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Research on CP2/P30 Bivalent DNA Vaccine and Subunit Vaccine Against Cryptosporidium

Author: LiChunYu
Tutor: ZhangXiChen
School: Jilin University
Course: Preventive Veterinary Medicine
Keywords: Cryptosporidium parvum DNA vaccine Subunit vaccine
CLC: R392
Type: Master's thesis
Year: 2010
Downloads: 60
Quote: 6
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Abstract

Cryptosporidium (Cryptosporidium) is a zoonotic parasitic protozoa, is not only one of the important pathogens of humans and animals diarrhea, with the occurrence of AIDS epidemic. Mainly parasitic in the epithelial cells of the gastrointestinal tract, poultry respiratory mucosa of humans and other mammals, which caused the disease called cryptosporidiosis. Cryptosporidiosis are widely distributed human and animal infection rate, and serious harm, clinically severe watery diarrhea as the main feature. Currently no effective drug for the treatment and prevention of cryptosporidiosis. Therefore, this disease is important parasitic disease of the future medical and veterinary academia prevention. Many scholars believe that the vaccine is the potential of the precautionary approach. Cryptosporidium parvum explore CP2/P30 gene immune protective effect of CP2, P30 gene primers were designed using recombinant DNA technology CP2, P30 gene and two genes gene eukaryotic expression vector pVAX-1 is connected in series construct recombinant eukaryotic expression vector, the two genes alone and tandem cloned into the prokaryotic expression vector the PET28-a in the Construction of prokaryotic expression vector. Recombinant eukaryotic expression plasmid transfected Hela cells, and its expression in eukaryotic cells by indirect immunofluorescence method validation; to express recombinant CP2, P30, and CP2/P30 protein prokaryotic expression vector transformed into E. coli. Nucleic acid vaccines and subunit vaccines goats were immunized mice and during pregnancy detection its immune protection force. The results show that, amplification the CP2, P30 open reading frame, show homology Blast than 99.41% and 99.91%, respectively, after restriction enzyme digestion showed that the vector was successfully constructed. Hela cells after recombinant plasmid in the the Hela fine after transfection by indirect immunofluorescence method detected a specific protein. SDS-PAGE protein bands were detected, consistent with the size of the target protein, total protein was extracted and confirmed by Western blot test resulting protein has reactogenicity; nucleic acid vaccine Protection Results showed that the experimental group compared with the control group, the specific antibody titers degrees, CD4 and CD8 T lymphocyte count differences were highly significant (p lt; 0.01), CD4 / CD8 T-lymphocyte count value of the difference was not statistically significant (p gt; 0.05). Experimental group of mice after the attack insect discharge oocysts time decreased by two days, the worm reduction rate and the control group was significantly different, with the empty vector control group were not statistically significant (p gt; 0.05), reduced oocyst maximum of 63.5% . The pregnant goat birth Lamb worm reduction rate of 47.3%. Recombinant CP2, P30 protein and two gene tandem subunit vaccine protection results show that, with the immune increase in the number of antibody titers gradually increased, the test group compared with the control group, a significant difference (p lt; 0.01). CD4 and CD8 T lymphocyte count with the control group compared to the significant difference (p lt; 0.01), CD4 / CD8 T-lymphocyte count value of the difference was not statistically significant (p gt; 0.05). The test mice worm reduction rate significantly, worm reduction rates of up to 61.5%. The pregnant goat birth Lamb worm reduction rate of 42.1%.

Research on CP2/P30 Bivalent DNA Vaccine and Subunit Vaccine Against Cryptosporidium

Abstract

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