ABSTRACT
Increasing evidence demonstrates that interleukin-10 (IL-10), known as an immunosuppressive cytokine, induces antitumor effects depending on CD8+ T cells. However, it remains elusive how immunosuppressive effects of IL-10 contribute to CD8+ T cell-mediated antitumor immunity. We generated Cetuximab-based IL-10 fusion protein (CmAb-(IL10)2) to prolong its half-life and allow tumor-targeted delivery of IL-10. Our results demonstrated potent antitumor effects of CmAb-(IL10)2 with reduced toxicity. Moreover, we revealed a mechanism of CmAb-(IL10)2 preventing dendritic cell (DC)-mediated CD8+ tumor-infiltrating lymphocyte apoptosis through regulating IFN-γ production. When combined with immune checkpoint blockade, CmAb-(IL10)2 significantly improves antitumor effects in mice with advanced tumors. Our findings reveal a DC-regulating role of IL-10 to potentiate CD8+ T cell-mediated antitumor immunity and provide a potential strategy to improve cancer immunotherapy.
Subject(s)
Antineoplastic Agents, Immunological/pharmacology , Apoptosis/drug effects , CD8-Positive T-Lymphocytes/drug effects , Cetuximab/pharmacology , Dendritic Cells/drug effects , Interleukin-10/pharmacology , Lymphocytes, Tumor-Infiltrating/drug effects , Neoplasms/drug therapy , Animals , Antineoplastic Agents, Immunological/pharmacokinetics , CD8-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/metabolism , CD8-Positive T-Lymphocytes/pathology , Cell Communication , Cell Line, Tumor , Cell Proliferation/drug effects , Cetuximab/pharmacokinetics , Coculture Techniques , Dendritic Cells/immunology , Dendritic Cells/metabolism , Dendritic Cells/pathology , Drug Resistance, Neoplasm , Female , Humans , Interleukin-10/pharmacokinetics , Lymphocytes, Tumor-Infiltrating/immunology , Lymphocytes, Tumor-Infiltrating/metabolism , Lymphocytes, Tumor-Infiltrating/pathology , Mice, Inbred C57BL , Mice, Inbred NOD , Mice, Knockout , Mice, SCID , Molecular Targeted Therapy , Neoplasms/immunology , Neoplasms/metabolism , Neoplasms/pathology , Recombinant Fusion Proteins/pharmacology , Signal Transduction , Tumor Burden/drug effects , Xenograft Model Antitumor AssaysABSTRACT
BACKGROUND: Programmed cell death ligand-1 (PD-L1) with its receptor PD-1 pathway is overactivated in many tumors. Inhibiting the interaction of PD-L1 and PD-1 is an attractive strategy to restore tumor-specific T cell immunity for tumor therapy. METHODS: A fully human anti-PD-L1 monoclonal antibody (mAb) B60-55 was identified by yeast surface display. The affinity, specificity, activity, and efficacy of mAb B60-55 were investigated in vitro or in vivo. RESULTS: mAb B60-55 (purity >99%) could bind to PD-L1 that is expressed on HEK293 cells with a dissociation constant of 0.2 nM, and specifically bind to human or cynomolgus macaque PD-L1 without a cross-reaction with murine PD-L1. Moreover, mAb B60-55 is an antagonistic antibody, which can block PD-L1 binding to its receptors, including PD-1 (PDCD1) and B7.1 (CD80). In vitro assays demonstrated the ability of mAb B60-55 to enhance T cell responses and cytokine production in the mixed lymphocyte reaction. In vivo studies showed that administration of mAb B60-55 exhibited a potent antitumor activity toward tumor cell carcinoma xenograft, with a mean half-life of 177.9h in cynomolgus monkeys. CONCLUSION: mAb B60-55 is a potential candidate for clinical development in cancer treatment.
Subject(s)
Antibodies, Blocking/immunology , Antibodies, Monoclonal/immunology , Antineoplastic Agents/immunology , B7-H1 Antigen/immunology , Lymphocyte Activation/drug effects , T-Lymphocytes/drug effects , Animals , Antibodies, Blocking/isolation & purification , Antibodies, Monoclonal/isolation & purification , Antineoplastic Agents/isolation & purification , Cell Proliferation/drug effects , Cross Reactions , Cytokines/metabolism , Female , HEK293 Cells , Humans , Lymphocyte Culture Test, Mixed , Macaca fascicularis , Mice , Mice, Inbred C57BL , Programmed Cell Death 1 Receptor/antagonists & inhibitors , Protein Binding/drug effects , T-Lymphocytes/immunology , Xenograft Model Antitumor AssaysSubject(s)
MicroRNAs/metabolism , Motor Activity/genetics , Peripheral Nervous System , Actins/metabolism , Age Factors , Animals , Animals, Genetically Modified , Animals, Newborn , Drosophila , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Embryo, Nonmammalian , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Horseradish Peroxidase/metabolism , MicroRNAs/genetics , Neuromuscular Junction/embryology , Neuromuscular Junction/growth & development , Neuromuscular Junction/metabolism , Peripheral Nervous System/embryology , Peripheral Nervous System/growth & development , Peripheral Nervous System/metabolism , Tumor Suppressor Proteins/genetics , Tumor Suppressor Proteins/metabolismABSTRACT
Chromosomal rearrangement (CR) events result from abnormal breaking and rejoining of the DNA molecules, or from crossing-over between repetitive DNA sequences, and they are involved in many tumor and non-tumor diseases. Investigations of disease-associated CR events can not only lead to important discoveries about DNA breakage and repair mechanisms, but also offer important clues about the pathologic causes and the diagnostic/therapeutic targets of these diseases. We have developed a database of Chromosomal Rearrangements In Diseases (dbCRID, http://dbCRID.biolead.org), a comprehensive database of human CR events and their associated diseases. For each reported CR event, dbCRID documents the type of the event, the disease or symptoms associated, and--when possible--detailed information about the CR event including precise breakpoint positions, junction sequences, genes and gene regions disrupted and experimental techniques applied to discover/analyze the CR event. With 2643 records of disease-associated CR events curated from 1172 original studies, dbCRID is a comprehensive and dynamic resource useful for studying DNA breakage and repair mechanisms, and for analyzing the genetic basis of human tumor and non-tumor diseases.
Subject(s)
Chromosome Aberrations , Databases, Factual , Disease/genetics , Chromosome Breakpoints , Humans , Neoplasms/genetics , User-Computer InterfaceABSTRACT
MicroRNAs (miRNAs) are a kind of endogenous non-coding small RNAs whose specific functions in animals are generally important. Although functions of some miRNAs have been identified, the role of miR-184 remains unknown. Here, we determined the temporal and spatial expression pattern of miR-184 during the different development stages and tissues in Drosophila. Strikingly, miR-184 is expressed ubiquitously in Drosophila embryos, larvae and adults, its expression pattern shows a dynamic changes during the development of embryo, especially in the central nervous system. This expression profile suggests that miR-184 may act important function in Drosophila development.
