ABSTRACT
In order to explore the possibility of human adenovirus infection with tree shrews,the neutralizing antibody ti-ters of five kinds of human adenoviruses (HAdv)in the serum of tree shrews were analyzed.The levels of Ad3,Ad4,Ad7, Ad14 and Ad55 neutralizing antibody were detected by virus neutralization test.The results showed that the positive rate of four adenoviruses in group B were higher than Ad4 in group E,and the positive rates respectively were Ad14 (55.88%),Ad3 (47.06%),Ad55 (29.71%),Ad7 (14.71%)and Ad4 (8.82%).The antiserum mainly mixed with Ad3,Ad14 and Ad55 anti-body.Five species of human adenovirus can be naturally infected with tree shrews.Tree shrews are used as experimental ani-mals to establish human adenovirus infection model is alternative.
ABSTRACT
<p><b>OBJECTIVE</b>To obtain the monoclonal antibody against hexon protein of human adenovirus.</p><p><b>METHODS</b>BALB/c mice were immunized with purified recombinant hexon protein, and the spleen cells of the mice were isolated and fused with myloma cells. Four hybridoma cell strains were screened by indirect ELISA and cultured, and the sensitivity, specificity and virus neutralizing activity were analyzed with ELISA, Western blotting and neutralizing test.</p><p><b>RESULTS</b>The mouse ascites produced by these hybridoma cells contained specific monoclonal antibodies against hexon protein of human adenovirus as identified by ELISA and Western blot, and the antibody generated by 4C6 strain showed human adenovirus type 3-neutralizing activity.</p><p><b>CONCLUSION</b>The monoclonal antibodies against hexon protein with high specificity have been successfully obtained, and these antibodies can be useful in developing assays for early diagnosis of HAdV3 infection and also in study of therapeutic drugs of the infection.</p>
Subject(s)
Animals , Humans , Mice , Adenoviruses, Human , Chemistry , Allergy and Immunology , Antibodies, Monoclonal , Allergy and Immunology , Antibodies, Viral , Allergy and Immunology , Blotting, Western , Capsid Proteins , Genetics , Allergy and Immunology , Enzyme-Linked Immunosorbent Assay , Escherichia coli , Genetics , Hybridomas , Bodily Secretions , Mice, Inbred BALB C , Recombinant Proteins , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To clone, express and characterize the capsid protein of human Norwalk virus Guangzhou strain NVgz01.</p><p><b>METHODS</b>On the basis of successful construction of full-genome clones and sequence analysis of human norovirus Guangzhou strain NVgz01, the full capsid gene was ligated into pET28a (+) for expression. After IPTG induction, the recombinant protein was purified through metal (Ni(2+)) chelating affinity chromatography. Western blotting and enzyme-linked immunosorbent assay (ELISA) were used to determine the antigenicity of the recombinant protein.</p><p><b>RESULTS</b>The recombinant capsid gene was overexpressed in E.coli, yielding the recombinant protein with relative molecular mass of 62x10(3) that was highly purified through metal (Ni(2+)) chelating affinity chromatography. IDEIA Norovirus Kit and immunoassay showed that the recombinant protein had good antigenicity.</p><p><b>CONCLUSION</b>The capsid gene of norovirus Guangzhou strain has been cloned and expressed, which can be useful for developing diagnostic reagents or vaccine of norovirus.</p>