Accumulation of multiple T cell clonotypes in lungs of healthy individuals and patients with pulmonary sarcoidosis.

Abstract

T cell accumulation and activation in lung may play a major role in the pathogenesis of immunologic lung diseases such as sarcoidosis. Using the combination of RT-PCR and subsequent single-strand conformation polymorphism analysis, we examined T cell clonality in lung and peripheral blood of healthy individuals (n = 5) and patients with active pulmonary sarcoidosis (n = 7). RNA was extracted from PBLs and bronchoalveolar lavage fluid cells, and converted to cDNA. PCR was performed using a set of V beta-C beta primers (V beta 1-20). Products were denatured and electrophoresed in nondenaturing 5% polyacrylamide gel. The existence of a distinct T cell clonotype was detected as a band on a smear. In both groups T cells in PBLs showed a number of clones that expanded. A significantly greater number of clones was detected in BAL compared with PBL in both groups (normal: 25.3 +/- 7.2 in PBL vs 62.8 +/- 5.2 in lung; sarcoid: 25.0 +/- 6.2 in PBL vs 90.0 +/- 6.6 in lung, mean value +/- SEM). In sarcoid lung greater numbers of clones were detected than in lungs of healthy controls (p < 0.012). These clonal expansions were observed over all the 20 V beta families examined, and were not restricted to certain V beta families. These results suggest that there are clonal expansions even in normal lung, and that in sarcoidosis, apparently, additional T cell clones using multiple V beta segments might be activated and accumulated at the site of the disease.