Melanoma B16-F1 cells coated with fusion protein of mouse calreticulin and virus G-protein coupled receptor induced the antitumor immune response in Balb/C mice

Abstract

In apoptotic progress of tumor cells stimulated by special agents, the calreticulin (CRT) was relocated from endoplasmic reticulum onto the cell surface. When used as cellular antigen to immunize experimental animals, these CRT-coated apoptotic tumor cells could initiate effective anti-tumor immunoresponse against homologous tumor cells, indicating the value of CRT in anti-tumor immunotherapy. In order to develop an universal technique that could make CRT-coating more efficiently in the tumor cells, in this study, a mouse CRT recombinant gene with virus G-protein coupled receptor (vGPCR) was constructed and cloned into vector pcDNA3.1(+). When resulted plasmid pcDNA3.1(+)-mCRT/ vGPCR was stably transfected into the mouse melanoma B16-F1 cells, the mCRT-vGPCR recombinant protein was synthesized. With the membrane-locating ability of vGPCR in the recombinant protein, mCRT-vGPCR was carried onto the surface of B16-F1 cells efficiently. Overexpression of mCRT-vGPCR on the cell surface could enhance the phagocytosis of B16-F1 by macrophages in vitro. When mCRT-vGPCR coated B16-F1 cells were used as a cell-antigen to immunize mice, the specific anti-tumor immune response against the homologous tumor cells was initiated efficiently. Our data in this study may provide a new possibility for CRT-mediated tumor immune prevention and treatment.

Toxicity and Immunogenicity in Murine Melanoma following Exposure to Physical Plasma‐Derived Oxidants

“…Exogenously added CRT also potentiates the immunogenicity of melanomas in patients [43]. A CRT fusion-protein added to B16 cells evoked an antitumor immune response in mice [44]. Intriguingly, therapeutic intervention associated with upregulation of CRT involves the generation of reactive species [45–47].…”

Antitumor Immunity Triggered by Melphalan Is Potentiated by Melanoma Cell Surface–Associated Calreticulin

“…Danger signaling-potentiating therapies have been recently shown to associate with favorable clinical outcome in cancer patients (5,12,13). Moreover, it has been proposed that, combinatorial therapy with exogenously supplied danger signals could hold great immunogenicity-promoting potential (14).…”

Whole-cell vaccine coated with recombinant calreticulin enhances activation of dendritic cells and induces tumour-specific immune responses

“…lines, a method that can easily coat CRT onto the surface of any tumour cell would aid in providing a potent research tool for antitumour immunotherapy. In our previous studies, we used a B16-F1 mouse melanoma cell line coated with mCRT-vGPCR as a whole-cell tumour vaccine to immunise experimental animals and found that this whole-cell vaccine could induce a strong antitumour effect against the homologous tumour (18). In this study, we further evaluated the immune responses induced by this mCRT-vGPCR-coated whole-cell vaccine both in vivo and in vitro.…”