Studies on effect enhancement of vegetal polysaccharide on schistosoma japonicum DNA vaccine PV1223
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OBJECTIVE To investigate the immunogenicity of schistosma japonicum DNA vaecine PV1223 enhanced by likage vegetal polysaccharide. METHODS BALB/c mice were immunized subcutaneously with 10~(8) CFU PV1223 combined with tea and mushroom polysaccharides in linkage,and then challenged with schistosoma japonica cercariae on the 8th week after immunization,and killed on the 6th week of infection.The rate of reduced worm burdens,reduced egg burdens,and the examination of the IgG level were determined.RESULTS The rates of reduced worm and egg burdens were 65.96% and 80.88%,respectively.There were remarkable difference between control and immunized group.But no significant differences among the IgG titres were observed after challenge.CONCLUSION This study indicates that the vegetal polysaccharide linked with vaceine PV1223 can promote the efficacy of vaccine PV1223 by the mechanism of cellular immune response.Keywords:
Schistosoma
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Aim To study the anti-fecundity immunity effect in mice induced by Schistosoma japonicum cathepsin B DNA vaccine(Sj31BIN) Methods The DNA Vaccine of S japonicum was constructed by inserting Sj 319 one into vector VR 1012 The mice were immunized with Sj31BIN and the anti-fecundity immunity was determined by counting worms,and egg burden in liver,intestine and faeces on the 46th day after challenge infection with carcariae of S japonicum Results The immnization with Sj31BIN reduced the worm discovery and egg burden in tissues and faeces The egg reduction rate was 59 3%～65 0% in the liver,57 6%～62 1% in the intestine and 86 5% in the faeces ln addition,the number of eggs in the uteri of female adults was obviously decreased in mice immuicized with Sj31BIN,Conclusion Sj31BIN immunization induces anti-fecundity immunity in mice
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Objective: To study the effect of recombinant BCG-Sj26GST vaccine of schistosoma japonicum on IL-6 produced by spleen cells of mice. Methods: In the first experiment, mice were immunized subcutaneously by the vaccine, with PBS serving as control, challenged with Sj cercariae after 8w of immunization and killed on 6w of the infection to separate spleens; in the second experiment, after subcutaneous and intravenous immunization by the vaccine respectively, 4 mice were killed to separate spleens on 0,4,8,10,14 and 16w of immunization, spleen cells were stimulated by Sj26 or PHA, and the level of IL-6 in supernatant of spleen cells were detected by ELISA. Results: The level of IL-6 had no obvious change by immunization with the vaccine against challenge with Sj cercariae;dynamic observation showed that IL-6 reached the highest level on 8~10w of immunization. Conclusion: IL-6 may not be related to the protective immunity elicited in mice by vaccination with recombinant BCG-Sj26GST vaccine of schistosoma japonicum.
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To evaluate the potential value of the novel gene of S. japonicum SjCWL01 as a nucleic acid candidate vaccine by the construction of pcDNA3/SjCWL01 followed by observation of its immunological protection effect on mice. A novel gene of S. japonicum SjCWL01 was subcloned into the eukaryotic expression vector pcDNA3. The positive recombinants were identified by PCR and restriction enzyme digestion. Preparation of the DNA vaccine was made by transforming the plasmid pcDNA3/SjCWL01 into E.coli DH5α. The experimental mice were immunized with purified pcDNA3/SjCWL01 DNA vaccine and challenged with 40±1 S. japonicum cercariae percutaneously at the 2nd week after the third immunization. Levels of the specific antibody were detected by ELISA before infection. The reduction rates of the worm burden, liver eggs per gram and fecal eggs per gram were examined 45 days after challenge infection. Compared with the control group, the reduction rates of the worm burden, liver eggs per gram and fecal eggs per gram were 27.6%, 39.5% and 45.9% in the experimental mice, respectively. The pcDNA3/SjCWL01 nucleic acid vaccine can induce partial protective immunity against S. japonicum infection in mice.
Eggs per gram
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Objective To study the immuno-protection of rSj14-3-3 and rSj14-3-3/SjGST against Schistasoma japonicum in mice. Methods The BALB/c mice were immunized 3 times with rSj14-3-3 and rSj14-3-3/SjGST and then challenged with S. japonicum cereariae via abdominal skin after 5 days last immunization. S. japonicum adult worms and eggs were collected after 6 weeks challenged,and the worm reduction rate and egg reduction rate were calculated respectively. By indirect ELISA,the A value of specific IgG antibodies in BALB/c mice before and after immunization were estimated. The two recombinants' effection to the formation of egg granuloma in liver were observed,the average diameter of single egg granulomas was estimated under microscope. Results The worm reduction rates of rSj14-3-3 and rSj14-3-3/GST groups were 31.93% and 34.39% respectively;The egg reduction rates per gram liver tissue were 53.24% and 60.06%,and the egg reduction rates per worm pair were 33.39% and 40.48%. Before immunization,the difference of the A value of specific IgG antibodies in every mouse was not apparent,but it was apparent after immunization. Compared with control group,the average diameter of egg granulomas in liver of immunized mice reducted 35.23% and 46.13 % respectively. Conclusion Signaling protein 14-3-3 has immuno-protection not only in anti-infectious immune but also in anti-schistosomiasis. The immuno-protection of combinant polyvalency vaccine is probably better than that of monovalency vaccine.
Schistosoma
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Abstract The aim of this study was to construct and evaluate the immunity efficacy of the DNA multivalent vaccine pVIVO2SjFABP-23. The vaccine was constructed and produced as follows. Forty BALB/c mice were divided into four groups designated pVIVO2, pVIVO2Sj23, pVIVO2SjFABP and pVIVO2SjFABP-23. Each mouse was immunized with 100 μg of the corresponding plasmid DNA by intramuscular injection. 28 days post-vaccination, the mice were challenged with S. japonicum cercariae, and the worm and egg burdens were determined 42 days post-challenge. Serum samples were collected from all the mice before and after vaccination and at the end of the experiment, and used for antibody detection. The IFN-γ and IL-4 levels were quantified in the supernatants of specifically stimulated spleen cells. The number of worms was reduced by 52%, 40% and 42% in mice respectively immunized with pVIVO2SjFABP-23, pVIVO2Sj23 or pVIVO2SjFABP. A respective 61%, 38% and 39% egg reduction was determined relative to those mice that only received the empty pVIVO2 plasmid. pVIVO2SjFABP-23 immunization increased IgG levels against SWAP and SEA. Increased IFN-γ levels were detected in the supernatant of specific stimulated spleen cells from mice immunized with the 3 different constructs. The multivalent DNA vaccine developed induced higher levels of protection than the two monovalent tested vaccines.
Intramuscular injection
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Aim To investigate the immune efficacy of DNA vaccination in mice against yolk ferritin of Chinese Schistosoma japonicum.Methods Set two-series of immunization experiments.Designed beforehand infection in group Ⅰ,but not in group Ⅱ.BALB/c mice were vaccinated by each quadriceps muscle of left leg with pL-Sj Ferl encoding yolk ferritin gene of Chinese Schistosoma japonicum.Each group was immunized twice with 2wk interval. Mice vaccinated with pLXSN blank vector served as control.Mice were challenged five weeks after final DNA boosting by percutaneous infection with cercariae.Six weeks after infection the mice were perfused,worm burden and eggs in liver,dead eggs were counted.Seta from mice were collected from tail vein at weeks 3,11,17 in group Ⅰand weeks 0,6,12 in group Ⅱ,respectively.Results In groupⅠ,mice produced predominantly IgG1 and IgG3,whereas in group Ⅱ,IgG2a and IgG2b antibodies.Mice vaccinated with pL-SjFerl also produced IgA and IFN-γ.immunization with pL-SjFerl in two group could confer significant worm reduction rate(26.47%,34.08%),egg reduction rate(40.02%,36.83%),and dead egg ratio(27.50%,30.13%).Conclusion DNA vaccination with pL-SjFerl could induce protective immunity in BALB/c mice significantly.It worth further research.
Yolk
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Objective To observe the protective immunity induced by the Calpain DNA vaccine of S.japonicum Chinese strain in infected Balb/c mice. Methods Preparation of plasmid pVAC DNA and plasmid pVAC-Calpain DNA.Fifty Balb/c mice were randomly divided into two groups;each mouse of control group was injected with 10μg plasmid pVAC DNA by i.m.route;the experimental group was injected with 10μg of recombinant plasmid pVAC-Calpain DNA.Each mouse was immunized at week 1 and 3 and challenged with 25 cercariae of S.japonicum Chinese strain by abdomen skin at week 4.All mice were killed and perfused 6 weeks after challenge;the numbers of recovered worms and hepatic eggs were counted.The sera IgG antibody level of mice before immunization,two weeks after immunization and two weeks after infection were determined with ELISA. Results The worm reduction rate of experimental group is 36.84%,its female worm reduction rate is 36.36% and its eggs reduction rate is 43.31% in comparison with the control group.The IgG antibody level is rising greater in the experimental group after infection than the control group.The numbers of T lymphocytes of the control group are significant greater than that of the experimental group. Conclusion The Calpain DNA vaccine could confer partial protection against a subsequnet challenge of S.japonicum Chinese strain in Balb/c mice and might be a potential DNA vaccine.
BALB/c
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Objective To observe the protective immunity induced by the DNA vaccines of VR1012 Sj31 and VR1012 SjGST Sj31 of Schistosoma japonicum (Sj)in BALB/c mice.Methods Thirty six BALB/c mice were divided into three groups.Of control group A,each mouse was injected at quadriceps femoris with 100μg VR1012 plasmid DNA;of group B,each mouse was injected intramuscularly with 100μg VR1012 Sj31 plasmid DNA;of group C,each mouse was injected intramuscularly with 100μg VR1012 SjGST Sj31 plasmid DNA.All mice were immunized three times with an interval of 2 weeks and were challenged each with 10 cercariae of Sj at the 21th days after final immunization.At day 42 after challenge,all mice were sacrificed,the numbers of worms and hepatic eggs were counted.Antibody level in the sera of mice before and three weeks after immunization was determined with ELISA.Results There are specific IgG in the sera of mice in the test groups after the 3rd immunization.Compared with the control group,the worm reduction were 25.0% and 30.0%( P 0.05);the egg reduction were 54.90% and 69.74%( P 0.001);the egg reduction rates in the liver per female worm were 47 96% and 54 62%( P 0 001)respectively.Compared with the B group,the egg reduction of C groups were 32 92%( P 0 001),the egg reduction rates in the liver per female worm were 12 79%( P 0 05).Conclusion The DNA vaccines,VR1012 Sj31 and VR1012 SjGST Sj31 could induce partial protective immunity against Sj in mice.The protective immunity induced by bi valent DNA vaccine was better than that induced by monovalent DNA vaccine.
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To investigate the protective immunity against Schistosoma japonicum in mice immunized with recombinant specific very low density lipoprotein binding protein (SVLBP) and its potential as vaccine candidate.Recombinant SVLBP antigen was over-expressed under IPTG induction and purified by Ni-NTA affinity chromatography. C57BL/6 mice were immunized three times with purified reSVLBP complexed with Freund's adjuvant, at biweekly intervals. Then 35+/-1 cercariae of S. japonicum were given to each mouse by abdominal skin 10 days after the 3rd immunization. 45 days later, all mice were sacrificed to collect adult worms and count liver eggs. serum samples were collected before immunization and after challenge respectively, and were probed the antigen-specific antibodies using a panel of ELISAs.The worm burden and the egg deposition in liver tissue were reduced by 33.4% and 47.6% respectively in the immunized group, in comparison with the adjuvant control group (P<0.05). Higher titer (>1:6 400) of total IgG was observed after challenge infection. The vaccinated mice developed significantly higher levels of IgG2a, IgG2b, IgG1 than those of control mice.The recombinant tegumental SVLBP antigen could induce partial protection against S. japonicum infection. These data demonstrate the potential of SVLBP as a schistosome vaccine candidate.
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Objective To enhance the immunogenicity of the recombinant pVIVO2,IL12,Sj23 vaccine of Schistosoma japonicum by using mixed vegetal polysaccharides as adjuvant.Methods The plasmid pVIVO2,IL12,Sj23 was constructed.3 groups of BALB/C mice were injected intramuscularly with normal saline(Group A),pVIVO2,IL12,Sj23 plasmid DNA(B),and pVIVO2,IL12,Sj23 plus mixed vegetal polysaccharides(C) respectively,and challenged with S.japonicum cercariae on the 4th week after immunization.Mice were killed to calculate the worm reduction rate and egg reduction rate in liver tissue on the 6th week after infection.Before and 4 weeks after immunization blood samples were collected.Results The worm reduction rate and egg reduction rate were 64.3% and 79.9%,respectively in group C,45.5% and 58.4%,respectively in group B,showing a remarkable difference between them(P0.05).ELISA analysis showed a significantly higher level of IgG specific for Sj23 4 weeks after vaccination in groups B and C(P0.05).However,there was no significant difference in IgG level between groups C and B(P0.05).Conclusion When mixed vegetal polysaccharides are used as adjuvant,the effect of the vaccine pVIVO2,IL12,Sj23 can be considerably enhanced.
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