title: Response of a murine epidermal V gamma 1/V delta 6-TCR+ hybridoma to heat shock protein HSP-60.
abstract: In the epidermis, the major population of T lymphocytes expresses a T-cell receptor (TCR) with V gamma 5 and V delta 1 variable regions, which is unique to this tissue. Roberts et al and Ezquerra et al, also describe a minor population of gamma delta-TCR+ cells in the epidermis that expresses a V gamma 1/V delta 6 TCR. These cells are different from other epidermal T cells in that they "spontaneously" produce cytokines, a result thought to be due to autoreactivity. Over the past 5 years, our laboratory has produced V gamma 1/V delta 6+ T-cell hybridomas from many tissue sources. These spontaneously produce cytokines but also are activated by heat shock protein (HSP-60)-derived peptides. Ezquerra et al report that none of their V gamma 1/V delta 6+ epidermal T-cell lines derived from C3H/HeN mice respond to HSP-60. Of the 99 gamma delta-TCR+ hybridomas we have produced from epidermal T cells of C57BL/6 mice, only one expressed the V gamma 1/V delta 6 TCR. This hybridoma, 70BET-2.12, not only spontaneously produces cytokines, but, unlike the V gamma 1/V delta 6-TCR+ epidermal T cells of Ezquerra et al, it also responds to the whole HSP-60 protein and a 17-mer HSP-60 peptide from M. leprae, producing increased levels of interleukin-2 of up to approximately ten-fold above the spontaneous level. This shows that V gamma 1/V delta 6-TCR+ epidermal T cells can respond to HSP-60. To confirm that 70BET-2.12 expresses TCR genes similar to those of cells that have HSP-60 reactivity, V gamma 1-C gamma 4 and V delta 6-C delta cDNA were produced from RNA isolated from this hybridoma, amplified by the polymerase chain reaction, and sequenced. The gamma and delta TCR gene sequences were similar but not identical to previously published sequences of HSP-60-reactive cells from lymphoid and other organs. No explanation can be found for the discrepancy between our findings and those of others at the level of TCR expression such that other strain-specific factors might be responsible for HSP-60 reactivity.
authors.affiliation: National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado.
authors.forename: C L
authors.initials: CL
authors.lastname: Reardon
authors.forename: M
authors.initials: M
authors.lastname: Vollmer
authors.forename: R
authors.initials: R
authors.lastname: Cranfill
authors.forename: R
authors.initials: R
authors.lastname: van der Zee
authors.forename: R L
authors.initials: RL
authors.lastname: O'Brien
authors.forename: W K
authors.initials: WK
authors.lastname: Born
mesh_terms: D000818:Animals; D001483:Base Sequence; D018834:Chaperonin 60 / Q000494:pharmacology*; D006825:Hybridomas / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000502:physiology*
chemical_list: D018834:Chaperonin 60; D016692:Receptors, Antigen, T-Cell, gamma-delta