Clonal heterogeneity of synovial fluid T lymphocytes from patients with rheumatoid arthritis (original) (raw)

Abstract

Although substantial evidence suggests that synovial T lymphocytes are critical in the pathogenesis of rheumatoid arthritis (RA), little is known regarding their antigenic specificities, antigen receptor gene rearrangements, and mechanisms of activation. To assess the extent ofexpansion of specific clones among RA synovial fluid T cells, Southern blot analyses of T-cell receptor (TCR) gene rearrangements were performed on 40 RA synovial fluid T-cell clones, as well as on both fresh and polydonally activated T cells from RA synovial fluid, RA peripheral blood, and normal peripheral blood. Two of the clones had identical TCR rearrangement patterns, but the remainder were unique. The nonclonal RA T-cell samples showed the same pattern of TCR fl-chain rearrangement that was observed among normal peripheral blood T cells, indicating no dominant clonal T-cell population in these samples. It was noted that with sufficient exposure of autoradiograms of the Southern blots, discrete TCR gene rearrangements, representing in some cases common DpJp (D, diversity; J, joining) rearrangements,. were evident in T cells from peripheral blood of normal individuals and patients with RA, as well as T cells from RA synovial fluid. Taken together, the findings indicate that only a minor degree of oligoclonality can be demonstrated among T lymphocytes from RA synovial fluid.

Figures (4)

To assess the degree of oligoclonality among RA synovial fluid T cells, T cells cloned under conditions of limiting Fic.1. Restriction enzyme map of the human TCR #-chain gene. The germ-line organization has been reported (43). The location of the 2 diversity (Dg), 13 functional joining (Jg), and 2 constant (Cg) gene segments is indicated by jagged vertical lines, vertical lines, and solid rectangles, respectively. The location of the EcoRI (R) and the distances between them in kilobase pairs (kb) is indicated. Probes are restriction enzyme DNA fragments. The size and location of the J,g1, Cg, Dg2, and Jg2 probes are indicated by stippled and hatched bars.

To assess the degree of oligoclonality among RA synovial fluid T cells, T cells cloned under conditions of limiting Fic.1. Restriction enzyme map of the human TCR #-chain gene. The germ-line organization has been reported (43). The location of the 2 diversity (Dg), 13 functional joining (Jg), and 2 constant (Cg) gene segments is indicated by jagged vertical lines, vertical lines, and solid rectangles, respectively. The location of the EcoRI (R) and the distances between them in kilobase pairs (kb) is indicated. Probes are restriction enzyme DNA fragments. The size and location of the J,g1, Cg, Dg2, and Jg2 probes are indicated by stippled and hatched bars.

Fic. 2. Southern blot analysis of B-chain gene rearrangements in T-cell clones derived from the synovial fluid of a patient with RA. DNA from a representative group of T-cell clones (lanes a—o) and from the B-cell line, LAZ509 (lane p), were digested with EcoRI, fractionated on a 0.7% agarose gel, transferred to nitrocellulose, hybridized to the Jg1 and Jg2 probes, and analyzed by autoradiog- raphy. The autoradiograms are purposely overexposed to detect faint bands. Clones that exhibited more than two rearranged fragments (lane o) were not included in the analysis. Jg1 rearrangements vary in intensity because the rearrangement process deletes various amounts of DNA that hybridizes to the Jgl probe (see Fig. 1 and text). The artifactual band described in Fig. 3 and the text does not appear in these blots. The sizes of the fragments in kilobase pairs (kb) are indicated on the right.

Fic. 2. Southern blot analysis of B-chain gene rearrangements in T-cell clones derived from the synovial fluid of a patient with RA. DNA from a representative group of T-cell clones (lanes a—o) and from the B-cell line, LAZ509 (lane p), were digested with EcoRI, fractionated on a 0.7% agarose gel, transferred to nitrocellulose, hybridized to the Jg1 and Jg2 probes, and analyzed by autoradiog- raphy. The autoradiograms are purposely overexposed to detect faint bands. Clones that exhibited more than two rearranged fragments (lane o) were not included in the analysis. Jg1 rearrangements vary in intensity because the rearrangement process deletes various amounts of DNA that hybridizes to the Jgl probe (see Fig. 1 and text). The artifactual band described in Fig. 3 and the text does not appear in these blots. The sizes of the fragments in kilobase pairs (kb) are indicated on the right.

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