Effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the anti-fertility efficacy of canine zona pellucida glycoprotein 3 (CZP3) DNA vaccine / 中华微生物学和免疫学杂志

ABSTRACT

Objective To investigate the effects of GM-CSF (granulocyte-macrophage colony-stimulating factor) as an immune adjuvant on the anti-fertility efficacy of canine zona pellucida glycoprotein 3 (CZP3) DNA vaccine.Methods RT-PCR and quantitative real-time PCR were performed to analyze the effects of GM-CSF on the maturation of antigen presenting cells (APCs) and the enrichment of APCs at injection site.Female BALB/c mice were immunized with CZP3 DNA vaccine pcDNA3-CZP3 alone or in combination with genetic adjuvant pcDNA3-GM-CSF by using the method of electrical impulses.ELISA (enzyme-linked immunosorbent assay) was used to detect the levels of IgG and sIgA (secretory IgA) antibodies in serum samples and genital tracts and the levels of IL-4 and IFN-γ in serum samples.MTT method was used to analyze the proliferation of spleen T cells in mice.The binding ability of serum anti-CZP3 antibody to native mouse oocytes was analyzed by indirect immunofluorescence assay.All of the female BALB/c mice were coupled with male mice of the same age six weeks after the first-dose vaccination.Litter size at birth in each group was counted and the differences between different groups were comparatively analyzed.ResultsImmunizing the mice with pcDNA3-CZP3 and GM-CSF significantly promoted the expression of CD80, CD83 and CD86 (P<0.05) and increased the sIgA antibody level in genital tract and IgG level in serum (P<0.01).Moreover, the levels of IL-4 and IFN-γ in serum samples were significantly up-regulated (P<0.05) and the proliferation of spleen T cells was significantly enhanced (P<0.05).Results of the indirect immunofluorescence assay showed that the fluorescence intensity and density on mouse egg surface were positively correlated with the level of antibody in serum.Results of the anti-fertility test suggested that GM-CSF significantly reduced the litter size in mice immunized with pcDNA3-CZP3 vaccine (P<0.05).Conclusion GM-CSF could be used as an effective adjuvant to enhance the anti-fertility efficacy of CZP3 DNA vaccine.