In Vitro and In Vivo Studies for Development of a Leishmaniasis Vaccine.
Annual rept. 1 Jul 85-30 Jun 86,
TUFTS UNIV BOSTON MA
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The purpose of the work supported by this contract is to identify antigens of Leishmania that could serve as candidates for an antileishmanisasis vaccine. To accomplish this goal, monoclonal antibodies against Leishmania promastigotes are being developed as analytic and preparative reagents. In the first year of the contract period covered by this report five anti-L. major monoclonal antibodies were obtained by fusion of spleen cells from a mouse immunized with fixed promastigotes in adjuvant, and over 60 were obtained from an infected mouse. Two monoclonals of interest were purified from ascites fluid. All were IgM, k isortype. Monoclonal 1B1 was chosen from those screened for its high titer, reactivity with the surface of infected macrophages, and with secretedexcreted products of promastigotes. 1B1 was purified from ascites and is being used in immunoprecipitation assays of Sulfur 35 methionine-labelled promastigote lysates. Preliminary results indicates it recognizes an epitope on proteins with Mr 95, 88, 68, and 43kd. Additional monoclonal antibodies are being produced to select ones with IgG isotype. Efforts to utilize these antibodies for affinity purification of antigens are being initiated to permit subsequent in vivo assessment of their vaccine potential. Keywords Antigens ELISA assay Immunofluorescence Macrophages Serodiagnosis Immunoglobin.