Objective. To determine if the T cell antigen receptor V-beta usage of
unstimulated rheumatoid arthritis (RA) synovial fluid (SF) T cells is
biased compared with those in peripheral blood (PB). Methods. Freshly
isolated, matched synovial fluid and peripheral blood T cells were an
alyzed for V(b)eta gene expression using quantitative polymerase chain
reaction (PCR) methods. Ten synovial fluid samples from the knees of
7 patients with RA were studied. The PCR assay used 26 V-beta primers
with a constant region C-beta primer, and 2 C-alpha primers that co-am
plified a product that served as an internal standard. Cycle number an
d complementary DNA content were controlled to ensure the linear accum
ulation of PCR products. Labeled products were separated on 10% polyac
rylamide gels and counted with a Betascope blot analyzer. Results. The
re were consistent differences between the V-beta gene usage of SF and
PB T cells directly isolated from patients with RA, regardless of HLA
-DR haplotype. In all synovial specimens, V(beta)2 was increased relat
ive to the peripheral blood, while V(beta)13.1 and V(beta)13.2 were de
creased. V(beta)6 and V(beta)21 were increased in 9 of the 10 synovial
samples. Analyses of bilateral SF specimens from 2 subjects and seria
l specimens from the same knee of 1 subject revealed virtually identic
al patterns in each patient. The SF V-beta bias was not solely due to
differences in the proportion of CD4+ and CD8+ cells, because the CD4:
CD8 ratios in SF and PB were similar. However, V-beta gene usage of se
parated CD4+ and CD8+ synovial T cells showed that V(beta)2 and V(beta
)6 were more highly expressed on CD4 cells. Conclusion. Freshly isolat
ed synovial T cells from inflamed (not end-stage) knees of patients wi
th RA have a remarkably consistent biased V(b)eta gene usage compared
with PB T cells. V(beta)2 and V(beta)6 are uniformly increased, and th
is increase is primarily in CD4+ T cells. The same V-beta bias in the
SF T cells of several RA patients suggests that shared antigens may be
stimulating the T cell response.