Reaction patterns of monoclonal antibodies to HLA-G in human tissues and on cell lines: a comparative study

Blaschitz, Astrid; Hutter, Heinz; Leitner, Verena; Pilz, Stefan; Wintersteiger, Reinhold; Dohr, Gottfried; Sedlmayr, Peter

doi:10.1016/s0198-8859(00)00207-x

Cited by 40 publications

(35 citation statements)

References 30 publications

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“…The pattern of soluble MHC class I expression in the rhesus testis was unexpected. HLA-G protein was not detected in human testes in previous studies (5,41), although mRNA expression has been reported in some, but not all, studies (42,43). In our studies with the rhesus, whereas there was only very rarely detectable membrane-bound Mamu-AG within the seminiferous tubules (as determined by 25D3 staining), it was clear that abundant soluble protein was present in selected germ cell populations, colocalized with Mamu-AG mRNA.…”

Section: Discussioncontrasting

confidence: 55%

“…In this situation the syncytiotrophoblasts are a more effective way to release a soluble molecule directly into the maternal circulation. The expression of soluble HLA-G by the syncytiotrophoblasts remains somewhat controversial (5,14,41), and detection may be dependent on the conditions used for fixation, embedding and processing as well as the Abs used. Indeed, conflicting results on placental and extraplacental localization of HLA-G support both further study as well as the development of appropriate animal models.…”

Section: Discussionmentioning

confidence: 99%

“…However, it seems unlikely that testicular soluble HLA-G may be the source of circulating serum HLA-G in men, since the patency of the blood-testis barrier would preclude this route of trafficking of the molecule. In addition, the assays used to detect soluble HLA-G typically detect the class I-␤ 2 -microglobulin complex (15)(16)(17)(18), and immunostaining of human testes with 16G1 did not detect soluble HLA-G protein (41). Thus, another site of expression of human soluble HLA-G seems likely.…”

Section: Discussionmentioning

confidence: 99%

“…The expression of HLA-G has been noted in macrophages invading psoriatic skin lesions (45) and in activated macrophages and dendritic cells in lung tumors (34,46). Although the expression of soluble or membrane-bound HLA-G in placental macrophages is not fully resolved (5,14,41), the rhesus monkey may be an experimental model in which to explore the functional significance of soluble nonclassical MHC class I molecules in vivo.…”

Section: Discussionmentioning

confidence: 99%

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A Soluble Isoform of the Rhesus Monkey Nonclassical MHC Class I Molecule Mamu-AG Is Expressed in the Placenta and the Testis

Ryan¹,

Grendell²,

Geraghty

et al. 2002

The Journal of Immunology

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The nonclassical MHC class I locus HLA-G is expressed primarily in the placenta, although other sites of expression have been noted in normal and pathological situations. In addition, soluble HLA-G isoforms have been detected in the serum of pregnant and nonpregnant women as well as men. The rhesus monkey placenta expresses a novel nonclassical MHC class I molecule Mamu-AG, which has features remarkably similar to those of HLA-G. We determined that the rhesus placenta expresses Mamu-AG mRNA (Mamu-AG5), retaining intron 4 as previously noted in HLA-G5. Immunostaining experiments with Ab 16G1 against the soluble HLA-G5 intron 4 peptide demonstrated that an immunoreactive protein(s) was present in the syncytiotrophoblasts of the chorionic villi of the rhesus placenta, within villous cytotrophoblasts, and occasionally within cells of the villous stroma. The Mamu-AG5 mRNA was readily detected in rhesus testis (although not in ejaculated sperm). Whereas an Ab against membrane-bound Mamu-AG stained few cells, primarily in the interstitium of the testis, there was consistent immunostaining for Mamu-AG5 in cells within the seminiferous tubules, which was corroborated by localization of Mamu-AG mRNA by in situ hybridization. While primary spermatocytes were negative, Sertoli cells, spermatocytes, and spermatids were consistently positive for 16G1 immunostaining. The specific recognition of the soluble Mamu-AG isoform was confirmed by Western blotting of Mamu-AG5 expressed in heterologous cells. The results demonstrate that a soluble nonclassical MHC class I molecule is expressed in the rhesus monkey placenta and testis, and confirm and extend the unique homology between HLA-G and the rhesus nonclassical molecule Mamu-AG.

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Section: Discussioncontrasting

confidence: 55%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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A Soluble Isoform of the Rhesus Monkey Nonclassical MHC Class I Molecule Mamu-AG Is Expressed in the Placenta and the Testis

Ryan¹,

Grendell²,

Geraghty

et al. 2002

The Journal of Immunology

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“…Importantly, HLA-G FHC are present in vivo only on EVT cells as demonstrated here by us (Fig. 5D, I) and by others (34,61). Further research is required to assess the physiological role of these FHC molecules and the process of their generation in the decidua.…”

Section: Discussionsupporting

confidence: 54%

The CD85J/Leukocyte Inhibitory Receptor-1 Distinguishes between Conformed and β2-Microglobulin-Free HLA-G Molecules

Gonen‐Gross¹,

Achdout²,

Arnon³

et al. 2005

The Journal of Immunology

109

124

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For a proper development of the placenta, maternal NK cells should not attack the fetal extravillous cytotrophoblast cells. This inhibition of maternal NK cells is partially mediated via the nonclassical MHC class I molecule HLA-G. Recently, we demonstrated that HLA-G forms disulfide-linked high molecular complexes on the surface of transfected cells. In the present study, we demonstrate that HLA-G must associate with β2m for its interaction with CD85J/leukocyte Ig-like receptor-1 (LIR-1). Although HLA-G free H chain complexes are expressed on the surface, they are not recognized and possibly interfere with CD85J/LIR-1 and HLA-G interaction. The formation of these complexes on the cell surface might represent a novel mechanism developed specifically by the HLA-G protein aimed to control the efficiency of the CD85J/LIR-1-mediated inhibition. We also show that endogenous HLA-G complexes are expressed on the cell surface. These findings provide novel insights into the delicate interaction between extravillous cytotrophoblast cells and NK cells in the decidua.

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Secretion of pro-apoptotic intron 4-retaining soluble HLA-G1 by human villous trophoblast

et al. 2002

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One major materno‐fetal interface in the human placenta is constituted by the syncytiotrophoblast, in contact with maternal blood of the intervillous space, which derives from differentiation and fusion of the villous cytotrophoblast (vct). In the present work, we purified vct from term placenta by depleting HLA class I‐ and class II‐positive cells. We found by RT‐PCR that both soluble intron 4‐retaining HLA‐G1 (sHLA‐G1) and HLA‐G2 isoforms were transcribed in purified vct. Using different HLA‐G‐specific mAb, we demonstrated by intracellular flow cytometry, Western blotting and ELISA, that sHLA‐G1 but no soluble HLA class Ia molecule was secreted by vct. We then purified sHLA‐G1 from vct culture supernatant and found that it exhibited an unusual glycosylation pattern. Finally, we showed that such trophoblast‐derived sHLA‐G1 triggered specific apoptosis of activated CD8+ T cells. Taken together, these results demonstrated that vct did secrete functional sHLA‐G1 in primary culture and suggested that, in vivo, sHLA‐G1 might be an important immunomodulatory molecule controlling the activity of maternal immune effector CD8+ cells circulating in the blood that immerses chorionic villi.

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Reaction patterns of monoclonal antibodies to HLA-G in human tissues and on cell lines: a comparative study

Cited by 40 publications

References 30 publications

A Soluble Isoform of the Rhesus Monkey Nonclassical MHC Class I Molecule Mamu-AG Is Expressed in the Placenta and the Testis

A Soluble Isoform of the Rhesus Monkey Nonclassical MHC Class I Molecule Mamu-AG Is Expressed in the Placenta and the Testis

The CD85J/Leukocyte Inhibitory Receptor-1 Distinguishes between Conformed and β2-Microglobulin-Free HLA-G Molecules

Secretion of pro-apoptotic intron 4-retaining soluble HLA-G1 by human villous trophoblast

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