Transfection of IL-2 Augments CTL Response to Human Melanoma Cells In Vitro

Elsas, Andrea van; Aarnoudse, Corlien A.; Minne, Carolien E. van der; Spek, Corry W. van der; Brouwenstijn, Nathalie; Osanto, S.; Schrier, Peter I.

doi:10.1097/00002371-199709000-00003

Cited by 14 publications

(6 citation statements)

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“…In the present study we have detected A2/CAMEL [1][2][3][4][5][6][7][8][9][10][11] ϩ tetramer ϩ CD8 ϩ T cells in peptide-stimulated PBMC from 3 of 33 (9%) A2 ϩ patients as well as in TILN. Thus, a response to this alternative ORF-derived epitope is not an exception resulting from a unique treatment/stimulation procedure (25). The proportion of patients with circulating CAMEL 1-11 -specific CD8 ϩ T cells appeared to be lower than that for HLA-A2 NY-ESO-1 ORF1-specific CD8 ϩ T cells observed by us and others (10,24).…”

Section: Discussionmentioning

confidence: 65%

Efficient Simultaneous Presentation of NY-ESO-1/LAGE-1 Primary and Nonprimary Open Reading Frame-Derived CTL Epitopes in Melanoma

Rimoldi

Rubio-Godoy²,

Dutoit³

et al. 2000

The Journal of Immunology

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Recent studies have shown that CTL epitopes derived from tumor-associated Ags can be encoded by both primary and nonprimary open reading frames (ORF). In this study we have analyzed the HLA-A2-restricted CD8+ T cell response to a recently identified CTL epitope derived from an alternative ORF product of gene LAGE-1 (named CAMEL), and the highly homologous gene NY-ESO-1 in melanoma patients. Using MHC/peptide tetramers we detected CAMEL1–11-specific CD8+ T cells in peptide-stimulated PBMC as well as among tumor-infiltrated lymph node cells from several patients. Sorting and expansion of tetramer+ CD8+ T cells allowed the isolation of tetramerbright and tetramerdull populations that specifically recognized the peptide Ag with high and low avidity, respectively. Remarkably, only high avidity CAMEL-specific CTL were able to recognize Ag-expressing tumor cells. A large series of HLA-A2-positive melanoma cell lines was characterized for the expression of LAGE-1 and NY-ESO-1 mRNA and protein and tested for recognition by CAMEL-specific CTL as well as CTL that recognize a peptide (NY-ESO-1157–165) encoded by the primary ORF products of the LAGE-1 and NY-ESO-1 genes. This analysis revealed that tumor-associated CD8+ T cell epitopes are simultaneously and efficiently generated from both primary and nonprimary ORF products of LAGE-1 and NY-ESO-1 genes and, importantly, that this occurs in the majority of melanoma tumors. These findings underscore the in vivo immunological relevance of CTL epitopes derived from nonprimary ORF products and support their use as candidate vaccines for inducing tumor specific cell-mediated immunity against cancer.

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Section: Discussionmentioning

confidence: 65%

Efficient Simultaneous Presentation of NY-ESO-1/LAGE-1 Primary and Nonprimary Open Reading Frame-Derived CTL Epitopes in Melanoma

Rimoldi

Rubio-Godoy²,

Dutoit³

et al. 2000

The Journal of Immunology

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“…This is in contrast to the CAMELspecific CTL clone 1/37, which was reactive with these tumor cells in both cytokine production and cytotoxicity assays. 8,26 Jrt-TCR␣3␤5 cells could be activated by melanoma cell line 518/ IL2.14 when either CD8␣ levels were increased on the Jurkat reporter cells (Fig. 3d) or the amount of antigen was enhanced on the melanoma cells (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…All these cell lines were well recognized by the CAMEL-specific CTL clone 1/37 in cytotoxicity 26 and TNF␣-release assays. 8 However, FM3, FM6 and MM415 did not induce any NFAT activation in Jrt-TCR␣3␤5 cells (data not shown) and only a minor activation of NFAT was induced in 2 clones upon stimulation with 518/IL2.14 ( Fig.…”

Section: Reactivity Of Jrt-tcr␣3␤5 With Tumor Cell Linesmentioning

confidence: 98%

TCR reconstitution in Jurkat reporter cells facilitates the identification of novel tumor antigens by cDNA expression cloning

Aarnoudse

Krüse

Konopitzky

et al. 2002

Intl Journal of Cancer

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The identification of novel tumor antigens is of extreme importance for effective immunotherapy against cancer. A major obstacle in this field is the limited life span of tumorspecific cytotoxic T lymphocytes (CTLs) in vitro. Therefore we searched for a method to isolate the tumor specificity of these CTLs, i.e., their T-cell receptors (TCRs) and transfer it to an immortalized T-cell line. For this purpose, a TCRnegative Jurkat T-cell line was equipped with a nuclear factor of activated T cells ( Key words: T-cell receptor; tumor antigen; NFAT; CAMEL; cDNA expression cloning; CD8; luciferase; retrovirus; JurkatSince the characterization of the first human tumor-specific antigen in 1991, MAGE-1, 1 a considerable number of other targets for tumor-specific cytotoxic T cells (CTLs) have been identified. Most of these antigens have been cloned from melanoma cells 2 and less frequently from other types of cancer. 3,4 For the development of effective immunotherapy against cancer, the identification of a larger variety of tumor-associated antigens is of great importance, to broaden the range of different tumors that can be treated as well as to prevent the outgrowth of antigen-loss tumor cells after vaccination.Several strategies have led to the identification of tumor-specific T-cell epitopes. Most frequently, cDNA expression cloning has been employed, in which the gene encoding the antigen is identified by transfecting pools of tumor-derived cDNA into recipient cells, followed by screening with the tumor-specific CTL. [5][6][7][8] In another approach, the peptide epitope for the tumor-specific CTL is identified, either by peptide elution and mass spectrometry 9 or by mimicry from designated peptide libraries, 10 although with the latter method no tumor antigens have been identified yet. Several tumor antigens have been found by the method of "reverse immunology." These antigens were initially cloned on the basis of their differential expression in tumors versus healthy tissues and antigenic peptides were subsequently identified by performing in vitro T-cell inductions. 11,12 For efficient elucidation of novel physiologically relevant tumor-specific antigens, large quantities of tumor-specific CTLs are required to allow for multiple screening rounds. In practice, the limited amount of CTLs available often constitutes the major obstacle for the cloning of new tumor antigens. We have sought to circumvent this issue by cloning and transferring the T-cell receptor (TCR) of a tumor-specific T cell to an immortalized T-cell line. In this way, an unlimited source of tumor-specific T cells can be obtained, which facilitates the identification of novel tumor antigens. The feasibility of TCR cloning and transfer has already been demonstrated by others. [13][14][15][16][17] We studied the properties of a TCR, isolated from a characterized CAMEL (CTL-recognized antigen on melanoma)-specific CTL clone, 8 which was transferred into a TCR-negative human leukemia Jurkat T-cell line. To detect TCRmediated activation, the cells were tr...

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“…The human melanoma cell line 518A2 [20] which expresses high levels of HMW-MAA and M14 [21], a human melanoma cell line with no detectable expression of HMW-MAA, were maintained in RPMI 1640 medium (Lonza, Verviers, Belgium) supplemented with 10% (v/v) FCS and 1% (v/v) antibiotic-antimycotic mix (both from Gibco, Paisley, UK). Both cell lines were cultured in a humidified atmosphere containing 5% CO 2 and 95% ambient air at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Specificity of Mimotope-Induced Anti-High Molecular Weight-Melanoma Associated Antigen (HMW-MAA) Antibodies Does Not Ensure Biological Activity

et al. 2011

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Vaccines based on peptide mimics (mimotopes) of conformational tumor antigen epitopes have been investigated for a variety of human tumors including breast cancer, tumors expressing the carcinoembryonic antigen, B cell lymphoma, neuroblastoma, and melanoma. In our previous work, we designed a vaccine based on a mimotope of the high molecular weight-melanoma associated antigen (HMW-MAA) that elicited HMW-MAA-specific antibodies (Abs) with anti-tumor activity in vitro and in vivo. In this study, we aimed to identify mimotopes of additional distinct HMW-MAA epitopes, since they could be used to construct a polymimotope melanoma vaccine. For this purpose, random peptide phage libraries were screened with the anti-HMW-MAA monoclonal antibodies (mAbs) VT80.12 and VF1-TP43 yielding one peptide ligand for each mAb. Both peptides inhibited the binding of the corresponding mAb to the HMW-MAA. Furthermore, when coupled to the carrier protein keyhole limpet hemocyanin (KLH), both HMW-MAA mimotopes elicited peptide-specific Abs in rabbits or BALB/c mice, but only the mimotope isolated with the mAb VT80.12 elicited HMW-MAA-specific Abs and only in mice. However, the latter Abs had no detectable effect on HMW-MAA expressing human melanoma cells in vitro. These results describe limitations related to the phage display technique and emphasize the need to characterize the functional properties of the mAb utilized to isolate mimotopes of the corresponding epitopes.

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Transfection of IL-2 Augments CTL Response to Human Melanoma Cells In Vitro

Cited by 14 publications

References 0 publications

Efficient Simultaneous Presentation of NY-ESO-1/LAGE-1 Primary and Nonprimary Open Reading Frame-Derived CTL Epitopes in Melanoma

Efficient Simultaneous Presentation of NY-ESO-1/LAGE-1 Primary and Nonprimary Open Reading Frame-Derived CTL Epitopes in Melanoma

TCR reconstitution in Jurkat reporter cells facilitates the identification of novel tumor antigens by cDNA expression cloning

Specificity of Mimotope-Induced Anti-High Molecular Weight-Melanoma Associated Antigen (HMW-MAA) Antibodies Does Not Ensure Biological Activity

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