Rearrangement and expression of Vzeta1, Vzeta2 and Vzeta3 TCR zeta genes in C57BL/6 mice.
We have recently described a mAb (2.11) that recognizes the Vgamma1-Jgamma4-Cgamma4 chain. With this mAb and an anti-delta mAb we separated gammadelta+ 2.11(+) and gammadelta+ 2.11(-) intraepithelial lymphocytes (i-IEL) by FACS. Transcripts of rearranged TCR Vgamma1 and Vdelta2 genes in both i-IEL populations were analyzed by PCR followed by sequence analysis of cDNA spanning the junction of variable (V) and joining (J) genes. Roughly the same number of Vgamma1 and Vgamma2 transcripts were found in the 2.11(+) population while >90% of the transcripts in the 2.11(-)population contained a Vgamma gene sequence. Furthermore, gamma transcripts in the 2.11+ population were functional, while only 30-40% of the Vgamma transcripts in either population contained an in-frame sequence. The observed frequency of transcripts is what would be expected from cell populations that have not gone through cellular selection mediated by the RCR. Expression of Vgamma mRNA in RCRalphabeta and TCRgammadelta thymocytes was studied by a technique that analyzed populations of transcripts of rearranged genes. In both T cell population similar levels of Vgamma transcripts were found and about two out of three transcripts were out-of-frame. During the cloning and sequence analysis, we indentified a clone that expresses the Vgamma segment rearranged to the Jgamma3-Cgamma region in C57BL/6 mice. Together with the PCR cloning and sequencing of the complete Cgamma region in C57BL/6 mice these data demonstrate that the Jgamma-Cgamma gene is functional in this strain. Taken together, these studies revealed that: (i) cells expressing the Vgamma1 chain are an important subset of the gammadelta i-IEL population and that they show extensive junctional diversity; (ii) there is no correlation between expression of in-frame Vgamma transcripts and expression of Vgamma2 chains at the cell surface; and (iii) cells expressing the Vgamma3 chain might be a minor subset of the gammadelta T cell population in C57BL/6 mice.
Pereira
P
P
Unite d'Immunobiologie CNRS URA 1961 andUnite de Biologie Moleculaire du Gene, U277 INSERM, Institut Pasteur, Paris, France.
Gerber
D
D
Regnault
A
A
Huang
S Y
SY
Hermitte
V
V
Coutinho
A
A
Tonegawa
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015870:Gene Expression; D015334:Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics*; D017421:Sequence Analysis; D013601:T-Lymphocytes / Q000737:chemistry* / Q000276:immunology
D016692:Receptors, Antigen, T-Cell, gamma-delta