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ngthen cellular immunity, especially Th1-type immune response to HPV11-E7 DNA vaccine in mice.
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Objective To obtain the specific IgE antibody related epitopes of Schistosoma japonicum from the phage display library. Methods Serum samples from 150 individials living in the epidemic regions of Schistosomiasis japonica were detected by ABC ELISA. 15 samples with high titer specific IgE antibodies were selected. Their pooled sera were absorbed with Protein G Sepharose beads to remove the IgG antibodies,then,it was used for immunoscreening of a phage display library of random peptide 12 mers. After 5 cycles of screening,DNA samples from 35 phage clones were sequenced. The phage clones with different inserted epitopes were identified immunologically. Results 4 independent phage clones of phage 3,phage 6,phage 8 and phage 15 were determined. Western blotting analysis showed that all of them could be recognized by specific IgE antibodies from the pooled sera. When they were used to immunize BALB/c mice,each clone could cause significant specific IgE antibody response. Conclusions The specific IgE antibody related epitopes of Schistosoma japonicum were screened successfully from the phage display library.
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Objective To identify the immunogenicity and the potential of using Schistosoma japonicum mitochondria related protein(rSj338) as a candidate vaccine antigen for Schistosomiasis japonica . Methods The expressed recombinant fusion protein(rSj338/26GST) was purified and recognized by S.japonicum heavily infected rabbit sera and rSj338/26GST immunized rabbit sera by Western blotting. The inclusion bodies,the revivified protein and the purified protein from S 12 gel filtration were injected twice into rabbits to produce anti rSj338/26GST antibody which titer was then measured by dot ELISA. Balb/c mice were immunized with the inclusion bodies and the purified protein from S 12 gel filtration and challenged with S. japonicum cercariae to identify the protective immunity produced by rSj338/26GST. Results The inclusion bodies, the revivified protein and the purified protein from S 12 gel filtration could stimulate the rabbits to produce high level of anti rSj338/26GST antibodies and could be recognized by S. japonicum heavily infected rabbit sera and rSj338/26GST immunized rabbit sera. In Balb/c mice, the inclusion bodies could induce 27.8%( P
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Objective To construct and express human papillomavirus type 11(HPV11) E7 gene with recombinant adenovirus vectors. Methods HPV11 E7 gene was amplified by PCR and directionally cloned into vector pENTR-TOPO to form TOPO-E7 plasmid. E7 gene was transferred into the pAD/CMV/V5-DESTTM gateway vector by LR recombination reaction with pAD/CMV/V5-DESTTM gateway vectors and TOPO-E7 plasmid. The recombination vector was digested by Pac I enzyme and transfected into 293A cell by Lipofectamine method to obtain recombinant adenovirus vectors pAD-E7. Expression of E7 on HaCaT cells infected with pAD-E7 vectors was analyzed by confocal microscopy. Results The recombinant plasmid TOPO-E7 was identified and confirmed with enzyme digestion and sequencing. Recombinant adenovirus vectors pAD-E7 were generated efficiently with a titer of 1.4 ? 107 pfu/mL in transfected 293A cells. E7 protein could be identified in HaCaT cells with confocal microscope 48 h after infected with recombinant adenovirus vector. Conclusions The results indicate efficient expression of HPV11 E7 gene in eukaryotic cells by recombinant adenovirus mediated transfer, which facilitates further research of its function.
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In this study, the recombinant plasmid pPFl4 labeled with photobiotin was used as a probe to detect the patients infected with Plasmodium falciparum ( P. f.) by dothybridiza-tion. The results showed that out of 35 cases with P. f., 29 were positive, 5 were negative and one was doubtful. One patient with P.f. and P. vivax mixed infection showed positive result. The total positive rate was 83. 3% (30/36). 3 out of 33 normal human blood samples were positive, so the false positive rate was 9%. In addition, there was a correlation between the positive rate of detection and parasitaemia level. The detection sensitivity was 5 ?10-5.
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A 21-base, Plasmodium falciparum specific, enzyme-conjugated synthetic DNA probe (PFRl-AP) was used for the diagnosis of falciparum malaria. The blood samples (53 P. falciparum, 5 P. vivax, and 3 P. f and P.v mixed infection cases ) collected from Hainan province were tested. The samples of 32 college students were used for normal control. The probe proved to be specific and sensitive. 10pg of purified P.f DNA could be always detected, and there was no cross reaction with the purified DNA of human leukocytes. When testing Hainan blood specimens, PFRl-AP specifically detected P.f infections. In dot blot, when Nytran membrane with 50 microliters of treated blood samples being used, 39 out of 52 P.f specimens hybridized with this probe positively. When the volume of blotted sample was increased to one hundred microliters, the accumulative total positive rate rose up to 88.46%. The samples of P.v and normal control showed negative reaction with this probe.