Quantitative analysis of T-cell receptor beta variable-gene usage in cutaneous late-phase reactions: implications for T-lymphocyte recruitment in cutaneous inflammation

Abstract

To determine if functionally distinct T-lymphocyte (T cell) subsets accumulate in late-phase immunoglobulin E-mediated reactions (LPR), we quantitatively analyzed the immunophenotype and the T-cell receptor beta variable-gene (Vbeta) repertoire of T cells in cutaneous LPR. Peripheral blood and skin biopsies were obtained 6 or 24 h after sensitive subjects were challenged with intradermal injections of grass pollen allergen (Ag) and control (C) solution. The frequency of cells expressing CD3, CD4, CD8, CD45RO, and CD25/mm2 was determined by immunohistochemistry in nine subjects. Vbeta usage was assessed by reverse transcription-PCR in five of nine subjects. A significantly greater frequency of CD3(+) and CD45RO+ (memory) T cells was detected in Ag sites than in C sites at 24 h after challenge but not at 6 h. The frequency of activated (CD25(+)) and helper (CD4(+)) T cells appeared to be increased in Ag sites as well, though not significantly. Vbeta6 was the most commonly expressed Vbeta detected in Ag sites, but it was also detected in accompanying C sites. Vbeta2 was the most commonly expressed Vbeta detected in C sites. Sequence analysis in one case revealed Vbeta expression in a 6-h Ag site to be essentially polyclonal. Our findings suggest that memory T cells with Vbeta expression similar to that in normal skin accumulate in developing cutaneous LPR. The limited usage of Vbeta suggests a preferential recruitment or retention of reactive T cells from an endogenous subset of skin-homing T cells with its own skewed Vbeta repertoire.