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9V
2 T Cell Response to Colon Carcinoma Cells1
* Institut National de la Santé et de la Recherche Médicale (INSERM), Unité 601, Nantes, France;
Faculté des Sciences et des Techniques, Nantes, France;
Centre René Gauducheau, Nantes, France;
INSERM, Unité 539, and Laboratoire d’Anatomopathologie, Hôpital Laënnec, St. Herblain, France
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Abstract |
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9V
2 T cell clone showing strong reactivity against autologous tumor cell lines. This clone killed a large fraction of allogeneic colon carcinoma and melanoma cell lines, but did not affect a normal colon cell line, colon fibroblasts, or melanocytes. Tumor cell recognition was TCR and NKG2D dependent and induced TNF-
and IFN- secretion by the clone; accordingly, tumor targets expressed several NKG2D ligands, such as MHC class I chain-related gene A and UL16-binding protein molecules. Colon tumor recognition by V9V2 T cells was highly dependent on isopentenyl pyrophosphate production and ICAM-1 expression by target cells. Finally, similar reactivity patterns against colon carcinoma cell lines were observed using polyclonal V9V2 T cells of various origins, and V9V2 lymphocytes were present in the majority of colon tumor samples studied. Together, these results suggest that V9V2 T cells contribute to the natural immune surveillance against colon cancers. Therefore, this study provides a strong rationale for the use of V9V2 T cell agonists in immunotherapies targeting colon tumors. |
Introduction |
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T lymphocytes, which represent the dominant TIL population in most tumors, the V1 subpopulation of T cells frequently infiltrates some solid tumors, such as lung, kidney, breast, prostate, colon, ovarian carcinomas, and cutaneous malignancies (1, 2, 3, 4, 5). In contrast, V9V2 T cells, which predominate among PBL in healthy adults, have been rarely described in tumors (6).
Both V1 and V2 subsets have been implicated in tumor immunity based on their capacity to kill tumor cell lines in vitro in a TCR-dependent manner. Nonetheless, the tumor ligands recognized by these subsets are likely to be distinct. Indeed, although V1 T cells derived from TIL kill a wide array of solid tumors (7, 8, 9, 10, 11), V9V2 T cells preferentially kill hemopoietic tumor cell lines, such as myeloma, lymphoma, and erythroleukemia (12, 13, 14, 15). However, more recent studies reported recognition of some melanoma and renal carcinoma cell lines by V9V2 T cells and killing of autologous metastatic renal carcinoma cells (RCC) by autologous PBL-derived V9V2 T cell lines (16, 17, 18).
Tumor cell recognition by V9V2 T lymphocytes is mostly MHC unrestricted and is controlled by both TCR-dependent and TCR-independent signals. Several nonpeptidic phosphorylated compounds produced through the isoprenoid biosynthetic pathway and classically referred to as phosphoantigens can activate V9V2 T cells in a TCR-dependent fashion. Although natural and synthetic phosphoantigens (such as isopentenyl pyrophosphate (IPP), hydroxydimethylallyl pyrophosphate, and bromohydrin-pyrophosphate) readily stimulate V9V2 T cells in vitro, other compounds, such as aminobiphosphonates, sensitize target cells to V9V2 killing by promoting the accumulation of V9V2 agonists (e.g., IPP) in treated cells (19, 20, 21, 22, 23, 24, 25, 26, 27). The mode of recognition of phosphoantigen by V9V2 cells remains unclear, but their presentation by as yet undefined receptors is likely. Various receptors, such as the LFA-1 integrin, CD2 (19, 20), and NK receptors, both activatory, such as NKG2D, and inhibitory, were also reported to be important for tumor cell line recognition and lysis by V9V2 T lymphocytes (14, 18, 21, 22, 23, 24, 25, 26).
The aim of the present study was to investigate the adaptive cellular immunity against colon carcinoma cells through analysis of colon tumor reactivity of CD8+ T cells derived from tumor patients. Two colon carcinoma cell lines (CCCL) and CD8+ T cells reactive to these tumor cells were derived from ascites of a metastatic colon carcinoma patient. Identification of V9V2 T cells within the tumor-reactive CD8+ population prompted us to perform an in-depth analysis of V9V2 T cell reactivity against various CCCL and to study the modalities of CCCL recognition by V9V2 T cells and the frequency of these lymphocytes present within colon tumors. Our results suggest that V9V2 T cells infiltrate and recognize a significant fraction of human colon cancers and, therefore, may contribute to the immune surveillance of these tumors.
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Materials and Methods |
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Tumor cell lines C4A (adherently growing) and C4S (growing in suspension) were established from an ascites of a stage IV colon cancer patient (C4). Other carcinoma cell lines used were SW620, SW1116, SW707, HT29, HTC116, LS180, SW480, CaCo2, Colo205, and SW1222 (gifts from J. Le Pendu, J. F. Chatal, C. Saï (Institut National de la Santé et de la Recherche Médicale, Unité 601, Nantes, France), and M. Denis (Unité Mixte de Recherche 539, Nantes, France)). The normal colon cell line CCL-241 was provided by American Type Culture Collection and was cultured in DMEM (Sigma-Aldrich) containing 10% FCS, 100 U/ml penicillin, 100 µg/ml streptomycin (Sigma-Aldrich), 2 mM L-glutamine (Sigma-Aldrich), and 30 ng/ml epidermal growth factor. Mouse fibrosarcoma WEHI 164 clone 13, used for TNF production assays, was obtained from T. Boon (Ludwig Institute for Cancer Research, Brussels, Belgium). The B-EBV-transformed cell line LAZ 338 was a gift from T. Hercend (Vertex Pharmaceutical, Abingdon, U.K.). All tumor cell lines and LAZ 338 were cultured in RPMI 1640 (Sigma-Aldrich) containing 10% FCS, 100 U/ml penicillin, 100 µg/ml streptomycin (Sigma-Aldrich), and 2 mM L-glutamine (Sigma-Aldrich).
Cell lines transfected by ICAM-1 cDNA
Tumor cell lines LS180 and Colo205 were transfected by electroporation using the AMAXA system according to the manufacturer’s instructions with a full-length human ICAM-1 cDNA (gift from T. Yoshimasa, Kyoto University, Kyoto, Japan) inserted in pEF-BOS neoSE expression vector. The transfected cell lines were cultured in medium containing 0.4 mg/ml G418 (Sigma-Aldrich) for colo205 and 0.6 mg/ml G418 for LS180. These cell lines were later cloned by limited dilution or were sorted using an anti-ICAM-1 mAb coupled with anti-mouse mAb-labeled microbeads, according to the manufacturer’s instructions (Miltenyi Biotec), to establish stable clones or cell lines highly expressing ICAM-1.
T lymphocytes
Polyclonal TIL were obtained from a 12-day culture of tumor fragments in RPMI 1640 (Sigma-Aldrich) containing 8% AB human serum (local production), antibiotics, glutamine (as mentioned above), and 150 U/ml rIL-2 (Chiron). Polyclonal TIL were expanded in 96-well multiplates at 1000 TIL/well in the presence of irradiated feeder cells (1 x 104 LAZ cells and 105 PBL/well), 1 µg/ml PHA-L (Sigma-Aldrich), and rIL-2. TIL clones were derived by limiting dilution and then were expanded under the same culture conditions (27). The T cell clone C4-112 was derived from CD8+ T cells present in the ascites of patient C4. The other V2 populations were sorted from the TIL from colon and RCC cancer patients (C1 and R9, respectively) or from the PBMC of a healthy donor. CD8+ T cells and V2 T cells were purified using magnetic beads coated with CD8 mAb-conjugated microbeads or with an anti-V2 mAb (IMMU389; Immunotech) and Ig anti-mouse-coated magnetic beads (Miltenyi Biotec).
Flow cytometry and blocking mAb
For T cell or tumor cell labeling, we used the following Abs: anti-CD94 (HP-3B1), anti-NKG2-A (Z199), anti-P70 or KIR3DL1/S1 (Z27), anti-KIR2DL2/S2 (Gl183), anti-KIR2DL1/S1 (EB6), and PE-conjugated or unconjugated anti-V2 TCR (389) from Immunotech; PE-conjugated anti-NKRP1 (191B8) and FITC-conjugated anti-IFN- from BD Pharmingen; PE-conjugated anti-ILT2 (HP-F1) from Beckman Coulter; anti-CD16 (Leo Ac1), anti-PEN5 (5H10), anti-KIR3DL2 (DEC66), and anti-KIR2DS4 (FSTR) from E. Vivier (Centre d’Immunologie de Marseille-Luminy, Marseille, France); pan--TCR (IMMU510) ascites; an anti-NKG2D (149810) from R&D Systems; and anti-ICAM-1 (MCA 1615), ICAM-2 (MCA 1140), ICAM-3 (MCA 1485), and VCAM (MCA 907) from Serotec. Anti-MHC chain-related gene A (MICA) (M673), anti-MICB (M362), anti-UL16-binding protein 1 (ULBP1) (M295), anti-ULBP2 (M311), and anti-ULBP3 (M551) were gifts from D. Cosman (Amgen, Seattle, WA). The anti-mouse FITC- or PE-conjugated rabbit F(ab')2 were obtained from Beckman Coulter, and the isotype controls (IgG or IgG2a) from Immunotech. Cell fluorescence was analyzed by flow cytometry using CellQuest software (BD Biosciences). The fluorescence intensity ratio (RFI) was calculated as follows: mean fluorescence intensity obtained with the test/mean fluorescence intensity obtained with the isotypic control. Intracellular cytokines were measured as previously described (28).
Analysis of T cell responses by measurement of TNF release
T lymphocytes were stimulated in duplicate or triplicate cultures by tumor cell lines at an E:T cell ratio of 3:1. Culture supernatants were tested 6 h later by a biological colorimetric assay using the TNF-sensitive mouse fibrosarcoma WEHI 164 clone 13, as previously described (29).
Analysis of T cell responses by cytotoxicity assay
Target cells were labeled with 100 µCi of Na51CrO4 (Oris Industries) for 1 h at 37°C. After washing, target cells were incubated with effector T cells for 4 h at 37°C. In blocking experiments, the effector or target cells were previously treated with various concentrations of specific or isotype control mAbs for 30 min at 4°C. The radioactivity released by target cells was measured, 4 h later, on a beta plate counter. The percentage of specific 51Cr lysis was calculated using the following equation: percent specific lysis = 100 x [(test release) – (spontaneous release)]/[(maximal release) – (spontaneous release)].
Pamidronate or mevastatin treatments
Colon tumor cell lines were cultured in 24-well, flat-bottom plates. Twenty-four hours later, the medium was changed to a medium containing pamidronate (Calbiochem) at various concentrations (50–150 µM) or containing 25 µM mevastatin (Sigma-Aldrich). After overnight incubation with pamidronate, cells were washed extensively and incubated with T lymphocytes. Mevastatin-treated tumor cells were mixed with T lymphocytes in the presence of 25 µM mevastatin, because the effect of this drug is rapidly reversible (12).
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Results |
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9V2 TIL C4-112 clone recognizes and kills autologous and allogeneic tumor cell lines in a TCR-dependent manner
Two CCCL, C4A (adherent) and C4S (suspension), and CD8+ T cells (C4 TIL) were derived from an ascites sample of a colon carcinoma patient. Because the CD8+ TIL population produced TNF in response to autologous CCCL C4A and C4S (Fig. 1A), we cloned these TIL to isolate and characterize the tumor-reactive T cells. Several tumor-reactive clones were obtained, among which was the C4-112 CD8+ clone. This clone expressed V9V2 TCR and CD8 (Fig. 1B and data not shown). As shown by TNF- secretion and cytolysis assays, clone C4-112 recognized and killed the autologous and 7 of 11 allogeneic CCCL, but not the normal colon cell line tested (Fig. 1C). It also produced IFN- in response to the autologous tumor cell line (Fig. 1D). It also recognized four melanoma and three RCC lines: erythroleukemia (K562), ovary, and pancreatic tumor cell lines. By contrast, this clone did not recognize the breast, lung, or neuroblastoma tumor cell lines tested or various nontumor cell targets, such as a normal colon cell line, normal colon fibroblasts, and melanocytes (Fig. 1C and data not shown).
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TCR but not by an anti-HLA-I mAb (Fig. 1, E and F). Together, these results indicate the presence of V9V2 CD8+ T cells in the ascites of colon carcinoma patient C4, which are able to recognize and kill a large fraction of CCCL in a TCR-dependent and HLA-I-independent fashion.
CCCL recognized by clone C4-112 are lysed by V9V2 T lymphocytes of various origins
To test whether CCCL recognition was a general property of V9V2 T cells, we generated several polyclonal V9V2 T cell lines after immunomagnetic sorting of V2 cells from PBL of a healthy donor (81.65% V2+), RCC-infiltrating T cells (R9; 99.78% V2+), and another colon carcinoma patient (C1; 99.77% V2+). These polyclonal V9V2 T cell lines showed CCCL reactivity patterns very similar to that of clone C4-112, because they killed the same set of CCCL and to a similar extent as clone C4-112 (Fig. 2). Therefore, all V9V2 T cells, regardless of their origin and clonality, efficiently recognize a large fraction of CCCL.
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V9V2 T cells can be activated by various low Mr nonpeptidic Ags (30, 31, 32, 33, 34, 35). Among these, metabolites of the mevalonate pathway, such as IPP, have been shown to be tumor cell ligands of the V9V2 TCR (12, 36, 37). To determine whether CCCL recognition by the C4-112 V9V2 clone was due to IPP expression by these cell lines, we used mevastatin, an inhibitor of the mevalonate pathway, to block IPP synthesis by these cell lines. After mevastatin treatment, lysis of all CCCL was decreased by 50–90% (Fig. 3A). This indicates that production of mevalonate metabolites is required for CCCL recognition by V9V2 T cells. Because some CCCL were not recognized by the C4-112 clone, we asked whether this was due to a defect in IPP or other isoprenoid pathway metabolite synthesis. To address this, we studied V9V2-mediated killing of V9V2-susceptible and -resistant CCCL after treatment with pamidronate, an aminobiphosphonate known to precede IPP accumulation in treated cells. As shown in Fig. 3B, this treatment increased the lysis of cell lines C4A and SW1116 by 10 and 30%, respectively, and induced that of cell lines HT29 and SW1222 by clone C4-112, suggesting that recognition of most CCCLs correlates with the activity of the mevalonate pathway. The limited increase in lysis might be due to an already suboptimal IPP production in the absence of treatment or to another factor that limits the activation that results from T cell/CCCL interaction. Two cell lines, LS180 and Colo205, were not sensitized to C4-112 clone lysis upon pamidronate treatment, suggesting that either they were unresponsive to this drug or they lacked other components required for V9V2 T cell clone activation.
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Accessory/adhesion receptors, such as CD2 and LFA-1, play an important role in V9V2 T lymphocyte activation (19, 20). We screened CCCL for LFA-3 (the CD2 counterreceptor); ICAM-1, ICAM-2, and ICAM-3 (LFA-1 counterreceptors); and VCAM (VLA-4 ligand) expression. None of these cell lines expressed ICAM-3 (data not shown). Most cell lines expressed LFA-3; although no clear correlation was found between LFA-3 expression levels and V9V2 lysis susceptibility, the most efficiently killed cell lines expressed LFA-3 at the highest levels. Most CCCL, including some of those killed by the C4-112 clone, expressed VCAM and ICAM-2 at low levels, and there was no clear correlation between the expression levels of these molecules and killing intensity. All CCCL recognized by the C4-112 clone, either spontaneously or after pamidronate treatment, expressed ICAM-1, and the correlation coefficient between this expression and V9V2 lysis was >0.75 (Fig. 4A and data not shown). The two cell lines that failed to be recognized even after pamidronate treatment (Colo205 and LS180) lacked ICAM-1 and VCAM (Fig. 4A). To address the contributions of ICAM-1 and VCAM to the recognition of colon tumor cell lines by the C4-112 clone, CCCLs were preincubated with blocking anti-ICAM-1, anti-VCAM mAb or an isotype control before the cytolysis assay. Anti-ICAM-1 mAb inhibited CCCL lysis to variable extents: >30% for four of seven colon tumor cell lines (C4A, SW1116, Caco2, and HTC116) and <10% for the other three cell lines (SW480, SW620, and SW707). The VCAM mAb, in contrast, had no or a very minor inhibitory effect on CCCL recognition (Fig. 4B). To determine whether the absence of lysis of LS180 and Colo205 CCCL by V9V2 T cells could be accounted for by lack of ICAM-1 expression, we transfected these cell lines with human ICAM-1 cDNA and established stable polyclonal and monoclonal transfectants expressing high levels of ICAM-1. ICAM-1 transfection induced both ICAM-1 and VCAM expression on Colo205 and LS180 (data not shown). Only ICAM-1 transfection in Colo205 dramatically increased its susceptibility to lysis by clone C4-112 after pamidronate treatment. In contrast, ICAM-1-transfected LS180 CCCL remained resistant to V9V2 T cell lysis even after pamidronate treatment (Fig. 5A). Lysis of pamidronate-treated ICAM-1-expressing colo205 cells was strongly inhibited by the anti-ICAM-1 mAb, but not by the anti-VCAM mAb (Fig. 5B). Together, these results demonstrate that ICAM-1 expression by CCCL contributes, although to a variable extent, to the recognition and lysis of these cells by V9V2 T cells.
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9V2 C4-112 T cell clone
NKR are expressed by V9V2 T cells and can modulate the functions of these cells (14, 21, 22). We therefore studied the expression of common NKR using available mAbs NKG2D, NKRP1-A, CD94/NKG2A, ILT2, PEN5, KIR2DS4, KIR3DL1, and KIR2DL2/L3 by the C4-112 clone. The C4-112 clone expressed NKG2D, CD94/NKG2A, and KIR3DL1 (Fig. 6). The staining patterns obtained with ILT2- and KIR2DL2/S2-specific mAb were more heterogeneous, because only a small fraction of C4-112 cells were stained by these Ab (Fig. 6A). No labeling was observed with mAb directed against NKRP1-A, PEN-5, and KIR2DS4 NKR (data not shown). The NKG2D receptor is known to positively modulate the cytotoxicity of NK and T cell effectors (38). To determine whether this receptor contributed to the lysis of colon tumor cell lines by the C4-112 clone, we screened CCCL for the expression of several NKG2D ligands (MICA, MICB, and ULBP1, -2, and -3). All tumor cell lines expressed various levels of MICA, ULBP2, and 3, and most of them also expressed ULBP1 (Fig. 6B). None was MICB+ (data not shown). The expression levels of NKG2D ligands correlated with V9V2 lysis susceptibility of CCCL. Furthermore, CCCL lysis by the C4-112 clone was significantly inhibited by blocking NKG2D-specific mAb (Fig. 6C). These data suggest that the TCR-dependent recognition of CCCL by V9V2 T cells is costimulated by NKG2D.
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9V2 T lymphocytes
Finally, because the C4-112 clone was isolated from ascites of a colon cancer patient, we asked whether solid colon carcinoma tumor samples also contained V9V2 T cells. TIL were isolated from fragments of colon tumor samples (C1–C38), cultured for a few days in IL-2-containing medium, then expanded using polyclonal T cell stimulators and screened for the presence of V2+ T lymphocytes (Fig. 7). V9V2 lymphocytes were present in the majority (11 of 15) of colon TIL populations at frequencies ranging from 0.1–3%.
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Discussion |
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TopAbstractIntroductionMaterials and MethodsResultsDiscussionDisclosuresReferences |
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9V2 TCR clone isolated from a colon cancer ascites, that efficiently kills, in a TCR-dependent and MHC-unrestricted fashion, autologous and allogeneic carcinoma cell lines, including a large fraction of CCCL. Similar tumor recognition patterns were observed with all V9V2 T cell lines tested regardless of their origins. Besides, CCCL killing efficiency by V9V2 T cells depended on both tumor expression levels of V9V2 TCR stimuli (such as mevalonate pathway metabolites), of tumor ligands engaging the NKG2D receptor, and of counter-receptors of adhesion molecules, such as ICAM-. We also showed that more than two-thirds of colon carcinoma tumors were infiltrated by V9V2 T cells. A previous study also reported the presence in these tumors of 1+ T cells recognizing autologous and allogeneic CCCL (7). This subpopulation was not investigated in our study.
To our knowledge, this is the first report describing the recognition and efficient killing of CCCL by V9V2 T cells. Among solid tumor cell lines, only some RCCs and melanomas had been shown to be targets for this T cell subset, either spontaneously or after pamidronate treatment (12, 17, 18). Because melanocytes were not killed by these cells, the lytic potential of V9V2 T cells appears selective for transformed epithelial cells, because it is for transformed hemopoietic cells, such as leukemia, myeloma, and lymphoma (12, 13, 14, 15).
Human V9V2 T cells are activated in a TCR-dependent manner by several phosphorylated (34, 39) or aminated (40) alkyl molecules. Furthermore, the expression of IPP has been shown to correlate with the susceptibility of some tumor cell lines to V9V2 T cell lysis (12, 30, 31, 32, 33, 34, 35). In this study data obtained through stimulation or inhibition of IPP biosynthesis by pamidronate and mevastatine, respectively, suggest that CCCL recognition might also correlate with IPP production. Consistent with the assumption that tumor cell recognition is due to increased expression and presentation of mevalonate metabolites is the finding that some tumor cell types show increased expression and function of the limiting enzyme 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (41, 42). Recently, new ligands of the V9V2 TCR were identified on the surface of hemopoietic tumors and RCC lines, namely, an F1-ATPase-related structure and one of its ligands, a delipidated form of apolipoprotein A1 (43). Although this work did not establish the relationships between recognition of these ligands and that of phosphoantigens on tumor cells by V9V2 T lymphocytes, it was hypothesized that a complex of these two molecules may bind phosphoantigens and present them to V9V2 TCR (43). It is possible that such a complex is exposed by colon carcinoma cells, making them susceptible to V9V2 TCR lysis.
It is known that V9V2 T cell activity is tightly regulated by NK-like receptors for MHC class Ia and Ib ligands. Accordingly, we showed that NKG2D contributed to CCCL lysis by V9V2 clones, and that, as previously described (18, 44, 45), CCCL expressed very high levels of several NKG2D ligands, such as MICA and ULBP1, -2, and -3. The V9V2 clone C4-112 also expressed several inhibitory receptors, such as KIR3DS1 and the C-type lectin receptor CD94/NKG2A. Because HLA-Bw4 and HLA-E, the respective ligands of these receptors, were expressed by the autologous tumor cell lines (data not shown), these two receptors might decrease the lysis efficiency of autologous CCCL. The fact that despite this, autologous CCCL were strongly lysed by clone C4-112 suggests that tumor cells expressed sufficiently high levels of activatory signals to overcome these inhibitory signals. It is still possible that variations in HLA-E and Bw4 expression by some other cell lines contribute to the varied susceptibility of CCCL to V9V2 T cell clone lysis. Nonetheless, we did not observe any correlation between the HLA-Bw4/Bw6 type of these cells and their lysis susceptibility.
A critical role of ICAM-1 molecule in the recognition of lung tumor and lymphoma cell lines by V9V2 T cells was previously suggested (19, 20). Our data show that ICAM-1 expression by colon carcinoma cells strongly contributes to the activation of V9V2 T cells, probably by strengthening effector/target cell interactions via LFA-1 engagement. We also addressed the roles of VCAM and ICAM-2, two other LFA-1 counterreceptors, and that of LFA-3, a ligand of the coactivatory TCR CD2, which was previously shown to be important in melanoma cell recognition by T cells (46). Our data do not suggest a key contribution of VCAM, ICAM-2, and LFA-3 molecules to CCCL recognition by V9V2 T cells, because several CCCL killed by the V9V2 T cells (spontaneously or after ICAM-1 expression) poorly expressed these molecules, and VCAM-specific mAb failed to inhibit CCCL killing by V9V2 T cells.
About one-third of CCCLs were not recognized by V9V2 T cells. For some of them, this could be explained by defective expression of the adherent molecule ICAM-1 (20) and/or by the inability to accumulate and/or present phosphoantigen ligands. However, one cell line (LS180) that spontaneously expressed NKG2D ligands remained resistant to lysis by the C4-112 clone even after ICAM-1, VCAM, and pamidronate treatment. This might be due to tumor cell unresponsiveness to pamidronate. Alternatively, it would suggest the existence of an additional requirement for CCCL recognition by V9V2 T cells, such as a minimal expression of LFA-3 or surface expression of ATP synthase or other V9V2 TCR ligands (43).
The presence of V9V2 T lymphocytes in the majority of colon cancer TIL populations and the response of this T cell subpopulation to many CCCL suggest that a natural immune response mediated by these lymphocytes contributes to the immunosurveillance of these tumors. Such observations may foster the development of novel alternative or adjuvant therapies targeting V9V2 T cells for the treatment of colon cancer patients. This might be achieved by stimulating V9V2 T cell activity in these patients through pamidronate and IL-2 injection, as recently performed in hematological malignancies (47). Alternatively, it could be achieved by passive transfer of ex vivo expanded autologous V9V2 T cells derived from cancer patients. These T cells could be obtained from patient blood, because PBL-derived V9V2 T cells exhibited a similar or even stronger reactivity than TIL to CCCL. Interestingly, because the mevalonate metabolites are required for cell survival and growth, their targeting on colon tumor cells will not have the risk of generating Ag loss tumor variants. In conclusion, our results show that colon carcinoma cancer patients are suitable candidates for active or passive immunotherapy targeting V9V2 T cells.
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Disclosures |
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Footnotes |
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1 This work was supported by the Cancer Research Institute (Cancer Antigen Discovery Collaborative) and the Ligue Nationale contre le Cancer (01-04). 
2 Address correspondence and reprint requests to Dr. Francine Jotereau at the current address: Institut de Biologie, 9 quai Moncousu, 44035 Nantes Cedex 1, France. E-mail address: jotereau{at}nantes.inserm.fr
3 Abbreviations used in this paper: TIL, tumor-infiltrating lymphocyte; CCCL, colon carcinoma cell line; IPP, inositol pyrophosphate; RCC, renal carcinoma cell; RFI, ratio fluorescence intensity; MICA, MHC class I chain-related gene A; ULBP, UL16-binding protein.
Received for publication April 14, 2005. Accepted for publication August 3, 2005.
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