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Immune evasion by murine melanoma mediated through CC chemokine receptor-10.

Murakami T, Cardones AR, Finkelstein SE, Restifo NP, Klaunberg BA, Nestle FO, Castillo SS, Dennis PA, Hwang ST - J. Exp. Med. (2003)

Bottom Line: Compared with B16 murine melanoma, cells rendered more immunogenic via overexpression of luciferase, B16 cells that overexpressed both luciferase and CCR10 resisted host immune responses and readily formed tumors.In vivo, cutaneous injection of neutralizing antibodies to endogenous CCL27 blocked growth of CCR10-expressing melanoma cells.We propose that CCR10 engagement by locally produced CCL27 allows melanoma cells to escape host immune antitumor killing mechanisms (possibly through activation of PI3K/Akt), thereby providing a means for tumor progression.

View Article: PubMed Central - PubMed

Affiliation: Dermatology Branch, CCR, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.

ABSTRACT
Human melanoma cells frequently express CC chemokine receptor (CCR)10, a receptor whose ligand (CCL27) is constitutively produced by keratinocytes. Compared with B16 murine melanoma, cells rendered more immunogenic via overexpression of luciferase, B16 cells that overexpressed both luciferase and CCR10 resisted host immune responses and readily formed tumors. In vitro, exposure of tumor cells to CCL27 led to rapid activation of Akt, resistance to cell death induced by melanoma antigen-specific cytotoxic T cells, and phosphatidylinositol-3-kinase (PI3K)-dependent protection from apoptosis induced by Fas cross-linking. In vivo, cutaneous injection of neutralizing antibodies to endogenous CCL27 blocked growth of CCR10-expressing melanoma cells. We propose that CCR10 engagement by locally produced CCL27 allows melanoma cells to escape host immune antitumor killing mechanisms (possibly through activation of PI3K/Akt), thereby providing a means for tumor progression.

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Growth and regional LN metastasis of CCR10- and pLNCX2-B16 tumor cells. CCR10- or pLNCX2-B16 cells were injected into the footpads of mice. (A) After 21 d, animals were killed and tumor sizes in the footpads were compared (P = 0.003, n = 5 per group). (B) Draining popliteal LN from injected footpads. (C–E) Representative ears from mice injected in the dermis of the ear with pLNCX2-B16 cells (C) showing a residual tumor macule indicated by the white arrow (D) and from mice injected at analogous sites with CCR10-B16 cells (E) are shown 20 d after inoculation. Dissection of the cervical region ipsilateral to the tumor injection sites reveals a large cervical LN metastasis in a CCR10-B16–injected mouse (F), but not in a pLNCX2-B16–injected mouse (G; representative animals are shown in F and G). Scale bar in B–E is calibrated in mm.
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fig2: Growth and regional LN metastasis of CCR10- and pLNCX2-B16 tumor cells. CCR10- or pLNCX2-B16 cells were injected into the footpads of mice. (A) After 21 d, animals were killed and tumor sizes in the footpads were compared (P = 0.003, n = 5 per group). (B) Draining popliteal LN from injected footpads. (C–E) Representative ears from mice injected in the dermis of the ear with pLNCX2-B16 cells (C) showing a residual tumor macule indicated by the white arrow (D) and from mice injected at analogous sites with CCR10-B16 cells (E) are shown 20 d after inoculation. Dissection of the cervical region ipsilateral to the tumor injection sites reveals a large cervical LN metastasis in a CCR10-B16–injected mouse (F), but not in a pLNCX2-B16–injected mouse (G; representative animals are shown in F and G). Scale bar in B–E is calibrated in mm.

Mentions: To characterize the role of CCR10 in a murine model of melanoma growth and metastasis, we transduced B16/F1 cells with the full-length cDNA-encoding murine CCR10 (12). As noted previously (10), B16 cells show negligible levels of mRNA expression for several chemokine receptors tested, including CCR10. Whereas CCR10 was not detectable by RT-PCR (not depicted) and Western blotting in lysates of B16 cells transduced with the vector alone (pLNCX2-B16) or with CXCR4 cDNA (Fig. S1 A, lanes 1 and 2), CCR10-transduced B16 cells (CCR10-B16) expressed a 42-kD protein that was recognized by a specific anti-CCR10 antibody. CCR10-B16 cells underwent a calcium flux in response to CCL27 but not to CXCL12 (Fig. S1 B). Footpad tumors of CCR10-B16–injected mice were moderately increased in size (P = 0.003) compared with the tumors from pLNCX2-B16–injected mice (Fig. 2 A). Strikingly, the popliteal LN from CCR10-B16–injected mice showed frequent gross metastases (Fig. 2 B). By contrast, pLNCX2-B16 cells rarely metastasized to the draining LN (Fig. 2 B), a phenotype of these cells that we had reported previously (10). Luciferase quantification revealed a mean luciferase activity of 13988 relative light units in the popliteal LN from CCR10-B16–injected mice versus 21 relative light units for pLNCX2-B16–injected mice (P = 0.003, n = 7, Mann-Whitney test). Thus, in addition to demonstrating a moderate advantage in growth of the primary tumor compared with control cells, CCR10-B16 cells display a striking increase in metastasis to the draining LN after injection into the footpads of mice.


Immune evasion by murine melanoma mediated through CC chemokine receptor-10.

Murakami T, Cardones AR, Finkelstein SE, Restifo NP, Klaunberg BA, Nestle FO, Castillo SS, Dennis PA, Hwang ST - J. Exp. Med. (2003)

Growth and regional LN metastasis of CCR10- and pLNCX2-B16 tumor cells. CCR10- or pLNCX2-B16 cells were injected into the footpads of mice. (A) After 21 d, animals were killed and tumor sizes in the footpads were compared (P = 0.003, n = 5 per group). (B) Draining popliteal LN from injected footpads. (C–E) Representative ears from mice injected in the dermis of the ear with pLNCX2-B16 cells (C) showing a residual tumor macule indicated by the white arrow (D) and from mice injected at analogous sites with CCR10-B16 cells (E) are shown 20 d after inoculation. Dissection of the cervical region ipsilateral to the tumor injection sites reveals a large cervical LN metastasis in a CCR10-B16–injected mouse (F), but not in a pLNCX2-B16–injected mouse (G; representative animals are shown in F and G). Scale bar in B–E is calibrated in mm.
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fig2: Growth and regional LN metastasis of CCR10- and pLNCX2-B16 tumor cells. CCR10- or pLNCX2-B16 cells were injected into the footpads of mice. (A) After 21 d, animals were killed and tumor sizes in the footpads were compared (P = 0.003, n = 5 per group). (B) Draining popliteal LN from injected footpads. (C–E) Representative ears from mice injected in the dermis of the ear with pLNCX2-B16 cells (C) showing a residual tumor macule indicated by the white arrow (D) and from mice injected at analogous sites with CCR10-B16 cells (E) are shown 20 d after inoculation. Dissection of the cervical region ipsilateral to the tumor injection sites reveals a large cervical LN metastasis in a CCR10-B16–injected mouse (F), but not in a pLNCX2-B16–injected mouse (G; representative animals are shown in F and G). Scale bar in B–E is calibrated in mm.
Mentions: To characterize the role of CCR10 in a murine model of melanoma growth and metastasis, we transduced B16/F1 cells with the full-length cDNA-encoding murine CCR10 (12). As noted previously (10), B16 cells show negligible levels of mRNA expression for several chemokine receptors tested, including CCR10. Whereas CCR10 was not detectable by RT-PCR (not depicted) and Western blotting in lysates of B16 cells transduced with the vector alone (pLNCX2-B16) or with CXCR4 cDNA (Fig. S1 A, lanes 1 and 2), CCR10-transduced B16 cells (CCR10-B16) expressed a 42-kD protein that was recognized by a specific anti-CCR10 antibody. CCR10-B16 cells underwent a calcium flux in response to CCL27 but not to CXCL12 (Fig. S1 B). Footpad tumors of CCR10-B16–injected mice were moderately increased in size (P = 0.003) compared with the tumors from pLNCX2-B16–injected mice (Fig. 2 A). Strikingly, the popliteal LN from CCR10-B16–injected mice showed frequent gross metastases (Fig. 2 B). By contrast, pLNCX2-B16 cells rarely metastasized to the draining LN (Fig. 2 B), a phenotype of these cells that we had reported previously (10). Luciferase quantification revealed a mean luciferase activity of 13988 relative light units in the popliteal LN from CCR10-B16–injected mice versus 21 relative light units for pLNCX2-B16–injected mice (P = 0.003, n = 7, Mann-Whitney test). Thus, in addition to demonstrating a moderate advantage in growth of the primary tumor compared with control cells, CCR10-B16 cells display a striking increase in metastasis to the draining LN after injection into the footpads of mice.

Bottom Line: Compared with B16 murine melanoma, cells rendered more immunogenic via overexpression of luciferase, B16 cells that overexpressed both luciferase and CCR10 resisted host immune responses and readily formed tumors.In vivo, cutaneous injection of neutralizing antibodies to endogenous CCL27 blocked growth of CCR10-expressing melanoma cells.We propose that CCR10 engagement by locally produced CCL27 allows melanoma cells to escape host immune antitumor killing mechanisms (possibly through activation of PI3K/Akt), thereby providing a means for tumor progression.

View Article: PubMed Central - PubMed

Affiliation: Dermatology Branch, CCR, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.

ABSTRACT
Human melanoma cells frequently express CC chemokine receptor (CCR)10, a receptor whose ligand (CCL27) is constitutively produced by keratinocytes. Compared with B16 murine melanoma, cells rendered more immunogenic via overexpression of luciferase, B16 cells that overexpressed both luciferase and CCR10 resisted host immune responses and readily formed tumors. In vitro, exposure of tumor cells to CCL27 led to rapid activation of Akt, resistance to cell death induced by melanoma antigen-specific cytotoxic T cells, and phosphatidylinositol-3-kinase (PI3K)-dependent protection from apoptosis induced by Fas cross-linking. In vivo, cutaneous injection of neutralizing antibodies to endogenous CCL27 blocked growth of CCR10-expressing melanoma cells. We propose that CCR10 engagement by locally produced CCL27 allows melanoma cells to escape host immune antitumor killing mechanisms (possibly through activation of PI3K/Akt), thereby providing a means for tumor progression.

Show MeSH
Related in: MedlinePlus