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The Journal of Immunology, 1972, 108: 1570-1577.
Copyright © 1972 by The American Association of Immunologists, Inc.
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Leishmania Donovani; Partial Characterization of Some Flagellate Cytoplasmic Immunogens1

Bruce A. Clinton2, Nicholas C. Palczuk and Leslie A. Stauber

From the Department of Zoology and Bureau of Biological Research, Rutgers, The State University, New Brunswick, New Jersey 08903

Abstract

Leishmania donovani promastigotes were cultured in Millipore-filtered medium and subjected to freeze-thaw and homogenization regimens. The resulting homogenate was centrifuged, and the supernatant, consisting mainly of the cytoplasmic portion, was injected into rabbits. The rabbit anti-promastigote antibody was used to study promastigote immunogens by immunoelectrophoresis and by our modification of thin-layer immunochromatography (ICG). Both procedures revealed nine distinct precipitin lines; however, ICG disclosed the approximate molecular weights of the cytoplasmic immunogens ranging from 8637 to 112,250. In other characterization studies, promastigote homogenate in CFA was used to sensitize 16 guinea pigs. After 7 days the animals were challenged with individual cytoplasmic fractions (3 to 4 µg protein) separated by G-200 Sephadex. Intradermal challenge sites were measured, for diameters of erythema at 10 min; 4, 7, 24, 48 and 96 hr. Erythema appeared at 4 hr post-challenge, peaking at 24 hr and thereafter decreasing in intensity. Two distinct fractions from the separated homogenate were responsible for the most intense delayed skin responses. Used culture medium produced delayed skin reactions; whereas, unused medium provoked no response.

Footnotes

1 This investigation was supported in part by United States Public Health Service Research (AI-00092) and Training (AI-00187) Grants from NIAID.

2 Present address: Department of Experimental Pathology, Scripps Clinic and Research Foundation, La Jolla, California 92037.






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