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Clin Chem. 2011 Jul;57(7):1013-22. doi: 10.1373/clinchem.2010.159673. Epub 2011 Apr 28.

Identification of circulating nonclassic human leukocyte antigen G (HLA-G)-like molecules in exudates.

Author information

1
Department of Biochemistry, University Clinic of Navarra, 31008 Pamplona, Spain. agonzaleh@unav.es

Abstract

BACKGROUND:

HLA-G in biological fluids has been proposed to be useful as a tumor marker as both a diagnostic and prognostic factor. Most HLA-G measurement procedures are based on ELISA methods using highly specific antibodies. However, results of published studies are in conflict regarding the clinical utility and even the nature of HLA-G present in circulation.

METHODS:

We collected 118 exudates, 94 from cancer patients and 24 from patients without tumors. We measured HLA-G concentrations by ELISA using MEM-G/9 or G233 as capture antibody. Samples were immunoprecipitated with an anti-HLA-G antibody and analyzed by Western blot using a different anti-HLA-G antibody.

RESULTS:

Discrepancies in HLA-G concentrations in exudates were observed depending on what capture anti-HLA-G antibody was used for ELISA (r = 0.376). These discrepancies were not observed when the ELISAs were performed using culture supernatants from HLA-G1-transfected cells (r = 0.983). Immunoprecipitation and Western blot of cell culture supernatants with 2 different anti-HLA-G antibodies produced the typical band at 39 kDa assigned to HLA-G. When the immunoprecipitation and western blot were performed with exudates, however, there were bands at 53 kDa and 70-76 kDa, higher molecular weights than those usually assigned to HLA-G. These HLA-G-like molecules were associated with β(2)-microglobulin and could also form disulfide bridges with other HLA-G-like molecules.

CONCLUSIONS:

The main HLA-G antigenic molecules in exudates are HLA-G-like complexes, a factor that should be considered when analyzing HLA-G in biological fluids.

PMID:
21527645
DOI:
10.1373/clinchem.2010.159673
[Indexed for MEDLINE]
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