Molecular analysis of the γδ T‐cell receptor repertoire in normal human skin and in Oriental Cutaneous Leishmaniasis

Abstract

The extent of diversity of the gamma delta T-cell receptor (TCR) in normal human skin and Oriental Cutaneous Leishmaniasis (OCL) was examined by molecular analysis of the variable (V) delta gene segment, junctional (J) delta gene segment and junctional regions. To examine the expression of TCR delta genes, segments of gamma delta T lymphocytes, DNA isolated from normal human skin and from OCL were subjected to enzymatic gene amplification by the polymerase chain reaction (PCR) method using TCR V delta- and J delta-specific oligonucleotides as primers. PCR amplification using these primers indicated that the V delta 2 gene segment was predominantly used by gamma delta T lymphocytes in both normal human skin and OCL. To determine the extent of junctional diversity in the delta gene of gamma delta T cells in normal human skin and OCL, we sequenced the nucleic acid sequences corresponding to the V delta 2/J delta 1 junctional regions. Sequence analysis of junctional regions demonstrated broad junctional diversity in normal skin but only limited diversity in OCL. Our findings support the hypothesis that skin gamma delta T lymphocytes may derive from a fetal subset of gamma delta T lymphocytes that leaves the thymus early and colonizes the periphery. The limited junctional diversity demonstrated in OCL lesions indicates that gamma delta T cells can undergo oligoclonal expansion following recognition of a specific ligand and supports the idea that junctional regions are important in the recognition of antigenic determinant.