Lall Mahima, Dar Lalit, Bhatla Neerja, Kumar Pankaj, Choudhary Aashish, Mathur Sandeep R, Gupta Rajiv M
Armed Forces Medical College, Pune, India.
All India Institute of Medical Sciences, New Delhi, India.
Front Reprod Health. 2021 Sep 13;3:695254. doi: 10.3389/frph.2021.695254. eCollection 2021.
Both human papillomavirus (HPV) and the human immunodeficiency virus (HIV) are sexually transmitted. High-risk (HR) HPV types are a causal factor in cervical cancer. Persistent HPV infection in this subset of immunocompromised women results in faster disease progression. The study determined the prevalence of HPV genotypes in cervicovaginal secretions of HIV seropositive women and the correlation with CD4 counts and cytology. One hundred, non-pregnant, HIV-positive women of 18 years of age and above were enrolled in this cross-sectional study following approval by the institutional ethical committee. A written consent, questionnaire, followed by sample collection including a Papanicolaou (Pap) smear for cytology was undertaken. Cervicovaginal secretion samples were collected in the Digene specimen transport medium (STM) (Qiagen Gaithersburg Inc., MD, USA). HPV genotyping was carried out with PCR amplification of a 65-base pair (bp) fragment in the L1 region of the HPV genome using the short PCR fragment (SPF10) primers followed by reverse hybridization by line probe assay (LPA) using the INNOLiPA HPV Genotyping Extra kit (Fujirebio, Belgium). Quantitation of HPV-16 and-18 viral loads (VLs) was done by real-time PCR. Results of Pap smear cytology were correlated with CD4 counts and HPV-16 and-18 VLs. Mean age of the subjects was 34.9 years ± 7.2 years (median 33.0 years, range 24-60 years). HPV was detected in 62 of 93 (66.6%) samples. Twenty (32.25%) of these 62 samples harbored a single HPV genotype. Multiple genotypes (more than two) were detected in 38 (61.3%) samples. HPV-16 was the commonest genotype detected in 26 (27.9%) of all samples and 41.9% of HPV positive samples. Pap smear cytology was reported for 93 women included in the study. Women who had normal cytology were reported as negative for intraepithelial malignancy or lesion (NILM; = 62; 71.36%), two women had a high-grade squamous intraepithelial lesion (HSIL), low-grade squamous intraepithelial lesion (LSIL; = 11), atypical squamous cells of undetermined significance (ASCUS; = 12). Those smears with inadequate material were reported as scant ( = 6). The median CD4 count was 363/cu.mm (range 39-787) in HPV-positive women compared to 423/cu.mm (range 141-996) in those HPV-negative women. Quantitation of HPV-16 and-18 VL was done in duplicate for samples positive by PCR reverse hybridization (INNOLiPA). Of these 20 samples (65%), 12 samples were positive by real-time PCR. The normalized HPV-16 VL ranged between 18 and 240,000 copies/cell. The normalized HPV-18 VL in cervical samples ranged between ~24 and 60,000 copies/cell. HIV-positive women may be infected with multiple genotypes other than HPV-16 and-18. This may have implications on the vaccines available currently which target few specific genotypes only. Studies are required to determine the predictive role of HR HPV genotypes, in significant copy numbers especially in HIV seropositive women. It would be clinically relevant if the HPV VLs, cervical cytology, and CD4 counts are considered into cervical cancer screening programs for triage and follow-up of these women.
人乳头瘤病毒(HPV)和人类免疫缺陷病毒(HIV)均通过性传播。高危(HR)HPV类型是宫颈癌的致病因素。免疫功能低下的女性持续感染HPV会导致疾病进展加快。本研究确定了HIV血清阳性女性宫颈阴道分泌物中HPV基因型的流行情况及其与CD4细胞计数和细胞学检查结果的相关性。在获得机构伦理委员会批准后,100名年龄在18岁及以上的非妊娠HIV阳性女性参与了这项横断面研究。研究人员获取了书面知情同意书并进行问卷调查,随后进行样本采集,包括用于细胞学检查的巴氏涂片。宫颈阴道分泌物样本采集于Digene标本运输培养基(STM)(美国马里兰州盖瑟斯堡的Qiagen公司)中。采用短PCR片段(SPF10)引物对HPV基因组L1区域的65个碱基对(bp)片段进行PCR扩增,然后使用INNOLiPA HPV基因分型额外试剂盒(比利时富士瑞必欧公司)通过线性探针分析(LPA)进行反向杂交,从而进行HPV基因分型。通过实时PCR对HPV-16和-18病毒载量(VL)进行定量分析。巴氏涂片细胞学检查结果与CD4细胞计数以及HPV-16和-18病毒载量相关联。研究对象的平均年龄为34.9岁±7.2岁(中位数33.0岁,范围24 - 60岁)。在93份样本中的62份(66.6%)检测到HPV。这62份样本中有20份(32.25%)携带单一HPV基因型。在38份(61.3%)样本中检测到多种基因型(超过两种)。HPV-16是在所有样本的26份(27.9%)以及HPV阳性样本的41.9%中检测到的最常见基因型。对纳入研究的93名女性进行了巴氏涂片细胞学检查报告。细胞学检查结果正常的女性报告为上皮内恶性肿瘤或病变阴性(未见上皮内病变或恶性病变;n = 62;71.36%),两名女性患有高级别鳞状上皮内病变(HSIL),低级别鳞状上皮内病变(LSIL;n = 11),意义不明确的非典型鳞状细胞(ASCUS;n = 12)。那些取材不足的涂片报告为样本量少(n = 6)。HPV阳性女性的CD4细胞计数中位数为363/立方毫米(范围39 - 787),而HPV阴性女性的CD4细胞计数中位数为423/立方毫米(范围141 - 996)。对PCR反向杂交(INNOLiPA)呈阳性的样本,对HPV-16和-18病毒载量进行了一式两份的定量分析。在这20份样本(65%)中,12份样本通过实时PCR检测为阳性。标准化后的HPV-16病毒载量范围在18至240,000拷贝/细胞之间。宫颈样本中标准化后的HPV-18病毒载量范围在约24至60,000拷贝/细胞之间。HIV阳性女性可能感染除HPV-16和-18之外的多种基因型。这可能对目前仅针对少数特定基因型的可用疫苗产生影响。需要开展研究以确定HR HPV基因型的预测作用,尤其是在HIV血清阳性女性中具有显著拷贝数的情况下。如果在宫颈癌筛查项目中将HPV病毒载量、宫颈细胞学检查结果和CD4细胞计数纳入对这些女性的分流和随访中,将具有临床相关性。