Establishment a Carcinoembryonic Antigen Stable Cell Line for Cancer Immunotherapy Study

Abstract

Introduction:Cancer immunotherapy is one of the effective treatment methods that provide a better quality of life with limited side effects for patients. Carcinoembryonic Antigen (CEA) can be an appropriate target for cancer immunotherapy. Methods: A lentivector expressing CEA antigen, pCDH-CEA, was constructed by cloning CEA cDNA downstream of the CMV promoter. The constructed plasmid was co-transfected with helper plasmids, into Lenti-X 293 T cells. The lentivector-containing supernatant was collected. Titers of the CEA-lentivector were estimated using the RT-PCR method. The CT26 cells were then infected by CEA-lentivector. Puromycin as a selective antibiotic was added to the culture for 2 weeks to select CEA-positive cells. The ability to produce tumors in BALB/c mice was investigated. Results: The results showed that CEA expressing lentivector plasmids and the two other helper plasmids could be transfected into Lenti-X 293T cells efficiently and packaged successfully as a pseudo-lentivector. The detection of CEA mRNA and protein expression in the 6th and 14th passages of CT26-CEA cells was confirmed in the engineered stable cell line. Tumor formation was confirmed in cell inoculated mice. Conclusion: CT26-CEA cell line with stable expression of CEA can be used as a suitable tumor model to facilitate research on colorectal cancer in vitro and in mice models; therefore, it could be served as a valuable tool for cancer immunotherapy.

Oncolytic reovirus enhances the effect of CEA immunotherapy when combined with PD1-PDL1 inhibitor by reducing the immunosuppressive tumor microenvironment in a colorectal cancer model

“…The stable expression of CEA was con rmed in different passage numbers of the cells. The ability of cells to establish tumors in mice was con rmed with GPF imaging of tumorized mice as described previously (21).…”