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Analysis of HLA-G in Maternal Plasma, Follicular Fluid, and Preimplantation Embryos Reveal an Asymmetric Pattern of Expression¹

Valerie R. Shaikly^*,, Ian E. G. Morrison^*, Mohamed Taranissi, Clare V. Noble, Anna D. Withey, Richard J. Cherry^*, Sandra M. Blois^* and Nelson Fernández^2,*

^* Department of Biological Sciences, University of Essex, Essex, United Kingdom; and Assisted Reproduction and Gynaecology Centre, London, United Kingdom

Soluble HLA-G (sHLA-G) secretion by human preimplantation embryos in culture has been associated with successful embryo development, and therefore has potential to serve as a noninvasive marker of embryo viability. We have examined the spatial and temporal expression of HLA-G in embryos of varying developmental competence and the role of maternal factors in human embryonic HLA-G expression. Embryos that reached blastocyst stage on day 5 showed a higher frequency of sHLA-G secretion than those at morula or arrested stages (p < 0.05). There was no significant difference in sHLA-G secretion between normal embryos and those diagnosed as chromosomally abnormal by preimplantation genetic diagnosis. HLA-G detected in maternal plasma and follicular fluid did not appear to correlate with HLA-G expressed in the embryo or embryo supernatants. Confocal microscopy analysis indicated that HLA-G protein expression in embryos was not homogeneous; mostly, it was confined to blastocysts localized on trophectoderm and trophectoderm projections. Single-particle fluorescent imaging analysis of HLA-G on the cell surface of JEG-3 cells showed that HLA-G particles were mostly monomeric, but dimeric and higher order oligomers were also observed. These results suggest that HLA-G play an important role in preimplantation embryo development. However, the observed expression of HLA-G in arrested and chromosomally abnormal embryos indicates that HLA-G testing should be used with caution and in conjunction with conventional methods of embryo screening and selection.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported by grants from the U.K. Wellcome Trust Foundation (to N.F.) and the C. N. Bennett Charitable Trust (to V.R.S.).

² Address correspondence and reprint requests to Prof. Nelson Fernández, Department of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, Essex, CO4 3SQ, U.K. E-mail address: nelson{at}essex.ac.uk

³ Abbreviations used in this paper: sHLA-G, soluble HLA-G; IVF, in vitro fertilization; PGD, preimplantation genetic diagnosis; SPFI, single-particle fluorescent imaging.