Targeting Heat Shock Proteins for Immunotherapy in Multiple Myeloma: Generation of Myeloma-Specific CTLs Using Dendritic Cells Pulsed with Tumor-Derived gp96

Qian, Jianfei; Wang, Siqing; Yang, Jing; Xie, Jingyuan; Lin, Pei; Freeman, Muta E.; Yi, Qing

doi:10.1158/1078-0432.ccr-05-1553

Cited by 58 publications

(42 citation statements)

References 39 publications

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“…These peptides were then bound to an endoplasmic reticulum-resident chaperone, GP96, and loaded onto major histocompatibility complex class I molecules in the endoplasmic reticulum (30 -32). It was also reported that peptides bound to Hsp70 in tumor cells were identified as tumor antigens and that the purified tumor-derived Hsp70-peptide complex elicited tumor-specific cytotoxic T cell immunity in vitro and in vivo (33)(34)(35)(36). We showed here that the new Hsp70-binding motif was a heptapeptide, LRIALRY.…”

Section: Discussionmentioning

confidence: 56%

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Biological Heterogeneity of the Peptide-binding Motif of the 70-kDa Heat Shock Protein by Surface Plasmon Resonance Analysis

Maeda

Sahara

Mori

et al. 2007

Journal of Biological Chemistry

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70-kDa heat shock protein family is a molecular chaperone that binds to a variety of client proteins and peptides in the cytoplasm. Several studies have revealed binding motifs between 70-kDa heat shock protein family and cytoplasmic proteins by conventional techniques such as phage display library screening. However, little is known about the binding motif based on kinetic parameters determined by surface plasmon resonance analysis. We investigated the major inducible cytosolic 70-kDa heat shock protein (Hsp70)-binding motif with the human leukocyte antigen B*2702-derived peptide Bw4 (RENL-RIALRY) by using a Biacore system based on surface plasmon resonance analysis. The K D value of Hsp70-Bw4 interaction was 1.8 ؋ 10 ؊6 M. Analyses with truncated Bw4 variant peptides showed the binding motif of Hsp70 to be seven residues, LRI-ALRY. To further study the characteristics of this motif, 126 peptides derived from Bw4, each with single amino acid substitution, were synthesized and analyzed for Hsp70 binding affinity. Interestingly, the Hsp70 binding affinity was abrogated when the residues were substituted for by acidic (Asp and Glu) ones at any position. In contrast, if the substitute residue was aromatic (Trp, Tyr, and Phe) or an Arg residue at any position, Hsp70 binding affinity was maintained. Thus, this study presents a new binding motif between Hsp70 and peptides derived from the natural protein human leukocyte antigen B*2702 and may also elucidate some characteristics of the Hsp70 binding characteristic, enhancing our understanding of Hsp70-binding determinants that may influence diverse cellular and physiological processes.Molecular chaperones of the 70-kDa heat shock protein family perform numerous functions in the quality control of cells that mediate protein folding, translocation, assembly/disassembly, and repair of unfolded proteins damaged by environmental stress (1). Chaperone activity of this protein family is necessary to recognize binding sites of a target native protein, referred to as the binding motif. It has been reported that the endoplasmic reticulum 70-kDa heat shock protein Bip (Grp78) binds to a peptide containing at least seven residues with maximal affinity in the presence of ATPase activity (2, 3). It was also suggested that peptide-binding sites of Bip show a preference for sequences rich in aliphatic residues. Subsequently, from a study employing a phage display library, the binding motif for Bip was revealed to be a heptapeptide with high contents of aromatic and hydrophobic residues (4).On the other hand, the bacterial cytoplasmic 70-kDa heat shock protein homolog DnaK has a substrate-binding region in the C terminus, and repeated binding/release to substrates is dependent on the ATPase cycle at an ATPase domain of the N terminus (5, 6). Zhu et al. (7) reported a crystal structure of the C-terminal substrate-binding domain of DnaK, which was located in a hydrophobic substrate-binding cleft with a central pocket placed between a ␤ sheet and two ␣ helices. The consensus motif recogni...

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Section: Discussionmentioning

confidence: 56%

“…Interestingly, in this motif, aromatic residues or an Arg residue could be substituted at any position. These data might help elucidate the interaction of this motif with tumor antigens and suggest the possibility of construction of immunomodulatory cancer vaccines (35,36).…”

Section: Discussionmentioning

confidence: 99%

Biological Heterogeneity of the Peptide-binding Motif of the 70-kDa Heat Shock Protein by Surface Plasmon Resonance Analysis

Maeda

Sahara

Mori

et al. 2007

Journal of Biological Chemistry

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“…On day 17, the number of AGR2-specific T cells was analyzed by IFN-c ELISPOT kit (BD Bioscience) 23,24 and the cytolytic activity of CTLs was examined by using a standard 51 Cr-release assay. 25 ELISPOT assay T2 cells pulsed with AGR2 peptide, HLA-A*0201 1 AGR2-positive CRC cell lines (HCT-8 and DLD1) and HLA-A*0201 1 AGR2-negative CRC cell line (HCT116) were used as target cells with or without anti-major histocompatibility complex (MHC) class I mAb (30 mg/ml) for the antigen-specific, IFN-c ELISPOT assay. The number of IFN-c spots was enumerated by an automatic CTL immunospot analyzer (Cellular Technology Ltd, Shaker Heights, OH, USA).…”

Section: Cells) Briefly Cd8mentioning

confidence: 99%

“…25 T2 pulsed with AGR2 peptide or HIV peptide, K562, HLA-A*0201 1 AGR2-positive CRC cell lines (HCT-8 and DLD1) and HLA-A*0201 1 AGR2-negative CRC cell line (HCT116) were used as target cells. The target cells were incubated with 100 mCi 51 Cr-sodium chromate for 1 h, washed extensively, seeded (1310 4 cells/well) into 96-well U-bottomed plates in T-cell medium and cocultured for 4 h with various numbers of CTLs.…”

Section: Cytotoxicity Assaymentioning

confidence: 99%

Identification of novel HLA-A*0201-restricted epitopes from anterior gradient-2 as a tumor-associated antigen against colorectal cancer

et al. 2012

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Anterior gradient-2 (AGR2) promotes tumor growth, cell migration and cellular transformation and its enhanced expression is almost completely restricted to malignant tissues, thus making AGR2 an interesting target for the development of immunotherapeutic strategies. We investigated whether the AGR2 molecule comprises human leukocyte antigen (HLA)-A*0201-binding epitopes recognized by human cytotoxic T lymphocytes (CTLs), which could be targeted in dendritic cell (DC)-based cancer immunotherapy against colorectal cancer (CRC). We reviewed the sequence of AGR2 for peptides that could potentially bind to HLA-A*0201 with the aid of a computer-based program. Five candidate peptides with different binding scores were synthesized and tested. These peptides were then assessed for their immunogenicity to elicit specific immune responses mediated by CTLs in vitro by means of enzyme-linked immunospot assays and CTL assays. AGR2 was highly expressed in several CRC cell lines, including DK01, DLD1, KM12C, HCT-8 and HT-29. DCs pulsed with AGR2-P2 (aa 11-19; LLVALSYTL) or AGR2-P4 (aa 127-135; RIMFVDPSL) generated potent CTLs that could lyse T2 cells pulsed with AGR2-P2 or AGR2-P4 and HLA-A0201 1 AGR2-positive CRC cell lines in a strong dose-dependent and HLA-A*0201-restricted manner. In conclusion, these novel epitopes derived from AGR2 protein may be attractive candidates for DC-based immunotherapy for CRC.

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“…47,55,56,81,182 Another solution to the problem could be the use of established cancer cell lines of the required tumor type expressing shared tumor antigens. Interestingly, pooled Gp96 preparations from various allogeneic myeloma cell lines have been shown to be as effective as the autologous counterpart in eliciting human-specific CD4 þ as well as CD8 þ T-cell responses in vitro 183 and to protecting mice from tumor challenge. 176 …”

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confidence: 99%

Active‐specific immunotherapy of human cancers with the heat shock protein Gp96—revisited

Randazzo

Terness

Opelz

et al. 2012

Intl Journal of Cancer

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The passive administration of specific antibodies that selectively target tumors is a well-known strategy in cancer treatment. Active immunotherapy using peptide vaccines, in contrast, is expected to induce specific, cytolytic T cells in the patient, which react against tumor antigens and destroy malignant cells. Although several concepts exist, the identification and low immunogenicity of tumor-specific peptides remain a serious problem. Heat shock proteins (HSPs), notably glycoprotein (Gp) 96, are of special interest, because they are able to take molecular peptide-fingerprints of the protein array characteristic for a particular cell. Association of Gp96 with peptides has been shown to be essential for crosspresentation and activation of T cells. Consequently, Gp96-peptide complexes extracted from cancer cells harbor the tumor-specific peptides and are immunogenic, thus offering a tool for active immunization against the tumor. Already, several immunotherapy studies of human cancers have been carried out, showing no severe adverse effects but unfortunately only limited improvement in the clinical outcome. Vitespen, a commercial HSP-peptide complex vaccine based on tumor-derived Gp96, seems to induce an improved overall survival for subsets of early stage melanoma and kidney cancer patients. The limited access to vaccine material derived from the autologous tumor requires the development of alternative protocols. Moreover, counteracting immunosuppressive mechanisms induced by the malignancy might further improve the efficacy of vaccinations. This review critically analyzes the current state of clinical immunotherapy with Gp96, with special attention to Vitespen.Heat shock proteins (HSPs) came to attention of immunologists, because they chaperone peptides and interact with antigen-presenting cells (APCs) in a specific manner. As molecular chaperones, HSPs are essential for maintaining cellular functions by preventing misfolding and aggregation of nascent polypeptides and by facilitating protein folding. This function has been described for every organism. 1In the 1980s, the potential importance of HSPs for tumor therapy emerged, when it was observed that preparations of certain HSPs isolated from cancer cells elicited immunity by capturing the antigenic fingerprint of a specific cancer, so that they could act as a vaccine against subsequent tumor challenge.2 Since then, the immunological properties of HSPs were studied intensively, particularly those of endoplasmic reticulum (ER)-resident HSPs glycoprotein (Gp)96 (Grp94)

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Targeting Heat Shock Proteins for Immunotherapy in Multiple Myeloma: Generation of Myeloma-Specific CTLs Using Dendritic Cells Pulsed with Tumor-Derived gp96

Abstract: Purpose: To develop effective immunotherapies for patients with multiple myeloma, it is important to use novel tumor antigens. Recent studies in solid tumors show that tumor-derived heat shock proteins

Cited by 58 publications

References 39 publications

Biological Heterogeneity of the Peptide-binding Motif of the 70-kDa Heat Shock Protein by Surface Plasmon Resonance Analysis

Biological Heterogeneity of the Peptide-binding Motif of the 70-kDa Heat Shock Protein by Surface Plasmon Resonance Analysis

Identification of novel HLA-A*0201-restricted epitopes from anterior gradient-2 as a tumor-associated antigen against colorectal cancer

Active‐specific immunotherapy of human cancers with the heat shock protein Gp96—revisited

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