The Construction Of Anti-Human Interleukin-13Vaccine And Its Immunological Characteristics

Interleukin-13(IL-13)is recognized as one of the most pivotal mediator in the pathological mechanisms of asthma, hence the need for deeper understanding on the pathological mechanisms of asthma and new efficient therapy strategies is urgent. At present, monoclonal antibodies against IL-13are undergoing phase Ⅱ clinical trials. However, the needs for repeatedly administrations make treatment with monoclonal antibodies very expensive and easy to become resistant. The strategy of anti-cytokine vaccine, which produces neutralizing antibodies by active immunization, will induce high level and persistent auto-antibodies and thus provide a long last intervening effect. And thus, the vaccine strategy shows a more promising prospect than the passive immunization with monoclonal antibodies.Potential antigenic epitopes of human IL-13molecule were predicted by bioinformatics methods; encoding sequence of predicted epitopes were inserted into the immunodominant domain of Hepatitis B core antigen (HBcAg) by using gene recombinant techniques, and the recombinant plasmids were transformed into E.coli and induced by IPTG for the expression of the chimeric HBcAg proteins; the recombinant proteins were purified by ammonium sulfate precipitation and washing, gel filtration chromatography and sucrose gradient ultracentrifugation, and the presence of virus-like particles (VLPs) were confirmed by electron microscopy; the antisera were collected form the mice immunized with purified VLPs; Assay based on TF-1cells or A549cells were set up for the detection of IL-13activity and assessment of the neutralizing potentials of antisera induced by the vaccine; the immunological characteristics of the VLPs vaccine were evaluated by immunizing mice with different doses, adjuvants, and inoculation routes, and the local and systematic IgA and IgG responses in different tissues, as well as the IgG subtypes responses including IgG1and IgG2a were investigated. In addition, the mature IL-13molecules were obtained by employing fusion and non-fusion expression patterns.