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Immunotherapeutic effect of Ag85A DNA vaccine and Ag85B DNA vaccine on bladder tumor in rats
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Abstract:
To explore the immunotherapeutic effect of Ag85A DNA vaccine and Ag85B DNA vaccine in treatment of bladder tumor in rats. Methods: The bladders of female Wistar rats were irrigated with carcinogen methyl nitrosourea (MNU) to create the model of bladder tumor. Totally 48 model rats were evenly randomized into 6 groups: normal saline (NS), pcDNA3.1, bacille Calmette Guérin(BCG), Ag85A DNA vaccine, Ag85B DNA vaccine and Ag85A+ Ag85B DNA vaccine groups. The corresponding drugs were injected into the right hind limbs of rats intramuscularly on day 7, 14, and 21 after model establishment. Animals were sacrificed on day 28, and the spleens were removed aseptically. The percentages of CD4 +T cells and CD8 +T cells in splenocytes were measured by flow cytometry and the value of CD4 +/CD8 + was calculated. Level of serum IFN γ was assayed by ELISA and pathological examination was done. Results: The rat model of bladder tumor was successfully constructed, with a tumorigenesis rate of 100%. The bladder tumor sizes in Ag85A DNA vaccine group, Ag85B DNA vaccine group and Ag85A+ Ag85B DNA vaccine group were decreased after treatment, and the tumor pathological grades were also improved, but the outcomes were not better than those of the BCG group. The percentages of CD4 +T cells in the 4 groups were (17.27±2.95)%， (23.15±156)%， (3080±183)%， (38.05±1.48)%，respectively; and the percentages of CD8 +T cells were (9.03±106)%, (1028±039)%, (11.29±0.74)%, (1314±1.24)%, respectively; the relative values of CD4 +/CD8 + were 1.90±0.10, 2.25±0.08, 2.73±0.19, 297±0.23, respectively; the production of IFN γ in Ag85A DNA vaccine group, Ag85B DNA vaccine group, Ag85A+ Ag85B DNA vaccine group, BCG group were (96.94±1238), (131.03±26.68) , (179.20±28.88)，(240.53±32.17) pg/ml, respectively. The above 4 parameters in the Ag85A DNA vaccine group, Ag85B DNA vaccine group and Ag85A+ Ag85B DNA vaccine group were obviously improved compared with the pcDNA3.1 and NS groups, but were still poorer than those of the BCG group ( P<0.01). The effects of Ag85A+ Ag85B DNA vaccine group were better than those of the Ag85A and Ag85B DNA vaccine group( P< 0.05). Conclusion: Ag85A DNA vaccine and Ag85B DNA vaccine can improve the immune response of rats with bladder tumor; a combination of both has better outcome than they are used alone, but even the combination can not reach the effect of BCG.
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Project Supported:
Supported by the Natural Science Foundation of Shanxi Province (NO.20051099)
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