Human Papillomavirus Load Research Articles

Research Progress of Immune Mechanisms Related to Persistent HPV Infection in CIN after Cervical Conization

Persistent human papillomavirus (HPV) infection is associated with the grading of cervicalintraepithelial neoplasia (CIN), high-risk HPV infection, multiple HPV infections, high HPV load,HPV infection of surgical margin, and age in CIN after conization. The immune mechanism iscomplex and is primarily related to vaginal microecology disorders, immune escape, immuneresponse impairment, and the release of regulatory cytokines. Currently, the treatment methods forpostoperative persistent HPV infection include surgical treatment, antiviral treatment, vaccination,and other approaches.

Polyhexamethylene Biguanide Reduces High-Risk Human Papilloma Virus Viral Load in Cervical Cell Samples Derived from ThinPrep Pap Test.

Human papilloma virus (HPV) infection and its progression still represent a great medical challenge worldwide. Clinical evidence has demonstrated the beneficial effects of polyhexamethylene biguanide (PHMB) on HPV clinical manifestations; however, evidence of the effect of this molecule on HPV viral load is still lacking. In this in vitro study, 13 ThinPrep Papanicolaou (Pap) tests were treated with a PHMB solution (0.10 g/100 mL) for 2 h. We observed no cytological changes but a significant reduction in the viral load of high-risk (HR) HPV after PHMB treatment, also revealing a dose-dependent antiviral effect. In addition, by stratifying the obtained results according to HR-HPV genotype, we observed a significant reduction in the viral load of HPV 16, P2 (56, 59, 66), 31, and P3 (35, 39, 68) and a strong decrease in the viral load of HPV 45, 52, and P1 (33, 58). Overall, 85% of the analyzed cervical cell samples exhibited an improvement in HPV viral load after PHMB exposure, while only 15% remain unchanged. For the first time, the data from this pilot study support the activity of PHMB on a specific phase of the HPV viral lifecycle, the one regarding the newly generated virions, reducing viral load and thus blocking the infection of other cervical cells.

Abstract 4352: A high rate of episomal HPV16 is present in head and neck squamous cell carcinoma tumors by long-read whole genome sequencing

Abstract Persistent Human Papillomavirus (HPV) infection causes an increased risk of Head and Neck Squamous Cell Carcinoma (HNSCC). However, a comprehensive understanding of the intricate molecular events within virus-positive tumors remains a critical knowledge gap. Therefore, we used long-read sequencing to study the intricacies of HPV integration events within the genomic landscape of HPV+ HNSCC tumors. A total of 16 tissue biopsies, including 10 tumors and 6 matched adjacent normal tissues from histopathologically confirmed HNSCC subjects were sequenced by LSK114 chemistry of Oxford Nanopore Technology with an average coverage of 30X and 10X, respectively. The predicted mitochondrial haplotype from Mitomaster confirmed that the tumor/normal pair was from the same individual. The HPV integration status was determined by aligning fastq_pass files at the Cancer Genomics Cloud and viewing the breakpoints in the Integrated Genome Viewer. HPV lineage and sub-lineage classification was determined using ClustalOmega. We detected HPV16 in 8/10 tumors, with sublineages A1 and A2 accounting for 38% (3/8) and D3 for 25% (2/8). One tumor sample had a very low HPV load with only one HPV16 sequence read and low detection by PCR. Interestingly, we found that 5/8 HPV positive tumors had episomal HPV16 with different states. So, two tumors had intact HPV episomes, two more had either a small deletion or an insertion in the episomal HPV, and the last one had a rearranged episomal multimer (Table 1). The remaining three tumors had integrated HPV16: one with type 1 integrations at 11q12.13 and two others with type 2 integrations at chr1q42.12 and chr14q21.1. Our results indicate that episomal HPV16 is more prevalent than integrated HPV16 in primary HPV+ HNSCC samples. These findings provide new insights into the molecular mechanisms and consequences of HPV infection in HNSCC. Table 1: HPV characterization in HNSCC studied tissue biopsies. Patient ID Race HPV status HPV sublineages Predicted Haplotype HPV pattern #PID_1 Caucasian HPV16 + A2 T2b (T2b) Type 2 Integration (chr1q42.12) #PID_2 - HPV- - I1a (I1a1b) - #PID_3 Caucasian HPV16 + A1 H1q (H1q) Episomal (rearranged multimer) PID_4 African American HPV16 + A1 - Episomal (deletion of 636bp from 4892 to 5528) #PID_5 - HPV16 + A2 H1 (H1+16189) Type 1 Integration (complex chr11q12.13) *#PID_6 Caucasian HPV16 + - T2b (T2b) - PID_7 Caucasian HPV16 + D3 - Episomal #PID_8 African American HPV16 + D3 L1c (L1c3a) Type 2 Integration (chr14q21.1) PID_9 African American HPV16 + A2 - Episomal (insertion of 257bp from 7310-7567) PID_10 Caucasian HPV16 + A1 - Episomal *Very weakly HPV16 positive; #Tumor-normal pair Citation Format: Sonam Tulsyan, Vera Mukhina, Isabel Rodriguez, Subhendu R. Choudhury, Erin Allor, Kyle Hatte, Michael Dean, Daria A. Gaykalova. A high rate of episomal HPV16 is present in head and neck squamous cell carcinoma tumors by long-read whole genome sequencing [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4352.

Human papillomavirus (HPV) load is higher in HPVDNA/p16 positive than in HPVDNA positive/p16 negative oropharyngeal squamous cell carcinoma but does not differ significantly between various subsites or correlate to survival

ObjectivePatients with human papillomavirus DNA positive (HPVDNA+) and p16ink4a overexpressing (p16+) oropharyngeal squamous cell carcinoma (OPSCC), especially those with cancer in the tonsillar and base of tongue subsites as compared to other OPSCC subsites have a better outcome than those with only HPVDNA+ or only p16+ cancer. Likewise having a high viral load has been suggested to be a positive prognostic factor. We therefore hypothesized, that HPV viral load could vary depending on OPSCC subsite, as well as with regard to whether the cancer was HPVDNA+ and p16+, or only HPVDNA+, or only p16+ and that this affected outcome. Material and methodsTo address these issues HPV viral load was determined by HPV digital droplet (dd) PCR in tumor biopsies with previously known HPVDNA/p16 status from 270 OPSCC patients diagnosed 2000–2016 in Stockholm, Sweden. More specifically, of these patients 235 had HPVDNA+/p16+, 10 had HPVDNA+/p16-, 13 had HPVDNA-/p16+ and 12 had HPVDNA-/p16- cancer. ResultsWe found that HPVDNA+/p16+ OPSCC had a significantly higher viral load than HPVDNA+/p16- OPSCC. Moreover, there was a tendency for a higher viral load in the tonsillar and base of tongue OPSCC subsites compared to the other subsites and for a low viral load to correlate to a better clinical outcome but none of these tendencies reached statistical significance. ConclusionTo conclude, the mean viral load in HPVDNA+/p16+ OPSCC was higher than in HPVDNA+/p16- OPSCC, but there was no statistically significant difference in viral load depending on OPSCC subsite or on clinical outcome.

Recombinant Human Interferon α-2b Nanoparticles in the Treatment of Human Papillomavirus Infection Model and Detection of LMX 1A Expression in Cervical Exhumation Cells

Interferon (INF) is a kind of multifunctional and highly active protein-like cytokines produced by animal cells after stimulation, which exerts the broad-spectrum antiviral, anti-tumor, and immunomodulatory effects. Preparing the nanoparticles (NPs) loaded drug can improve the stability of protein polypeptides in vivo, prolong the biological half-life period, and improve the bioavailability. In this research, recombinant human interferon (RHUINF) α-2b NPs were prepared to explore their therapeutic effect on human papillomavirus (HPV) infection model. Firstly, the RHUINF α-2b poly (lactic acid-glycolic acid) copolymer (PLGA) of composite bioactive glass was prepared by electrostatic spray technology. The in vitro quality, stability, and pharmacokinetic characteristics in vivo of the NPs were measured. Then, a mouse model infected with HPV16 pseudovirus infection (NC group) was prepared. Next, they were treated by painting imiquimod (IMQ) (IMQ group), intramuscularly injecting the RHUINF α-2b (Intron A), intramuscularly injecting the RHUINF α-2b-Plga NP suspension (Intra/PLGA), and intramuscularly injecting the RHUINF α-2B-bioactive glass-PLGA NP suspension (Intron A/BAG/PLGA), respectively. In addition, the differences in serum inflammatory cytokines, vaginal biofluorescence activity (VBA), HPV load, and LMX 1A expressions in cervical exhumation cells (CECs) were analyzed. The average particle sizes (PSs) of Intron A/PLGA and Intron A/BAG/PLGA NPs were 352.3 nm and 385.1 nm, respectively, and no great differences were observed in encapsulation rate (ER), drug loading (DL), and in vivo release (IVR). The maximum peak times (MPTs) of Intron A, Intron A/PLGA, and Intron A/BAG/PLGA were 1.3 h, 6.2 h, and 6.5 h, respectively, and their maintenance durations were 5.5 h, 120.7 h, and 245.6 h, respectively. Based on the NC group, IFN-γ, IL-2, and TNF-α in the IMQ group, Intron A group, Intron A/PLGA group, Intron A/PLGA group, and Intron A/BAG/PLGA group were decreased, VBA and HPV load were decreased, and mRNA of LMX 1A in ECEs was increased (p < 0.05). Based on the Intron A group, the Intron A/PLGA and Intron A/BAG/PLGA groups exhibited decreased serum inflammatory factors (IFFs), VBA, HPV load, and LMX 1A mRNA in the CECs were increased (p < 0.05). By taking the Intron A/PLGA group as reference, the Intron A/BAG/PLGA group showed greatly reduced serum IFFs, VBA, HPV load, and LMX 1A mRNA expression in CECs (p<0.05). Preparing RHUINF α-2b NPs could prolong the half-life period of RHUINF α-2b in vivo, reduce the inflammatory response and load of HPV-DNA in HPV infection models, and upregulate LMX 1A in CECs. In addition, RHUINF α-2b NPs could improve the therapeutic effect of RHUINF α-2b.

Effects of two and three vaccinations with the bivalent HPV vaccine on the prevalence and load of HPV in clearing and persistent infections in young women.

Human papillomavirus viral load (HPV VL) is associated with persistence, which increases cervical cancer risk. The bivalent vaccine protects against oncogenic HPV16/18 and cross-protects against several non-vaccine types. We examined the effect of two-dose (2D) and three-dose (3D) vaccination on HPV prevalence and viral load in clearing (CIs) and persistent infections (PIs), 6y and 12y post-vaccination, respectively. Vaginal swabs collected from HPV Among Vaccinated And Non-vaccinated Adolescents (HAVANA, 3D-eligible) and HAVANA-2 (2D-eligble) participants were genotyped for HPV with the SPF10-DEIA-LiPA25. HPV VL was measured with type-specific qPCRs. HPV16, 18, 31, 33 and 45 CI and/or PI prevalence and HPV16, 18, and 31 VLs in CIs were significantly reduced in 3D vaccinated women compared to unvaccinated women. Except for HPV11 and HPV59 CIs, no significant VL differences were observed among vaccinated women, ≤6y and >6y post-vaccination. Infection numbers were low in 2D-eligible women, with no HPV16 and 18 in vaccinated women. No VL differences for the remaining types were found. 3D vaccination reduces HPV prevalence in CIs and PIs and decreases HPV VLs in CIs, 12y post-vaccination for vaccine and several non-vaccine types. 2D-eligible women had low infection numbers, with no HPV16/18 among vaccinated women.

Association of MicroRNA-10b Expression and P16 with Pathological Features in Various Stages of Cervical Precancerous Lesions.

Cervical cancer is the fourth most prevalent cancer for females with 14,100 new cases each year globally. Efficient screening and intervention at the precancerous stage is the key point to the prevention and treatment of cervical cancer. However, no widely recognized biomarkers have been discovered yet. We investigated the expression of miR-10b in cervical cells and its correlation with clinicopathological features in different pathological grades of cervical precancerous lesions. The expression of miR-10b in cervical cytology samples from 20 cases of LSIL, 22 cases of HSIL, 18 cases of early-stage cervical cancer, and 20 cases of cervicitis controls were assessed using qPCR. From the same cervical cytology samples, the human papillomavirus (HPV) load was assessed using semi-PCR and the lesion size, and gland involvement levels from the same subjects were assessed during the cervical examination. The correlation between miR-10b expression and different pathological grades of cervical lesions was analyzed. We also calculated the correlation between HPV load, lesion size, gland involvement, P16 expression, and different pathological grades. The expression of miR-10b exhibited a step-decreasing manner from cervicitis control (4.23(4.00,4.71)) to LSIL (2.67(2.52,2.90)), HSIL (1.49(1.30,1.80)) and cervical cancer group (0.65(0.55,0.80)). There is a significant difference (P<0.001) between cervicitis and HSIL, cervicitis and cervical cancer, ISIL and HSIL, as well as ISIL and cervical cancer but not between the cervicitis group and the LSIL group. In addition, more severe pathological grades were correlated with a bigger rate of gland involvement (P＜0.001). We also found that different pathological grades were correlated with the intensity of P16 expression (P=0.001), and the intensity of P16 expression is positively correlated with different pathological grades (P<0.05). Repressed expression of miR-10b is related to the progression of cervical precancerous lesions. Increased gland involvement rate and increased intensity of P16 expression are risk factors for developing cervical cancers. Our result showed that miR-10b may be a potential biomarker for the screening and ranking of cervical precancerous lesions.

Changes in the Cervical Microbiota of Women with Different High-Risk Human Papillomavirus Loads

The cervical microbiota is essential in female sexual health, and its altered states seem to have a central role in the dynamic of high-risk papillomavirus (hrHPV) infections. This study aimed to evaluate the variation in bacterial communities' compositions according to hrHPV. We collected two samples per woman, with a difference of 12 ± 1 months between them, and performed a follow-up on 66 of these women. The viral load (VL) of the hrHPV was estimated by quantitative PCR (qPCR), then it was normalized (using the HMBS gene as reference) and transformed to the Log10 scale to facilitate the interpretation. The VL was categorized as Negative, without hrHPV copies; Low, less than 100 hrHPV copies; Medium, between 100 to 102 hrHPV copies; and High, >102 hrHPV copies. The microbiota composition was described through the Illumina Novaseq PE250 platform. The diversity analyses revealed changes regarding the hrHPV VL, where women with low VL (<100 hrHPV copies) presented high diversity. The community state type (CST) IV was the most common. However, in women with high VL, a lower association with Lactobacillus depletion was found. Lactobacillus gallinarum and L. iners were the most abundant species in women with high VL, whereas women with low VL had a 6.06 greater probability of exhibiting Lactobacillus dominance. We identified conspicuous differences in the abundance of 78 bacterial genera between women with low and high VL, where 26 were depleted (e.g., Gardnerella) and 52 increased (e.g., Mycoplasma). A multilevel mixed-effects linear regression showed changes in the diversity due to the interaction between the measurement time and the VL, with a decrease in diversity in the second follow-up in women with low VL (Coeff. = 0.47), whereas the women with medium VL displayed an increase in diversity (Coeff. = 0.58). Here, we report for the first time that the cervical microbiota is influenced by the number of copies of hrHPV, where a decrease in the abundance of Lactobacillus, greater diversity, and enrichment of bacterial taxa is relevant in women with low VL.

Quantification of Tumor-Associated Cell-Free HPV DNA in Surgical Drain Fluid Equates to Molecular Residual Disease and Correlates with High-Risk Pathology

<h3>Purpose/Objective(s)</h3> Multi-institutional efforts aimed at safely de-escalating treatment in human papillomavirus (HPV) associated oropharyngeal squamous cell carcinoma (OPC) patients rely on subjective pathology for risk stratification. While plasma HPV assays are emerging for post-treatment disease surveillance, they have poor sensitivity to detect postoperative molecular residual disease (MRD). Here we present a liquid biopsy assay of the serosanguinous fluid removed from surgical drains, proximal to the surgical site, to sensitively measure postoperative MRD. We hypothesize that HPV quantified in surgical drain fluid (SDF), 1) represents post-operative regional MRD, and 2) could improve patient risk-stratification to guide precision radiotherapy. <h3>Materials/Methods</h3> We collected SDF at 24-48 hours post-surgery for cell-free (cf) DNA extraction. Our 87-patient head and neck cancer cohort included 74 HPV(+) and 8 HPV(-) OPCs, and 5 benign tumors. Paired plasma and tumor samples were collected from approximately half of all patients. We used the HPV16 E6T2aE7 plasmid to compare our Taqman quantitative PCR (qPCR) to digital droplet PCR (ddPCR). We validated HPV(+) OPC PCR results using next-generation HPV sequencing (NGS) in 9 patients. Then we correlated SDF HPV copies to extranodal extension (ENE), AJCC 8^th edition pathological stage, smoking pack-years, recurrence-free survival (RFS), and composite risk scores derived from OPC de-escalation clinical trials. Statistical methods included Fisher's Exact test, Mann-Whitney (MW) test, Kruskal-Wallis (KW) test, and Kaplan-Meier (KM) analysis. <h3>Results</h3> Our HPV qPCR assay's limit of detection was 9.7 copies, similar to what we saw with ddPCR. HPV genotyping was concordant between NGS and PCR, and all SDF cfDNA samples with nondetectable (ND) HPV via PCR were ND via NGS. HPV was detected in 63 of 74 (85%) SDF samples compared to only 7 of 48 (15%) paired plasma samples. Median HPV copies in ENE+ patients' SDF was 15-fold higher than SDF from ENE- patients (P = 0.002). HPV copies also increased with increasing nodal (P = 0.01) but not primary tumor stage (P = 0.4), suggesting post-operative SDF HPV load reflects invasive nodal disease and is a proxy for regional MRD. When we stratified our cohort according to high-risk and low-risk scores drawn from four clinical trials (RTOG 9501, EORTC 22931, ECOG 3311, MC1273), SDF HPV load was consistently higher in patients with trial-defined high-risk pathology (mean AUC = 0.73). Lastly, KM RFS analysis of HPV-high and HPV-low patients, defined by an RTOG 9501-derived SDF classifier, showed HPV-high patients trended toward worse RFS (HR = 3.1; P = 0.1). <h3>Conclusion</h3> Liquid biopsy assessment of SDF, the biofluid most proximal to the nodal environment, enables postoperative regional MRD detection in HPV(+) OPC, and correlates with high-risk nodal pathology and worse RFS. If confirmed, SDF cfDNA analysis could aid traditional pathology and imaging to personalize adjuvant radiotherapy for surgically treated HPV(+) OPC.

Hookworm infection associates with a vaginal Type 1/Type 2 immune signature and increased HPV load.

Helminth infection-driven changes to immunity in the female reproductive tract (FRT) is an immune axis that is currently understudied but can have major implications for the control of FRT infections. Here we address how human hookworm infection associates with vaginal immune profile and risk of Human papillomavirus (HPV) infection. Stool, blood, cervical swabs and vaginal flushes were collected from women from the Central region of Togo to screen for hookworms (Ancylostoma duodenale) and high carcinogenic risk HPV types, via Kato Katz and PCR, respectively. Cytokine, chemokine and immunoglobulin levels were analysed in cervicovaginal lavages and plasma samples. A pronounced mixed Type 1/Type 2 immune response was detected in the vaginal fluids of women with hookworm infection and this immune signature was a notable feature in hookworm-HPV co-infected women. Moreover, hookworm infection is positively associated with increased risk and load of HPV infection. These findings highlight helminth infection as a significant risk factor for acquiring a sexually transmitted viral infection and potentially raising the risk of subsequent pathology.

Strategic Significance of Low Viral Load of Human Papillomavirus in Uterine Cervical Cytology Specimens.

Infection with high-risk (HR) Human Papillomavirus (HPV) is associated with the development of precancerous lesions or invasive carcinoma of the uterine cervix. Thus, the high viral load (VL) of HR-HPV DNA currently serves as a representative quantitative marker for cervical cancer. However, the clinical significance of low HPV DNA VL remains undetermined. This study aimed to evaluate the clinical association between the low HPV DNA VL and cytology/histologic diagnosis of cervical samples. We searched the electronic medical databases for the resultant analyses of HPV genotyping among patients who underwent treatment for any cervical lesion or who had undergone gynecological examinations with any positive HPV results according to the national cancer screening service between 2015 and 2016. HPV testing with genotyping and semi-quantitative VL measurement was conducted using an AnyplexTM II H28 Detection assay (H28 assay, Seegene, Seoul, Republic of Korea). The H28 assay is a multiplex semi-quantitative real-time PCR test using the tagging of oligonucleotide cleavage and extension (TOCE) technology. The VL was semi-quantified as high (3+; positive signal before 31 PCR cycles), intermediate (2+; positive between 31 and 39 PCR cycles), or low (1+; positive after 40 PCR cycles). Out of 5940 HPV VL analyses, 356 assays (5.99%) were reported as low VL (1+) of HPV DNA. Matched cytology diagnoses were mostly negative findings (n = 347, 97.5%), except for seven cases of atypical squamous cells of undetermined significance (1.9%) and two cases of atypical glandular cells (0.6%). During the follow-up periods, abnormal cytologic diagnoses were identified, including one case of high-grade squamous intraepithelial lesion (HSIL) and two low-grade squamous intraepithelial lesions (LSILs). The matched, confirmative histologic diagnosis of HSIL cytology was compatible with chronic inflammation, wherein the two LSILs had regular check-ups. None revealed clinically concerned outcomes associated with HPV-related squamous lesions. The cytology was most likely negative for malignancy when the VL of HPV DNA was low (1+). Additional strategic monitoring and management may thus be unnecessary.

A new option for the treatment of condyloma acuminatum in the male urethra: Multimodal ultrasound image-guided scraping and photodynamic therapy (USP)

ObjectivesCondyloma acuminatum (CA) is a sexually transmitted disease with a high recurrence rate due to the rapid replication of human papillomavirus (HPV) and its subtle immune escape mechanism, which makes the diagnosis and treatment of CA in the male urethra particularly difficult. This study aims to evaluate the efficacy of comprehensive treatments for male urethral CA after accurate localization of warts under ultrasound guidance. MethodsThe study included 15 men with intraurethral CA. Before treatment, the urethra was examined by ultrasonography and HPV-PCR. After examination of the invisible urethral warts, wart curettage (penetrating operation with a special stainless steel medical curettage tool) combined with ALA-PDT was used for treatment. The ultrasound and HPV load were reviewed 1 week after treatment, and again at 1, 3, and 6 months after treatment. ResultsAll patients achieved satisfactory results 1 week after the last treatment. The viral load of human papilloma was significantly reduced or turned negative, ultrasound imaging exploration showed no neoplasm in the urethra, and no obvious intraoperative or postoperative complications were observed. The side effects in patients included a mild burning or tingling sensation confined to the treated area. After a 6 month follow-up period, only 2 patient relapsed. ConclusionThe combined diagnosis and treatment of CA in the male urethra under the guidance of multi-mode ultrasound imaging is an effective, economical, safe, and well-tolerated treatment method.

Comprehensive Analysis of the Carcinogenic Process, Tumor Microenvironment, and Drug Response in HPV-Positive Cancers.

Human papillomavirus (HPV) is a common virus, and about 5% of all cancers worldwide is caused by persistent high-risk HPV infections. Here, we reported a comprehensive analysis of the molecular features for HPV-related cancer types using TCGA (The Cancer Genome Atlas) data with HPV status. We found that the HPV-positive cancer patients had a unique oncogenic process, tumor microenvironment, and drug response compared with HPV-negative patients. In addition, HPV improved overall survival for the four cancer types, namely, cervical squamous cell carcinoma (CESC), head and neck squamous cell carcinoma (HNSC), stomach adenocarcinoma (STAD), and uterine corpus endometrial carcinoma (UCEC). The stronger activity of cell-cycle pathways and lower driver gene mutation rates were observed in HPV-positive patients, which implied the different carcinogenic processes between HPV-positive and HPV-negative groups. The increased activities of immune cells and differences in metabolic pathways helped explain the heterogeneity of prognosis between the two groups. Furthermore, we constructed HPV prediction models for different cancers by the virus infection score (VIS) which was linearly correlated with HPV load and found that VIS was associated with drug response. Altogether, our study reveals that HPV-positive cancer patients have unique molecular characteristics which help the development of precision medicine in HPV-positive cancers.

Association between ТР53, MDM2 and NQO1 gene polymorphisms and viral load among women with human papillomavirus.

The risk of cervical cancer is caused by persistent human papillomavirus (HPV) infection. Cervical cancer is the most frequent cancer among women. Our purpose was to investigate the association between TP53 215C>G (Pro72Arg), MDM2 -410T>G, and NQO1 609C>T gene polymorphisms with a high HPV load and the inf luence of gene-gene interactions on prolonged HPV infection. Eighty-nine women with a high HPV viral load and 114 healthy women were involved in a case–control study. Genotyping for TP53 215C>G (Pro72Arg) and MDM2 -410T>G SNPs was carried out by allele-specif ic PCR and genotyping for NQO1 609C>T was performed by a TaqMan assay. Quantitative analysis of HPV DNA was performed by AmpliSens® HPV HCR screen-titer-FRT test system. Gene-gene interactions were analyzed using the multifactor dimensionality reduction (MDR) method. The study of separate SNPs of MDM2 -410T>G and NQO1 609C>T genes did not reveal any statistically signif icant difference in genotype and allele frequencies among women within the two groups. The frequency of the 215G (72Arg) allele and 215GG (72Arg/ Arg) genotype of the TP53 gene was signif icantly higher in the case group than in the control group (OR = 1.74, 95 % CI = 1.10–2.73; p = 0.02 and OR = 1.97, 95 % CI = 1.13–3.46; p = 0.04, respectively). MDR analysis showed the signif icance of intergenic interactions of the three studied loci TP53 (rs1042522) – MDM2 (rs2279744) – NQO1 (rs1800566) for the formation of a high HPV load (OR = 3.05, 95 % CI = 1.73–5.46; p = 0.0001).

Expediency of immunocytochemical study as a marker of proliferative activity in cervical intraepithelial neoplasia

Aim. To assess the feasibility of using immunocytochemical studies as a marker of proliferative activity in cervical intraepithelial neoplasia.&#x0D; Materials and methods. A comprehensive examination of 78 women, aged 25 to 45 years, with the presence of cervical intraepithelial neoplasia was carried out. Verification of the diagnosis was carried out on the basis of data from clinical, cytological, colposcopic, histological studies. Human papillomavirus typing was carried out using the polymerase chain reaction method (Kvant 21 test).&#x0D; Results. The results obtained in our study indicate that in patients with CIN II, III, chronic cervicitis associated with human papillomavirus (HPV) is an obligatory background pathology. Regardless of the severity of cervical dysplasia (CIN IIIII), among the various types of HPV, HPV type 16 and associations of various HPV types of high oncological risk prevailed (41.03 and 25.64%, respectively). The HPV load result was expressed in decimal logarithms (Lg). Co-expression of p16ink4a oncoprotein and proliferative activity marker Ki-67 in epithelial cells was observed at viral load parameters of 35 Lg in 64.10% of cases, and at values of 5 Lg in 100% of cases.&#x0D; Conclusion. Our data indicate the need for wider use of non-invasive determination of markers р16ink4a and Ki-67 during primary screening, which will improve the tactics of managing patients with cervical precancer. Persistent course of human papillomavirus in patients with cervical intraepithelial neoplasia II in the presence of signs of coexpression of p16ink4a and Ki-67 dictates the inexpediency of performing cervical excision with subsequent control after 6 months.

Analysis of related factors of cervical intraepithelial neoplasia complicated with vaginal intraepithelial neoplasia.

To explore the underlying risk factors and to prevent misdiagnosis of cervical intraepithelial neoplasia (CIN) coexisted with vaginal intraepithelial neoplasia (VaIN). Clinical data of patients pathologically diagnosed with CIN were collected from January 2017 to December 2018. A total of 446 cases were analyzed, including 406 cases of single lesions ('CIN single' group) and 40 cases complicated with VAIN ('VAIN concurrent' group). The median age of the VAIN concurrent group was 53years (46.25-59years), and the median age of the CIN single group was 44years (36-50years). Regarding menopausal status, there were 28 cases (70.0%) in the VAIN concurrent group and 89 cases (21.9%) in the CIN single group (P < 0.005). The median load of high-risk human papillomavirus (Hr-HPV) in the VAIN concurrent and CIN single group was 923.4 relative light units/cutoff (RLU/CO) (145-2172.2 RLU/CO) and 229.155 RLU/CO (18.615-638.1275 RLU/CO), respectively (P = 0.037). The results revealed that the menopausal status was an independent risk factor for VAIN occurrence in CIN patients. The risk of VAIN in menopausal patients was higher than that in non-menopausal CIN patients (OR = 8.311, 95% CI 4.062-17.005). Age and HPV load were also related to the concurrence of VAIN and CIN. Examinations regarding vaginal screening are of great importance in the diagnosis of perimenopausal and postmenopausal CIN patients, especially patients with Hr-HPV load. Colposcopy and tissue biopsy should also be performed, when necessary, to avoid misdiagnosis and the appearance of vaginal lesions.

Polymorphism of TP53 gene and the risk of high human papillomavirus load in cervical epithelial cells

BackgroundCervical cancer is mostly caused by persistent human papillomavirus infection. Host genetic variables, such as polymorphisms, may play a role in the development of cervical cancer and may be a risk factor for HPV-induced carcinogenesis. AimsTo investigate the association between TP53 Pro72Arg polymorphism and the high HPV loads in cervical epithelial cells of women older than 30 years. Subjects and methods154 women infected with HPV and 156 healthy women were recruited into a case–control study. Total DNA was isolated from scrapings of epithelial cells from the cervical canal. Genotyping of the TP53 Pro72Arg polymorphism was carried out using allele-specific PCR. Quantitative analysis of human papillomavirus DNA was performed by the AmpliSens®HPV HCR screen- titre-FRT test system. ResultsNo statistically significant difference was detected between allele and genotype frequencies (p = 0.54 and p = 0.67 respectively) in either women with a medium viral load (3–4 log HPV genomes per 100 thousand human cells) or the control group. The frequencies of the ТР53 Arg72 allele and ТР53 72Arg/Arg genotype were significantly higher in women with a high viral load (above 4 log HPV genomes per 100 thousand human cells) than in the control group (OR = 2.36, p = 0.001 and OR = 2.63, p = 0.002 respectively). The ТР53 Pro72 allele and ТР53 72Pro/Pro genotype are associated with a decreased risk of high HPV load group (OR = 0.42, p = 0.001 and OR = 0.14, p = 0.002 respectively). ConclusionThe risk of developing a high viral load in cervical epithelial cells is associated with the TP53 72Arg allele and the TP53 72Arg/Arg genotype in HPV-positive women.

Viral load as a factor of persistence of papillomavirus infection in cervical intraepithelial neoplasia grade I

Introduction. The viral antigenic load can influence the nature of the infectious response, leading to elimination of the virus or to chronicity of the process, and in some cases to the progressive course of the disease. The aim of the study was to investigate the relationship between the types of viral load of human papillomavirus and the age of a patient with cervical cervical intraepithelial neoplasia grade I. Material and methods. 86 patients with cervical papillomavirus-induced cervical intraepithelial neoplasia grade I by liquid oncocytology were examined. The frequency of human papillomavirus genotypes of high carcinogenic risk in this group of women was analyzed taking into account the viral load by polymerase chain reaction with real-time hybridization-fluorescence detection. The correlation between the viral load of the human papillomavirus and the age of the patient with further prediction of the regression or progression of cervical pathology was studied. Results. This study found that female patients under 30 years of age with cervical intraepithelial neoplasia grade I associated with papillomavirus infection showed a decrease in viral load after 12 months of follow-up in 30.2%, and in patients with cervical intraepithelial neoplasia grade I of older age — in 9.3%. Discussion. The management tactics for grade I cervical intraepithelial neoplasia associated with papillomavirus infection depends largely on the age of the patients and the degree of viral load. In young patients with immediate reproductive plans, a wait-and-see approach with dynamic observation for 12 months without drug therapy is possible in view of the probable spontaneous elimination of the human papillomavirus within a year. The results of the study suggest a higher rate of human papillomavirus elimination in cervical intraepithelial neoplasia grade I in women under 30 years of age. Conclusion. In cervical intraepithelial neoplasia of degree I in women under 40, infection with several types of the human papilloma virus (16, 18, 31, 51) with a clinically significant viral load (over 4.69±0.07 HPV per 100,000 cells) was found most frequently. In the group of the older reproductive age (40-49 years old), HPV HCC is less frequent in 6.90%. And the predominant role belongs to type 16, which was spread in 69.7% of cases.

Association of Relative Telomere Length and Risk of High Human Papillomavirus Load in Cervical Epithelial Cells.

Importunate high-risk HPV (HR-HPV) infection is the most common trigger for the cervical carcinogenesis process. In this respect, the presence of cancer can be imputed to telomere lengthening or shortening. This paper explores the possible correlation between relative telomere length and viral load in two groups of women, namely: those with high-risk HPV infection and those who do not have this infection. Thus, samples comprising of 50 women in each group were evaluated for this research. The Amplisens HPV HCR screen-titre-FRT PCR kite was employed for quantitative analysis. Relative telomere length was quantified by real-time PCR. In each of the two HPV load groups, there was no correlation between age and telomere length. Telomere shortening was found in the cervical cell samples of women with high HPV loads, compared with women in the control group. Telomere shortening is associated with elevated HPV loads.

Preliminary study of the use of E6/E7mRNA detection in screening and triage management of HR-HPV infection during pregnancy.

BackgroundDetection of E6 and E7 mRNA load of high-risk human papillomavirus (HR-HPV) infection during pregnancy was compared with conventional cytopathology and DNA detection by pathological examination as colposcopy to evaluate the application of E6 and E7 mRNA detection in the diagnosis and management in HR-HPV infection for high -grade cervical lesions during pregnancy.MethodsFrom January 2014 to June 2019, 1,058 pregnant women of childbearing age who were filed for regular obstetrics in the Department of Obstetrics and Gynecology, Xuanwu Hospital of Capital Medical University, were separately assessed using cervical liquid-based cytology and HPV DNA detection. If the results were abnormal, colposcopy was performed as a follow-up. The presence of HR-HPV E6/E7 mRNA fragments was detected through the HR-HPV E6/E7 mRNA test, and monitored at the same time as colposcopy. The diagnostic efficacy of the HR-HPV DNA test versus the HR-HPV E6/E7 mRNA test for high-grade cervical lesions during pregnancy was compared.ResultsThe positive rate of HR-HPV E6/E7 mRNA detection in the overall cervical intraepithelial neoplasia (CIN) and above during pregnancy was lower than that with HR-HPV DNA detection, and there was a significant statistical difference between the two methods. In CIN I and normal or inflammatory results, the positive rate of HR-HPV E6/E7 mRNA detection was lower than that of HR-HPV DNA detection, while in the results of CIN II and CIN III, the positive rate of the two was not significantly different. HR-HPV E6/E7 mRNA detection is the same as HR-HPV DNA detection, both of which increased with the severity of cervical lesions, and the positive rate increased. In cases of maintenance or progression of cervical lesions, the positive rate of HR-HPV E6/E7 mRNA detection during pregnancy can reach 81.8%. High-grade cervical lesions during pregnancy had a higher rate of reversal to a lower level after delivery.ConclusionsThe results suggested that the use of HR-HPV E6/E7 mRNA detection in cases of positive HR-HPV DNA detection can significantly improve the diagnostic specificity of CIN II and above high-grade cervical lesions.