Preparing Human TNF-α Monoclonal Antibody To Treat Allergic Rhinitis

Objective Preparing human TNF-αmonoclonal antibody successfully to study the preventive effect of anti- TNF-αon mouse model of allergic rhinitis.Methods The VH/K21 gene with NcoⅠand NotⅠsites was amplified by PCR and cloned into the prokaryotic expression vector pET-22b(+). E.coli.BL21(DE3) was transformed with the recombinant expression plasmid pET-VH/K21 and protein expression was induced in Luria-Bertani medium by addition of IPTG..The products were identified by western-bloting and ELISA. Twenty-four BALB/C mice were evenly randomized into allergic rhinitis group(group A),TNF-αtreatment group(group B) and normal control group(group C).Ovalbumin sensitization was used to create allergic rhinitis model in mice.Mice in group B were given nasal dropping of 5μg TNF-αhalf an hour before challenge.The symptom score,count of eosinophils in nasal mucosa were compared between the 3 groups.Results The recombinant expression plasmid pET-22b(+)-VH/K21 successfully expressed VH/K21 protein.The protein showed specific immunological reaction with human TNF-αin ELISA,which can neutralize toxicity of L929.The symptom scores in group A(7.625±1.061)were significantly higher than those in group B(0.875±0.835)and C(0.375±0.518)(both P< 0.01),no significant difference was observed between the latter two groups.The counts of eosinophils in group A (9.125±1.553) was also significantly higher than those of group B(1.500±0.926)and C(0.500±0.756)(both P < 0.01),no significant difference was observed between the latter two groups.Conclusion Prepared human TNF-αantibody may be used as a new immunotherapy agent against allergic rhinitis in mice and established a solid basis for further studying the biological function of the antibody to TNF-α..