In vitro Studies of Ablastin, the Reproduction‐Inhibiting Antibody to Trypanosoma lewisi*

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Abstract
SYNOPSIS. Previously, the reproduction‐inhibiting effects of ablastin could only be shown in vivo. The present report describes techniques for the in vitro demonstration and titration of this antibody. With a medium composed of Hanks' balanced salt solution, rat serum, lactalbumin hydrolysate, yeast extract and rat blood lysate, blood stream forms of Trypanosoma lewisi can be grown for approximately 24 hours at 37d̀C. Starting with the medium containing normal rat serum and inoculated with adult (inhibited) trypanosomes from infected rat blood, 50% or more of the parasites are in various stages of division after incubation overnight. Under similar conditions with ablastic rat serum, the parasites do not reproduce but remain as adults. If the medium is inoculated with reproducing trypanosomes from the blood, parasites in the presence of normal serum continue to reproduce, whereas those exposed to ablastin are almost completely converted to adult, non‐reproducing forms. Similar results are obtained when the immune sera used are first adsorbed with living parasites to remove all trypanocidal antibodies. Ablastic serum inactivated at 56d̀C for 20 minutes does not lose its inhibitory activity indicating that ablastin is not complement dependent, and parasites grown on media at room temperature are not affected by the antibody suggesting that basic antigenic differences exist between blood stream forms at 37d̀C and culture forms at room temperature. Studies of the conversion of blood stream forms to culture forms indicate that the critical temperature range for the conversion lies between 28d̀ and 30d̀C. The significance of these results is discussed, and possible applications of the techniques described to studies of the mechanism of ablastic action are considered.