Mobile website
Monographic Report
Changes in immune function of dendritic cells infected by recombinant adenovirus containing PSMA and 4-1BBL genes
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective : To observe changes in immune function of dendritic cells(DCs) infected by recombinant adenovirus containing human prostate specific membrane antigen (PSMA) and 4-1BB ligand [STBX〗 (4-1BBL)〖STBZ〗 genes. Methods: The empty Ad-GFP infected immature DCs at different multiplicities of infection (MOI), and the optimal MOI was determined. DCs from peripheral blood of healthy volunteers were infected with recombinant adenoviruses at the optimal MOI. The DCs were allocated into five groups: co-infected group (Ad-PSMA/4-1BBL-DC), PSMA-infected group (Ad-PSMA-DC), 4-1BBL-infected group (Ad-4-1BBL-DC), negative control group (Ad-GFP-DC) and non-infected group. Morphological changes of DCs were observed under a fluorescence microscope; and the phenotype changes of CD80, CD83, CD86 of tranfected DCs were detected by FACS, and the expressions of PSMA and 4-1BBL proteins was detected by Western blotting. IL-12 concentrations of DC supernatant in different groups were measured by ELISA. Allogenic T cell proliferations stimulated with DCs in different groups and the cytotoxic effect on prostate cancer cell lines LNCap, Du145 and 22RV were analyzed by CCK-8 assay. Results: The optimal MOI was 200. The immunophenotype expressed in the co-infected DC group, such as CD80, CD83, CD86, HLA-DR, was significantly higher than that expressed in the other DC groups (\[38.72±0.99\]%，\[44.65±0.77\]%，\[63.60±0.75\]%，\[62.25±0.58\]% vs \[28.27±104\]%，\[ 2808± 116%\]，\[41.05±1.33\]%，\[46.87±1.12\]%； P<0.05). The secretion of IL-12 in supernatant was greatly higher in the co-infected group than that in single-infected group or non-infected DC group (\[134.29±222\] vs \[ 79.51± 160\], \[70.33±1.13\], \[69.67±1.43\], \[28.88±2.97\] pg；P<0.05). At the same rate of DCs to T, the co-infected group had a more significant increase in allogenic T cell proliferation than other single-infected group or non-infected group (P<0.05). The cytotoxic rate of PSMA/4-1BBL-DC-CTL against PSMA positive target LNCap cells was significantly stronger than that against PSMA negative target cells such as DU145 and 22RV cells (P<0.05), and also higher than that of PSMA-DC-CTL and 4-1BBL-DC-CTL (P<0.05). Conclusion: Ad-PSMA/4-1BBL-infected DCs not only own high secretion of IL-12, but also stimulate and enhance the cytotoxic effect of tumor specific CTLs against PSMA positive prostate cancer cells which highly express PSMA protein. In addition, the DCs infected with Ad-PSMA and Ad-4-1BBL induce more effective tumor specific CTLs than single-infected DCs.
Keywords:
Project Supported:
Project supported by the Science and Technology Program from Science and Technology Bureau of Liaoning Province (No.2010225034)
Clc Number:
