Typing Of Human Papillomavirus And Preliminary Study For Integrin Antibody Inhibiting The Recurrence Of Condyloma Acuminatum

Condyloma acuminatum (CA), caused by human papilloma virus (HPV), is a sexually transmitted disease (STD) which usually occurs in the genital and perianal areas. There is a high incidence and recurrence rate in CA patients. Many methods may be used to treat CA, however the recurrence is unavoidable. Different types of HPV can cause different clinical manifestations, so the typing of HPV is useful for the clinical diagnosis and prognosis.Integrin is an important member of the cell adhesive molecule family. Much research in the relationship between integrins and virus infection has shown that integrins may act as viral receptors or co-receptors to infect host cells. Integrinα6 is expressed specifically in normal basal layer of epidermis. HPV causes damage to the host cell through infecting the basal layer of epidermis. Is there any correlation between them? Does integrin a6 play an important role in the process of HPV infection? If blocking the initial stage of the expression of integrins, can HPV be inhibited to infect host cells? Further studies of these problems may be helpful to reduce the HPV infection and the recurrence of CA fundamentally.Objective①To identify and assess the feasibility of HPV DNA testing and typing is done through polymerase chain reaction (PCR) with joint restriction fragment length polymorphism (RFLP) and gene sequencing technologies. Testing was performed on CA and four kinds of skin disease of perianal and genitals.②To investigate the expression of integrin a6 mRNA in CA lesions of clinical patients and to reveal the significance of integrinα6 in the pathogenesis of primary and recurrent CA.③To investigate the role of integrin a6 monoclonal antibody (GoH3) in the infection of HPV 6/11 virus particles on the immortalized human keratinocyte cell line (HaCaT cells) in vitro.Methods①HPV DNA from the organizations of CA, Bowenoid papulosis (BP), squamous cell carcinoma (SCC), lichen sclerosus et atrophicus (LSA) and extramammary Paget's disease (EMPD) testing and typing was detected by PCR-RFLP with joint gene sequencing technologies.②The expression of integrin a6 mRNA in primary and recurrent CA lesions were determined by RT-PCR.③The DNA loads of clinical separated HPV 6/11 virions which used to infect HaCaT cells in vitro were detected by fluorescence quantitative polymerase chain reaction (FQ-PCR).Results①50 cases of clinical samples diagnosed by pathological examination.35 patients were detected HPV DNA positive. The positive rates of HPV DNA in CA, BP, SCC, LSA and EMPD were 100.0%(26/26),47.1%(8/17), 50.0%(1/2),0.0%(0/4) and 0.0%(0/1) respectively. In the HPV DNA positive samples, there were 19 cases of HPV 6,3 cases of HPV 11,8 cases of HPV 16,4 cases of HPV 6+11,1 case of HPV 62. Gene sequencing results correspond to the PCR-RFLP's results.②The relative contents of integrin a6 mRNA in primary and recurrent CA lesions (0.79±0.16 and 1.07±0.29 respectively) were obviously higher than that in normal controls (0.25±0.10). The content of integrin a6 mRNA in recurrent CA lesions were higher than that in primary CA lesions (P<0.05).③The viral load of 4 different concentrations of HPV 6/11 virions which was used to infect HaCaT cells were different (P=0.001). At the concentration of 106copies/ml of HPV 6/11 virions, the infection rate of HaCaT cells was 100%, and the HPV DNA load was higher than other 3 groups (P<0.05). When the dilution of the GoH3 was>1:100, its inhibition rate to HPV 6/11 infecting HaCaT cells is proportional to its concentration. However, when the dilution of GoH3 was<1:100, even increasing the concentration of GoH3, there was no significant increased inhibition rate. The average viral load of 8 clinical separated HPV 6/11 virions infecting HaCaT cells in vitro before and after being added GoH3 were (5.81±2.51)×104 copies/ml and (3.02±1.21)×104 copies/ml, respectively. The average inhibition rate of GoH3 was (46.9±4.7)%.Conclusion①PCR-RFLP method is used in the detection and typing of HPV DNA.②Integrinα6 maybe play some role in the incidence and recurrence of CA.③GoH3 may partially inhibit the infection of HPV 6/11 to HaCaT cells.