The 17XNL strain of Plasmodium yoelii induces a highly effective and permanent T-cell dependent immunity in mice of the CBA strain; the lethal variant P. yoelii 17XL and P. berghei (ANKA) fail to activate an effective immune response in the same host. These differences in immunogenicity are unexplained. We recently observed that in CBA/CaJ mice the intracellular blood stages of P. yoelii 17XNL were almost exclusively within reticulocytes whereas lethal P. yoelii 17XL and P. berghei (ANKA), at comparable stages of infection, were predominantly erythrocytic. Induction of a reticulocytosis converted the normally lethal P. yoelii 17XL infection into a nonlethal one, and reticulocytic P. yoelii was shown to be more immunogenic than the erythrocytic form. Since one of the differences between reticulocytes and erythrocytes that might have influenced the development of immunity was greater expression of MHC antigens of the former cell type we examined the expression of H-2K, H-2D and Ia on reticulocytes infected with P. yoelii 17XNL. These cells showed a very marked increase in H-2K and D antigen expression compared to normal reticulocytes or erythrocytes. No Ia was detected. Red blood cells (RBC) infected with lethal P. yoelii 17XL or P. berghei showed no increase in H-2K or H-2D antigen expression. Finally, the level of expression of H-2K on P. yoelii 17XNL parasitized red blood cells from different strains of mice correlated closely with the ability of these strains to control the infection.
Comparisons made among various procedures leading to the isolation of variant antigen from Trypanosoma brucei rhodesiense bloodstream trypomastigotes. As a means of parasite disruption, freeze-thawing solubilized 36% more variant antigen than did sonication. Protease inhibitors were important additions to the suspension prior to cellular disruption. If trypanosomal extracts were frozen for at least 1 wk prior to chromatographic isolation of variant antigen, recovery of the antigen was reduced by 70%. Ion exchange chromatography was more efficient in the isolation of variant antigen than either lentil-lectin or antibody-affinity columns. All three methods yielded qualitatively similar variant antigen preparations. Using the most efficient isolation procedure tested, about 4 mg of variant antigen was isolated per 10(10) bloodstream trypomastigotes. The most efficient means of isolating variant antigen from plasma of infected rats began with passage of fresh plasma with protease inhibitors through an ion exchange column followed by antibody-affinity chromatography. This resulted in a preparation that was 52% variant antigen, a 370-fold concentration over plasma levels.
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