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Identification and molecular characterization of NKp30, a novel triggering receptor involved in natural cytotoxicity mediated by human natural killer cells.

Pende D, Parolini S, Pessino A, Sivori S, Augugliaro R, Morelli L, Marcenaro E, Accame L, Malaspina A, Biassoni R, Bottino C, Moretta L, Moretta A - J. Exp. Med. (1999)

Bottom Line: This novel receptor is associated with CD3zeta chains that become tyrosine phosphorylated upon sodium pervanadate treatment of NK cells.Molecular cloning of NKp30 cDNA revealed a member of the immunoglobulin superfamily, characterized by a single V-type domain and a charged residue in the transmembrane portion.Moreover, we show that NKp30 is encoded by the previously identified 1C7 gene, for which the function and the cellular distribution of the putative product were not identified in previous studies.

View Article: PubMed Central - PubMed

Affiliation: Istituto Nazionale per la Ricerca sul Cancro, 16132 Genova, Italy.

ABSTRACT
Two major receptors involved in human natural cytotoxicity, NKp46 and NKp44, have recently been identified. However, experimental evidence suggested the existence of additional such receptor(s). In this study, by the generation of monoclonal antibodies (mAbs), we identified NKp30, a novel 30-kD triggering receptor selectively expressed by all resting and activated human natural killer (NK) cells. Although mAb-mediated cross-linking of NKp30 induces strong NK cell activation, mAb-mediated masking inhibits the NK cytotoxicity against normal or tumor target cells. NKp30 cooperates with NKp46 and/or NKp44 in the induction of NK-mediated cytotoxicity against the majority of target cells, whereas it represents the major triggering receptor in the killing of certain tumors. This novel receptor is associated with CD3zeta chains that become tyrosine phosphorylated upon sodium pervanadate treatment of NK cells. Molecular cloning of NKp30 cDNA revealed a member of the immunoglobulin superfamily, characterized by a single V-type domain and a charged residue in the transmembrane portion. Moreover, we show that NKp30 is encoded by the previously identified 1C7 gene, for which the function and the cellular distribution of the putative product were not identified in previous studies.

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NKp30 functions as an activating receptor in fresh NK cells and is involved in their natural cytotoxicity. (A) Freshly isolated peripheral blood NK lymphocytes, derived from a representative donor, were analyzed for cytolytic activity in a redirected killing assay against the FcγR-positive P815 target cell line in the absence or presence of c127 (anti-CD16), BAB281 (anti-NKp46), AZ20, A76, Z25, and c218 (anti-CD56) mAbs. The E/T ratio used was 20:1. (B) Freshly isolated peripheral blood NK cells were analyzed for cytolytic activity against the indicated FcγR-negative/HLA class I–negative melanoma cell lines either in the absence or presence of mAbs to the indicated molecules. c218 (anti-CD56), AZ20 (anti-NKp30), and BAB281 (anti-NKp46) mAbs were used. The E/T ratio was 20:1.
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Figure 4: NKp30 functions as an activating receptor in fresh NK cells and is involved in their natural cytotoxicity. (A) Freshly isolated peripheral blood NK lymphocytes, derived from a representative donor, were analyzed for cytolytic activity in a redirected killing assay against the FcγR-positive P815 target cell line in the absence or presence of c127 (anti-CD16), BAB281 (anti-NKp46), AZ20, A76, Z25, and c218 (anti-CD56) mAbs. The E/T ratio used was 20:1. (B) Freshly isolated peripheral blood NK cells were analyzed for cytolytic activity against the indicated FcγR-negative/HLA class I–negative melanoma cell lines either in the absence or presence of mAbs to the indicated molecules. c218 (anti-CD56), AZ20 (anti-NKp30), and BAB281 (anti-NKp46) mAbs were used. The E/T ratio was 20:1.

Mentions: Since NKp30 molecule, like NKp46, was expressed on fresh NK cells, we analyzed whether it could trigger the cytolytic activity of these cells as demonstrated previously for NKp46. As shown in Fig. 4 A, AZ20, A76, and Z25 mAbs induced a strong increase of cytolytic activity against P815 target cells, whereas the isotype–matched anti-CD56 mAb had no effect. This triggering effect was comparable to that obtained with anti-NKp46 mAb. Moreover, in these experiments, the use of AZ20 F(ab′)2 fragments did not induce triggering of cytolytic activity, indicating that mAb-dependent NKp30 stimulation requires efficient cross-linking mediated by FcγR on target cells (data not shown).


Identification and molecular characterization of NKp30, a novel triggering receptor involved in natural cytotoxicity mediated by human natural killer cells.

Pende D, Parolini S, Pessino A, Sivori S, Augugliaro R, Morelli L, Marcenaro E, Accame L, Malaspina A, Biassoni R, Bottino C, Moretta L, Moretta A - J. Exp. Med. (1999)

NKp30 functions as an activating receptor in fresh NK cells and is involved in their natural cytotoxicity. (A) Freshly isolated peripheral blood NK lymphocytes, derived from a representative donor, were analyzed for cytolytic activity in a redirected killing assay against the FcγR-positive P815 target cell line in the absence or presence of c127 (anti-CD16), BAB281 (anti-NKp46), AZ20, A76, Z25, and c218 (anti-CD56) mAbs. The E/T ratio used was 20:1. (B) Freshly isolated peripheral blood NK cells were analyzed for cytolytic activity against the indicated FcγR-negative/HLA class I–negative melanoma cell lines either in the absence or presence of mAbs to the indicated molecules. c218 (anti-CD56), AZ20 (anti-NKp30), and BAB281 (anti-NKp46) mAbs were used. The E/T ratio was 20:1.
© Copyright Policy
Related In: Results  -  Collection

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Figure 4: NKp30 functions as an activating receptor in fresh NK cells and is involved in their natural cytotoxicity. (A) Freshly isolated peripheral blood NK lymphocytes, derived from a representative donor, were analyzed for cytolytic activity in a redirected killing assay against the FcγR-positive P815 target cell line in the absence or presence of c127 (anti-CD16), BAB281 (anti-NKp46), AZ20, A76, Z25, and c218 (anti-CD56) mAbs. The E/T ratio used was 20:1. (B) Freshly isolated peripheral blood NK cells were analyzed for cytolytic activity against the indicated FcγR-negative/HLA class I–negative melanoma cell lines either in the absence or presence of mAbs to the indicated molecules. c218 (anti-CD56), AZ20 (anti-NKp30), and BAB281 (anti-NKp46) mAbs were used. The E/T ratio was 20:1.
Mentions: Since NKp30 molecule, like NKp46, was expressed on fresh NK cells, we analyzed whether it could trigger the cytolytic activity of these cells as demonstrated previously for NKp46. As shown in Fig. 4 A, AZ20, A76, and Z25 mAbs induced a strong increase of cytolytic activity against P815 target cells, whereas the isotype–matched anti-CD56 mAb had no effect. This triggering effect was comparable to that obtained with anti-NKp46 mAb. Moreover, in these experiments, the use of AZ20 F(ab′)2 fragments did not induce triggering of cytolytic activity, indicating that mAb-dependent NKp30 stimulation requires efficient cross-linking mediated by FcγR on target cells (data not shown).

Bottom Line: This novel receptor is associated with CD3zeta chains that become tyrosine phosphorylated upon sodium pervanadate treatment of NK cells.Molecular cloning of NKp30 cDNA revealed a member of the immunoglobulin superfamily, characterized by a single V-type domain and a charged residue in the transmembrane portion.Moreover, we show that NKp30 is encoded by the previously identified 1C7 gene, for which the function and the cellular distribution of the putative product were not identified in previous studies.

View Article: PubMed Central - PubMed

Affiliation: Istituto Nazionale per la Ricerca sul Cancro, 16132 Genova, Italy.

ABSTRACT
Two major receptors involved in human natural cytotoxicity, NKp46 and NKp44, have recently been identified. However, experimental evidence suggested the existence of additional such receptor(s). In this study, by the generation of monoclonal antibodies (mAbs), we identified NKp30, a novel 30-kD triggering receptor selectively expressed by all resting and activated human natural killer (NK) cells. Although mAb-mediated cross-linking of NKp30 induces strong NK cell activation, mAb-mediated masking inhibits the NK cytotoxicity against normal or tumor target cells. NKp30 cooperates with NKp46 and/or NKp44 in the induction of NK-mediated cytotoxicity against the majority of target cells, whereas it represents the major triggering receptor in the killing of certain tumors. This novel receptor is associated with CD3zeta chains that become tyrosine phosphorylated upon sodium pervanadate treatment of NK cells. Molecular cloning of NKp30 cDNA revealed a member of the immunoglobulin superfamily, characterized by a single V-type domain and a charged residue in the transmembrane portion. Moreover, we show that NKp30 is encoded by the previously identified 1C7 gene, for which the function and the cellular distribution of the putative product were not identified in previous studies.

Show MeSH
Related in: MedlinePlus