The pathogen Leishmania is a protozoan parasite causing serious worldwide morbidity affecting over 200 million individuals. Regulation of the cellular immune response directed against Leishmania is critical for the establishment of effective control of the pathogen, while limiting the development of pathology. Our long-term goal is to identify immunomodulatory leukocyte populations and cytokine networks that are involved in the development of protective or pathogenic immune responses in human leishmaniasis, while at the same time strengthening research capacities in Brazil. We have examined T cell responses in human CL and have determined that a/? CD4-CD8-, double negative (DN) T cells, are the second most prevalent cellular source of Th1 type cytokine producing cells. Thus, our aim is to obtain a functional and structural vision of a/? DN T cells from patients with CL. Specifically, studying PBMC and/or lesions from CL patients (in aim 4 from mucosal (ML) patients) from an endemic area near Salvador, Brazil, we will determine: 1) What is the relative contribution of a/? and ?/d DN T cells to the overall DN T cell profile seen in CL patients? PBMC derived a/? vs. ?/d DN T cells will be studied both ex vivo and following antigen stimulation to determine: a) their relative activation state and proliferative capacity (by CFSE staining and flow cytometry), b) their inflammatory and regulatory cytokine production, and c) their cytotoxic capacity (by expression of granzyme or perforin). Multiparameter flow cytometry will be used to address these aims. 2) Do a/? DN T cells from CL patients recognize antigens presented via CD1? We will determine: a) which CD1 molecules (CD1a, b, c, or d) are involved in the activation of a/? DN T cells (by blocking with mAbs), and b) are a/? DN T cells stimulated by CD1/antigen conjugates loaded with artificial (Gal-Cer) or Leishmania derived lipids and/or extracts (using CD1 trasfected cell lines). 3) What is the representation of a/? DN T cells in the lesion of CL patients? We will determine: a) the frequency of a/? DN T cells in the lesion as compared to the blood, b) the activation state of DN T cells in the lesion as compared to those in the blood, c) the functional profile of DN T cells from the lesion as compared to the blood. We will use flow cytometry and confocal microscopy to address this aim. 4) Does the DN T cell population arise in ML patients? We will determine if mucosal patients express the DN T cell populations in the same frequencies, and with similar or different functional activities to that seen in CL. Public health relevance: Leishmaniasis is a devastating disease that causes suffering and the death of millions worldwide. Thus, determining what factors control the development of protective or pathogenic immune responses could lead to new treatments and vaccines for this parasitic disease. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Small Research Grants (R03)
Project #
1R03AI066253-01A1
Application #
7093751
Study Section
Immunity and Host Defense Study Section (IHD)
Program Officer
Rao, Malla R
Project Start
2008-09-30
Project End
2011-09-29
Budget Start
2008-09-30
Budget End
2009-09-29
Support Year
1
Fiscal Year
2008
Total Cost
$54,000
Indirect Cost
Name
Universidade Federal de Minas Gerais
Department
Type
DUNS #
899644116
City
Belo Horizonte
State
Country
Brazil
Zip Code
31270--901
Gollob, K J; Viana, A G; Dutra, W O (2014) Immunoregulation in human American leishmaniasis: balancing pathology and protection. Parasite Immunol 36:367-76
Vieira, É L M; Keesen, T S L; Machado, P R et al. (2013) Immunoregulatory profile of monocytes from cutaneous leishmaniasis patients and association with lesion size. Parasite Immunol 35:65-72
Dutra, Walderez O; de Faria, Daniela Rodrigues; Lima Machado, Paulo Roberto et al. (2011) Immunoregulatory and Effector Activities in Human Cutaneous and Mucosal Leishmaniasis: Understanding Mechanisms of Pathology. Drug Dev Res 72:430-436
Keesen, T S L; Antonelli, L R V; Faria, D R et al. (2011) CD4(+) T cells defined by their V? T cell receptor expression are associated with immunoregulatory profiles and lesion size in human leishmaniasis. Clin Exp Immunol 165:338-51
Antonelli, Lis R V; Dutra, Walderez O; Oliveira, Ricardo R et al. (2006) Disparate immunoregulatory potentials for double-negative (CD4- CD8-) alpha beta and gamma delta T cells from human patients with cutaneous leishmaniasis. Infect Immun 74:6317-23