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The Journal of Immunology, 2000, 164: 4025-4031.
Copyright © 2000 by The American Association of Immunologists

The CC Chemokine CKß-11/MIP-3ß/ELC/Exodus 3 Mediates Tumor Rejection of Murine Breast Cancer Cells Through NK Cells1

Stephen E. Braun2,*,{ddagger},§, Keyue Chen{dagger}, Richard G. Foster*,{ddagger},§, Chang H. Kim*,{ddagger},§, Robert Hromas*,{ddagger},§,{dagger}, Mark H. Kaplan*,{ddagger},§, Hal E. Broxmeyer*,{ddagger},§,{dagger} and Kenneth Cornetta*,{dagger}

Departments of * Microbiology/Immunology and {dagger} Medicine (Hematology/Oncology), and {ddagger} Walther Oncology Center, Indiana University School of Medicine, Indianapolis, IN 46202; and § Walther Cancer Institute, Indianapolis, IN 46208


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
CKß-11 chemoattracts T cells, B cells, dendritic cells, macrophage progenitors, and NK cells and facilitates dendritic cell and T cell interactions in secondary lymphoid tissues. We hypothesized that expression of CKß-11 in tumor cells may generate antitumor immunity through these interactions. After transduction with the retroviral vector L(CKß11)SN, the murine breast cancer cell line C3L5 (C3L5-CKß11) showed expression of retroviral mRNA by Northern analysis and production of functional CKß-11 by chemotaxis of human NK cells to C3L5-CKß11 supernatant. Only 10% of mice injected with C3L5-CKß11 developed tumors, compared with 100% of mice injected with a transduced control C3L5 line (C3L5-G1N). Importantly, the in vitro growth characteristics of the CKß-11-transduced cell line were unaffected, suggesting the difference in growth in vivo was a result of chemokine production. Vaccination with C3L5-CKß11 partially protected animals from parental C3L5 challenge. Immunodepletion with anti-asialo-GM1 or anti-CD4 during C3L5-CKß11 vaccination significantly reduced CKß-11 antitumor activity compared with control and anti-CD8-treated groups. Splenocytes from NK-depleted animals transferred the acquired immunity generated with C3L5-CKß11 vaccination, while splenocytes from the CD4-depleted animals did not. These results indicate, for the first time, that expression of CKß-11 in a breast cancer cell line mediates rejection of the transduced tumor through a mechanism involving NK and CD4+ cells. Furthermore, CKß-11-transduced tumor cells generate long-term antitumor immunity that requires CD4+ cells. These studies demonstrate the potential role of CKß-11 as an adjuvant in stimulating antitumor responses.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Breast cancer is the most common cancer in American women and the second most common cause of cancer death (1). Adjuvant chemotherapy and radiation have improved the survival of women presenting with localized disease, but subsets of women continue to have a poor prognosis (2, 3, 4). Metastatic breast cancer has a uniform poor outcome with a median survival rate of ~2 years (3).

Various cytokines and chemokines are known to have antitumor activity, and cytokine-expressing autologous tumor cell vaccines can enhance antitumor immune responses against malignant cells in vivo. Rejection of tumor cells has been noted in various murine tumor models in which tumor cells have been modified with early acting cytokines (GM-CSF, Flt3L, G-CSF) (5, 6, 7, 8), immunoregulatory cytokines (IL-2, IL-4, IL-7, IL-10, IL-12) (9, 10, 11, 12, 13), inhibitory cytokines (TNF-{alpha}, IFN-{gamma}) (14, 15), chemokines (MIP-1{alpha},3 RANTES, lymphotactin, TCA3, JE/MCP-1/MCAF) (16, 17, 18, 19, 20, 21, 22, 23), and costimulatory molecules (CD40L, B7.1) (24, 25).

Chemokines are a large family of cytokines with four subgroups (CC, CXC, CX3C, and C) based on the N-terminal cysteine motifs (26, 27). The CC chemokine CKß-11/MIP-3ß/ELC/Exodus 3 (CKß-11) chemoattracts T cells (28, 29, 30, 31), B cells (29, 30), dendritic cells (DC) (32), macrophage progenitor cells (33), and NK cells (29), and may be involved in the interactions of DC and T cells in secondary lymphoid tissues (28, 34).

Since CKß-11, expressed in the secondary lymphoid tissues, has the potential to induce specific homing of most lymphoid cells and of DC, we hypothesized that in vivo expression of CKß-11 in the murine breast cancer cell line C3L5 may generate antitumor immunity by facilitating localization of lymphocytes and DC at the site of tumorigenesis. After vaccinating animals with CKß11-transduced C3L5 cells, we found rejection of the CKß-11-transduced tumor was mediated through NK cells and CD4+ cells (including CD4+ NKT cells), while antitumor immunity in a subsequent challenge with parental C3L5 cells required CD4+ cells at the time of C3L5-CKß11 vaccination. These results suggest, for the first time, two levels of antitumor activity for CKß-11; one level is mediated mostly through NK cells, and another is mediated through CD4+ cells.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Construction of the retroviral vector

The cDNA for CKß-11 was generated by RT-PCR of total mRNA from human PBMC. After reverse transcription with oligo(dT) primer and Superscript II (Life Technologies, Rockville, MD), the forward primer (5'-GTT CGG TAC CTG CCT CTG TTC ACC CTC CAT G-3') and the reverse primer (5'-AGT GCT CGA GTT ACT TGT CAT CAT CGT CCT TGT AGT CAC TGC TGC GGC GCT TCA TCT T-3') were used to amplify the complete coding sequence. PCR product was digested with XhoI and KpnI and cloned into the expression vector pREP4 (Invitrogen, Carlsbad, CA). Cloning of CKß-11 cDNA was confirmed by sequencing (35).

pREP4-CKß-11 was transfected into 293-EBNA cells (Invitrogen) by a standard electoporation method, and hygromycin-resistant cells were selected at 400 µg/ml of hygromycin. Supernatants from 293-EBNA cells and 293-EBNA-CKß-11 cells were tested for chemotactic activity for NKL cells, as previously described (29). Three-day culture supernatants from 293-EBNA-CKß-11, but not 293-EBNA cells, showed a chemotactic activity equal to that of 50–100 ng/ml control CKß-11 protein (30).

The retroviral vector L(CKß11)SN (see Fig. 1GoB) was generated by cloning the 350-bp cDNA fragment from EBNA-CKß-11 into the EcoRI and XhoI sites of pLXSN (36) with CKß-11 transcriptionally regulated by the Moloney murine leukemia virus (Mo-MLV) long terminal repeat (LTR). Correct orientation was verified by sequence analysis (Biochemistry Biotechnology Facility, Indiana University School of Medicine, Indianapolis, IN).



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  		FIGURE 1. Schematic diagrams of the retroviral vectors. A, G1Na vector containing the neomycin phosphotransferase (neo) gene transcriptionally regulated by the Mo-MLV LTR. B, L(CKß11)SN vector containing the human CKß11 cDNA transcriptionally regulated by the Mo-MLV LTR and the neo gene regulated by the SV40 early promoter. {Psi}, The RNA packaging signal.

 
Cell culture

C3L5 (37, 38), AM12 (39), and GP+E86 (40) cells were cultured in DMEM (BioWhittaker, Walkersville, MD) supplemented with 10% FBS (HyClone, Logan, UT), 100 U/ml penicillin, and 100 µg/ml streptomycin (BioWhittaker) at 37°C with 5% CO2. The NK cell line NKL was cultured in RPMI 1640 (BioWhittaker) plus 15% FBS, 100 U/ml penicillin, 100 µg/ml streptomycin, 1 mM sodium pyruvate, and 100 U/ml IL-2 (R&D Systems, Minneapolis, MN) at 37°C with 5% CO2.

Gene transfer

The retroviral vector L(CKß11)SN was shuttle packaged through the ecotropic packaging cell line and GP+E86 into the amphotropic packaging cell line AM12. Supernatant (0.45 µm filtered) from these cells plus Polybrene (8 µg/ml; Sigma, St. Louis, MO) was used to transduce the murine breast cancer cell line C3L5 (C3L5-CKß11), as previously described (6, 7), before selection in media containing G418 (400 µg/ml; Life Technologies/BRL, Grand Island, NY). Fig. 1Go represents a schematic of the retro- viral vectors used in these studies. G1N (neo control)-transduced murine breast cancer cells (C3L5-G1N) were described in our previous study (6). The vector G1Na (see Fig. 1GoA) was provided by Genetic Therapy (Gaithersburg, MD).

Chemotaxis assay

Cultures of untransduced, C3L5-G1N and C3L5-CKß-11 cells (1.5 x 106) were plated in 3 ml DMEM without serum or with 10% FCS for 48 or 72 h. Cellular debris was removed from the conditioned media by filtering or centrifugation. NKL cells were washed in PBS and resuspended in DMEM plus 0.5% BSA chemotaxis media. Costar Transwells (6.5 mm diameter, 5 µm pore size, polycarbonate membrane; Cambridge, MA) were used to separate the conditioned media from NKL cells for 3–4 h at 37°C with 5% CO2. The number of cells that transmigrated through the membrane were counted using the FACScan cell analyzer (Becton Dickinson, San Jose, CA), and chemotaxis was calculated as the percentage of the input cells.

In vivo studies

To determine the protection from tumor growth by CKß-11, mice were vaccinated with 1 x 104 transduced cells injected s.c. into the anterior chest wall of female C3H/HeN mice (tumor vaccine). Tumor growth was measured weekly and tumor volume was calculated (vol = width2 x length/2). Animals tumor free at 4 wk were rechallenged with 1 x 104 parental C3L5 cells in the contralateral chest wall (parental tumor challenge). Animal studies were reviewed and approved by the Institutional Review Board.

In adoptive transfer studies, 1 x 107 splenocytes from animals tumor free after 4 wk were harvested and injected i.v. into previously untreated (naive) mice that simultaneously received 1 x 104 untransduced C3L5 cells in the anterior chest wall.

To understand the mechanism and cellular mediators of antitumor activity, mice were injected with PBS, control rabbit serum (100 µl i.p. per mouse on days -3, 0, and +3) (Sigma), rabbit anti-asialo-GM1 to immunodeplete NK cells (20 µl i.p. per mouse every 4 days during the primary tumor challenge or on days -3, 0, and +3) (Wako Pure Chemical Industries, Richmond, VA), anti-CD4 (clone GK1.5; PharMingen, San Diego, CA), and anti-CD8 (clone 53-6.7; PharMingen) Abs (0.1 mg per mouse on days -3, 0, and +3).

Statistical analysis

Differences in tumor growth were evaluated over 4 wk by the number of animals with tumor and by the tumor volume in these animals. Tumor growth was determined using the Mann-Whitney statistic for nonparametric analysis and the one-tailed critical values of Us, as described (41). Differences in chemotactic responses and splenocyte percentages were evaluated by Student’s t test.

Flow-cytometric analysis

Single-cell suspensions of splenocytes were resuspended in PBS with 0.5% BSA and dual stained by incubation with FITC-conjugated anti-CD3{epsilon} (clone 145-2c11; PharMingen) or the appropriate isotype control (PharMingen), and with PE-conjugated anti-CD4 (clone RM4-5; PharMingen), PE-conjugated anti-CD8 (clone 53-6.7; PharMingen), PE-conjugated antipan NK cells (clone DX5; PharMingen), or the appropriate isotype control (PharMingen) for 1 h at 4°C. C3L5 cells were stained with FITC-conjugated anti-CD1{delta} (clone 1B1; PharMingen) and the appropriate isotype control (PharMingen). The cells were washed and analyzed for the percentage of cells in each population by flow cytometry using the FACScan cell analyzer (Becton Dickinson).


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Transduction and expression of the chemokine CKß-11 in the murine breast cancer cell line C3L5

To assess the effectiveness of CKß-11 as a tumor vaccine, we generated the full-length cDNA by RT-PCR and constructed the retroviral vector L(CKß11)SN with CKß-11 transcriptionally regulated by the Mo-MLV LTR and the neo gene transcriptionally regulated by the SV40 early promoter (Fig. 1GoB). After shuttle packaging this retroviral vector was used to transduced the murine breast cancer cell line C3L5 before selection in medium containing G418 (C3L5-CKß11). The control vector G1Na, containing the neo gene transcriptionally regulated by the Mo-MLV LTR (Fig. 1GoA), was also used to generate a control cell line (C3L5-G1N), as previously described (6, 7).

To measure the level of mRNA expression from the integrated provirus, Northern analysis was performed on total cellular RNA isolated from C3L5, C3L5-G1N, and C3L5-CKß11 cell populations (Fig. 2Go). Because the vectors contain the neomycin phosphotransferase gene as a common sequence, a neo-specific probe was used for hybridization. The parental C3L5 cells do not hybridize to the neo probe (Fig. 2Go, lane 1). In contrast, the C3L5-G1N- and C3L5-CKß11-transduced populations hybridized to the neo-specific probe with higher levels of neo expression in C3L5-G1N than in C3L5-CKß11 (Fig. 2Go, lanes 2 and 3). With the C3L5-CKß11-transduced cells, two transcript lengths are evident (Fig. 2Go, lane 3), consistent with expression from the LTR and the SV40 promoters. This molecular analysis demonstrates expression of the retroviral messenger RNA from the integrated provirus in transduced cells.



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  		FIGURE 2. Molecular analysis of retroviral mRNA expression. Total RNA from C3L5, C3L5-G1N, and C3L5-CKß11 cells was isolated, separated by gel electrophoreses, and blotted to nylon membranes. Membranes were probed with a radiolabeled neomycin phosphotransferase (Neo)-specific probe. After washing, the membrane was exposed to film for 6 h. Bottom frame, Ethidium bromide (EtBr) staining of RNA gel shows equal loading of samples. Top frame, Neo-specific hybridization to C3L5-G1N and C3L5-CKß11. The arrows show multiple transcript lengths in C3L5-CKß11 cells resulting from the two promoters.

 
To demonstrate production of the chemokine CKß-11, we measured the functional activity of CKß-11 in conditioned media, as Abs against murine CKß-11 are not commercially available. Supernatant from parental C3L5 and C3L5-CKß11-transduced populations and a soluble rCKß-11 positive control (100 ng/ml) were studied for chemoattractant potential in chemotaxis assay with the human NK cell line NKL. As shown in Table IGo, a previously determined optimal concentration of CKß-11 (100 ng/ml) attracted 42 ± 11% of input NKL cells through the membrane (mean for three separate experiments). From previous studies, the maximum sensitivity of this assay is ~1 ng/ml and the responsiveness of NKL cells to CKß-11 is bell shaped, with concentrations higher than 100 ng/ml desensitizing chemotaxis (29). Supernatant from the C3L5-CKß11 cells attracted 24 ± 3.4% of the NKL cells through the membrane, while only 5 ± 2.2% of cells transversed the membrane when supernatant from the parental C3L5 cells was tested (Table IGo). Although we cannot determine the precise concentration, from these results, we concluded that C3L5-CKß11 supernatants contain between 1 and 10 ng/ml of CKß-11 (0.5–10 ng/106 cells/24 h) and that these cells produce biologically relevant levels of CKß-11 protein.


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  		Table I. Chemotaxis of NKL cells with conditioned medium from transduced cells or recombinant CKß-111

 
To determine the effect of CKß-11 expression on the growth of C3L5 cells, the growth rate of transduced cells was studied. As shown in Fig. 3Go, expression of the chemokines in C3L5 cells did not significantly (p > 0.05) alter the in vitro growth rate of these cells compared with parental C3L5 and with G1N-transduced C3L5 cells.



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  		FIGURE 3. In vitro growth curve of C3L5, C3L5-G1N, and C3L5-CKß11 cells. Cells were plated at 1 x 104 cells/well and cultured for 6 days. Viable cell number was determined using a hemocytometer every 2 days. Results are given as means ± SD from triplicate cultures.

 
Tumor vaccine and parental tumor challenge

To determine whether chemokine expression prevented tumor formation, female C3H/HeN mice were vaccinated with 1 x 104 C3L5-G1N or C3L5-CKß11 cells s.c. in the anterior chest wall (tumor vaccine). Animals were followed for 4 wk, and tumor volume and the percentage of tumor-free animals (TFA) were determined. After inoculation with the tumor vaccine (Fig. 4Go), C3L5-CKß11-vaccinated mice (90% TFA, vol = 1.9 cm3) had significantly decreased tumor formation (p < 0.05) when compared with control C3L5-G1N-vaccinated animals (0% TFA, vol = 1.6 ± 0.29 cm3).



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  		FIGURE 4. In vivo tumor size of C3L5-G1N and C3L5-CKß11 vaccine. From the results of two independent experiments, the percentage of animals that developed tumor 4 wk after vaccination (1 x 104 transduced cells per animal) or after parental C3L5 challenge (1 x 104 cells per animal) is represented under the figure. The animals without tumor ({circ}) are represented below the ordinate, and the animals with tumor (•) are represented by the tumor volume.

 
To determine whether C3L5-CKß11-vaccinated animals had generated tumor immunity, TFA at 4 wk were rechallenged with 1 x 104 parental C3L5 cells in the contralateral chest wall (parental tumor challenge). During this secondary challenge with parental C3L5 cells (Fig. 4Go), vaccination with C3L5-CKß11 significantly reduced (p < 0.05) the tumor growth, but did not prevent tumor formation (11% TFA, vol = 0.87 ± 0.45 cm3). These results indicate that CKß-11-transduced cells can mediate tumor rejection and generate a low level of antitumor immunity.

Immunodepletion of lymphoid subsets with Ab injections

Several mechanisms may be involved in generating antitumor activity by CKß-11-transduced C3L5 cells. Because CKß-11 is known to chemoattract effector cells, such as naive and memory T cells and NK cells, we studied the role of CD4+, CD8+, and NK cells in CKß-11 antitumor activity by immunodepleting these cells in vivo with Abs. As shown in Fig. 5Go, immunodepletion of NK cells with anti-asialo-GM1 and of CD4+ cells with anti-CD4 during the C3L5-CKß11 vaccination period, resulted in reduction of CKß-11 antitumor activity (p < 0.05) (0% TFA, vol = 1.3 ± 0.25 cm3; 40% TFA, vol = 1.3 ± 0.25 cm3, respectively) compared with vaccinated animals treated with control serum (100% TFA) and of CD8+ cells immunodepleted with anti-CD8 (100% TFA).



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  		FIGURE 5. In vivo tumor size of C3L5 and C3L5-CKß11 vaccine. From the results of two independent experiments, the percentage of animals that developed tumor 4 wk after vaccination (1 x 104 transduced cells per animal) and after immunodepletion of specific lymphocyte subsets with Abs (anti-asialo-GM1 for NK cells, anti-CD4 for Th cells, anti-CD8 for CTL) is represented under the figure. The animals without tumor ({circ}) are represented below the ordinate, and the animals with tumor (•) are represented by the tumor volume.

 
To confirm the immunodepletion of each cellular subset by in vivo Ab treatments, splenocytes were isolated 1 or 4 days after the final Ab treatment and stained for CD3+, CD4+, CD8+, and DX5+ cells (Table IIGo). The CD3+ T cells are divided into CD4+ and CD8+ fractions. NK cells are divided into CD3- NK cells and the recently described CD3+ NKT subset, which also coexpress CD4+ (42). As shown in Table IIGo, normal spleen cells are 19% DX5+ cells and 43–45% CD3+ T cells. This CD3+ population contains 31 ± 3.7% CD4+ and 15 ± 2.4% CD8+ T cells.


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  		Table II. Analysis of splenic CD3, CD4, CD8, and NK subsets after immunodepletion with specific Abs1

 
After immunodepletion of the NK cells with anti-asialo-GM1, the percentage of pan CD3- NK splenocytes (3.6%) was significantly reduced (72%; p < 0.05) compared with control animals receiving no treatment (PBS) or rabbit serum (13 ± 3.7%). After immunodepletion of the CD8+ cells with anti-CD8, the percentage of CD3+ CD8+ splenocytes (1.4%) was significantly reduced (90%; p < 0.05) compared with the normal controls (15 ± 2.4%). Additionally, the percentage of CD3+CD4- splenocytes (3.6%) was also significantly reduced (71%; p < 0.05) compared with the normal controls (12 ± 1.5%), which independently confirms the reduction of CD8+ T cells. After immunodepletion of the CD4+ cells with anti-CD4, the percentage of CD3+CD4+ splenocytes (3.2%) was significantly reduced (90%; p < 0.05) compared with the normal controls (31 ± 3.7%). Additionally, the CD3+CD8- splenocytes (5.8%) were significantly decreased (81%; p < 0.05), and the CD3+CD4- splenocytes (21%) were significantly increased (69%; p < 0.05) compared with normal controls (30 ± 3.1%; 12 ± 1.5%, respectively), again independently confirming the immunodepletion of CD4+ cells and the compensatory increase in CD3+CD8+ splenocytes. Interestingly, because the NKT cells are CD3+CD4+ NK cells, during immunodepletion with anti-CD4, the percentage of DX5+ CD3+ splenocytes (2.5%) was also significantly reduced (58%; p < 0.05) compared with normal controls (6 ± 1.3%). These results indicate that the target subpopulations were substantially reduced by the Ab treatments.

Transfer of splenocytes from vaccinated and immunodepleted animals

To determine the role of the various subpopulations in the generation of acquired immunity, splenocytes from animals immunodepleted of lymphocyte subsets during C3L5-CKß11 vaccination were transferred to naive animals during challenge with parental C3L5 cells. Transfer of splenocytes from C3L5-CKß11-vaccinated animals significantly reduced (p < 0.05) the tumor volume of the parental tumor challenge compared with PBS-treated or naive splenocyte-treated animals (Table IIIGo). Treatment with anti-asialo-GM1 and anti-CD8 did not inhibit the acquired immunity generated during the C3L5-CKß11 vaccination, as the adoptive transfer of splenocytes from these animals significantly reduced (p < 0.05) tumor volume compared with control animals (Table IIIGo). In contrast, adoptive transfer of splenocytes from the anti-CD4-depleted animals lost this acquired immunity, as tumor volume was not significantly different (p > 0.05) than controls (Table IIIGo). Although adoptive immunity was apparent with the transfer of splenocytes, no in vitro antitumor activity was detectable (data not shown). These results indicate that C3L5-CKß11 vaccination generates some level of transferable immunity and that this response is not generated through NK+ or CD8+ cells, but through CD4+ cells.


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  		Table III. Protection of animals with splenocytes transferred from C3L5-CKß11-vaccinated animals1

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Chemokines were originally identified and described by their involvement in the inflammatory response and their capacity to chemoattract leukocytes. However, it has recently been observed that some chemokines influence the trafficking of lymphocytes to primary (thymus), secondary (lymph nodes), and tertiary (periphery) lymphoid tissues (34). CKß-11, a ligand for CCR7 (35), is expressed at high level in the thymus and in the lymph nodes, constitutively in DC within the T cell zone of the lymph node, and is also known to strongly chemoattract CD8+ T cells, naive CD4+ T cells, mature DC, and NK cells. CCR7 was found recently to coordinate the primary immune response in receptor knockout mice, as naive T cells and activated skin DC are unable to migrate into the lymph nodes (43). We hypothesized that expression of CKß-11 in tumor cells would facilitate the formation of antitumor immunity, an effect possibly mediated by NK cells and lymphocytes.

These results indicate, for the first time, that expression of CKß-11 in a tumor model inhibited formation of the transduced tumor (even though in vitro growth of the transduced tumor cell was not effected) and that CKß-11-transduced tumor vaccine generated antitumor immunity in vivo. These results also suggest that NK cells and CD4+ cells are involved in the antitumor activity elicited by CKß-11 and that CD4+ cells are necessary for establishing antitumor immunity.

Many immunoregulatory molecules have been shown to mediate antitumor activity in a variety of tumor models. The murine breast cancer cell line C3L5 used in these studies is poorly or nonimmunogenic, as 106 irradiated C3L5 cells offer no protection against future tumor challenges (data not shown). We have previously shown that both Flt3L- and GM-CSF-transduced C3L5 cells generate antitumor immunity (6, 7). Flt3L and GM-CSF are both known to stimulate proliferation and differentiation of DC (44, 45, 46), although the immunity generated with Flt3L-transduced tumor vaccine was stronger than GM-CSF- and CKß-11-transduced tumor vaccines ( (7) and these data). Flt3L is also known to stimulate proliferation of NK cells in vivo (47, 48), and we showed that NK cells are involved in mediating antitumor activity of Flt3L-transduced tumor cells (7). CKß-11 does not activate NK cells (data not shown), but does chemoattract both DC and NK cells (29, 32).

Several groups have shown antitumor activity with a variety of chemokines. With nine known CC chemokine receptors and five known CXC chemokine receptors, expression of receptors in the lymphoid cells varies widely. Additionally, the specificity of chemokine receptors is promiscuous, allowing for redundant activity of several chemokines on leukocytes that express the receptor. Therefore, the type of infiltrating leukocytes is determined by the expression profile of chemokines (34).

Studies have shown antitumor activity for MIP-1{alpha}, RANTES, lymphotactin, and TCA3 (16, 17, 18, 19). MIP-1{alpha} expression in adenocarcinoma cells, but not IL-8 expression, led to reduced tumor formation and increased infiltration of macrophages and neutrophils (18). After subsequent challenges with MIP-1{alpha}-expressing and parental tumor cells, protective immunity was also observed (18). With TCA3 expression, tumor formation was inhibited and tumor-specific immunity was generated; however, histological analysis revealed mostly neutrophils around the TCA3-expressing tumor (17). RANTES also inhibited tumor formation and generated tumor immunity (16). Additionally, immunodepleting CD8+ T cells, blocking macrophage migration with Abs to adhesion receptors, and, to a lesser extent, immunodepleting CD4+ T cells all restored RANTES-transduced tumor formation (16). Lymphotactin, by itself, reduced tumor growth; however, in combination with IL-2, it enhanced antitumor immunity and inhibited an established tumor (19). These interactions were mediated through CD4+ and CD8+ T cells (19).

JE/MCP-1/MCAF (MCP-1) was shown to reduce in vivo growth of tumor cells and increase infiltration of macrophages/monocytes to the tumor site (20, 21, 22, 23). Localization of fibroblasts engineered to express MCP-1 recruits migration of macrophages to the lungs and suppresses lung metastasis of renal adenocarcinoma (21). This may occur because MCP-1 and LPS are synergistic in activating cytotoxicity of macrophages against tumor cells (49). Additionally, the combined expression of MCP-1 and the addition of Abs against P-glycoprotein inhibited tumor formation of multi-drug-resistant lung cancer cells in vivo (22).

In the normal host immune response, DC acquire Ag and present them to CD4+ and CD8+ T cells through MHC class I and II molecules. NK cells will kill a target cell unless a signal from the target cell MHC class I is received through the NK cell inhibitory receptor Ly-49A (50). Expression of cytokines or chemokines in the tumor cells stimulates the normal immune response to mediate the antitumor response. Host immune cells may infiltrate the tumor site, proliferate, differentiate, secrete other secondary cytokines or chemokines, recruit other effector cells, or become activated, eventually targeting tumor cells for destruction.

Because DC are chemoattracted by CKß-11, these cells may be involved in presenting tumor-specific Ag to the CD4+ cells that are mediating the antitumor immunity. Studies have demonstrated that Ag-activated CD4+ NKT cells express CD40 ligand (CD154) and engage the CD40 receptor on APC, which then produce IL-12 (51) for the activation of NKT cells (52). The transduced tumor cells are probably not presenting the tumor-specific Ag to the NKT cells, as we found that C3L5-CKß11 cells had equivalent levels of CD1{delta} expression as C3L5-G1N cells (data not shown). As noted previously, IFN-{gamma} expression in C3L5, although it raises the level of MHC class I and II expression, does not mediate antitumor responses as strong as that of Flt3L expression (7). Although Flt3L is known to stimulate DC and NK cells, we found a predominate role for NK cells in antitumor activity in this tumor model.

It has been suggested that effector cells in innate immunity play a pivotal role in shaping initial T cell activation (51), possibly by conditioning DCs for subsequent immune responses (52). NKT cells are CD3+ T cells that also express NK cell markers and usually the invariant V{alpha}14-J{alpha}281 chain of the TCR (53) to interact with CD1{delta} (MHC-like) molecules on APC (54). NKT cells are important in IL-12-mediated antitumor activities, as mice with a deletion of CD1 gene or the J{alpha}281 gene segment, and the subsequent loss of NKT cells could no longer mediate the IL-12-induced rejection of tumors (13). These studies have not separated the effects of the NK+ CD3- cells or NK+ CD3+ NKT cells from the CD4+CD3+ T cells in generating antitumor activity or priming immunity against the transduced tumor cells.

Our results with CKß-11 suggest two levels of responsiveness: antitumor activity against the transduced cells mediated mostly by NK+ cells, and antitumor immunity mediated through CD4+ cells. More recently, several groups showed that MCP-1, MIP-1{alpha}, and RANTES chemoattract immature DC migration (32, 55), and that MCP-1 also activate CTL and NK cytolytic responses (56). Our results with CKß-11 and Flt3L support the notion that NK cells activate adaptive immunity and may suggest a common mechanism for antitumor activity with other stimulatory molecules. Whether these same interactions are responsible for the increased antitumor response we observed with Flt3L compared with GM-CSF or CKß-11 needs further studies.

Because chemokines have important functions in host defenses, they may be used to modulate other activities, such as antitumor immunotherapy. Local expression of chemokines by transduced tumor cells may augment other therapeutic modalities, particularly those that stimulate DC, NK cells, or T cells, such as GM-CSF, Flt3L, IL-2, IFN-{gamma}, or IL-12. Future studies are necessary to support its role in clinical immunotherapeutic trials.


   Acknowledgments
 
We thank Dr. Randy Brutkiewicz for anti-CD1{delta} and isotype control Abs used in flow-cytometric analysis, and Dr. Young-June Kim for help with in vitro cytotoxic assays.


   Footnotes
 
1 These studies were supported by grants from Catherine Peachy Foundation (K.C.), the Centers for Excellence in Molecular Hematology (National Institute of Diabetes and Digestive and Kidney Diseases, Grant P50DK 49218), National Institutes of Health Grants RO1 DK 53674 and HL 56416, and a project in PO1 HL 53386 (to H.E.B.), National Institute of Allergy and Infectious Diseases Grant RO1 AI 45515 (to M.H.K.), and National Gene Vector Laboratory Grant NCRR V42 RR11148 (to K.C.). M.H.K. is a Special Fellow and C.H.K. is a post doctoral fellow of the Leukemia Society of America. Back

2 Address correspondence and reprint requests to Dr. Stephen E. Braun at his current address: Division of Immunology, Harvard Medical School, One Pine Hill Drive, Southborough, MA 01772. Back

3 Abbreviations used in this paper: MIP, macrophage-inhibitory protein; DC, dendritic cell; Flt3L, Flt3/Flk2 ligand; LTR, long terminal repeat; MCP, monocyte chemoattractant protein; Mo-MLV, Moloney murine leukemia virus; TCA3, T cell activation-specific gene 3; TFA, tumor-free animals. Back

Received for publication November 12, 1999. Accepted for publication February 3, 2000.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Wingo, P., T. Tong, S. Bolden. 1995. Cancer statistics. Cancer J. Clin. 45:8.[Abstract/Free Full Text]
  2. Fisher, B., C. Redmond, R. Poisson, R. Margolese, N. Wolmark, L. Wickerham, E. Fisher, M. Deutsch, R. Caplan, Y. Pilch, et al 1989. Five year results of a randomized clinical trial comparing total mastectomy and segmental mastectomy and lymphectomy with or without irradiation in the treatment of breast cancer. N. Engl. J. Med. 320:822.[Abstract]
  3. Gradishar, W. J., M. S. Tallman, J. S. Abrams. 1996. High dose chemotherapy for breast cancer. Ann. Intern. Med. 125:599.[Abstract/Free Full Text]
  4. Wood, W. C., D. R. Budman, A. H. Korzun, M. R. Cooper, J. Younger, R. D. Hart, A. Moore, J. A. Ellerton, L. Norton, C. R. Ferree, et al 1994. Dose and dose intensity of adjuvant chemotherapy for stage II, node-positive breast cancer. N. Engl. J. Med. 330:1253.[Abstract/Free Full Text]
  5. Dranoff, G., E. Jaffee, A. Lazenby, P. Golumbek, H. Levitsky, K. Brose, V. Jackson, H. Hamada, D. Pardoll, R. C. Mulligan. 1993. Vaccination with irradiated tumor cells engineered to secrete murine granulocyte-macrophage colony-stimulating factor stimulates potent, specific, and long-lasting anti-tumor immunity. Proc. Natl. Acad. Sci. USA 90:3539.[Abstract/Free Full Text]
  6. Chen, K., S. Braun, S. Lyman, Y. Fan, C. M. Traycoff, E. A. Wiebke, J. Gaddy, G. Sledge, H. E. Broxmeyer, K. Cornetta. 1997. Antitumor activity and immunotherapeutic properties of Flt3-ligand in a murine breast cancer model. Cancer Res. 57:3511.[Abstract/Free Full Text]
  7. Braun, S. E., K. Chen, B. R. Blazar, P. J. Orchard, G. Sledge, M. J. Robertson, H. E. Broxmeyer, K. Cornetta. 1999. Flt3 ligand antitumor activity in a murine breast cancer model: a comparison with granulocyte-macrophage colony-simulating factor and a potential mechanism of action. Hum. Gene Ther. 10:2141.[Medline]
  8. Colombo, M. P., G. Ferrari, A. Stoppacciaro, M. Parenza, M. Rodolfo, F. Mavilio, G. Parmiani. 1991. Granulocyte colony-stimulating factor gene transfer suppresses tumorigenicity of a murine adenocarcinoma in vivo. J. Exp. Med. 173:889.[Abstract/Free Full Text]
  9. Mackensen, A., A. Lindemann, R. Mertelsmann. 1997. Immunostimulatory cytokines in somatic cells and gene therapy of cancer. Cytokine Growth Factor Rev. 8:119.[Medline]
  10. Fearon, E. R., D. M. Pardoll, T. Itaya, P. Golumbek, H. I. Levitsky, J. W. Simons, H. Karasuyama, B. Vogelstein, P. Frost. 1990. Interleukin-2 production by tumor cells bypasses T helper function in the generation of an antitumor response. Cell 60:397.[Medline]
  11. Gansbacher, B., K. Zier, B. Daniels, K. Cronin, R. Bannerji, E. Gilboa. 1990. Interleukin-2 gene transfer into tumor cells abrogates tumorigenicity and induces protective immunity. J. Exp. Med. 172:1217.[Abstract/Free Full Text]
  12. Golumbek, P. T., A. J. Lazenby, H. I. Levitsky, L. M. Jaffee, H. Karasuyama, M. Baker, D. M. Pardoll. 1991. Treatment of established renal cancer by tumor cells engineered to secrete interleukin-4. Science 254:713.[Abstract/Free Full Text]
  13. Cui, J., T. Shin, T. Kawano, H. Sato, E. Kondo, I. Toura, Y. Kaneko, H. Koseki, M. Kanno, M. Taniguchi. 1997. Requirement for V{alpha}14 NKT cells in IL-12-mediated rejection of tumors. Science 278:1623.[Abstract/Free Full Text]
  14. Asher, A. L., J. J. Mule, A. Kasid, N. P. Restifo, J. C. Salo, C. M. Reichert, G. Jaffe, B. Fendly, M. Kriegler, S. A. Rosenberg. 1991. Murine tumor cells transduced with the gene for tumor necrosis factor-{alpha}: evidence for paracrine immune effects of tumor necrosis factor against tumors. J. Immunol. 146:3327.
  15. Rosenthal, F. M., K. Cronin, R. Bannerji, D. W. Golde, B. Gansbacher. 1994. Augmentation of antitumor immunity by tumor cells transduced with a retroviral vector carrying the interleukin-2 and interferon-{gamma} cDNAs. Blood 83:1289.[Abstract/Free Full Text]
  16. Mule, J. J., M. Custer, B. Averbook, J. C. Yang, J. S. Weber, D. V. Goeddel, S. A. Rosenberg, T. J. Schall. 1996. RANTES secretion by gene-modified tumor cells results in loss of tumorigenicity in vivo: role of immune cell subpopulations. Hum. Gene Ther. 7:1545.[Medline]
  17. Laning, J., H. Kawasaki, E. Tanaka, Y. Luo, M. E. Dorf. 1994. Inhibition of in vivo tumor growth by the ß chemokine, TCA3. J. Immunol. 153:4625.[Abstract]
  18. Nakashima, E., A. Oya, Y. Kubota, N. Kanada, R. Matsushita, K. Takeda, F. Ichimura, K. Kuno, N. Mukaida, K. Hirose, et al 1996. A candidate for cancer gene therapy: MIP-1{alpha} gene transfer to an adenocarcinoma cell line reduced tumorigenicity and induced protective immunity in immunocompetent mice. Pharm. Res. 13:1896.[Medline]
  19. Dilloo, D., K. Bacon, W. Holden, W. Zhong, S. Burdach, A. Zlotnick, M. Brenner. 1996. Combined chemokine and cytokine gene transfer enhances antitumor immunity. Nat. Med. 2:1090.[Medline]
  20. Rollins, B. J., M. E. Sunday. 1991. Suppression of tumor formation in vivo by expression of the JE gene in malignant cells. Mol. Cell. Biol. 11:3125.[Abstract/Free Full Text]
  21. Huang, S., K. Xie, R. K. Singh, M. Gutman, M. Bar-Eli. 1995. Suppression of tumor growth and metastasis of murine renal adenocarcinoma by syngeneic fibroblasts genetically engineered to secrete the JE/MCP-1 cytokine. J. Interferon Cytokine Res. 15:655.[Medline]
  22. Nishioka, Y., S. Yano, F. Fujiki, N. Nukaida, K. Matsushima, T. Tsuruo, S. Sone. 1997. Combined therapy of multidrug-resistant human lung cancer with anti-P-glycoprotein antibody and monocyte chemoattractant protein-1 gene transduction: the possibility of immunological overcoming of multidrug resistance. Int. J. Cancer 71:179.
  23. Bottazzi, B., S. Walter, D. Govoni, F. Colotta, A. Mantovani. 1992. Monocyte chemotactic cytokine gene transfer modulates macrophage infiltration, growth, and susceptibility to IL-2 therapy of a murine melanoma. J. Immunol. 148:1280.[Abstract]
  24. Nakajima, A., T. Kodama, S. Morimoto, M. Azuma, K. Takeda, H. Oshima, S. Yoshino, H. Yagita, K. Okumura. 1998. Antitumor effect of CD40 ligand: elicitation of local and systemic antitumor responses by IL-12 and B7. J. Immunol. 161:1901.[Abstract/Free Full Text]
  25. Baskar, S., L. Glimcher, N. Nabavi, R. T. Jones, S. Ostrans-Rosenberg. 1995. Major histocompatibility complex class II+ B7-1+ tumor cells are potent vaccines for stimulating tumor rejection in tumor-bearing mice. J. Exp. Med. 181:619.[Abstract/Free Full Text]
  26. Broxmeyer, H. E., C. H. Kim. 1999. Regulation of hematopoiesis in a sea of chemokine family members with a plethora of redundant activities. Exp. Hematol. 27:1113.[Medline]
  27. Broxmeyer, H. E., and C. H. Kim. 1999. Chemokines and hematopoiesis. In Chemokines and Cancer. R. B. J. Humana Press, Totowa, pp. 263–291.
  28. Ngo, V. N., H. L. Tang, J. G. Cyster. 1998. Epstein-Barr virus-induced molecule 1 ligand chemokine is expressed by dendritic cells in lymphoid tissues and strongly attracts T cells and activated B cells. J. Exp. Med. 188:181.[Abstract/Free Full Text]
  29. Kim, C. H., L. M. Pelus, E. Appelbaum, K. Johanson, N. Anzai, H. E. Broxmeyer. 1999. CCR7 ligands, SLC/6Ckine/Exodus2/TCA4 and CKß-11/MIP-3ß/ELC, are chemoattractants for CD56+CD16- NK cells and late stage lymphoid progenitors. Cell. Immunol. 193:226.[Medline]
  30. Kim, C. H., L. M. Pelus, J. R. White, E. Applebaum, K. Johanson, H. E. Broxmeyer. 1998. CK ß-11/macrophage inflammatory protein-3 ß/EBI1-ligand chemokine is an efficacious chemoattractant for T and B cells. J. Immunol. 160:2418.[Abstract/Free Full Text]
  31. Kim, C. H., L. M. Pelus, J. R. White, H. E. Broxmeyer. 1998. Differential chemotactic behavior of developing T cells in response to thymic chemokines. Blood 91:4434.[Abstract/Free Full Text]
  32. Dieu, M.-C., B. Vanbervliet, A. Vicari, J.-M. Bridon, E. Oldham, S. Ait-Yahia, F. Briere, A. Zlotnik, S. Lebecque, C. Caux. 1998. Selective recruitment of immature and mature dendritic cells by distinct chemokines expressed in different anatomic sites. J. Exp. Med. 188:373.[Abstract/Free Full Text]
  33. Kim, C. H., L. M. Pelus, J. R. White, H. E. Broxmeyer. 1998. Macrophage-inflammatory protein-3 ß/EBI1-ligand chemokine/CK ß-11, a CC chemokine, is a chemoattractant with a specificity for macrophage progenitors among myeloid progenitor cells. J. Immunol. 161:2580.[Abstract/Free Full Text]
  34. Kim, C. H., H. E. Broxmeyer. 1999. Chemokines: signal lamps for trafficking of T and B cells for development and effector function. J. Leukocyte Biol. 65:6.[Abstract]
  35. Yoshida, R., T. Imai, K. Hieshima, J. Kusuda, M. Baba, M. Kitaura, M. Nishimura, M. Kakizaki, H. Nomiyama, O. Yoshie. 1997. Molecular cloning of a novel human CC chemokine EBI1-ligand chemokine that is a specific functional ligand for EBI1, CCR7. J. Biol. Chem. 272:13803.[Abstract/Free Full Text]
  36. Miller, A. D., G. J. Rosman. 1989. Improved retroviral vectors for gene transfer and expression. BioTechniques 7:980.[Medline]
  37. Khoo, N., F. Chan, M. Saarloos, P. Lala. 1992. Immunotherapy of mammary adenocarcinoma metastases in C3H/HeN mice with chronic administration of cyclooxygenase inhibitors alone or in combination with IL-2. Clin. Exp. Metastasis 10:239.[Medline]
  38. Saarloos, M., N. Khoo, P. Lala. 1992. Effects of cancer immunotherapy with indomethacin and interleukin-2 on murine hematopoietic stem cells. Cancer Res. 52:6452.[Abstract/Free Full Text]
  39. Markowitz, D., S. Goff, A. Bank. 1988. Construction and use of a safe and efficient amphotropic packaging cell line. Virology 167:400.[Medline]
  40. Markowitz, D., S. Goff, A. Bank. 1988. A safe packaging line for gene transfer: separating viral genes on two different plasmids. J. Virol. 62:1120.[Abstract/Free Full Text]
  41. Sokal, R. R., F. J. Rohlf. 1981. Biometry: The Principles and Practice of Statistics in Biological Research 2nd Ed. W. H. Freeman and Company, San Francisco.
  42. Yoshimoto, T., W. E. Paul. 1994. CD4+, NK1.1+ T cells promptly produce interleukin 4 in response to in vivo challenge with anti-CD3. J. Exp. Med. 179:1285.[Abstract/Free Full Text]
  43. Forster, R., A. Schubel, D. Breitfeld, E. Kremmer, I. Renner-Muller, E. Wolf, M. Lipp. 1999. CCR7 coordinates the primary immune response by establishing functional microenvironments in secondary lymphoid organs. Cell 99:23.[Medline]
  44. Shah, A. J., E. M. Smogorzewska, C. Hannum, G. M. Crooks. 1996. Flt3 ligand induces proliferation of quiescent human bone marrow CD34+ CD38- cells and maintains progenitor cells in vitro. Blood 87:3563.[Abstract/Free Full Text]
  45. Maraskovsky, E., K. Brasel, M. Teepe, E. R. Roux, S. D. Lyman, K. Shortman, H. J. McKenna. 1996. Dramatic increase in the numbers of functionally mature dendritic cells in Flt3 ligand-treated mice: multiple dendritic cell subpopulations identified. J. Exp. Med. 184:1953.[Abstract/Free Full Text]
  46. Drakes, M. L., L. Lu, V. M. Subbotin, A. W. Thomson. 1997. In vivo administration of flt3 ligand markedly stimulates generation of dendritic cell progenitors from mouse liver. J. Immunol. 159:4268.[Abstract]
  47. Peron, J.-M., C. Esche, V. M. Subbotin, C. Maliszewski, M. T. Lotze, M. R. Shurin. 1998. FLT3- ligand administration inhibits liver metastases: role of NK cells. J. Immunol. 161:6164.[Abstract/Free Full Text]
  48. Shaw, S. G., A. A. Maung, R. J. Steptoe, A. W. Thomson, N. L. Vujanovic. 1998. Expansion of functional NK cells in multiple tissue compartments of mice treated with Flt3-ligand: implications for anti-cancer and anti-viral therapy. J. Immunol. 161:2817.[Abstract/Free Full Text]
  49. Singh, R. K., K. Berry, K. Matsushima, K. Yasumoto, I. J. Fidler. 1993. Synergism between human monocyte chemotactic and activating factor and bacterial products for activation of tumoricidal properties in murine macrophages. J. Immunol. 151:2786.[Abstract]
  50. William, N. S., T. A. Moore, J. D. Schatzle, I. J. Puzanov, P. V. Sivakumar, A. Zlotnik, M. Bennett, V. Kumar. 1997. Generation of lytic natural killer 1.1+ Ly-49- cells from multipotential murine bone marrow progenitors in stroma-free culture: definition f cytokine requirements and developmental intermediates. J. Exp. Med. 186:1609.[Abstract/Free Full Text]
  51. Tomura, M., W. G. Yu, H. J. Ahn, M. Yamashita, Y. F. Yang, S. Ono, T. Hamaoka, T. Kawano, M. Taniguchi, Y. Koezuka, H. Fujiwara. 1999. A novel function of V{alpha}14 + CD4 + NKT cells: stimulation of IL-12 production by antigen-presenting cells in the innate immune system. J. Immunol. 163:93.[Abstract/Free Full Text]
  52. Kitamura, H., K. Iwakabe, T. Yahata, S. Nishimura, A. Ohta, Y. Ohmi, M. Sato, K. Takeda, K. Okumura, L. Van Kaer, et al 1999. The natural killer T (NKT) cell ligand {alpha}-galactosylceramide demonstrates its immunopotentiating effect by inducing interleukin (IL)-12 production by dendritic cells and IL-12 receptor expression on NKT cells. J. Exp. Med. 189:1121.[Abstract/Free Full Text]
  53. Lantz, O., A. Bendelac. 1994. An invariant T cell receptor {alpha} chain is used by a unique subset of major histocompatibility complex class I-specific CD4+ and CD4-8- T cells in mice and humans. J. Exp. Med. 180:1097.[Abstract/Free Full Text]
  54. Bendelac, A.. 1995. Positive selection of mouse NK1+ T cells by CD1-expressing cortical thymocytes. J. Exp. Med. 182:2091.[Abstract/Free Full Text]
  55. Taub, D. D., J. R. Ortaldo, S. M. Turcovski-Corrales, M. L. Key, D. L. Longo, W. J. Murphy. 1996. ß chemokines costimulate lymphocyte cytolysis, proliferation, and lymphokine production. J. Leukocyte Biol. 59:81.[Abstract]
  56. Xu, L. L., M. K. Warren, W. L. Rose, W. Gong, J. M. Wang. 1996. Human recombinant monocyte chemotactic protein and other c-c chemokines bind and induce directional migration of dendritic cells in vitro. J. Leukocyte Biol. 60:365.[Abstract]



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