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Oligoclonality of Rat Intestinal Intraepithelial T Lymphocytes: Overlapping TCR ß-Chain Repertoires in the CD4 Single-Positive and CD4/CD8 Double-Positive Subsets¹

Lars Helgeland^2,*, Finn-Eirik Johansen^*, Jon O. Utgaard^*, John T. Vaage and Per Brandtzaeg^*

^* Laboratory for Immunohistochemistry and Immunopathology, Institute of Pathology, and Department of Anatomy, University of Oslo, Oslo, Norway

Previous studies in humans and mice have shown that gut intraepithelial lymphocytes (IELs) express oligoclonal TCR ß-chain repertoires. These studies have either employed unseparated IEL preparations or focused on the CD8⁺ subsets. Here, we have analyzed the TCR ß-chain repertoire of small intestinal IELs in PVG rats, in sorted CD4⁺ as well as CD8⁺ subpopulations, and important differences were noted. CD8 and CD8ß single-positive (SP) IELs used most Vß genes, but relative Vß usage as determined by quantitative PCR analysis differed markedly between the two subsets and among individual rats. By contrast, CD4⁺ IELs showed consistent skewing toward Vß17 and Vß19; these two genes accounted collectively for more than half the Vß repertoire in the CD4/CD8 double-positive (DP) subset and were likewise predominant in CD4 SP IELs. Complementarity-determining region 3 length displays and TCR sequencing demonstrated oligoclonal expansions in both the CD4⁺ and CD8⁺ IEL subpopulations. These studies also revealed that the CD4 SP and CD4/CD8 DP IEL subsets expressed overlapping ß-chain repertoires. In conclusion, our results show that rat TCR-ß⁺ IELs of both the CD8⁺ and CD4⁺ subpopulations are oligoclonal. The limited Vß usage and overlapping TCR repertoire expressed by CD4 SP and CD4/CD8 DP cells suggest that these two IEL populations recognize restricted intestinal ligands and are developmentally and functionally related.

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