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The pathogenicity, immunogenicity, and morphological stability of a knobless clone of strain FCR-3 of the human malaria parasite Plasmodium falciparum was investigated in Aotus monkeys. An early knob-bearing (K+), wild-type isolate of strain FCR-3 and the D3 knobless (K-) clone were adapted to Aotus monkey erythrocytes in continuous culture, establishing the parasites in Aotus cells without exposure to in vivo cellular or humoral immune responses. All monkeys, intact or splenectomized, which were infected with wild-type FCR-3 adapted to Aotus cells in vitro, developed virulent infections and had to be drug treated. The intact nonsplenectomized animals which received knobless D3 cloned parasites did not develop virulent infections even after multiple infections. The splenectomized monkeys which received the K- D3 clone had virulent infections. Late-stage wild-type K+ parasites sequestered in both intact and splenectomized monkeys, whereas late-stage D3 K- parasites did not sequester in the splenectomized animals. These results suggest that two elements affected the pathogenicity of the malaria parasites in these experiments. Knobs on K+-infected erythrocytes enabled these parasites to sequester, presumably by attachment to capillary endothelium. When present, the spleen eliminated circulating K- late-stage erythrocytes, presumably by selection on the basis of their nondeformability. Although clone D3 K- parasites are nonvirulent in intact monkeys, they induced some immunological protection against challenge with wild-type K+ parasites. The surface morphology of K--infected erythrocytes remains unaltered throughout these experiments, suggesting that loss of knobs is a stable condition.

Pathogenicity, stability, and immunogenicity of a knobless clone of Plasmodium falciparum in Colombian owl monkeys