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.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Intramural Research (Z01)
Project #
1Z01AI000494-03
Application #
3818268
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Niaid Extramural Activities
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Peters, Nathan C; Kimblin, Nicola; Secundino, Nagila et al. (2009) Vector transmission of leishmania abrogates vaccine-induced protective immunity. PLoS Pathog 5:e1000484
Peters, Nathan C; Egen, Jackson G; Secundino, Nagila et al. (2008) In vivo imaging reveals an essential role for neutrophils in leishmaniasis transmitted by sand flies. Science 321:970-4
Nylen, Susanne; Sacks, David (2007) Interleukin-10 and the pathogenesis of human visceral leishmaniasis. Trends Immunol 28:378-84
Huynh, Chau; Sacks, David L; Andrews, Norma W (2006) A Leishmania amazonensis ZIP family iron transporter is essential for parasite replication within macrophage phagolysosomes. J Exp Med 203:2363-75
Iborra, Salvador; Carrion, Javier; Anderson, Charles et al. (2005) Vaccination with the Leishmania infantum acidic ribosomal P0 protein plus CpG oligodeoxynucleotides induces protection against cutaneous leishmaniasis in C57BL/6 mice but does not prevent progressive disease in BALB/c mice. Infect Immun 73:5842-52
Tabbara, Khaled S; Peters, Nathan C; Afrin, Farhat et al. (2005) Conditions influencing the efficacy of vaccination with live organisms against Leishmania major infection. Infect Immun 73:4714-22
Anderson, Charles F; Mendez, Susana; Sacks, David L (2005) Nonhealing infection despite Th1 polarization produced by a strain of Leishmania major in C57BL/6 mice. J Immunol 174:2934-41
Flynn, Barbara; Wang, Vivian; Sacks, David L et al. (2005) Prevention and treatment of cutaneous leishmaniasis in primates by using synthetic type D/A oligodeoxynucleotides expressing CpG motifs. Infect Immun 73:4948-54
Seder, Robert A; Sacks, David L (2004) Memory may not need reminding. Nat Med 10:1045-7
Sacks, David; Anderson, Charles (2004) Re-examination of the immunosuppressive mechanisms mediating non-cure of Leishmania infection in mice. Immunol Rev 201:225-38

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