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The Journal of Immunology, Vol 156, Issue 6 2068-2076, Copyright © 1996 by American Association of Immunologists
ARTICLES |
A King, C Boocock, AM Sharkey, L Gardner, A Beretta, AG Siccardi and YW Loke
Research Group in Human Reproductive Immunobiology, Department of Pathology, University of Cambridge, United Kingdom.
In this study, evidence is provided that normal human first trimester extravillous trophoblast expresses class I HLA-C molecules in addition to HLA-G. cDNA from highly purified trophoblast cells obtained by flow cytometric sorting was amplified by reverse-transcriptase PCR using HLA locus-specific primers. The identity of the product was confirmed by Southern blotting and hybridization by a second HLA-C-specific oligonucleotide. HLA-C mRNA was clearly demonstrated in all trophoblast samples as well as in JEG-3 and BeWo choriocarcinoma cells. JAR choriocarcinoma cells did not express HLA-C. The presence of HLA-C protein in extravillous trophoblast was investigated using a panel of Abs: L31 is specific for heavy chains of all HLA-C alleles; Q1/28 reacts with all HLA class I products except HLA-G; HC-10 has preferential reactivity with HLA-B and HLA-C heavy chains. We performed 35S metabolic and 125I surface labeling of normal first trimester trophoblast and found abundant HLA-C intracellularly together with low levels of expression of both the beta 2m-associated forms and free heavy chains on the surface. Flow cytometric analysis of normal trophoblast confirmed the expression of a class I HLA molecule distinct from HLA-G by positive reactivity with Q1/28. Immunohistologic studies of first trimester placenta and the implantation site clearly showed expression of HLA-C in all extravillous trophoblast populations. Our results demonstrate the presence of two HLA class I molecules, HLA-G and HLA-C, on the surface of extravillous trophoblast. These results have implications in understanding how maternal uterine lymphocytes, notably the abundant NK-like cells, might recognize the implanting placenta.
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