Restricted usage of T cell receptor V alpha/J alpha gene segments with different nucleotide but identical amino acid sequences in HLA-DR3+ sarcoidosis patients.
BACKGROUND
Sarcoidosis is a granulomatous disease characterized by the accumulation of activated T cells in the lungs. We previously showed that sarcoidosis patients expressing the HLA haplotype DR3(17),DQ2 had increased numbers of lung CD4+ T cells using the T cell receptor (TCR) variable region (V) alpha 2.3 gene segment product. In the present study, the composition of both the TCR alpha- and beta-chains of the expanded CD4+ lung T cells from four DR3(17),DQ2+ sarcoidosis patients was examined.


METHODS
TCR alpha-chains were analyzed by cDNA cloning and nucleotide sequencing. TCR beta-chains were analyzed for V beta usage by flow cytometry using TCR V-specific monoclonal antibodies or by the polymerase chain reaction (PCR) using V beta- and C beta-specific primers. J beta usage was analyzed by Southern blotting of PCR products and subsequent hybridization with radiolabeled J beta-specific probes.


RESULTS
Evidence of biased J alpha gene segment usage by the alpha-chains of V alpha 2.3+ CD4+ lung T cells was found in four out of four patients. Both different alpha-chain nucleotide sequences coding for identical amino acid sequences and a number of identically repeated alpha-chain sequences were identified. In contrast, the TCR beta-chains of FACS-sorted V alpha 2.3+ CD4+ lung T cells were found, with one exception, to have a nonrestricted TCR V beta usage.


CONCLUSIONS
The finding of V alpha 2.3+ CD4+ lung T cells with identical TCR alpha-chain amino acid sequences but with different nucleotide sequences strongly suggests that different T cell clones have been selected to interact with a specific sarcoidosis associated antigen(s). The identification of T cells with restricted TCR usage, which may play an important role in the development of sarcoidosis, and the possibility of selectively manipulating these cells should have important implications for the treatment of the disease.
Microbiology and Tumorbiology Center (MTC), Karolinska Institute, Stockholm, Sweden.
J
J
Grunewald
T
T
Hultman
A
A
Bucht
A
A
Eklund
H
H
Wigzell
D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D015704:CD4 Antigens / Q000032:analysis; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D003001:Cloning, Molecular; D005260:Female; D005434:Flow Cytometry; D015803:HLA-DR3 Antigen / Q000032:analysis*; D006239:Haplotypes; D006801:Humans; D008168:Lung / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry / Q000235:genetics*; D012507:Sarcoidosis / Q000235:genetics / Q000276:immunology*; D017421:Sequence Analysis
D015704:CD4 Antigens; D015803:HLA-DR3 Antigen; D016693:Receptors, Antigen, T-Cell, alpha-beta