Work

CCR2b-Transduced Cytotoxic T Lymphocytes Show Enhanced Homing In Vivo Toward CCL2-Secreting Neuroblastoma: Implications for Enhanced Adoptive Immunotherapy

Year: 2009

Type: article

Abstract: The chemokine CCL2 (MCP-1) is produced by a variety of cancers including neuroblastoma, melanoma, breast cancer and hematological malignancies. However, cytotoxic T lymphocytes (CTL) used in clinical trials to treat these diseases do not express CCR2, the receptor for CCL2. This deficiency may contribute to suboptimal trafficking to tumor sites and reduced clinical efficacy. Therefore, we examined whether genetically modifying CTL with CCR2 could increase their homing towards tumor cells that produced CCL2. CTL were first examined by flow cytometry for chemokine receptor expression using antibodies to CCR2, CCR4, CCR5, CCR7 and CXCR4. These CTL showed nominal expression of CCR2 (<1%), low expression of CCR4 (10%), CXCR4 (25%), CCR7 (5%), and high expression of CCR5 (70%). ELISA for CCL2 was performed on established neuroblastoma cell lines (IMR-32, LAN-1, SK-N-SH, JF) and on neuroblastoma cell lines derived from patients primary tumors. All neuroblastoma cell lines secreted CCL2 in variation but notably, all patient-derived primary tumor cell lines secreted high levels of CCL2. T cells were activated by anti-CD3/anti-CD28 antibody stimulation for 3 days in the presence of IL-2, and then transduced using a CCR2b retroviral vector. Expression of CCR2b was detected by flow cytometry producing up to 78% transduction. In vitro chemotaxis of CCR2b-transduced T cells showed enhance migration to soluble CCL2 (>3 fold) as well as to supernatants from CCL2-secreting cell lines (>2 fold) compared to non-transduced T cells. To study in vivo migration, we co-transduced T cells with EGFPluciferase and CCR2b. Subsequent tail vein infusion of these co-transduced T cells into mice bearing CCL2 secreting tumors (SK-N-SH) showed preferential migration of only CCR2b-transduced T cells toward the tumor site when compared to non-transduced T cells. These results demonstrate the feasibility of influencing the trafficking of adoptively transferred T cells toward CCL2 secreting neuroblastoma and thereby suggesting a potential means to enhance future adoptive immunotherapies targeting neuroblastoma.

Source: Transplantation and Cellular Therapy

DOI: 10.1016/j.bbmt.2008.12.257

Authors: J.A. Craddock, A. Lu, C.M. Rooney, A.E. Foster

Institution: Baylor College of Medicine

Language: English

Topic: Immunotherapy and Immune Responses

Subfield: Immunology

Field: Immunology and Microbiology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: hybrid

Estimated APC paid: $3,650