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Enhanced recruitment of genetically modified CX3CR1-positive human T cells into Fractalkine/CX3CL1 expressing tumors: importance of the chemokine gradient.

Siddiqui I, Erreni M, van Brakel M, Debets R, Allavena P - J Immunother Cancer (2016)

Bottom Line: Adoptive T-cell based immunotherapies constitute a promising approach to treat cancer, however, a major problem is to obtain effective and long-lasting anti-tumor responses.Furthermore, significant inhibition of tumor growth was found in mice receiving modified CX3CR1-T cells.Our results also establish that a correct chemokine gradient between the systemic circulation and the tumor is an essential requirement in adoptive T-cell based immunotherapy to efficiently recruit T cell effectors at the correct sites.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Inflammation, Humanitas Clinical and Research Center, 20089 Rozzano, Milan Italy ; Ludwig Center for Cancer Research, Department of Oncology, University of Lausanne, 1066 Epalinges, Switzerland.

ABSTRACT

Background: Adoptive T-cell based immunotherapies constitute a promising approach to treat cancer, however, a major problem is to obtain effective and long-lasting anti-tumor responses. Lack of response may be due to insufficient trafficking of specific T cells to tumors. A key requirement for efficient migration of cytotoxic T cells is that they express chemokine receptors that match the chemokines produced by tumor or tumor-associated cells.

Methods: In this study, we investigated whether the in vivo tumor trafficking of activated T cells could be enhanced by the expression of the chemokine receptor CX3CR1. Two human colorectal cancer cell lines were used to set up a xenograft tumor model in immunodeficient mice; the NCI-H630, constitutively expressing the chemokine ligand CX3CL1 (Fractalkine), and the RKO cell line, transduced to express CX3CL1.

Results: Human primary T cells were transduced with the receptor CX3CR1-eGFP. Upon in vivo adoptive transfer of genetically modified CX3CR1-T cells in mice bearing NCI-H630 tumors, enhanced lymphocyte migration and tumor trafficking were observed, compared to mice receiving Mock-T cells, indicating improved homing ability towards ligand-expressing tumor cells. Furthermore, significant inhibition of tumor growth was found in mice receiving modified CX3CR1-T cells. In contrast, tumors formed by RKO cells transduced with the ligand (RKO-CX3CL1) were not affected, nor more infiltrated upon transfer of CX3CR1-T lymphocytes, likely because high levels of the chemokine were shed by tumor cells in the systemic circulation, thus ifying the blood-tissue chemokine gradient.

Conclusions: This study demonstrates that ectopic expression of CX3CR1 enhanced the homing of adoptively transferred T cells towards CX3CL1-producing tumors, resulting in increased T cell infiltration in tumor tissues and decreased tumor growth. Our results also establish that a correct chemokine gradient between the systemic circulation and the tumor is an essential requirement in adoptive T-cell based immunotherapy to efficiently recruit T cell effectors at the correct sites.

No MeSH data available.


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Expression of Fractalkine/CX3CL1 in human colon cancer tissues and cell lines. a Immunostaining score of Fractalkine/CX3CL1 in 171 patients with colorectal cancer; numbers on top of bars define exact number of patients with a given score; b Immunostaining of Fractalkine/CX3CL1 in human normal colonic mucosa (top) and in a representative human colon cancer tissue (bottom); c Immunofluorescence by confocal microscopy of Fractalkine/CX3CL1 protein expression in NCI-H630 cells and d in RKO cells upon transduction of the Fractalkine cDNA (CX3CL1-cherry) or mock-transduced (mock-cherry); e Quantification by ELISA of Fractalkine/CX3CL1 levels in the lysate of NCI-H630 tumors grown in NSG mice, normal skin tissue was used as reference control; f ELISA of Fractalkine/CX3CL1 levels in tumor lysates obtained from RKO tumors (RKO-MOCK and RKO-CX3CL1) grown in NSG mice
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Fig1: Expression of Fractalkine/CX3CL1 in human colon cancer tissues and cell lines. a Immunostaining score of Fractalkine/CX3CL1 in 171 patients with colorectal cancer; numbers on top of bars define exact number of patients with a given score; b Immunostaining of Fractalkine/CX3CL1 in human normal colonic mucosa (top) and in a representative human colon cancer tissue (bottom); c Immunofluorescence by confocal microscopy of Fractalkine/CX3CL1 protein expression in NCI-H630 cells and d in RKO cells upon transduction of the Fractalkine cDNA (CX3CL1-cherry) or mock-transduced (mock-cherry); e Quantification by ELISA of Fractalkine/CX3CL1 levels in the lysate of NCI-H630 tumors grown in NSG mice, normal skin tissue was used as reference control; f ELISA of Fractalkine/CX3CL1 levels in tumor lysates obtained from RKO tumors (RKO-MOCK and RKO-CX3CL1) grown in NSG mice

Mentions: The chemokine Fractalkine/CX3CL1 is expressed in several types of cancers and strongly induces chemotaxis and migration of Th1 lymphocytes, NK cells and macrophages [30, 33, 34]. We and others reported that this chemokine is highly expressed in tumor cells of human colorectal cancer (CRC) [30–32, 35, 36]. Figure 1a shows that from a series of 171 CRC samples studied by immunohistochemistry, 83.6 % were positive or strongly positive for CX3CL1 and only 16.4 % had faint or negative immunostaining. Of note, the normal non-involved colonic mucosa was negative or weakly positive, as shown in Fig. 1b, which also depicts a representative picture of a CX3CL1-positive tumor sample.Fig. 1


Enhanced recruitment of genetically modified CX3CR1-positive human T cells into Fractalkine/CX3CL1 expressing tumors: importance of the chemokine gradient.

Siddiqui I, Erreni M, van Brakel M, Debets R, Allavena P - J Immunother Cancer (2016)

Expression of Fractalkine/CX3CL1 in human colon cancer tissues and cell lines. a Immunostaining score of Fractalkine/CX3CL1 in 171 patients with colorectal cancer; numbers on top of bars define exact number of patients with a given score; b Immunostaining of Fractalkine/CX3CL1 in human normal colonic mucosa (top) and in a representative human colon cancer tissue (bottom); c Immunofluorescence by confocal microscopy of Fractalkine/CX3CL1 protein expression in NCI-H630 cells and d in RKO cells upon transduction of the Fractalkine cDNA (CX3CL1-cherry) or mock-transduced (mock-cherry); e Quantification by ELISA of Fractalkine/CX3CL1 levels in the lysate of NCI-H630 tumors grown in NSG mice, normal skin tissue was used as reference control; f ELISA of Fractalkine/CX3CL1 levels in tumor lysates obtained from RKO tumors (RKO-MOCK and RKO-CX3CL1) grown in NSG mice
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4836203&req=5

Fig1: Expression of Fractalkine/CX3CL1 in human colon cancer tissues and cell lines. a Immunostaining score of Fractalkine/CX3CL1 in 171 patients with colorectal cancer; numbers on top of bars define exact number of patients with a given score; b Immunostaining of Fractalkine/CX3CL1 in human normal colonic mucosa (top) and in a representative human colon cancer tissue (bottom); c Immunofluorescence by confocal microscopy of Fractalkine/CX3CL1 protein expression in NCI-H630 cells and d in RKO cells upon transduction of the Fractalkine cDNA (CX3CL1-cherry) or mock-transduced (mock-cherry); e Quantification by ELISA of Fractalkine/CX3CL1 levels in the lysate of NCI-H630 tumors grown in NSG mice, normal skin tissue was used as reference control; f ELISA of Fractalkine/CX3CL1 levels in tumor lysates obtained from RKO tumors (RKO-MOCK and RKO-CX3CL1) grown in NSG mice
Mentions: The chemokine Fractalkine/CX3CL1 is expressed in several types of cancers and strongly induces chemotaxis and migration of Th1 lymphocytes, NK cells and macrophages [30, 33, 34]. We and others reported that this chemokine is highly expressed in tumor cells of human colorectal cancer (CRC) [30–32, 35, 36]. Figure 1a shows that from a series of 171 CRC samples studied by immunohistochemistry, 83.6 % were positive or strongly positive for CX3CL1 and only 16.4 % had faint or negative immunostaining. Of note, the normal non-involved colonic mucosa was negative or weakly positive, as shown in Fig. 1b, which also depicts a representative picture of a CX3CL1-positive tumor sample.Fig. 1

Bottom Line: Adoptive T-cell based immunotherapies constitute a promising approach to treat cancer, however, a major problem is to obtain effective and long-lasting anti-tumor responses.Furthermore, significant inhibition of tumor growth was found in mice receiving modified CX3CR1-T cells.Our results also establish that a correct chemokine gradient between the systemic circulation and the tumor is an essential requirement in adoptive T-cell based immunotherapy to efficiently recruit T cell effectors at the correct sites.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Inflammation, Humanitas Clinical and Research Center, 20089 Rozzano, Milan Italy ; Ludwig Center for Cancer Research, Department of Oncology, University of Lausanne, 1066 Epalinges, Switzerland.

ABSTRACT

Background: Adoptive T-cell based immunotherapies constitute a promising approach to treat cancer, however, a major problem is to obtain effective and long-lasting anti-tumor responses. Lack of response may be due to insufficient trafficking of specific T cells to tumors. A key requirement for efficient migration of cytotoxic T cells is that they express chemokine receptors that match the chemokines produced by tumor or tumor-associated cells.

Methods: In this study, we investigated whether the in vivo tumor trafficking of activated T cells could be enhanced by the expression of the chemokine receptor CX3CR1. Two human colorectal cancer cell lines were used to set up a xenograft tumor model in immunodeficient mice; the NCI-H630, constitutively expressing the chemokine ligand CX3CL1 (Fractalkine), and the RKO cell line, transduced to express CX3CL1.

Results: Human primary T cells were transduced with the receptor CX3CR1-eGFP. Upon in vivo adoptive transfer of genetically modified CX3CR1-T cells in mice bearing NCI-H630 tumors, enhanced lymphocyte migration and tumor trafficking were observed, compared to mice receiving Mock-T cells, indicating improved homing ability towards ligand-expressing tumor cells. Furthermore, significant inhibition of tumor growth was found in mice receiving modified CX3CR1-T cells. In contrast, tumors formed by RKO cells transduced with the ligand (RKO-CX3CL1) were not affected, nor more infiltrated upon transfer of CX3CR1-T lymphocytes, likely because high levels of the chemokine were shed by tumor cells in the systemic circulation, thus ifying the blood-tissue chemokine gradient.

Conclusions: This study demonstrates that ectopic expression of CX3CR1 enhanced the homing of adoptively transferred T cells towards CX3CL1-producing tumors, resulting in increased T cell infiltration in tumor tissues and decreased tumor growth. Our results also establish that a correct chemokine gradient between the systemic circulation and the tumor is an essential requirement in adoptive T-cell based immunotherapy to efficiently recruit T cell effectors at the correct sites.

No MeSH data available.


Related in: MedlinePlus