人乳头瘤病毒16型（HPV16）E7 DNA疫苗及微针阵列透皮给药的研究
发布时间：2018-08-04 21:07
【摘要】：人乳头瘤病毒(human papillomavirus,HPV)是乳多空病毒的一种,生殖道HPV感染是当今流行最广的性传播疾病之一。HPV在临床上主要引起尖锐湿疣和宫颈癌等疾病,它的型别繁多,现已发现200多个,根据其致病力的大小分为低危型和高危型。前者包括HPV6,11,30,9等,主要导致生殖道乳头状瘤,尖锐湿疣,外生性湿疣类病变,低度子宫颈上皮内瘤变等。后者包括HPV16,18,31,33,35等,主要导致高度子宫颈上皮内瘤变和子宫颈癌的发生。根据研究发现子宫颈癌的发生与HPV的感染密切相关,90%以上的宫颈癌组织中可以检测到HPV,其中HPV16的检测率超过50%。所以可以说,HPV特别是HPV16是引起宫颈癌的主要因素。 近来有一种新的给药方式,它利用在显微镜下才看得见的针组成的微针阵列(microneedle array,简称微针,)在皮肤上产生许多微米级的通道。这些通道可以让大分子物质通过,甚至一些超分子化合物和微粒也能通过。微针仅穿透对药物扩散起屏障作用的皮肤角质层(15～25μm),并打通上百个给药通道,药物可经通道迅速扩散到毛细血管进入体循环。而且微针透皮给药方便,无明显创伤,病人痛苦小,可减少副作用,同时设备操作简单,价格便宜。作为抵御外界病源的第一道屏障,皮肤系统的免疫功能十分发达,能否通过透皮免疫引起有效的免疫反应,以建立一种新型的免疫途径,正是我们的研究目的。 本文构建了HPV16E7—pcDNA3.1(-)重组质粒,并以之作为DNA疫苗。在体外透皮的条件下,我们观察了30个小时,试验结果发现DNA疫苗可以透过皮肤,而且透过量随着时间的延长而逐步增加,透过量也比较大,在第30h时可以达到0.74mg/cm~2。再结合微针给药的方式将该DNA疫苗免疫BALB/c小鼠,每两周免疫一次,共给药三次,每次免疫剂量为200μg/只,阴性对照组用等体积生理盐水代替。最后一次免疫后两周采血,分别分离血清和淋巴细胞,用间接免疫荧光试验检测小鼠的体液免疫功能,用淋巴细胞转化实验来检测小鼠的细胞免疫功能。实验结果表明,微针免疫DNA疫苗可以诱导小鼠产生体液免疫和细胞免疫。 本文通过HPV16 E7的DNA疫苗的研制为HPV的治疗寻找一种有效的方法,同
[Abstract]:Human papillomavirus (human) is a kind of lactobacillus virus. Genital tract HPV infection is one of the most prevalent sexually transmitted diseases. HPVs mainly cause condyloma acuminatum and cervical cancer in clinic. According to its pathogenicity, it can be divided into low risk type and high risk type. The former included HPV6, 1130T9, which mainly resulted in genital papilloma, condyloma acuminatum, ectogenic condyloma, low grade cervical intraepithelial neoplasia and so on. The latter, including HPV16, 18, 31, 33, 35, mainly lead to high cervical intraepithelial neoplasia and cervical cancer. According to the study, HPV can be detected in more than 90% of cervical cancer tissues, and the detection rate of HPV16 is more than 50%. So it can be said that HPV, especially HPV16, is the main factor causing cervical cancer. Recently, a new method of drug delivery has been developed, which uses microneedle array (microneedle array, which is visible under microscope, to produce many micron channels in the skin. These channels allow macromolecules to pass through, even supramolecular compounds and particles. The microneedle penetrates only the cuticle (1525 渭 m), which acts as a barrier to drug diffusion, and opens hundreds of drug delivery channels, through which the drug can rapidly spread to the capillary and enter the systemic circulation. And the microneedle transdermal drug is convenient, no obvious trauma, the patient pain is small, can reduce the side effect, at the same time the equipment is simple to operate, the price is cheap. As the first barrier to resist external diseases, the immune function of skin system is very developed. Whether transdermal immunity can induce an effective immune response to establish a new immune pathway is our research goal. HPV16E7-pcDNA3.1 (-) recombinant plasmid was constructed and used as DNA vaccine. Under the condition of in vitro transdermal penetration, we observed for 30 hours, the results showed that the DNA vaccine could penetrate the skin, and the transmissivity increased gradually with time, and the transmissivity reached 0.74 mg / cm ~ (-2) at 30 h. BALB/c mice were immunized with the DNA vaccine once every two weeks with a dose of 200 渭 g / mouse. The negative control group was replaced by the same volume of normal saline. Blood samples were collected two weeks after the last immunization. Serum and lymphocytes were separated respectively. The humoral immune function of mice was detected by indirect immunofluorescence test and the cellular immune function of mice was detected by lymphocyte transformation test. The results showed that microneedle immunized DNA vaccine could induce humoral and cellular immunity in mice. Through the development of DNA vaccine of HPV16 E7, an effective method for the treatment of HPV was found.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392
本文编号：2165147
[Abstract]:Human papillomavirus (human) is a kind of lactobacillus virus. Genital tract HPV infection is one of the most prevalent sexually transmitted diseases. HPVs mainly cause condyloma acuminatum and cervical cancer in clinic. According to its pathogenicity, it can be divided into low risk type and high risk type. The former included HPV6, 1130T9, which mainly resulted in genital papilloma, condyloma acuminatum, ectogenic condyloma, low grade cervical intraepithelial neoplasia and so on. The latter, including HPV16, 18, 31, 33, 35, mainly lead to high cervical intraepithelial neoplasia and cervical cancer. According to the study, HPV can be detected in more than 90% of cervical cancer tissues, and the detection rate of HPV16 is more than 50%. So it can be said that HPV, especially HPV16, is the main factor causing cervical cancer. Recently, a new method of drug delivery has been developed, which uses microneedle array (microneedle array, which is visible under microscope, to produce many micron channels in the skin. These channels allow macromolecules to pass through, even supramolecular compounds and particles. The microneedle penetrates only the cuticle (1525 渭 m), which acts as a barrier to drug diffusion, and opens hundreds of drug delivery channels, through which the drug can rapidly spread to the capillary and enter the systemic circulation. And the microneedle transdermal drug is convenient, no obvious trauma, the patient pain is small, can reduce the side effect, at the same time the equipment is simple to operate, the price is cheap. As the first barrier to resist external diseases, the immune function of skin system is very developed. Whether transdermal immunity can induce an effective immune response to establish a new immune pathway is our research goal. HPV16E7-pcDNA3.1 (-) recombinant plasmid was constructed and used as DNA vaccine. Under the condition of in vitro transdermal penetration, we observed for 30 hours, the results showed that the DNA vaccine could penetrate the skin, and the transmissivity increased gradually with time, and the transmissivity reached 0.74 mg / cm ~ (-2) at 30 h. BALB/c mice were immunized with the DNA vaccine once every two weeks with a dose of 200 渭 g / mouse. The negative control group was replaced by the same volume of normal saline. Blood samples were collected two weeks after the last immunization. Serum and lymphocytes were separated respectively. The humoral immune function of mice was detected by indirect immunofluorescence test and the cellular immune function of mice was detected by lymphocyte transformation test. The results showed that microneedle immunized DNA vaccine could induce humoral and cellular immunity in mice. Through the development of DNA vaccine of HPV16 E7, an effective method for the treatment of HPV was found.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392
【参考文献】
相关期刊论文 前1条
1 解洪业;基因疫苗的研究近况[J];青海畜牧兽医杂志;2004年02期
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