Expression Of IgG1 And IgG2 Subclasses Toward Human Papillomavirus-like Particles Is Associated With Cervical Intraepithelial Neoplasia

Correlation of human papillomavirus genotypes and cervical intraepithelial neoplasiaObjective: To study the genotype distribution of human papillomavirus (HPV) infection in different grade of cervical lesions and its relation with cervical intraepithelial neoplasia (CIN), furthermore discuss the correlation between the multiple HPV infection and the development of CIN.Methods: During September 2006 to December 2007, sixty women were examined with cytological test(liquid-based Thin Prep cytology test), and high-risk HPV-DNA test (hybrid captureⅡ), and electrocolposcope, and diagnosed with HPV infection and CIN by cervical biopsies or cervicectomy, including HPV infection, CINⅠ, CINⅡ-Ⅲ, each of which has 20 patients, besides normal control group in 20 cases. DNA flow-through hybridization genotyping technique was applied to determine the HPV-DNA types.Results: The negative rate of HPV-DNA was 21.25%, The positive rate of HPV-DNA was 78.75%, The infection rate of HPV16, 58, 33, 31, 18-DNA were 43.75%, 13.75%, 7.50%, 6.25%, 3.75%, respectively. The multiple HPV infection was found in 7 cases(11.67%) in CINⅡ-Ⅲgroup, 2 case in HPV infection group (3.33%), but each of them was not distinctively different compared with control group (5%),(P>0.05). The positive rate of HPV16-DNA from cervical precancerous lesions group (58.33%)was significantly higher compared with that from control group (P<0.05). However, the positive rate of HPV 58, 33, 31, 18-DNA from cervical precancerous lesions group (13.33%, 10%, 8.33%, 5%)and control group (5%, 10%, 0, 0) was similar(P>0.05). There was a moderate correlation between the HPV16-DNA positivity and cervical precancerous lesions (r =0.479, P<0.05).Conclusion:①There is a close correlation between the infection of HPV16 and the occurrence of cervical precancerous lesions, while the multiple HPV infection was nearly independent of the grade of cervical lesions.②Normal subjects has a certain carry rate.The association between IgG1, IgG2 subclasses toward human papillom vavirus-like particles in cervix and cervical intraepithelial neoplasiaObjective: To investigate expression of IgG1 and IgG2 subclasses toward human papillomavirus-like particles (HPV VLPs) in cervix of patients with various grade of cervical lesions, and the relationship between expression of IgG subclasses and different grade of cervical lesions.Methods: During September 2006 to December 2007, sixty women were examined with cytological test(liquid-based Thin Prep cytology test), and high-risk HPV-DNA test (hybrid captureⅡ), and electrocolposcope, and diagnosed with HPV infection and CIN by cervical biopsies or cervicectomy, including HPV infection, CINⅠ, CINⅡ-Ⅲ, each of which has 20 patients, besides normal control group in 20 cases. The expression of IgG1 and IgG2 subclasses toward human papillomavirus-like particle in cervix from above patients were detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical staining.Results:①RT-PCR showed expression rate of HPV VLPs-IgG1 and IgG2mRNA from study group were 90%, 87%, respectively. The level from control group were 85%, 90%, respectively. The expression rate of HPV VLPs-IgG1,IgG2mRNA from CINⅠgroup was not significantly differently compared with that from HPV infection group. The density of HPV VLPs-IgG1mRNA expression in cervix from CINⅠpatients (1.306±0.022) was significantly higher compared with that in HPV infection patients (0.611±0.033), but was significantly fewer compared with that in CINⅡ-Ⅲpatients (1.956±0.041), (P<0.05).The density of HPV VLPs-IgG2mRNA expression in cervix from HPV infection patients (0.458±0.076) was significantly fewer compared with that in control(1.540±0.055), CINⅠpatients(1.311±0.065) and CINⅡ-Ⅲpatients(1.512±0.037), but there was no significant difference among control, CINⅠand CINⅡ-Ⅲpatients(P<0.05). The level of HPV VLPs-IgG1mRNA from HPV infection group and normal control group was similar(0.611±0.033, 0.403±0.005), so does the expression rate of HPV VLPs-IgG1 (P>0.05). However, The density of HPVVLPs-IgG2mRNA(1.540±0.055) was significantly higher than that of IgG1mRNA expression from control group (P<0.05). The IgG2 dominance (IgG2/IgG1 ration >1)from control group, HPV infection group, CINⅠgroup were much greater than that from CINⅡ-Ⅲgroup (P<0.05), but there was no significant difference between HPV infection group and control group, so does CINⅠgroup and control group(P >0.05).②Immunohistochemical analysis demonstrated that the expression rate of HPV VLPs-IgG1 in cervix from study group(62%)was significantly higher compared with that in control group(5%), (P<0.05). On the contrary, the expression rate of HPV VLPs-IgG2 in cervix from study group(65%)was obviously fewer than that in control group (100%). The expression rate of HPV VLPs-IgG1 increased with the grade of CIN, significant difference was found among all grades (P<0.05). The positive rate of HPV VLPs-IgG1 expression from HPV infection group(20%), CINⅠgroup(65%), and CINⅡ-Ⅲgroup (100%) were significantly higher than that from control group(5%), (P <0.05). The positive rate of HPV VLPs-IgG2 expression from control group, CINⅠgroup and CINⅡ-Ⅲgroup were 100%, 75%, 90%, significant difference was not found among all groups, but they were significantly higher compared with that from HPV infection group (30%), (P<0.05).The positive rate of HPV VLPs-IgG2 expression was significantly higher than that of IgG1 expression from control group (P<0.05). In control group, the expression of HPV VLPs-IgG1 was negative, while the IgG2 positive cells were mostly distributed throughout the stroma. In HPV infection group, HPV VLPs-IgG1, IgG2 positive cells were scattering distribution in the stroma, almost no expression of IgG1, IgG2 were found in the epithelium. In CINⅠgroup, the expression of HPV VLPs-IgG1, IgG2 positive cells presented widespread distribution in the stroma, only a small quantity of them was observed in the epithelium. In CINⅡ-Ⅲgroup, the expression of HPV VLPs-IgG1, IgG2 positive cells were not only distributed throughout the stroma, but also distributed above two thirds of the subepithelial or involved the whole layer of the epithelium(P<0.05).Conclusion:①An increase of the density of HPV VLPs-IgG1, IgG2mRNA expression and its positive rate in high grade cervical lesions might associated with persistent HPV infection and in pregression of CIN.②An increase of the IgG2 dominance (IgG2/IgG1 ration>1) of local immunity in cervix from low grade cervical lesions group and normal control group might indicating the clearance of HPV infection and the regression of cervical lesions.③There is a correlation between the distribution of HPV VLPs-IgG1 positive cells and the grade of cervical lesions. gThe association between IgG1, IgG2 subclasses toward human papillomavirus16-like particles in the serum and cervical intraepithelial neoplasiaObjective: To investigate expression of IgG1 and IgG2 subclasses toward human papillomavirus 16-like particles (HPV16VLPs) in the serum of patients with various grade of cervical lesions, and discuss the relationship between expression of IgG subclasses and different grade of cervical lesions.Methods: During September 2006 to December 2007, one hundred and five women were examined with cytological test(liquid-based Thin Prep cytology test), and high-risk HPV DNA test (hybrid captureⅡ), and electrocolposcope, and diagnosed with HPV infection and CIN by cervical biopsies or cervicectomy, including HPV infection in 32 cases, CINⅠin 30 cases, CINⅡ-Ⅲin 43 cases, besides normal control group in 24 cases. The expression of IgG1 and IgG2 toward human papillomavirus16-like particles in the serum of patients was measured using ELISA.Results:①The HPV16 IgG seroprevalence was 35.24%, the HPV16 IgG1 and IgG2 were all 29.52% in cervical precancerous lesions women, and these levels were significantly higher than control women (8.33%), (P<0.05). There was no difference with the HPV16 IgG, IgG1 and IgG2 seroprevalence between HPV infection group(25.00%, 8.75% and 25.00%, respectively) and CINⅠgroup (26.67%, 3.33% and 20.00%, respectively), but there has significant difference when compared with control group and CINⅡ-Ⅲgroup (48.84%), (P<0.05).The optical density (OD) value of IgG, IgG1 toward human papillomavirus16-likes particle increased with the grade of CIN, significant difference was found among HPV infection group(0.485±0.03, 0.311±0.020), CINⅠgroup (1.531±0.013, 1.104±0.029) and CINⅡ-Ⅲgroup(2.421±0.002,1.941±0.038). But the OD value of IgG, IgG1 toward human papillomavirus16 -like particles from HPV infection group and control group was similar (0.328±0.017,0.308±0.019), (P>0.05). The OD value of IgG2 toward human papillomavirus16-like particles from control group was significantly higher than that of IgG1 expression from control group (P<0.05). The OD value of IgG2 toward human papillomavirus16-like particles from CINⅠgroup and CINⅡ-Ⅲgroup (1.686±0.013,1.831±0.064) were quite close to that from control group(1.611±0.021), but they were much higher than that HPV infection group (0.551±0.042), (P<0.05).②The IgG2 dominance (IgG2/IgG1 ratio>1) from control group(100%), HPV infection group (87.50%), CINⅠgroup (75%) were much greater than that from CINⅡ-Ⅲpatients(9.52%), (P<0.05). But there was no significant difference among HPV infection group, CINⅠgroup and control group (P>0.05).③The HPV16-IgG and its subclasses seroprevalence from non-HPV16-DNA positive group (15.71% and 7.14%, respectively) were significantly fewer than that from HPV16-DNA positive group (74.29%), (P<0.05).But the OD value of IgG and its subclass toward human papillomavirus16-like particles from HPV16-DNA positive group (1.564±0.062 and 1.365±0.037, respectively) was significantly higher than that of from non-HPV16-DNA positive group(0.453±0.016 and 0.351±0.022, respectively), (P<0.05). There was a moderate correlation between the HPV16-DNA testing and detection of HPV16-IgG toward human papillomavirus16-like particles.Conclusion:①The detection of IgG and its subclasses toward human papillomavirus16-likeparticles is basically in accordance with the HPV-DNA genotyping.②An increase of the expression of IgG and its subclasses toward human papillomavirus16-like particles in the serum of the patients with cervical precancerous lesions especially those with CINⅡ-Ⅲmight associated with duration of HPV infection and severity of cervical lesions.③An increase of the IgG2 dominance (IgG2/IgG1>1) in serum from low grade cervical lesions group and normal control group, might indicating the clearance of HPV infection and the regression of cervical lesions.