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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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(a) 51Cr release assay of NKG7 against LCL721.221 (•) and  LCL721.221.G (○) in the presence of 2.5 μg/ml 5.133 (▾) or HP-3E4  (▿). (b) Flow cytometry analysis of NKAT3 and/or NKAT4 expression  of NKG7. Staining with the 5.133 antibody and a goat α–mouse FITClabeled secondary antibody (broken line). Staining with the secondary antibody only served as the negative control (solid line).
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Figure 5: (a) 51Cr release assay of NKG7 against LCL721.221 (•) and LCL721.221.G (○) in the presence of 2.5 μg/ml 5.133 (▾) or HP-3E4 (▿). (b) Flow cytometry analysis of NKAT3 and/or NKAT4 expression of NKG7. Staining with the 5.133 antibody and a goat α–mouse FITClabeled secondary antibody (broken line). Staining with the secondary antibody only served as the negative control (solid line).

Mentions: This was done in two sets of experiments. First, blocking of the receptor during 51Cr release assay with mAbs was used to prevent inhibition by HLA-G. Second, transfectants, as well as PHA blasts of typed donors, were surveyed for coinhibition of NKG1, NKG2, and NKG7. The mAbs 5.133, recognizing NKAT3 as well as NKAT4, and HP-3E4, recognizing NKAT1, were used at a final concentration of 2.5 μg/ml in the medium during 51Cr release assay. Addition of 5.133 prevented inhibition of LCL721.221 lysis by HLA-G, while addition of HP-3E4 did not. This effect was observed with all three NK lines. Representative data for NKG7 are given in Fig. 5 a. 5.133 Fab fragments and HP-3E4 antibodies were previously used to restore NK lysis by blocking KIRs (33, 34). In agreement with this finding, the 5.133 antibody stains the NK lines efficiently in flow cytometry (Fig. 5 b).


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)

(a) 51Cr release assay of NKG7 against LCL721.221 (•) and  LCL721.221.G (○) in the presence of 2.5 μg/ml 5.133 (▾) or HP-3E4  (▿). (b) Flow cytometry analysis of NKAT3 and/or NKAT4 expression  of NKG7. Staining with the 5.133 antibody and a goat α–mouse FITClabeled secondary antibody (broken line). Staining with the secondary antibody only served as the negative control (solid line).
© Copyright Policy
Related In: Results  -  Collection

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

Figure 5: (a) 51Cr release assay of NKG7 against LCL721.221 (•) and LCL721.221.G (○) in the presence of 2.5 μg/ml 5.133 (▾) or HP-3E4 (▿). (b) Flow cytometry analysis of NKAT3 and/or NKAT4 expression of NKG7. Staining with the 5.133 antibody and a goat α–mouse FITClabeled secondary antibody (broken line). Staining with the secondary antibody only served as the negative control (solid line).
Mentions: This was done in two sets of experiments. First, blocking of the receptor during 51Cr release assay with mAbs was used to prevent inhibition by HLA-G. Second, transfectants, as well as PHA blasts of typed donors, were surveyed for coinhibition of NKG1, NKG2, and NKG7. The mAbs 5.133, recognizing NKAT3 as well as NKAT4, and HP-3E4, recognizing NKAT1, were used at a final concentration of 2.5 μg/ml in the medium during 51Cr release assay. Addition of 5.133 prevented inhibition of LCL721.221 lysis by HLA-G, while addition of HP-3E4 did not. This effect was observed with all three NK lines. Representative data for NKG7 are given in Fig. 5 a. 5.133 Fab fragments and HP-3E4 antibodies were previously used to restore NK lysis by blocking KIRs (33, 34). In agreement with this finding, the 5.133 antibody stains the NK lines efficiently in flow cytometry (Fig. 5 b).

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