Experimental And Pre-clinical Study On Allogeneic Dendritic Cell Vaccine Of Lung Cancer

Objective: A variety of approaches have been developed to deliver tumor-associatedantigens (TAA) in conjunction with dendritic cells (DC) as cellular adjuvants. Theobjective of the study is to explore whether the allogeneic dendritic cells-pulsed withheat shocked, tumor cell lysates was able to induce antitumor T cell immunity.Methods: Heat-shocked tumor cell lysates (hTCL) was prepared from C57K~b micederived Lewis lung cancer cells (LLC) which were heated at 43℃for 1 hour/incubating at 37℃for 3 hours and then frozen-thawed. Allogeneic inbred mice(BALB/CK~d or 615K~k) bone marrow-derived DCs (Bm-DC) pulsed with LLC-hTCL(HLTCL) were used as vaccine. Expression of HSP70 was detected afterimmunoprecipitation with mAb (hsp70 Sc-24). The ability of taking up HLTCL byimmature DCs (Ira DCs) was evaluated by phagocytosis experiment in vitro and theuptake of CFSE labeled allogeneic DCs by recipient DCs was evaluated by tracingexperiment in vivo. Cytokins secretion was analyzed by ELISA. Double stained DCswas confirmed by FCM and Confocal fluorescence microscopy after co-staining withPE-conjugated mAb against mouse CD11c. The cytotoxicity of spleen cellsimmunized by Bm-DC was determined by LDH release assay. Immunological effectof allogeneic DC vaccine pulsed with HLTCL was evaluated by immunoprophylaxisin C57BL mice LLC cells transplanted model and therapeutic experiment oftumor-bearing mice of early stage with 1 day vaccination.Result:1. 43℃heating can significantly increase the expression of inducible HSP70 inLLC-TCL.2. Allogeneic mice ImDCs have full ability to take up HLTCL.3. Allogeneic mice ImDCS can take up HLTCL and subsequently promotematuration of ImDCs (enhancing expression of co-stimulatory molecules on DCsurface and its IL-12 secretion).4. In vivo subcutaneous injection of HLTCL pulsed with allogeneic DCs canenhance the uptake of allogeneic DCs by recipient (C57BL)DCs.5. Spleen cells from C57BL mice immunized with allogeneic DCs/HLTCL can specifically kill viable LLC, secrete Th-1 related cytokine (high level of INF-γandlow level of IL-4/IL-10).6. Experiment of immunoprophylaxis of LLC tumor transplant and therapy oftumor-carrying mice of early stage in C57BL mice, both confirm that immunizationwith allogeneic DCs/HLTCL can prevent or delay the growth of LLC tumor, whichelicit higher immunological effect than that with syngeneic DCs/HLTCL.Conclusion: For the first time our data indicates that immunization with allogeneicDCs/HLTCL is an alternative strategy to initiate a stronger immune responsefollowing DC vaccination for cancer therapy. Objective: To evaluate expression of CA-9, CDla, CD208 and CD3 in Stage Inon-small cell lung cancer (NSCLC), and analyze their clinical and prognosticsignificance. Methods: Expression of CA-9, HIF-1α, CDla, CD208 and CD3 weremeasured in 59 stage I NSCLCs by immunohistochemistry. The correlation betweenthese markers and the characteristics of the tumors, as well as overall survival wasanalyzed. Results: CDla, CD3 expression in adenocarcinoma were higher than insquamous carcinoma. Infiltration by CDla was significantly lower in patients withsevere tumor necrosis and with CA-9 high expression. The patients with severe tumornecrosis and with negative CD208 expression had lower overall survival thanminimal necrosis and positive expression. There wasn't overall survival differencefound between the patients with chemotherapy and without chemotherapy. Thefurther strata analysis indicated: the overall survival of patients with chemotherapywas shorter than without chemotherapy in positive CA-9 expression and in negativemature CD208 DCs infiltration. Conclusion: Infiltration of DCs and T lymphocytescan be observed in early-stage NSCLCs. Tumor hypoxia, histopathology and theextent of tumor necrosis perhaps affect the infiltration of DCs and T cells. The extentof tumor necrosis and mature CD208 DCs infiltration were strongly correlated withoverall survival. The expression of CA-9 and CD208 perhaps play an instructive rolein postoperative therapy of NSCLCs especially in chemotherapy.