TCRB CLONOTYPES ARE PRESENT IN CD4-CELL POPULATIONS PREPARED DIRECTLYFROM RHEUMATOID SYNOVIUM( T)

The identification of clonal T cells at sites of inflammation is hampe red by the large number of polyclonal T cells that nonspecifically acc umulate. In this report, we combine the use of T cell sorting with spe ctratyping of the third complementarity determining region (CDR3) and direct sequence analysis to rapidly screen for and identify clonal exp ansions of T cells from synovial tissue specimens from patients with r heumatoid arthritis (RA). Initially, we used a polymerase chain reacti on specific for the variable region gene of the T cell receptor beta c hain (TCRBV) to compare the TCRBV repertoire expressed by CD4+ T cells from the peripheral blood and synovium of five patients with long-sta nding RA. Each patient had several TCRBV genes that were amplified to a greater degree from synovium. Extensive sequence analysis (n > 170) showed that each patient contained junctional sequences that occurred more than once, implying che presence of T cell clones within the star ting CD4+ T cell population. To assess a more straightforward approach to identifying clones, six additional patients were recruited and CD4 +, TCRBV2+ synovial T cells were positively selected and analyzed by C DR3 spectratyping. Bands deviating from a normal distribution were exc ised from the gel and sequenced directly. Clones were detected in half of the patients. These data are consistent with the possibility of an antigen-driven T cell response in RA that remains present in the sett ing of advanced disease. Published by Elsevier Science Inc.