Ramirez A D, Rocha E M, Krettli A U
Departamento de Parasitologia, Universidade Federal de Minas Gerais, Brazil.
J Protozool. 1991 Jan-Feb;38(1):40-4. doi: 10.1111/j.1550-7408.1991.tb04796.x.
Exoerythrocytic forms of Plasmodium gallinaceum were cultured in vitro using salivary gland sporozoites extracted from experimentally infected Aedes fluviatilis mosquitoes. The host cells were macrophage precursors from chicken bone marrow. At various times after introduction of sporozoites, the cultures were stained by Giemsa or by immunofluorescence assay (IFA) using anti-sporozoite-specific monoclonal antibodies (MAb). The time to complete parasite development in vitro was 50-70 h. By 70 h, ruptured segmenters and free merozoites were visible within the cells. Inoculation of normal chickens with infected cultures induced parasitemia after a pre-patent period of 10-11 days. In vitro young exoerythrocytic forms, late schizonts that include the matured segmenters, and free merozoites shared common antigens with the sporozoites as revealed by IFA using anti-sporozoite-specific MAbs. Our data indicate that macrophages support development of P. gallinaceum sporozoites and that the circumsporozoite proteins are present until the end of the primary exoerythrocytic schizogony.
利用从实验感染的黄热病伊蚊中提取的唾液腺子孢子，在体外培养鸡疟原虫的红细胞外期。宿主细胞是来自鸡骨髓的巨噬细胞前体。在引入子孢子后的不同时间，使用吉姆萨染色或使用抗子孢子特异性单克隆抗体（MAb）通过免疫荧光测定（IFA）对培养物进行染色。体外完成寄生虫发育的时间为50 - 70小时。到70小时时，细胞内可见破裂的裂殖体和游离的裂殖子。用感染的培养物接种正常鸡，在10 - 11天的潜伏期后诱导出寄生虫血症。使用抗子孢子特异性MAb的IFA显示，体外年轻的红细胞外期形式、包括成熟裂殖体的晚期裂殖体以及游离的裂殖子与子孢子具有共同抗原。我们的数据表明，巨噬细胞支持鸡疟原虫子孢子的发育，并且环子孢子蛋白一直存在到初次红细胞外裂体增殖结束。
