Isolation and functional characterization of T cells from human sputum

Abstract

T cells play a central role in the control of inflammation in the bronchial mucosa through the elaboration of proinflammatory cytokines. This study describes a method for the isolation and cloning of T cells from sputum of adult subjects. In sputum, T cells were of a minor population (<2% of total cells), and not all expressed activation markers for CD29 (very late antigen-1 (VLA-1)), IL-2R and HLA-DR. When cultured in the presence of rIL-2 for 7 days and then cloned by limiting dilution, the ratios of CD4+ and CD8+ T cell clones (TCC) generated reflected those of CD4+ and CD8+ T cells found in sputum. CD4+ TCC and primary CD4+ T cell populations produced a range of proinflammatory cytokines when stimulated with immobilized anti-CD3 MoAb. Analysis of mRNA messages by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot showed good correlation with the production of cytokine in culture supernatants. A correlation existed between the pattern of cell infiltrate in sputum and the cytokine profile.