| Vaccination is one of the main apparoaches to the control of animal infectious diseases.However,intensive breeding and stress in the modern livestock industry usually impair the immune system and decrease the immune response of animals,resulting in the failure of immune system in response to vaccination.The present study was to evaluate the effects of oral administration astragalus polysaccharides(APS)and ginseng stem-leaf saponins(GSLS)on the immune response to foot-and-mouth(FMD)vaccine in a mouse model.Because traditional Chinese medicine is always orally administrated and the drugs act directly on the digestive tract after oral adminitration,the effect of APS and GSLS on the intestinal mucosal immunity were also investigated.The objectives of this study was to provide a basis for improvement of the animal immunity using herb-originated polysaccharides and saponins,and preliminarily discuss the mechanism of ASP and GSLS in immune enhancement.1 The effect of oral administration of APS on immunization with FMD vaccineTo investigate the effect of oral administration of APS on the mice immunized with FMD vaccine,we randomly divided 24 mice into four groups with 6 mice in each.Group 1 was treated with saline as a control.Mice of groups 2 to 4 were orally administrated solution containing APS 25,50 or 100 mg for 4 days.At 24 hour after final treatment,all mice received first subcutaneous injections of FMDV vaccine(OS/99+OZK/93)and boost immunization at two-week intervals.Blood samples were weekly collected in 5 weeks after the boost immunization for analysis of IgG and the IgG subclasses.Results showed that the serum level of IgG,IgG subtypes in APS group was significantly increased compared with control group during 3 weeks of post boost immunization(P<0.05),suggesting that the immune response was improved by oral administration of APS on FMD vaccine.2 The effect of oral administration of APS on splenocytes proliferation and intestinal mucosal immuneTo investigate the effect of oral administration APS on splenic lymphocytes proliferation and intestinal mucosal immunity,40 mice were randomly divided into two groups and orally administrated saline solution(control)or APS(25 mg)solution for 4 days.On 0,1,2,3 weeks after the treatment,5 mice were sacrificed in each group,splenocytes were prepared for lymphocyte proliferation test and intestinal tissues were sampled for immunohistochemistry test of IgA positive cells and intestinal intraepithelial lymphocytes(IELs).Results indicated that the stimulation index(SI)was significantly higher in APS group than the control group within2 weeks(P<0.05).Moreover,the numbers of IELs and IgA positive cells in mice administered GSLS were significantly higher than the control during 0 to 2 weeks after APS administration(P<0.05).The results suggested that oral administration of APS activated cellular and intestinal mucosal immune responses.3 The effect of oral administration of GSLS on immunization of mice with FMD vaccineTo investigatethe effect of oral administration of GSLS on mice immunized with FMD vaccine,24 mice were randomly divided into four groups with 6 mice in each.Group 1 was treated with saline solution as a control.Mice of groups 2 to 4 were orally administrated solution containing GSLS 0.05,0.5 or 5 mg for 4 days.At 24 hour after final treatment,all mice received first subcutaneous injections of FMDV vaccine(OS/99+OZK/93)and boost immunization at two-week intervals.Blood samples were weekly collected within 5 weeks after the booster immunization for analysis of IgG and the IgG subclasses.Results showed that the serum IgG and the IgG subtypes were significantly increased in GSLS group compared to the control group during the first 3 weeks post boost immunization.The peak value was obversed at week 3(P<0.05).Therefore,oral administration of GSLS significantly increased the immune response induced by FMD vaccine.4 The effect of oral administration of GSLS on splenocytes proliferation and intestinal mucosal immuneTo investigate the effect of oral administration of GSLS on spleen lymphocytes proliferation and intestinal mucosal immunity,40 mice were randomly divided into two groups and orally administrated saline solution or GSLS(0.5 mg)solution for 4 days.On 0,1,2,3 weeks after the treatment,5 mice were sacrificed in each group,splenocytes were prepared for lymphocyte proliferation test and intestinal tissues were sampled for immunohistochemistry test of IELs and IgA positive cells.Results showed that the stimulation index(SI)in GSLS group was significantly higher than the control group during 1 to 2 weeks after administration APS(P<0.05).Moreover,the numbers of IELs and IgA positive cells in mice administered GSLS were significantly higher than the control during 0 to 2 weeks after GSLS administration(P<0.05).The results suggested that oral administration of GSLS activated cellular and intestinal mucosal immune responses.5 Comparison of APS and GSLS for their ability to increase the response to FMD vaccineTo compare APS and GSLS for their ability to increase the response to FMD vaccine,40 mice were randomly divided into four groups with 6 mice in each.Groups 1 to 3 were received administration of saline solution,containing GSLS(0.5 mg)solution or containing APS(25 mg)solution for 4 days.Group 4 was no treatment and no immunization as negative control.At 24 hour after final treatment,all mice except for group 4 received first subcutaneous injections of FMDV vaccine(O/My98/XJ/2010+0/GX/09-7)and boost immunization at two-week intervals.Blood samples were collected at weeks 1 and 2 after the boost for analysis of serum IgG,IgG titer,IFN-γ.Additionally,mice were sacrificed at 3 week after boost immunization and the duodenum tissue were been sampled for measure mRNA expression by RT-PCR.The results showed that oral administration of APS and GSLS significantly increased the level of serum specific-IgG,IgG subtypes and IgG titer.The expression of mucosal chemokines(CCL25 and CCL28)and receptors(CCR9 and CCR10)mRNA were significantly up-regulated compare with control group.Those results showed that oral administration APS and GSLS enhanced the intestinal mucosal immunity through up-regulation expression of mucosal chemokines and receptors mRNA,CD80,CD86,MHC-Ⅱ,TLR-4 and NF-κB p65 mRNA in peyer’s patches.It hinted that oral administration APS and GSLS promoted dendritic cells mature,improve antigen presentation ability and activated immune cells(T and B lymphocyte),resulted in enhancement of intestinal mucosal immunity and adaptive immunity.The results of this paper showed that oral administration of APS and GSLS significantly increased the level of serum specific IgG,IgG subtypes and IgG titer,promoted splenocyte proliferation response stimulation with ConA and LPS.The immunohistochemistry staining examination showed that the mucosal immunity was enhanced on account of increase number of intraepithelial lymphocyte and IgA positive cells.The results of RT-PCR hinted that enhancement of mucosal immunity by oral administration of APS and GSLS was associated with up-regulation expression of intestinal mucosal some relevant immune genes.To sum up,APS and GSLS had oral adjuvant,served as new tradition medicine immunopotentiator improvement vaccination efficacy and prevent intestinal pathogen infection deserved further study. |
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