Study On DNA Vaccine Of Human Papillomavirus 16 (HPV16) E7 And Transdermal Delivery With Microneedle Array

Human papillomavirus (HPV) belongs to papovaviridae, and it is a kind of small DNA virus that usually produce benign lesions on cutaneous and mucosal surfaces. HPV infection is widespread in the population and associated with a broad spectrum of human diseases ranging from benign condyloma, common skin warts, laryngeal papillomas to anogenital cancers and cervical cancer. There is no doubt that infection with certain types of HPV is the main cause of cervical cancer. Initially, HPV DNA was detected in 93% of the cervical cancer specimens using a PCR assay, and HPV 16 was more than 50% of all. Therefore, HPV, especially HPV16 is the main factor to induce cervical cancer.Recently, the use of micron-scale needles in increasing skin permeability has been proposed and shown to dramatically increase transdermal delivery, especially for macromolecules. Microneedle array can only penetrate skin cutin(15-25μm),which is barrier for drug pervasion, and get through hundreds of channels. But the needlepoints can not run into cutis, which contain abundant nerve and capillary vessel. Therefore, drugs can diffuse rapidly via channels into capillary vessels and then into body cycle. Acting as the first barrier of resisting environment pathogen, the immunity function of skin system is quite strong, therefore, transdermal immunization can arouse efficient and strong immunoreaction. This newfashioned immune approach can increase drug effect, and reduce side-effect. Furthermore, the price of the equipment is cheap and can easily operate.In this article, in order to find an effective method to cure HPV, we prepared HPV16 E7 DNA vaccine and delivered with microneedle array. We establishedHPV16E7-pcDNA3.1 (-) recombinant vector, which acted as gene vaccine. The DNA vaccine can easily get across the microchannels generated by microneedle arrays in vitro. Moreover, the course was permanent and the whole infiltration quantity was comparatively big, reaching 0.74 mg/cm2 in the 30~th h. We immunized BALB/c mouse with a dose of 200Mg with microneedle array every two weeks. Two weeks after the third immunization, we separated serum and lymphocytes separately to detect the functions of humoral immunity with indirect immunofluorescence test, and detect the functions of cellular immunity with lymphocyte transformation test. As a result, the functions of humoral immunity and cellular immunity were obviously increased.This article was looking for an effective method to cure HPV by preparing the DNA vaccine of HPV16 E7, combined administer with microneedle array to make DNA vaccine more effective absorption and expression. We hope it's a beneficial attempt to open out DNA vaccine of HPV 16 and generalize this newfashioned immune approach - administer with microneedle array.