title: Calreticulin acts as an adjuvant to promote dendritic cell maturation and enhances antigen-specific cytotoxic T lymphocyte responses against non-small cell lung cancer cells.
abstract: Dendritic cell (DC)-based immunotherapy has promising for treatment of non-small cell lung cancer (NSCLC). Melanoma-associated antigen 3 (MAGE-A3) is a tumor-specific antigen and expressed in approximately 35-40% of NSCLC tissues. Calreticulin (CALR) is a protein chaperone and can enhance DC maturation and antigen presentation. In this study, we evaluated the adjuvant activity of CALR in human DC maturation and their capacity to induce MAGE-A3-specific CD8+ cytotoxic T lymphocyte (CTL) responses to NSCLC in vitro. Infection with recombinant Ad-CALR and/or Ad-MAGE-A3, but not with control Ads, induced CALR and/or MAGE-A3 expression in DCs. Infection with Ad-CALR significantly increased the percentages of CD80+, CD83+, CD86+ and HLA-DR+ DCs and IL-12 secretion, but reduced IL-10 production in DCs. Co-culture of autologous lymphocytes with DC-Ad-CALR or DC-Ad-CM significantly increased the numbers of induced CD8+ CTLs. The percentages of IFNγ-secreting CTLs responding to SK-LU-1 and NCI-H522 NSCLC, but not to non-tumor NL-20 cells in Ad-C-CTL, Ad-M-CTL and Ad-CM-CTL were significantly higher than that of DC-CTL and Ad-null-CTL. Ad-C-CTL, Ad-M-CTL and Ad-CM-CTL, but not control DC-CTL and Ad-null-CTL, induced higher frequency of MAGE-A3+HLA-A2+ NCI-H-522 cell apoptosis, but did not affect the survival of MAGE-A3+HLA-A2- SK-LU-1 and non-tumor NL20 cells in vitro. Treatment with anti-HLA-I antibody, but not with anti-HLA-II, dramatically diminished the cytotoxicity of Ad-CM-CTLs against NCI-H522 cells. Our data indicated that CALR acted as an adjuvant to promote DC maturation, which induced CTL development and enhanced MAGE-A3-specific CTL cytotoxicity against NSCLC.
authors.affiliation: Department of Thoracic Surgery, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China; Department of Medical Oncology, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China. Electronic address: cinlylau@hotmail.com.
authors.forename: Xinli
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Thoracic Surgery, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China. Electronic address: lijijia1982@163.com.
authors.forename: Jijia
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Thoracic Surgery, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China. Electronic address: liuyu1984@126.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Thoracic Surgery, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China. Electronic address: djq1988@126.com.
authors.forename: Jianqiao
authors.initials: J
authors.lastname: Ding
authors.affiliation: Department of Thoracic Surgery, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China. Electronic address: 910397562@qq.com.
authors.forename: Zhuang
authors.initials: Z
authors.lastname: Tong
authors.affiliation: Department of Medical Oncology, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China. Electronic address: lyylegend@hotmail.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Thoracic Surgery, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China. Electronic address: janezy1987@163.com.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Thoracic Surgery, Liaoning Cancer Hospital & Institute, Clinical Oncology College of Dalian Medical University, Shenyang 110042, China. Electronic address: lyymot@hotmail.com.
authors.forename: Yongyu
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000328:Adult; D017951:Antigen Presentation / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology; D015153:Blotting, Western; D037282:Calreticulin / Q000276:immunology*; D002289:Carcinoma, Non-Small-Cell Lung / Q000276:immunology*; D002454:Cell Differentiation / Q000276:immunology; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D058501:Enzyme-Linked Immunospot Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D008175:Lung Neoplasms / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology; D009363:Neoplasm Proteins / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D055815:Young Adult
chemical_list: D000276:Adjuvants, Immunologic; D000951:Antigens, Neoplasm; C576483:CALR protein, human; D037282:Calreticulin; C072013:MAGEA3 protein, human; D009363:Neoplasm Proteins
keywords: Adjuvant; Calreticulin; Cytotoxic T lymphocyte; Dendritic cells; MAGE-A3; Non-small cell lung cancer