Development of a new monoclonal antibody specific to mouse Vγ6 chain.

Shinya Hatano,Naoto Noguchi,Xun Sun,Xin Tun,Hisakata Yamada,Dan Yue,Mitsuru Matsumoto,Yasunobu Yoshikai

doi:10.26508/lsa.201900363

Abstract

There are seven Vγ gene segments in the TCR γ chain loci of mice. We developed monoclonal antibodies (mAbs) specific to the Vγ6 chain (Heilig & Tonegawa nomenclature). By immunizing Vγ4/6 KO mice with complementarity-determining region peptides in Vγ6 chains, we generated three hybridomas. These hybridomas produced mAbs capable of cell surface staining of Vγ6/Vδ1 gene-transfected T-cell line lacking TCR as well as of Vγ1- Vγ4- Vγ5- Vγ7- γδ T cells and the CD3high TCRδint γδ T cells in various organs. The location of Vγ6+ γδ T cells, which peaked in the newborn thymus, was associated with mTEC. In vivo administration of clone 1C10-1F7 mAb impaired protection against Klebsiella pneumoniae infection but ameliorated psoriasis-like dermatitis induced by imiquimod treatment. These new mAbs are useful to elucidate the development, location, and functions of Vγ6 γδ T cells in mice.

Highlights

TCR γ chain loci have three functional Cγ genes (Cγ1, Cγ2, and Cγ4) and one nonfunctional pseudo Cγ gene, four joining segments, including one pseudogene (Jγ1, Jγ2, φJγ3, and Jγ4), and seven variable (Vγ) gene segments (Saito et al, 1984)
The first T cells to appear from approximately embryonic day 12 (E12) to E16 carry γδ TCR composed of Vγ5 and Vδ1 chains (Vγ5Jγ1 and Vδ1Dδ2Jδ2), which populate the epidermis, and these T cells, which become wedged among keratinocytes and adopt a dendritic-like form, are termed dendritic epidermal T cells (Asarnow et al, 1988; Havran et al, 1989; Havran & Allison, 1990)
We consistently found that Vγ5+ γδ T cells were abundant in the fetal thymus at earlier stages of development, and the percentage decreased from E16 onward during embryonic development. γδ T-cell waves from E16 onward were Vγ4+ cells

Summary

Introduction

TCR γ chain loci have three functional Cγ genes (Cγ1, Cγ2, and Cγ4) and one nonfunctional pseudo Cγ gene (φCγ3), four joining segments, including one pseudogene (Jγ1, Jγ2, φJγ3, and Jγ4), and seven variable (Vγ) gene segments (Saito et al, 1984). The Vγ6+ γδ T cells in the PEC, reproductive organs, and c-LPL from BALB/c mice were stained with 1C10-1F7 mAb (Fig 2C).

Results

Conclusion