Deoxypodophyllotoxin Induces a Th1 Response and Enhances the Antitumor Efficacy of a Dendritic Cell-based Vaccine

BACKGROUND: Dendritic cell (DC)-based vaccines are currently being evaluated as a novel strategy for tumor vaccination and immunotherapy. However, inducing long-term regression in established tumor-implanted mice is difficult. Here, we show that deoxypohophyllotoxin (DPT) induces maturation and activation of bone marrow-derived DCs via Toll-like receptor (TLR) 4 activation of MAPK and NF-kappaB. METHODS: The phenotypic and functional maturation of DPT-treated DCs was assessed by flow cytometric analysis and cytokine production, respectively. DPT-treated DCs was also used for mixed leukocyte reaction to evaluate T cell-priming capacity and for tumor regression against melanoma. RESULTS: DPT promoted the activation of CD8+ T cells and the Th1 immune response by inducing IL-12 production in DCs. In a B16F10 melanoma-implanted mouse model, we demonstrated that DPT-treated DCs (DPT-DCs) enhance immune priming and regression of an established tumor in vivo. Furthermore, migration of DPT-DCs to the draining lymph nodes was induced via CCR7 upregulation. Mice that received DPT-DCs displayed enhanced antitumor therapeutic efficacy, which was associated with increased IFN-gamma production and induction of cytotoxic T lymphocyte activity. CONCLUSION: These findings strongly suggest that the adjuvant effect of DPT in DC vaccination is associated with the polarization of T effector cells toward a Th1 phenotype and provides a potential therapeutic antitumor immunity.

In vivo CTL Activity Induced by Prime-boost Vaccination using Recombinant Vaccinia Virus and DNA Vaccine Expressing Epitope Specific for CD8+ T Cells

DNA vaccine approaches have been applied to generate the protective immunity against various pathogens. However, the strength of immune responses induced by DNA vaccine is weak compared with conventional vaccines. The primeboost vaccination using DNA vaccine and other viral vector has been suggested as one way to circumvent this limitation. In the present study, we used in vivo CTL activity assay to determine CD8+ T cell-mediated immunity induced by prime-boost vaccination with a DNA vaccine (gB498-505 DNA) and recombinant vaccinia virus (VVgB498-505) expressing gB498-505 epitope peptide (SSIEFARL) of herpes simplex virus type 1 (HSV-1) glycoprotein B (gB). The most potent in vivo CTL activity was induced in mice received VVgB498-505 when both gB498-505 and VVgB498-505 were used at priming step and boosted with the alternative vaccine vector expressing whole antigen protein (gBw). Priming with vaccine vector expressing gBw followed by the use of VVgB498-505 at boosting step also induced strong in vivo CTL activity. We also examined in vivo CTL activity after immunization of mice with epitope-expressing vaccine vector at both priming and boosting step. Curiously, in vivo CTL activity mediated by CD8+ T cells was strongly elicited at memory stage when animals were primed with VVgB498-505 and subsequently boosted with gB498-505 DNA. Because the use of VVgB498-505 at priming followed by boosting with gB498-505 DNA induced most optimal immunity, these results suggest that the order of vaccine type should be carefully considered when used vaccine type expressing only epitope for prime-boost vaccination.

Expression of recombinant human MUC1-MBP fusion protein in E coli DH5?,and its purification and detection of immune responses in mice / 中国免疫学杂志

Objective:To study MUC1 based cancer vaccine.Methods:MUC1 gene was inserted into pMAL-p2 vector and constructed recombinant pMAL-MUC1. MUC1-MBP fusion protein expression was induced by IPTG in E coli DH5? transformed by the recombinant pMAL-MUC1 .The fusion protein was analyzed by Western blot and purified by amylose affinity chromatography. The antiserum,T cell proliferation and CTL activity of spleen from C57 mice immunized by MUC1-MBP were determined respectively by ELISA,adding 3H-TdR and MTT.Results:Had successfully constructed pMAL-MUC1 expression vector,and purified MUC1-MBP and MBP. C57 mice immunized by MUC-MBP generated MUC1 specific antibody and CTL.The titer of polyclonal antibody to MUC1 was about 1∶5 760?3 221. CTL cytotoxicity to the MCF7 and lewis lung cancer cells respectively were at 47.7%?4.3% and 67.5%?6.5%.Conclusion:Human recombinant MUC1-MBP fusion protein activated T and B cell response in mice.The results suggested that the recombinant Muc1 may be used to develop protein vaccine against carcinoma.

The Anti-Tumor Effect of Recombinant Human MUC1-MBP Fusion Protein / 中国肿瘤生物治疗杂志

Objective: To study the anti-tumor effect of recombinant human MUC1-MBP. Methods: The C57BL/6 mice were in oculated with MUC1-MBP by subcutaneous. MUC1 specific CTL activity of spleen were determined by MTT; The effects on prevention and treatment of tumor were observed by establishing lewis lung cancer-carrying mice. Results: The cytotoxicity of CTL from immunized mice to the MCF7 and Lewis lung cancer cells respectively was (47.7?4.3) % and (67.5 ?6.5) %; 5?10 5 lewis lung cancer cells following immunization were injected iv into C57BL/6 mice, after three weeks, the number of lung and tail tumor colonies was 51 and 5 for PBS and MUC1-MBP groups respectively and the suvival time was significantly delayed in immunized mice. The average volume of tumors in mice with MUC1-MBP was 386 mm 3 wherea control group was 4 000 mm 3 at tumor treating experiment. Conclusions: Recombinant human MUC1-MBP have significantly effects on prevention, treatment and inhibiting metastases of tumor. Our results suggested that the recombinant MUC1-MBP might be used to develop protein vaccine against human carcinoma.

The study of antitumor effect of tumor-draining lymph node cells from lung cancer patients induced by different stimulus in vitro / 中国免疫学杂志

Objective:To find a feasible method to stimulate tumor-draining lymph node(TDLN) cells in clinic.Methods:CTL activity of TDLN cells induced by different stimulus (IL-2 group, IL-2+autologous tumor antigen group, IL-2+GM-CSF+IL-4+autologous tumor antigen group) was measured by the method of maximal LDH enzyme release. The mechanisms were explored by observation in morphology and detection of the CD83 positive rate of TDLN cells.Results:The level of growth of TDLN cells induced by (IL-2+GM-CSF+IL-4+autologous tumor antigen) was significantly higher than TDLN cells induced by IL-2 and (IL-2+autologous tumor antigen)(P

CTL activity detection of immune response to hepatitis Cviral nucleic acid vaccine / 中国免疫学杂志

Objective:To discuss mice cellular immune response to the hepatitis C viral nucleic acid vaccine.Methods:Hepatitis C viral nucleic acid vaccine pSVL HCV/C+E 1 was inoculated BALB/C mice by subcutaneous injection.To detect the CTL activity of immune mice,Made the standard 4 hours lysis test by labeling the target cells with (Na) 2 51 CrO 4,which was prepared from the syngentic BALB/C mouse myeloma derived cell strain SP2/0 transfected by the eukaryotic expression recombinant plasmid pEGFP N 1 HCV/C+E 1.Results:The kill ratio of CTL is up to 40.7%.Conclusion:The result suggested hepatitis C viral nucleic acid vaccine could induce strong cellular immune response