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

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(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.