Impact of tumor-derived CCL2 on T cell effector function

doi:10.1016/j.imlet.2003.12.009

Immunology Letters

Volume 91, Issues 2–3, 15 February 2004, Pages 239-245

https://doi.org/10.1016/j.imlet.2003.12.009 Get rights and content

Abstract

To study the effects of tumor-derived monocyte chemoattractant protein-1 (MCP-1, CCL2) on the anti-tumor immune response we used the 4T1 murine mammary carcinoma which constitutively expresses CCL2. We generated 4T1 that do not express detectable levels of CCL2 and found that the T cell response to the tumors were altered. Lymph nodes draining the CCL2- tumor contained CD62L^lo cells that produced greater levels of INF-γ in response to the tumor than CD62L^lo cells from lymph nodes draining a tumor that produced CCL2. Moreover, exposure of splenic T cells to recombinant CCL2 in vitro decreased the ability of the T cells to produce IFN-γ. However, despite the enhanced effector function evident in the absence of CCL2, vaccination/challenge experiments failed to reveal an increase in immunogencity of the CCL2 null cells relative to the CCL2⁺ cells. Collectively, these data indicate that tumor-derived CCL2 could decrease T cell effector function, yet not the overall immunogenicity of the tumor.

Introduction

In 1983 Botazzi et al. [1] reported that the amount of tumor derived chemotactic factor (TDCF) produced by tumors correlated with the level of macrophage infiltration. Subsequently, monocyte chemoattractant protein-1 (MCP-1, CCL2) was found to be responsible for most of the biological effects of TDCF [2]. CCL2 is a CC chemokine that is capable of acting upon macrophages as well as T cells and basophils [3]. Following these initial studies multiple reports documented that additional human and murine tumors constitutively express CCL2 [4], [5], [6]. Yet the role of tumor-derived CCL2 in tumorigenicity and anti-tumor immunity has not been resolved. In fact, the field has been complicated by contrasting results.

Several reports have indicated that tumor-derived CCL2 can enhance progression of breast cancer in humans and increase malignancy of murine mammary tumors [5], [6]. The ability of CCL2 to recruit macrophages may be one mechanism by which CCL2 can mediate some of these effects [7]. In contrast, several studies have reported that introduction of the CCL2 gene into tumor cells correlated with decreased tumorigenicity and facilitated immune mediated tumor rejection [8], [9], [10]. Thus, the precise role of CCL2 in anti-tumor immunity remains unsettled. While many investigators have examined the influence of tumor-derived CCL2 on macrophages, the impact of CCL2 on the T cell response to cancer has for the most part been overlooked. One study that did focus on T cells reported that MCA205-derived CCL2 impaired T cell effector function [11].

In this study we were interested in determining how tumor-derived CCL2 influenced the T cell response to the 4T1 murine mammary carcinoma model. 4T1 is a relatively weakly immunogenic tumor cell line that constitutively expresses CCL2 [12]. Previously we reported that mice bearing the 4T1 tumor possess splenic T cells with altered chemokine receptor function [12]. Here we wanted to know whether inhibiting the tumor cells from producing CCL2 would influence the T cell response and consequently the immunogenicity of the tumor. For this reason we generated cells that lacked detectable levels messenger RNA encoding CCL2 and CCL2 protein. Next, we compared the T cell response elicited by the CCL2 null cells to the response elicited by the cells that produced CCL2. Analysis revealed that blockade of tumor-derived CCL2 could increase T cell effector function, but not the immunogenicity of the tumor.

Section snippets

Mice and tumor cell lines

For these studies 6–8 week old female BALB/c mice were used. The BALB/c mice were purchased from The Jackson Laboratory (Bar Harbor, ME). Food and water were provided ad libitum. All of the tumor cells used in this study (4T1, 4T1-9, 4T1-A4, 4T1-G7, and SM1) were maintained in complete RPMI (cRPMI) (RPMI 1640, BioWhittaker, Walkersvile, MD) supplemented with 10% heat-inactivated fetal bovine serum (Gibco BRL, Gaithersburg, MD), glutamine (2 mM, BioWhittaker), penicillin (100 U/mL, BioWhittaker),

Characterization of CCL2- and CCL2+ tumor cells

In order to investigate the role of tumor-derived CCL2 in the T cell response to cancer we compared CCL2- and CCL2+ tumors. For this purpose we used cells derived from a clone of 4T1. The A4 cells expressed similar levels of mRNA for CCL2 compared to the parental tumor, whereas no message was detected from the CCL2- tumor; G7 (Fig. 1A). Analysis of supernatants, collected from the tumor cells, for CCL2 protein revealed a similar pattern. A4 expressed high levels of CCL2 whereas G7 produced no

Discussion

Due to contrasting results in different animal models the role of CCL2 in anti-tumor immunity has been difficult to unravel. Some investigators have reported that tumor-derived CCL2 decreased tumorigenicity. For instance, Chinese Hamster Ovary (CHO) cells transfected with either the human or murine CCL2 gene lose their ability to form tumors in nude mice [8]. Similarly, a delay in tumor growth occurred when a B16 derived cell line, B78HI, was transfected with the CCL2 gene [16]. However, in

Acknowledgements

This project was funded by the Department of Defense Breast Cancer Research Program DAMD17-01-1-0288.

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Impact of tumor-derived CCL2 on T cell effector function

Abstract

Introduction

Section snippets

Mice and tumor cell lines

Characterization of CCL2- and CCL2+ tumor cells

Discussion

Acknowledgements

Blood

Immunol. Lett.

Cytokine

Cell Immunol.

Blood

Immunity

Regulation of the macrophage content of neoplasms by chemoattractants

Science

A chemoattractant expressed in human sarcoma cells (tumor-derived chemotactic factor, TDCF) is identical to monocyte chemoattractant protein-1/monocyte chemotactic and activating factor (MCP-1/MCAF)

Int. J. Cancer

The detection and localization of monocyte chemoattractant protein-1 (MCP-1) in human ovarian cancer

J. Clin. Invest.

Significance of macrophage chemoattractant protein-1 in macrophage recruitment, angiogenesis, and survival in human breast cancer

Clin. Cancer Res.

Suppression of tumor formation in vivo by expression of JE gene in malignant cells

Mol. Cell Biol.

Expression of the JE/MCP-1 gene suppresses metastatic potential in murine colon carcinoma cells

Cancer Immunol. Immunother.