Journal
AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE
Volume 166, Issue 6, Pages 843-848Publisher
AMER THORACIC SOC
DOI: 10.1164/rccm.2110094
Keywords
CD8-positive T lymphocytes; dendritic cells; tuberculosis
Categories
Funding
- NIAID NIH HHS [1KO8AI01645-03, 1K08AI01644] Funding Source: Medline
- NICHD NIH HHS [HD33703] Funding Source: Medline
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Previous studies in murine and human models have suggested an important role for CD8(+) T cells in host defense to Mycobacterium tuberculosis (Mtb). Consequently, a successful tuberculosis vaccine may require the elicitation of sustained CD4(+) and CD8(+) T cell responses. We tested the hypothesis that the potent CD4(+) T cell antigen Mtb39 is also a CD8(+) T cell antigen. A recombinant adenovirus-expressing Mtb39 (adenoMtb39) was used to infect monocyte-derived dendritic cells. Using interferon-gamma enzyme-linked immunospot, Mtb39-specific CD8(+) T lymphocytes were detected in three healthy individuals with latent tuberculosis infection who also had strong anti-Mtb39-specific CD4(+) T cell responses. An Mtb39-specific CD8(+) T cell line was generated using Mtb39-expressing dendritic cells. Mtb39-specific T cell clones were obtained by limiting dilution cloning. All seven T cell clones obtained were HLA-B44 restricted. Using a panel of synthetic overlapping peptides representative of Mtb39, the peptide epitope was identified for two clones. Furthermore, all T cell clones recognized Mtb-infected dendritic cells and were cytolytic. We conclude that infection of dendritic cells with adenoviral vectors expressing Mtb proteins allows for measurement of antigen-specific CD8(+) T cell responses from peripheral blood mononuclear cells. The technique will be useful in defining CD8(+) T cell antigens and in measuring immunogenicity of tuberculosis vaccines.
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