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Contenido archivado el 2024-06-10

Identification of Mycobacterial Antigens as Candidates for a Subunit Vaccine Against Tuberculosis in Healthy and HIV-Infected Individuals

Objetivo



Tuberculosis (TB) is a global health problem which causes 3 million deaths yearly. In industrialized countries, the incidence of the disease is rising because of the AIDS epidemic and immigration from developing countries. The available vaccine, Mycobacterium bovis bacillus Calmette-Guerin, is of questionable efficacy, therefore the development of a subunit vaccine is needed. The project proposed aims at identifying immunodominant antigens as candidate components of such a vaccine, with a special focus on proteins secreted by mycobacteria during growth in vitro or produced intracellularly. It has been shown that immune CD4+T cells play a key role in immunity to M. tuberculosis in animal models. Other cell populations (natural killer cells, CD8+ and gammadelta+ T cells) also seem to be involved. Although there is evidence that T cell immunity is essential to recover from human infection too, the antigens responsible for its induction and the role of each T cell subset are still unclear. The project proposed aims at: 1) identifying M. tuberculosis, M. av um or BCG antigens involved in protective immunity. This would proceed by isolating sets of mycobacterial proteins produced in vitro (membrane, cytosol, cell wall and secretory fractions) and in vivo, inside cells, and investigating their ability to induce proliferation of human immune lymphocytes in vitro and/or release of cytokines.
The final goal of this phase would be to molecularly clone them from a DNA expression library to test whether they might induce, a10ne or in combination, an immune status against M. tuberculosis in mice. 2) Dissection of human immune Iymphocyte proliferation to mycobacterial antigen fractions in vitro. The response of peripheral blood mononuclear cells from different human subjects (BCG-vaccinated or unvaccinated PPD+, PPD-, HIV+ and HIVtuberculous individuals) will be analyzed: i) to assess whether alfabeta+, gammadelta+ T cells or natural killer cells are mostly responsible for proliferation to the antigen fractions of choice or to cloned proteins, and whether a particular antigen is capable of inducing the release of interferon y or other immunologically relevant lymphokines; ii) to assess whether the reduction of CD4+ T cells in HIV+ patients is compensated by other lymphocyte subsets; iii) to determine whether any carbohydrate antigen accounts for proliferation of natural killer or gammadelta+ T cells. 3) Study of a rapid method for early detection of mycobacterial infection. A new method based on detection of lipoarabinomannan in urine will be set up and evaluated for the diagnosis of mycobacterial infection in HIV+ and HIV- tuberculous patients and for its ability to detect infection before active disease develops. The method will be also evaluated by monitoring infection in experimental animals

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Coordinador

Università degli Studi di Pisa
Aportación de la UE
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Dirección
Via San Zeno 35-39
56100 Pisa
Italia

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