Control and resolution of leishmanial infection depends primary on T cell mediated immune mechanisms. The nature of the leishmanial antigens involved in eliciting T cell immunity has been studied by the recently developed method of cellular immunoblotting. In the past year, we have modified this method to include the analysis of human T cell responses to nitrocellulose-bound antigens highly resolved by two-dimensional electrophoresis. The proliferative responses of cells from patients with mucosal, and cutaneous leishmaniasis were remarkably heterogeneous and occurred to as many as 50-70 distinct antigens, many of which were associated with gamma-interferon production. The method has also been used to identify specific 2-D resolved antigens which stimulate T cell clones derived from patients with acquired resistance to leishmaniasis. We are now attempting to obtain sequences off of these immobilon blotted proteins in order to construct oligonucleotide probes so that genes containing these T cell epitopes might be cloned. Recently, intralesional T lymphocyte lines and clones have been produced and their antigen specificity will be profiled and compared with that displayed by peripheral cells from the same patient. Finally, studies are soon to be initiated using cDNA probes for cytokine mRNA expressed by activated lymphocytes from patients with mucosal and cutaneous leishmaniasis were remarkably heterogeneous and occurred to as many as 50-70 distinct antigens, many of which were associated with gamma-interferon production. The method has also been used to identify specific 2-D resolved antigens which stimulate T cell clones derived from patients with acquired resistance to leishmaniasis. We are now attempting to obtain sequences off of these immobilon blotted proteins in order to construct oligonucleotide probes so that genes containing these T cell epitoptes might be clones. Recently, intralesional T lymphocyte lines and clones have been produced and their antigen specificity will be profiled and compared with that displayed by peripheral cells from the same patient.
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