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制备16型人乳头瘤病毒mE6Δ/mE7蛋白与I型人单纯疱疹病毒VP22Δ蛋白的治疗型分子内佐剂融合蛋白疫苗,并检测其免疫原性和抗肿瘤相关生物活性。通过克隆HSV-1 VP22Δ及HPV-16 mE6Δ/mE7基因,构建pET28a-VP22Δ-mE6Δ/mE7原核表达载体。重组质粒在Rosetta(DE3)宿主菌中进行诱导表达,表达蛋白经分离、复性后,通过镍离子亲和层析进行纯化,纯化蛋白经SDS-PAGE、Western blot鉴定,并免疫BalB/C及C57BL/6小鼠,检测其免疫原性和抗肿瘤活性。结果显示,VP22Δ-mE6Δ/mE7蛋白以包涵体形式表达,分子量约为34kDa,表达量约占菌体总蛋白的45%。该蛋白免疫小鼠后血清特异性IgG、特异性淋巴细胞增殖效果及对TC-1致瘤小鼠的肿瘤治疗效果均高于无佐剂单一重组蛋白疫苗。以上结果说明,所获得的重组融合蛋白具有较好的免疫原性和抗肿瘤活性,为治疗型HPV分子内佐剂疫苗的进一步研究奠定了基础。
A therapeutic intramolecular adjuvant fusion protein vaccine of type 16 human papillomavirus mE6Δ / mE7 protein and human herpes simplex virus type 1 VP22Δ protein was prepared and its immunogenicity and anti-tumor related biological activity were tested. The prokaryotic expression vector pET28a-VP22Δ-mE6Δ / mE7 was constructed by cloning HSV-1 VP22Δ and HPV-16 mE6Δ / mE7 genes. Recombinant plasmids were induced in Rosetta (DE3) host strain. The expressed proteins were separated and renatured. The expressed proteins were purified by nickel ion affinity chromatography. The purified proteins were identified by SDS-PAGE and Western blot and immunized BalB / C and C57BL / 6 mice were tested for their immunogenicity and antitumor activity. The results showed that the VP22Δ-mE6Δ / mE7 protein expressed in inclusion bodies, the molecular weight of about 34kDa, the expression of about 45% of the total bacterial protein. The protein immunized mice serum-specific IgG, specific lymphocyte proliferation and TC-1 tumor-causing mice tumors were higher than the effect of non-adjuvant single recombinant protein vaccine. The above results show that the obtained recombinant fusion protein has good immunogenicity and anti-tumor activity, which lays the foundation for further research on therapeutic HPV intramolecular adjuvant vaccine.