Analysis of antigens recognized on human melanoma cells by A2‐restricted cytolytic t lymphocytes (CTL)

Wölfel, Thomas; Hauer, M.; Klehmann, E.; Brichard, Vincent G.; Ackermann, Birgit; Knuth, Alexander; Boon, Thierry; Büschenfelde, K H Meyer zum

doi:10.1002/ijc.2910550212

Cited by 85 publications

(41 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The CTL clone IVSB was maintained according to the protocol published by Wölfel et al 28 Briefly, IVSB was restimulated weekly with irradiated autologous melanoma cells (SK29-Mel-1) and allogeneic feeder cells (AK-EBV-B) in RPMI 1640 medium supplemented with 10% human sera, 25 U/mL rhIL-2 (Proleukin; Chiron, Ratingen, Germany) and 5 U/mL rhIL-4 (PBH).…”

Section: Culture Of the Tyrosinase-specific Ctl Clone Ivsbmentioning

confidence: 99%

CD83+ human dendritic cells transfected with tumor peptide cDNA by electroporation induce specific T-cell responses: A potential tool for gene immunotherapy

et al. 2000

View full text Add to dashboard Cite

Dendritic cells (DC) are the most potent immunostimulatory cells, with the capacity to induce primary T-cell responses. Functional autologous DC can be generated from fetal calf serum-free peripheral blood mononuclear cells in the presence of interleukin-4 and granulocyte-macrophage colony-stimulating factor and are stimulated with a defined cytokine cocktail for terminal maturation. We were able to establish a nonviral transfection protocol for these DC by electroporation. Using enhanced green fluorescent protein as a reporter gene, we achieved transfection efficiencies of up to 10%. FACScan analyses revealed a stable phenotype, and the expression of major histocompatibility complex class II and CD83 was not affected by the transfection conditions used. Like their untransfected counterparts, DC that were functionally transfected with green fluorescent protein were potent inducers of allogeneic T cells. To assess whether cDNAs transfected into DC are functionally expressed, human tyrosinase cDNA was transfected into DC. Tyrosinase-transfected DC, but not controls, resulted in antigen-specific tumor necrosis factor-␣ release of the tyrosinase-specific cytolytic T-cell clone IVSB. Taken together, the data show that genuine (CD83 ϩ ) mature DC can be transfected using a nonviral method, and that the DC retain their functionality. These DC are ideal candidates for immunotherapy (e.g., cancer therapy). Cancer Gene Therapy (2000) D endritic cells (DC) are the most potent antigen (Ag)-presenting cells (APCs) of the immune system, with the capacity to activate resting naive T cells most efficiently and to initiate primary immune responses. Due to their functional properties, DC are ideal adjuvants for immunotherapeutic strategies using DC pulsed with tumor-associated Ags (TAA) for vaccination. 1,2 Recently several TAA have been characterized, and they are expressed by the tumor cells. TAA peptides in association with major histocompatibility complex (MHC) are presented on the tumor cell surface, are recognized by Ag-specific T cells, and provide a potential target for immunotherapeutic vaccination approaches. [3][4][5] In various murine studies, DC pulsed with TAA peptide can induce Ag-specific antitumoral responses in vivo. 6,7 There are even promising reports about clinical trials in which patients with B-cell lymphoma were successfully vaccinated using autologous Ag-pulsed DC. 8 Recently, a clinical pilot study was published that reports on the successful vaccination of melanoma patients with peptide-or tumor lysate-pulsed DC. 9 The use of genetically engineered DC as tumor vaccines combines the advantages of immunotherapy with those of gene therapy. The application of peptide-pulsed DC is limited by the human histocompatibility leukocyte Ag (HLA) restriction of characterized TAA epitopes. 3 Using recombinant techniques, the full-length protein Ag is expressed in the genetically engineered APCs and is accessible to the cellular Ag-processing machinery. Therefore, the repertoire of the epitopes presented in context w...

show abstract

Section: Culture Of the Tyrosinase-specific Ctl Clone Ivsbmentioning

confidence: 99%

CD83+ human dendritic cells transfected with tumor peptide cDNA by electroporation induce specific T-cell responses: A potential tool for gene immunotherapy

et al. 2000

View full text Add to dashboard Cite

show abstract

“…CD8 ϩ MLTC responders were regularly frozen in aliquots at different time points 3-4 days after a final stimulation with irradiated tumor cells. Some MLTCs were cloned by limiting dilution, and T cell clones were propagated as described (8). Notably, all functional assays in this study were carried out with CD8 ϩ MLTC or CD8 ϩ T cell clones (cytotoxic T lymphocytes, CTLs) stimulated with autologous tumor cells.…”

mentioning

confidence: 99%

The response of autologous T cells to a human melanoma is dominated by mutated neoantigens

Lennerz

Fatho²,

Gentilini³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

418

318

View full text Add to dashboard Cite

Our understanding of pathways leading to antitumor immunity may depend on an undistorted knowledge of the primary antigenic targets of patients' autologous T cell responses. In the melanoma model derived from patient DT, we applied cryopreserved shortterm autologous mixed lymphocyte-tumor cell cultures (MLTCs) in combination with an IFN-␥ enzyme-linked immunospot (ELISPOT) assay to cDNA expression screening. We identified three previously unknown peptides processed from melanosomal proteins tyrosinase (presented by HLA-A*2601 and -B*3801) and gp100 (presented by HLA-B*07021) and five neoantigens generated by somatic point mutations in the patient's melanoma. The mutations were found in the genes SIRT2, GPNMB, SNRP116, SNRPD1, and RBAF600. Peptides containing the mutated residues were presented by HLA-A*03011, -B*07021, and -B*3801. Mutation-induced functional impairment was so far demonstrated for SIRT2. Within MLTC responder populations that were independently expanded from the patient's peripheral blood lymphocytes of different years, T cells against mutated epitopes clearly predominated. These results document a high degree of individuality for the cellular antitumor response and support the need for individualizing the monitoring and therapeutic approaches to the primary targets of the autologous T cell response, which may finally lead to a more effective cancer immunotherapy.expression cloning ͉ peptide ͉ tumor antigen ͉ mixed lymphocyte-tumor cell culture ͉ cytotoxic T lymphocytes

show abstract

“…Recently, melanoma-specific HLA-A2-restricted CTL clones have been shown to recognize cultured normal melanocytes as well as their malignant counterparts, suggesting that shared melanoma antigens may be lineage specific (6). The multiplicity of melanoma-associated antigens present within isolated melanoma lesions, and even within individual melanoma cells, has been suggested on the cellular level (7,8) and confirmed on the peptide level (9,10). Five MHC class I-restricted melanoma-associated antigens have been molecularly defined (11)(12)(13)(14)(15)(16), encoding only a subset of these commonly expressed antigens.…”

mentioning

confidence: 99%

Human CD4+ T cells specifically recognize a shared melanoma-associated antigen encoded by the tyrosinase gene.

Topalian

Rivoltini

Mancini

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

244

101

View full text Add to dashboard Cite

Although commonly expressed human melanoma-associated antigens r g d by CD8+ cytolytic T cells have been described, little is known about CD4+ T-cell recognition of melanoma-aated antigens. Epstein-Barr virustransformed B cells were used to present antigens derived from whole cell lysates of autologous and allogeneic melanomas for recognition by melanoma-specific CD4+ T-cell lines and clones cultured from tumor-infiltrating lymphocytes. HLA-DRrestricted antigens were detected in the lysates on the basis of specific release of cytokines from the responding T cells. Antigen sharing was demonstrated in the majority of melanomas tested, as well as in cultured normal melanocytes, but not in other normal tissues or nonmelanoma tumors. T-cell clones manifested a single recognition pattern, suggesting the presence of an immunodominant epitope. This epitope was identified as a product of the tyrosinase gene, which has also been shown to encode class I-restricted epitopes recognized by CD8+ T cells from melanoma patients. Identification of commonly expressed tumor-associated protein molecules containing epitopes presented by both class I and class H major histocompatibility molecules may provide optimal reagents for cancer immunization strategies.Commonly expressed melanoma-associated antigens can be recognized by CD8+ cytotoxic T lymphocytes (CTLs) derived from melanoma patients. In short-term lysis assays, CTLs grown from in vitro sensitized peripheral blood lymphocytes (PBLs) or lymph node lymphocytes or from lymphocytes infiltrating metastatic melanoma lesions have been shown to recognize autologous and major histocompatibility complex (MHC) class I-compatible allogeneic melanomas but not HLA-matched nonmelanoma tumors, lymphoblasts, or cultured fibroblasts (1, 2). Similar recognition patterns have been observed by measuring cytokine secretion from tumor infiltrating lymphocytes (TILs) cocultivated with autologous or HLA-matched allogeneic tumor stimulators (3). Expression of shared melanoma antigens even in tumors of discrepant HLA phenotypes has been shown by genetically modifying these tumors to express the HLA-A2.1 molecule: HLA-A2-restricted melanoma-specific CTLs lysed the majority of gene-modified melanomas tested, but not HLA-A2+ nonmelanoma tumors or normal tissues (4, 5). Recently, melanoma-specific HLA-A2-restricted CTL clones have been shown to recognize cultured normal melanocytes as well as their malignant counterparts, suggesting that shared melanoma antigens may be lineage specific (6). The multiplicity of melanoma-associated antigens present within isolated melanoma lesions, and even within individual melanoma cells, has been suggested on the cellular level (7, 8) and confirmed on the peptide level (9, 10). Five MHC class I-restricted melanoma-associated antigens have been molecularly defined (11)(12)(13)(14)(15)(16), encoding only a subset of these commonly expressed antigens. Therapeutic vaccination strategies are evolving based on these CD8-recognized epitopes.While animal models of maligna...

show abstract

Analysis of antigens recognized on human melanoma cells by A2‐restricted cytolytic t lymphocytes (CTL)

Cited by 85 publications

References 28 publications

CD83+ human dendritic cells transfected with tumor peptide cDNA by electroporation induce specific T-cell responses: A potential tool for gene immunotherapy

CD83+ human dendritic cells transfected with tumor peptide cDNA by electroporation induce specific T-cell responses: A potential tool for gene immunotherapy

The response of autologous T cells to a human melanoma is dominated by mutated neoantigens

Human CD4+ T cells specifically recognize a shared melanoma-associated antigen encoded by the tyrosinase gene.

Contact Info

Product

Resources

About