Analysis of mRNA for class I HLA on human gametogenic cells.
We have studied mRNA expression for Class I HLA (human leukocyte antigen) on male germ cells by amplification of gene fragments in PCR technique and by Northern hybridization. RNA was extracted from fractionated gametogenic cells (isolated from testis) and reversely transcribed. Then, cDNA was amplified for specific HLA sequence (HLA, -A, -B, -C). The specificity of this product was confirmed in "nested" PCR of 400 bp gene fragment coding for alpha 2 domain, alpha 3 domain, and the transmembrane portion of Class I HLA. The results indicate minimal expression of classical Class I HLA on gametogenic cells. Northern hybridization with 669 bp cDNA fragment (spanning for alpha 3 domain, transmembrane, cytoplasmic, and 3' untranslated region) resulted in a low intensity signal from gametogenic cell fractions and confirmed our findings obtained by PCR. The minimal expression of classical HLA antigens may create a neutral cover for the male reproductive system, thereby preventing an immunological response during germ cell differentiation.
Janitz
M
M
Institute of Human Genetics, Polish Academy of Sciences, Poznan.
Fiszer
D
D
Michalczak-Janitz
K
K
Lukaszyk
A
A
Fernandez
N
N
Skorupski
W
W
Kurpisz
M
M
D015152:Blotting, Northern; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D006680:HLA Antigens / Q000235:genetics*; D015395:Histocompatibility Antigens Class I / Q000235:genetics*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics*; D013087:Spermatids / Q000737:chemistry / Q000276:immunology; D013094:Spermatozoa / Q000737:chemistry / Q000276:immunology*; D013737:Testis / Q000737:chemistry / Q000166:cytology / Q000276:immunology
D018076:DNA, Complementary; D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I; D012333:RNA, Messenger