人乳头瘤病毒58型L1/E7“假病毒”疫苗的制备及免疫效果评价

目的构建人乳头瘤病毒58型L1/E7"假病毒"疫苗,并研究其免疫学特性。方法采用sf9昆虫-杆状病毒表达系统表达HPV58L1蛋白,体外解离重组病毒颗粒,向已经解离的衣壳中加入mE7-pcDNA3.1+质粒,CaCl2室温透析促使病毒样颗粒(VLP)重新聚合,用"假病毒"颗粒滴鼻免疫小鼠,检测小鼠血清IgG,IgA中和抗体以及脾淋巴细胞IFN-γ的分泌水平和细胞毒性T淋巴细胞(CTL)反应。结果 SDS-PAGE,Western blot和红细胞凝集实验证实sf9昆虫-杆状病毒表达系统有效表达有功能活性的HPV58L1蛋白,HPV58L1蛋白能够有效的折叠、解聚以及再折叠为VLP。小鼠免疫保护实验发现,用包含有mE7-pcDNA3.1~+质粒的HPV58L1"假病毒"免疫小鼠,抗HPV58L1特异性IgG,IgA明显升高(P0.001),"假病毒"免疫组产生的IFN-γ明显高于VLP免疫组(P0.001);"假病毒"免疫过的小鼠脾淋巴细胞可以特异性杀伤Tc-1细胞,引起CTL反应。结论成功制备了人乳头瘤病毒58型L1/E7"假病毒"疫苗,能诱发免疫动物较强的体液免疫反应,激发保护性抗体的产生。     

GⅡ型诺如病毒衣壳蛋白抗原表位预测及单克隆抗体的制备

目的原核表达诺如病毒衣壳蛋白VP1,制备抗诺如病毒衣壳蛋白VP1的单克隆抗体,并研究其抗原表位特性。方法 PCR扩增GⅡ型湖州株诺如病毒VP1蛋白基因,将目的基因克隆至载体pET28a(+),转化至大肠埃希菌原核表达。纯化表达产物免疫BALB/c小鼠,将成功免疫的小鼠脾细胞与骨髓瘤SP2/0细胞融合。用Modeller9.9为诺如病毒GⅡ型湖州株VP1蛋白构建三维模型,构建好的三维模型提交到SEPPA在线B-cell空间表位预测软件,对GⅡ型湖州株VP1蛋白进行空间表位预测。用预测的表位序列来筛选阳性克隆,得到对应这些表位的单克隆抗体。结果成功构建重组表达载体pET28a(+)-Noro-VP1并在大肠埃希菌中获得表达,重组蛋白免疫的BALB/c小鼠的脾细胞与SP2/0细胞融合,用预测的抗原表位多肽筛选,获得10株杂交瘤细胞株。与重组蛋白的ELISA结果显示1E8、1P10、2C15和2L16四株反应性最强,其他几株反应性较弱。Western blotting结果显示只有2K10、1K16、1E8三株能与重组蛋白很好的结合。结论成功制备了抗GⅡ型湖州株诺如病毒VP1蛋白的单克隆抗体,抗原表位位于衣壳蛋白的P2区,为制备诺如病毒快速免疫诊断试剂盒及抗原表位的研究提供基础。     

增强型Trop2-CD40L病毒样颗粒疫苗的制备、纯化及活性研究

目的 获得嵌合免疫增强佐剂CD40配体(CD40L)的Trop2-CD40L病毒样颗粒(VLPs)疫苗,为后续探究VLPs在肿瘤治疗中的免疫原性及免疫保护作用奠定基础。方法 通过重叠延伸PCR方法扩增以Trop2为靶点、嵌合免疫佐剂CD40L的Trop2-CD40L融合基因,构建杆状病毒表达载体pFastbac^TM /Trop2-CD40L,并转化大肠埃希菌E.coli DH10bac,提取杆粒并经PCR鉴定成功命名为Trop2-CD40L Bacmid,后者转染Tn5昆虫细胞获得重组病毒Trop2-CD40L rBV,与Gag rBV共感染昆虫细胞制备重组病毒样颗粒,透析浓缩后蔗糖密度梯度离心纯化,Western Blot鉴定VLPs目的蛋白表达、电镜观察所制备VLPs形态,Cytokine ELISA方法检测Trop2-CD40L VLPs体外生物学活性。结果 成功扩增了Trop2-CD40L融合基因,通过杆状病毒表达系统、借助SIV Gag衣壳蛋白装配特点成功制备了增强型Trop2-CD40L 病毒样颗粒,与Trop2 VLPs比较具有更好的生物学活性,差异具有统计学意义(t=7.266,,P<0.05)。结论 制备出具有一定生物学活性的增强型Trop2-CD40L病毒样颗粒,证实了CD40L作为免疫佐剂使用有免疫增强效应。     

HPV16型L1蛋白优势表位原核表达及其血清学验证

目的利用原核系统对人乳头瘤病毒(HPV)16型L1蛋白优势抗原表位进行重组表达与纯化,并检测血清学反应。方法对HPV16型L1蛋白优势抗原表位进行全基因合成,将核酸片段克隆至pET-DsbC原核表达载体,重组质粒转化大肠埃希菌Rosetta(DE3),经异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,采用亲和层析对表达产物进行纯化,采用免疫印迹法、酶联免疫吸附试验(ELISA)验证重组蛋白的免疫学活性。结果成功构建了pET-DsbC-HPV16 L1表达载体,并在大肠埃希菌Rosetta(DE3)中实现了稳定的可溶性表达。重组DsbC-HPV16 L1蛋白经亲和层析进行纯化,相对分子质量为45 000,纯度为95%,重组蛋白纯化得率为22%。免疫印迹法结果显示,重组DsbC-HPV16 L1蛋白可与HPV16型阳性血清产生特异的抗原-抗体反应,与重组DsbC蛋白或健康人血清无交叉反应,具有良好的特异性。ELISA结果显示,重组DsbC-HPV16 L1蛋白与HPV16型阳性血清有较强的免疫反应性,A450 nm均值为0.56,高于健康人血清和磷酸盐缓冲液(PBS)阴性对照(A450nm均值分别为0.24和0.21)。结论采用原核表达系统实现了HPV16型L1蛋白优势抗原表位的可溶性高表达,获得的重组蛋白具有良好的免疫原性和免疫学活性。     

HPV L1衣壳蛋白在不同宫颈病变的表达及临床意义

目的 探讨人乳头状瘤病毒（human papillomavirus,HPV）L1衣壳蛋白在不同宫颈病变脱落细胞中的表达及临床意义.方法 收集2012年1月至2013年7月在我院经第二代杂交捕获法检测HPV DNA阳性患者的宫颈脱落细胞标本312份,采用CytoReact HPV L1试剂盒检测壳蛋白在宫颈脱落细胞中的表达,并追踪其组织病理结果.结果 HPV L1衣壳蛋白定位于细胞核,阳性细胞其细胞核着色为红褐色颗粒,位于鳞状细胞的表层,基底细胞无表达.312份宫颈液基细胞学标本中有105份HPV L1衣壳蛋白呈阳性表达,总阳性率为33.65％（105/312）,其中未见上皮内病变或恶性细胞（negative for intraepithelial lesion or malignancy,NILM）为27.33％ （47/172）;未明确意义的非典型鳞状上皮细胞（atypical squamous cell of undermined significance,ASC-US）为36.92％（24/65）;不排除高度鳞状上皮病变的非典型鳞状上皮细胞（ASC of can not exclude high-grade squamous intraepithelial lesion,ASC-H）为21.43％（3/14）;低度鳞状上皮内病变（low-grade squamous intraepithelial lesion,LSIL）为69.44％（25/36）;高度鳞状上皮内病变（high-grade squamous intraepithelial lesion,HSIL）为28.57％（6/21）;鳞癌（squamous cell carcinoma,SCC）为0.00％ （0/4）.六组间HPV L1衣壳蛋白阳性表达率差异有统计学意义（P＜0.05）,其中LSIL与HSIL相比,ASC-US/LSIL与ASC-H/HSIL相比,LSIL与HSIL/SCC相比,LSIL/ASC-H与HSIL/SCC相比,NILM与LSIL相比,HPV L1衣壳蛋白阳性表达率差异均有统计学意义（P均＜ 0.05）,且随宫颈细胞学病变程度的加重,HPV L1衣壳蛋白表达呈下降趋势.312例患者中有143例追踪到组织学结果,其中HPV L1衣壳蛋白阳性表达的有46例,总阳性率为32.17％ （46/143）,其中慢性宫颈炎为27.08％（13/48）,宫颈上皮内瘤样病变Ⅰ级（cervical intraepithelial neoplasia Ⅰ,CIN Ⅰ）为48.84％（21/43）,CINⅡ为33.33％ （8/24）,CINⅢ为17.39％ （4/23）,SCC为0.00％（0/5）.五组间HPVL1衣壳蛋白阳性表达率差异有统计学意义（P＜0.05）.CIN Ⅰ与CINⅡ/CINⅢ相比,慢性宫颈炎与CIN Ⅰ相比HPV L1衣壳蛋白阳性率差异均有统计学意义（P均＜0.05）,且随宫颈组织学病变程度加重,HPVL1衣壳蛋白表达呈下降趋势.以30岁为界将患者分为两组,其中≤30岁组HPV L1衣壳蛋白阳性率为44.00％（44/100）;＞30岁组HPV L1衣壳蛋白阳性率为28.77％（61/212）,两组间差异有统计学意义（P＜0.05）.结论 随宫颈病变程度加重,HPV L1衣壳蛋白表达呈下降趋势,HPV L1衣壳蛋白检测对宫颈鳞状上皮内病变有一定的预测作用.     

人偏肺病毒F1、F2亚单位原核表达系统的构建及多克隆抗体制备

目的 构建人偏肺病毒融合蛋白亚单位F1、F2原核表达系统,初步获得抗F1、F2重组蛋白的多克隆抗体,为相关疫苗的研究奠定基础。 方法 聚合酶链反应(PCR)法扩增F1、F2基因片段,经T-A克隆确保其准确性,双酶切后插入原核表达载体pET32a(+)中,转化大肠埃希菌BL21(DE3),按优化后条件大量诱导表达,纯化后经Western Blot检测特异性。取纯化蛋白免疫BALB/c小鼠制备多克隆抗体,ELISA法检测效价,间接免疫荧光法(IFA)检测多克隆抗体是否能与人偏肺病毒发生特异性反应。 结果 F1、F2基因均正确插入pET32a(+)中,并具有正确的读码框架。0.5 mmol/L IPTG 37 ℃诱导培养5 h可获得大量带His标签的重组蛋白,纯化后浓度分别达到200 g/ml和300 g/ml。Western Blot结果显示,重组蛋白可被抗His标签抗体特异识别;免疫小鼠获得多克隆抗体效价分别为抗pET32a-F1最高1 : 640,抗pET32a-F2最高1 : 40 960。IFA显示抗pET32a-F1、抗pET32a-F2多克隆抗体均可与人偏肺病毒作用产生特异性荧光。 结论 成功构建了F1、F2的原核表达质粒, 并在大肠埃希菌中获得高效表达,该蛋白具有良好的抗原性;免疫小鼠获得特异性多克隆抗体,有利于人偏肺病毒的深入研究。     

抗HPV18 E6多肽单克隆抗体的制备及鉴定

目的 制备特异性抗人乳头瘤病毒(human papillomavirus,HPV)18 E6多肽抗体。方法 通过在线分析软件ABCpred和Bcepred预测HPV 18E6特异性多肽序列,将多肽序列与牛血清蛋白(bovine albumin,BSA)偶联,以偶联后多肽为抗原免疫BALB/c小鼠,采用杂交瘤技术制备抗HPV18 E6多肽单克隆抗体; 以HPV18阳性分泌物标本DNA为模板,聚合酶链式反应(polymerase chain reaction,PCR)获得HPV 18 E6基因,将该基因插入表达质粒pET-28a,转化大肠埃希菌BL21(DE3),异丙基硫代-β-D-半乳糖苷诱导目的蛋白表达; 采用Western Blot方法鉴定抗HPV18 E6多肽抗体与目的蛋白的反应性。结果 建立的杂交瘤细胞株能稳定产生HPV18 E6多肽单克隆抗体; 重组表达质粒pET-28a-HPV18 E6测序证明构建正确,成功表达HPV18 E6蛋白; 纯化后的抗HPV 18 E6多肽抗体可特异性识别原核表达HPV 18 E6蛋白。结论成功制备抗HPV18 E6多肽序列,该抗体具有较好的特异性。     

人乳头瘤病毒16型和18型E6蛋白单抗制备及其鉴定

目的开发能够用于检测HPV16、HPV18感染的诊断技术,原核表达HPV16E6、18E6重组蛋白,制备相应的单克隆抗体,为建立HPV的诊断方法提供物质基础。方法BL21-PET28a-16E6、BL21-PET28a-18E6工程菌株由该实验室前期构建,异丙基-β-D-硫代半乳糖苷(IPTG)诱导HPV16E6、18E6重组蛋白表达,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)、蛋白质免疫印迹法(Western blot)对HPV16E6、18E6重组蛋白进行鉴定,His亲和层析柱对目的蛋白进行纯化。以纯化的重组蛋白HPV16E6、18E6为免疫原免疫BALB/c小鼠,杂交瘤技术制备相应单克隆抗体,间接法测抗体,酶联免疫吸附测定(ELISA)方法检测抗体效价,Western blot检测抗体特异性。结果表达并纯化获得了HPV16E6、18E6蛋白,杂交瘤技术获得HPV16E6-2-G9、HPV16E6-2-F2、HPV18E6-3-H1杂交瘤细胞株,Western bolt结果显示,其分泌抗体可分别与HPV16E6、18E6特异性结合,用HPV16E6-2-G9、HPV18E6-3-H1制备的腹腔积液效价分别可达106。结论成功制备抗HPV16E6、18E6单克隆抗体,该抗体效价高,特异度好,为建立血清学诊断方法打下了基础。     

HPV6晚期表达基因L1的克隆并在杆状病毒表达系统的表达及其抗血清制备

目的 获得具有正确序列的人乳头瘤病毒HPV6型晚期表达基因L1，并获得其高活性表达L1基因特异性抗血清，为进一步研究及临床检验应用奠定物质基础。方法 用PCR法从感染人乳头瘤病毒患者尖锐湿疣组织中获得HPV6晚期表达基因L1，测序验证后，将正确的HPV6晚期表达基因L1序列，插入Bac-to-Bac杆状病毒表达系统，转洒昆虫细胞Sf9，表达外壳蛋白L1，并以之作为抗原免疫兔，获得特异性抗血清。结果 获得了正确序列HPV6晚期表达基因L1，在Bac-to-Bac杆状病毒系统活性表达并获得其特异性抗血清。结论 获得HPV6晚期表达基因L1，及有抗原活性的表达和相应抗血清。     

人乳头瘤病毒16型L1基因真核表达载体的构建及表达

目的从感染人乳头瘤病毒（HPV）的新鲜宫颈癌组织中扩增HPV16型晚期蛋白L1基因全长，构建表达载体，并验证目的蛋白的表达。方法根据基因全长设计一对特异引物，用PCR的方法从宫颈癌组织DNA，中获得L1基因，以pMD18T为克隆载体，构建重组质粒进行亚克隆，再以pcDNA3．1（＋）为载体构建表达质粒，转染至人肝细胞系H7702，提取蛋白，采用Western Blot方法检测HPV16-L1蛋白的表达。结果从长春地区宫颈癌临床标本克隆到的HPV16型L1蛋白编码序列与Genebank报道序列高度同源，其真核表达产物与特异性抗体能够特异性结合。结论成功构建重组表达质粒pcDNA3．1（＋）-HPV16-L1，为进一步研制HPV预防性疫苗创造基础条件。     

Comparison of human papillomavirus type 16 L1 chimeric virus-like particles versus L1/L2 chimeric virus-like particles in tumor prevention

Chimeric human papillomavirus (HPV) virus-like particles (cVLPs) with the HPV16 E7 antigen fused to either the major capsid protein, L1, or the minor capsid protein, L2, have been used independently to protect against the formation of HPV-induced tumors in animal models. However, the advantages and disadvantages of both types of particles with respect to production and vaccine efficacy have never been analyzed. Therefore, in this study, we compared cVLPs with the HPV16 E7 antigen fused to L1 versus cVLPs with E7 fused to L2 with respect to their ability to protect mice from tumor challenge. The first 57 amino acids of E7 were used to overcome the size limitation and limited VLP production imposed by inserting polypeptides into L1 cVLPs. C57BL/6 mice were immunized with the above cVLPs at various doses. Tumor challenge was then performed with HPV16 E7-positive TC-1 cells. HPV16 L1-E7((1-57)) was superior to HPV16 L1/L2-E7((1-57)) in eliciting tumor protection at equivalent doses, although both types of particles were able to protect mice. Both cVLPs induced a specific cytotoxic T lymphocyte (CTL) response to the H2-D(b)-restricted E7 peptide (E7(49-57)) as determined by an ELISPOT assay and tetramer staining; however, immunization with the L1-E7((1-57)) cVLPs resulted in twofold higher CTL precursor frequencies. Our results demonstrate that cVLPs with the antigen fused to L1 are a more efficient vaccine with respect to tumor prevention than cVLPs with the antigen fused to L2. At the same time, however, L1 cVLPs are limited by the size of the antigen that can be incorporated and in the amount of cVLP that can be obtained from cultures when compared to L1/L2 cVLPs. This balances out their superior ability to induce protective immunity.     

HPV L1壳蛋白在宫颈病变中的表达及与HR-HPV DNA的关系

目的研究人乳头瘤病毒(HPV)L1壳蛋白在宫颈病变中的表达及与高危型人乳头瘤病毒(HR-HPV)DNA的关系,为宫颈病变的早期诊断提供预测指标,为HPV疫苗的研制提供理论依据。方法采用免疫细胞化学方法和第二代杂交捕获法(HC-Ⅱ),分别检测272例女性HPV L1壳蛋白的表达和高危型HPV(HR-HPV)病毒载量,组织活检复查宫颈病变情况。结果 HR-HPV DNA感染者中,随着宫颈病变级别(CINⅠ及以上)的升高,HPV L1壳蛋白阳性表达率增高(x~2=35.176,P0.001),HPV L1壳蛋白诊断CINⅡ及以上的灵敏度为89.77%,特异度为45.12%;按年龄分层后,HPV L1壳蛋白表达与宫颈病变的OR均小于1;小于35岁研究对象中,病毒载量与HPV L1壳蛋白阳性表达率之间存在剂量反应关系(x~2=12.915,P0.001)。结论 HPV L1壳蛋白的缺失与宫颈癌及癌前病变的发生、发展密切相关,年轻女性HPV L1壳蛋白的表达与HPVDNA载量存在剂量反应关系。     

Activation of dendritic cells and induction of T cell responses by HPV 16 L1/E7 chimeric virus-like particles are enhanced by CpG ODN or sorbitol

Chimeric human papillomavirus-like particles, consisting of human papillomavirus (HPV) 16 L1-E7 fusion proteins [HPV 16 L1/E7 chimeric virus-like particles (CVLP)], are a vaccine candidate for treatment and prevention of cervical cancer. Although in preclinical studies CVLPs were shown to induce neutralizing antibodies and L1- and E7-specific T cell responses, the results of a recent clinical trial emphasized the need of improved immunogenicity of CVLPs. Here we studied the interaction of HPV 16 L1/E7 CVLPs with mouse bone marrow-derived dendritic cells (BMDCs) activated with different immune adjuvants. We found that lipopolysaccharides (LPS), unmethylated CpG motifs (CpG ODN) and sorbitol enhanced CVLP-induced stimulation of C57BL/6 mouse BMDCs as revealed by increased levels of CD40, CD80, MHC II and CD54 at the cell surface. CpG ODN and sorbitol also enhanced the presentation of Db-restricted cytotoxic T lymphocyte epitopes to HPV 16 L1- or E7-specific T lymphocytes after loading of CVLPs onto BMDCs. Treatment of BMDCs with CpG ODN in combination with CVLPs improved in vitro priming of naive T lymphocytes by CVLP-loaded BMDCs. In vivo, CVLP-loaded BMDCs were more immunogenic as compared with injection of CVLPs alone. CpG ODN and sorbitol further enhanced priming of antigen-specific T cell responses. Our data demonstrate that CpG ODN- or sorbitol-activated BMDCs substantially increase the immunogenicity of CVLPs. Implementing our results in clinical trial protocols may lead to improved activity of therapeutic HPV vaccines for the treatment of HPV-induced cancer.     

HPV L1壳蛋白在诊断宫颈上皮内瘤变及预测其转归中的临床价值

目的 研究HPV L1壳蛋白检测在宫颈上皮内瘤变（CIN）诊断中的临床意义以及对CIN转归的预测价值,探索更加合理有效诊断处理CIN的方案.方法 选择在南京大学医学院附属鼓楼医院行官颈HPV-HC2检测阳性的妇女共187例,同时检查TCT、HPV L1壳蛋白检测及宫颈活检,以病理组织结果为金标准进行对照研究,并对其中继续在鼓楼医院治疗并随访的114例妇女行跟踪研究2年.结果 （1）根据宫颈活检病理结果分组,CIN1、CIN2、≥CIN3的L1壳蛋白阳性率分别是49.12％、30.00％、20.45％,运用卡方检验,x2=10.437,P=0.015 ＜0.05,有统计学差异CIN1组的L1阳性率大于其他各组,而其他组别的L1阳性率之间均无统计学差异,CIN2和CIN3合并与C1NI组进行卡方检验,x2=9.443,P=0.002 ＜0.05,有统计学差异.（2）在2年随访结果中,CIN1未手术组L1（＋）和L1（-）的HC2转阴率比较,P=0.022 ＜0.05,有统计学差异.（3） HPV L1壳蛋白对CIN1的转归有较好的预测作用,CIN1患者中L1（＋）的HPV转阴率较L1（-）的高.结论 （1） CIN1组的L1阳性率大于其他各组,而其他组别的L1阳性率之间均无统计学差异.（2）随访2年结果：未手术组L1（＋）和L1（-）的HC2转阴率比较,有统计学差异（P＜0.05）.（3）HPV L1壳蛋白可作为CIN1的临床预后指标.     

Prophylactic human papillomavirus vaccines

Human papillomavirus (HPV) infection causes virtually all cases of cervical cancer, the second most common cause of death from cancer among women worldwide. This Review examines prophylactic HPV subunit vaccines based on the ability of the viral L1 capsid protein to form virus-like particles (VLPs) that induce high levels of neutralizing antibodies. Following preclinical research by laboratories in the nonprofit sector, Merck and GlaxoSmithKline are developing commercial versions of the vaccine. Both vaccines target HPV16 and HPV18, which account for approximately 70% of cervical cancer. The Merck vaccine also targets HPV6 and HPV11, which account for approximately 90% of external genital warts. The vaccines have an excellent safety profile, are highly immunogenic, and have conferred complete type-specific protection against persistent infection and associated lesions in fully vaccinated women. Unresolved issues include the most critical groups to vaccinate and when the vaccine's cost may be low enough for widespread implementation in the developing world, where 80% of cervical cancer occurs.     

HPV L1壳蛋白表达与宫颈上皮瘤变及HPV分型的关系

目的分析HPV L1壳蛋白的表达与宫颈上皮病变的关系及其在HPV分型中的分布情况。方法对4 320例妇女进行宫颈液基细胞学(LCT)和HPV分型检测,对LCT≥无明确诊断意义的非典型鳞状上皮细胞(ASC-US)或HPV阳性者行HPV L1检测,并对LCT≥ASC-US且HPV阳性者行阴道镜下组织病理学检查。结果对LCT≥ASC-US或HPV阳性者(共401例)进行HPV L1检测,HPV L1阳性率为34.9%(140/401)。对LCT≥ASC-US且HPV阳性者129例进行组织病理学检查,LCT与组织病理学诊断符合率为79.2%。HPV L1在LCT诊断为ASC-US、低度鳞状上皮内病变(LSIL)、高度鳞状上皮内病变(HSIL)的患者中阳性率分别为40.2%(29/72)、69.2%(18/26)、9.0%(2/22)。386例HPV感染者中,单一型别感染率居前四位的是HPV52、58、16、18型,其HPV L1阳性率分别为53.3%、28.9%、12.2%、5.6%。结论 HPV L1阳性率随宫颈病变级别增高呈下降趋势,LCT联合HPV L1检测可为宫颈病变预后判定提供参考依据。     

Papillomavirus virus-like particles as anticancer vaccines

Papillomavirus virus-like particles (VLPs) are empty, non-replicative, non-infectious particles that retain conformationally correct epitopes for the generation of antibody responses to the viral capsid proteins. Chimeric human papillomavirus (HPV) virus-like particles incorporating non-structural virus proteins offer an exciting approach for combined prophylactic and therapeutic vaccines against HPV-induced lesions. Both HPV VLPs and chimeric VLPs can induce potent humoral and cellular immune responses when injected into mice, leading to the generation of virus-neutralizing antibodies, priming of CD8+ T-cells and activation of cytotoxic T-cell effector functions. This review summarizes recent advances in the production of chimeric VLPs, the immune response elicited by VLPs and chimeric VLPs, and their ability to generate strong protective and therapeutic antitumor immune responses.     

Human papillomavirus type 16 L1E7 chimeric capsomeres have prophylactic and therapeutic efficacy against papillomavirus in mice

Genital human papillomavirus (HPV) infection is the primary cause of cervical cancer in women. Although the HPV recombinant L1 protein was recently licensed as an available vaccine, it has numerous shortcomings. New vaccination strategies should be considered. To enable the design of a prophylactic and therapeutic low-cost vaccine candidate, chimeric HPV16 L1DeltaC34E7N1-60 capsomeres were produced in Escherichia coli. The immune characteristics and potential prophylactic and therapeutic effects of these capsomeres were examined in C57BL/6 mice. Following protein purification and renaturation, the majority of the recombinant chimeric proteins (L1DeltaC34E7N1-60) assembled into capsomeres. These capsomeres were able to induce conformational and neutralizing antibodies against HPV virus-like particles and trigger cell-mediated specific immune responses against the L1 and E7 peptides. In vivo tumor challenge assays showed that mice immunized with the capsomeres were protected against a challenge with both C3 and TC-1 tumor cells. Furthermore, in vivo tumor rejection assays showed that capsomeres have therapeutic efficacy in mice following inoculation with C3 and TC-1 tumor cells. Chimeric capsomeres are capable of preventing and eliminating HPV16 infection. Therefore, our study has provided an economical vaccine candidate.