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Human histocompatibility leukocyte antigen (HLA)-G molecules inhibit NKAT3 expressing natural killer cells.

Münz C, Holmes N, King A, Loke YW, Colonna M, Schild H, Rammensee HG - J. Exp. Med. (1997)

Bottom Line: We present three NK lines that are inhibited via the interaction of their NKAT3 receptor with HLA-G and with HLA-Bw4 molecules.Inhibition can be blocked by the anti-NKAT3 antibody 5.133.In conclusion, NK inhibition by HLA-G via NKAT3 may contribute to the survival of the fetal semiallograft in the mother during pregnancy.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, University of Tübingen, Federal Republic of Germany.

ABSTRACT
The crucial immunological function of the classical human major histocompatibility complex (MHC) class I molecules, human histocompatibility leukocyte antigen (HLA)-A, -B, and -C, is the presentation of peptides to T cells. A secondary function is the inhibition of natural killer (NK) cells, mediated by binding of class I molecules to NK receptors. In contrast, the function of the nonclassical human MHC class I molecules, HLA-E, -F, and -G, is still a mystery. The specific expression of HLA-G in placental trophoblast suggests an important role for this molecule in the immunological interaction between mother and child. The fetus, semiallograft by its genotype, escapes maternal allorecognition by downregulation of HLA-A and HLA-B molecules at this interface. It has been suggested that the maternal NK recognition of this downregulation is balanced by the expression of HLA-G, thus preventing damage to the placenta. Here, we describe the partial inhibition of NK lysis of the MHC class I negative cell line LCL721.221 upon HLA-G transfection. We present three NK lines that are inhibited via the interaction of their NKAT3 receptor with HLA-G and with HLA-Bw4 molecules. Inhibition can be blocked by the anti-NKAT3 antibody 5.133. In conclusion, NK inhibition by HLA-G via NKAT3 may contribute to the survival of the fetal semiallograft in the mother during pregnancy.

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Flow cytometry analysis of the surface molecules of NKG7  (broken line) by staining of (a) CD8 with a directly FITC-labeled αCD8  antibody; (b) CD4 with a directly FITC-labeled αCD4 antibody; (c)CD16 with the B73.1 antibody and a FITC-labeled goatα-mouse antibody; (d) CD56 with a directly FITC-labeled αCD56 antibody. The negative control represents NKG7 cells incubated with the secondary goat  α-mouse antibody (solid line).
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Figure 3: Flow cytometry analysis of the surface molecules of NKG7 (broken line) by staining of (a) CD8 with a directly FITC-labeled αCD8 antibody; (b) CD4 with a directly FITC-labeled αCD4 antibody; (c)CD16 with the B73.1 antibody and a FITC-labeled goatα-mouse antibody; (d) CD56 with a directly FITC-labeled αCD56 antibody. The negative control represents NKG7 cells incubated with the secondary goat α-mouse antibody (solid line).

Mentions: Positively MACSselected CD56+ PBL from healthy donors were diluted in 96-well plates, split into three sets of plates after 7 d, and assayed for killing of LCL721.221 and LCL721.221.G after another 7 d. 50% of the NK-containing wells showed inhibition of killing upon HLA-G transfection of the targets. Seven wells that showed 50–90% higher specific lysis of LCL721.221 compared with specific lysis of LCL721.221.G were expanded (NKG1–7; Fig. 2). Of the seven cultures, three cultures, NKG1, NKG2, and NKG7, preserved recognition of HLA-G in our culture conditions, and killing of LCL721.221 by these lines was inhibited to background levels upon HLA-G surface expression. FACS® analysis of the three NK lines showed homogeneous surface expression of the NK surface markers CD16 and CD56. CD4 and CD8 expression could not be detected, as shown in Fig. 3 for NKG7. NKG1 and NKG2 showed the same phenotype of tested surface markers: CD16+, CD56dim, CD4−, CD8− (data not shown). The ratios of specific lysis of LCL721.221 to the specific lysis of LCL721.221.G for the three lines after 4 wk in culture at a E/T ratio of 1:1 were the following: NKG1, 50/18; NKG2, 35/5; NKG7, 62/21 (Fig. 4).


Human histocompatibility leukocyte antigen (HLA)-G molecules inhibit NKAT3 expressing natural killer cells.

Münz C, Holmes N, King A, Loke YW, Colonna M, Schild H, Rammensee HG - J. Exp. Med. (1997)

Flow cytometry analysis of the surface molecules of NKG7  (broken line) by staining of (a) CD8 with a directly FITC-labeled αCD8  antibody; (b) CD4 with a directly FITC-labeled αCD4 antibody; (c)CD16 with the B73.1 antibody and a FITC-labeled goatα-mouse antibody; (d) CD56 with a directly FITC-labeled αCD56 antibody. The negative control represents NKG7 cells incubated with the secondary goat  α-mouse antibody (solid line).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196038&req=5

Figure 3: Flow cytometry analysis of the surface molecules of NKG7 (broken line) by staining of (a) CD8 with a directly FITC-labeled αCD8 antibody; (b) CD4 with a directly FITC-labeled αCD4 antibody; (c)CD16 with the B73.1 antibody and a FITC-labeled goatα-mouse antibody; (d) CD56 with a directly FITC-labeled αCD56 antibody. The negative control represents NKG7 cells incubated with the secondary goat α-mouse antibody (solid line).
Mentions: Positively MACSselected CD56+ PBL from healthy donors were diluted in 96-well plates, split into three sets of plates after 7 d, and assayed for killing of LCL721.221 and LCL721.221.G after another 7 d. 50% of the NK-containing wells showed inhibition of killing upon HLA-G transfection of the targets. Seven wells that showed 50–90% higher specific lysis of LCL721.221 compared with specific lysis of LCL721.221.G were expanded (NKG1–7; Fig. 2). Of the seven cultures, three cultures, NKG1, NKG2, and NKG7, preserved recognition of HLA-G in our culture conditions, and killing of LCL721.221 by these lines was inhibited to background levels upon HLA-G surface expression. FACS® analysis of the three NK lines showed homogeneous surface expression of the NK surface markers CD16 and CD56. CD4 and CD8 expression could not be detected, as shown in Fig. 3 for NKG7. NKG1 and NKG2 showed the same phenotype of tested surface markers: CD16+, CD56dim, CD4−, CD8− (data not shown). The ratios of specific lysis of LCL721.221 to the specific lysis of LCL721.221.G for the three lines after 4 wk in culture at a E/T ratio of 1:1 were the following: NKG1, 50/18; NKG2, 35/5; NKG7, 62/21 (Fig. 4).

Bottom Line: We present three NK lines that are inhibited via the interaction of their NKAT3 receptor with HLA-G and with HLA-Bw4 molecules.Inhibition can be blocked by the anti-NKAT3 antibody 5.133.In conclusion, NK inhibition by HLA-G via NKAT3 may contribute to the survival of the fetal semiallograft in the mother during pregnancy.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, University of Tübingen, Federal Republic of Germany.

ABSTRACT
The crucial immunological function of the classical human major histocompatibility complex (MHC) class I molecules, human histocompatibility leukocyte antigen (HLA)-A, -B, and -C, is the presentation of peptides to T cells. A secondary function is the inhibition of natural killer (NK) cells, mediated by binding of class I molecules to NK receptors. In contrast, the function of the nonclassical human MHC class I molecules, HLA-E, -F, and -G, is still a mystery. The specific expression of HLA-G in placental trophoblast suggests an important role for this molecule in the immunological interaction between mother and child. The fetus, semiallograft by its genotype, escapes maternal allorecognition by downregulation of HLA-A and HLA-B molecules at this interface. It has been suggested that the maternal NK recognition of this downregulation is balanced by the expression of HLA-G, thus preventing damage to the placenta. Here, we describe the partial inhibition of NK lysis of the MHC class I negative cell line LCL721.221 upon HLA-G transfection. We present three NK lines that are inhibited via the interaction of their NKAT3 receptor with HLA-G and with HLA-Bw4 molecules. Inhibition can be blocked by the anti-NKAT3 antibody 5.133. In conclusion, NK inhibition by HLA-G via NKAT3 may contribute to the survival of the fetal semiallograft in the mother during pregnancy.

Show MeSH
Related in: MedlinePlus