Preparation of Yersinia enterocolitica 0:8 Detoxified LPS-TetanusToxoid Conjugate and Evaluation Its Induced Antibodies in Mous

Yersinia enterocolitica is known to cause human enteric infections frequently isolated from these animals. The serum immunoglobulin G (IgG) to the lipopolysaccharide (LPS) may confer protective immunity to enteric pathogens by inducing bactericidal activity as well as polysaccharide-protein conjugates induce higher level of serum IgG. The aim of present study is immunological evaluation of Yersinia enterocolitica 0:8detoxified LPS -tetanus toxoid (TT) conjugate in mice as a candidate vaccine. Yersinia enterocolitica 0:8LPS, was extracted by hot phenol method, then LPS was detoxified by alkaline method.Detoxified LPS (D-LPS) coupled to TT with ADH as a spacer and EDAC as a linker and then purified by size exclusion chromatography. Then four group of BALB/c mice were selected (each group included 15 mice). Injection was performed by three doses (I.P) in two weeks interval and induced sera antibodies against LPS of Yersinia enterocolitica0:8measured by indirect ELISA to IgG, IgA, IgM. Results showed that conjugate could elicit serum antibodies to the Yersinia enterocolitica0:8LPS. The largest increase in LPS antibodies was of the IgG class. After second and third doses, conjugate showed significant increasing in all types of antibodies titer in comparison to non-conjugated LPS received group. These results shows that preparedYersinia enterocolitica0:8detoxified LPS-tetanustoxoid conjugate was antigenicand could increase the anti-LPS antibodies in the form of conjugate with tetanus toxoid and can be an appropriate as a effective candidate vaccine

Keywords:

Yersinia enterocolitica0:8 , Tetanus Toxoid , Conjugate , Antibodies