lymphocytes reacted with anti-Fas monoclonal antibody (mouse) was then incubated with microbeads-conjugated anti mouse lgs (rat), and subjected to the Magnetic Cell Separator. Electron microgram of the harvested cells revealed fragmentation of the nucleus or abnormal distribution of the chromatin in the nucleus in mononuclear cells, with the attachment of microbeads.