Analysis of HLA class Ib gene expression in male gametogenic cells.
We have investigated mRNA expression for nonclassical MHC class I genes (HLA-E,-F,-G) in human gametogenic cells. Testicular tissue was treated by collagenase and the resulting cell suspension was further purified by fractionation on Percoll gradients in a two-step procedure. Three gametogenic cell fractions were analyzed: purified heterogenous suspension of gametogenic cells, fraction of round spermatids and fraction of elongated spermatids. Total RNA isolated from each cell population was subjected to both reverse transcriptase/polymerase chain reaction and Northern blot analysis using oligonucleotides specific for HLA-E, -F and -G. Both method gave similar results. We have found a considerable level of HLA-E mRNA, very low amounts of reamplified cDNA for HLA-F and both a complete lack of mRNA and reamplified cDNA for the HLA-G gene in the analyzed gametogenic cell fractions. Additionally, we have localized HLA-E molecules on the cells of the adluminal compartment within seminiferous tubules using immunostaining with monoclonal antibodies specific for HLA-E heavy chain followed by confocal microscopy analysis. The unique expression pattern of HLA class I antigens in the male gonad could play an important role in an efficient protection against an autoimmunological attack toward germ cells.
Fiszer
D
D
Institute of Human Genetics, Polish Academy of Sciences, Poznań.
Ulbrecht
M
M
Fernandez
N
N
Johnson
J P
JP
Weiss
E H
EH
Kurpisz
M
M
D015152:Blotting, Northern; D002469:Cell Separation; D015870:Gene Expression / Q000276:immunology*; D005805:Genes, MHC Class I / Q000276:immunology*; D006680:HLA Antigens / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis; D013094:Spermatozoa / Q000276:immunology / Q000378:metabolism; D000097625:HLA-E Antigens
D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I; D012313:RNA; D012333:RNA, Messenger; D000097625:HLA-E Antigens