Development and Clinical Evaluation of a Highly Sensitive PCR-Reverse Hybridization Line Probe Assay for Detection and Identification of Anogenital Human Papillomavirus

Kleter, Bernhard; Doorn, Leen Jan Van; Schrauwen, Lianne; Molijn, Anco; Sastrowijoto, SH; Schegget, Jan ter; Lindeman, J.; Harmsel, Bram ter; Burger, M. P. M.; Quint, Wim

doi:10.1128/jcm.37.8.2508-2517.1999

Cited by 601 publications

(300 citation statements)

References 39 publications

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“…31 For reasons explained in the results section, 2 samples were also analyzed with the SPF10 DEIA/LiPA version 1 test, which is a more sensitive HPV test. 32 The HPV detection limit of the SPF10 DEIA/LiPa is 65 bp, and it detects 25 different HPV genotypes.…”

Section: Patientsmentioning

confidence: 99%

Imiquimod‐induced clearance of HPV is associated with normalization of immune cell counts in usual type vulvar intraepithelial neoplasia

Terlou

Seters

KleinJan

et al. 2010

Intl Journal of Cancer

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Recently, we reported on the efficacy of imiquimod for treatment of usual type vulvar intraepithelial neoplasia (uVIN). A histologic regression of uVIN to normal tissue was observed in 58% of patients. As success of treatment is related to clearance of high-risk human papilloma virus (HPV), the aim of our study was to assess differences in immune cell counts and in the expression of p16 INK4a in VIN tissue before and after imiquimod treatment, in relation to HPV clearance and clinical response. Vulvar tissue samples taken prior to imiquimod treatment and 4 weeks after treatment were tested for the presence of HPV. Previously determined immune cell counts (CD1a, CD207, CD208, CD123/CD11c, CD94, CD4, CD8 and CD25/HLA-DR) in epidermis and dermis of 25 VIN patients and 19 healthy controls were completed with the counts for CD14 and CD68. The expression of p16 INK4a was investigated by immunohistochemistry in 15 patients. Before imiquimod treatment, both HPV cleared and HPV noncleared patients showed mainly in the dermis significantly upregulated immune cell counts compared to healthy controls. However, in patients that cleared HPV and showed histologic regression already 4 weeks after imiquimod treatment, immune cell counts and p16 INK4a expression were normalized. In conclusion, our data indicate that imiquimodinduced clearance of HPV results in normalization of counts for certain immune cells and is strongly correlated with histologic regression of the disease.Vulvar intraepithelial neoplasia (VIN) is a premalignant disorder that is classified into differentiated type VIN, which is associated with lichen sclerosus, and usual type VIN (uVIN), 1 which is caused by a persistent infection with a high-risk or oncogenic HPV (hrHPV, usually HPV-type 16, 18 or 33). 2 Over the last decades, the incidence of uVIN has increased, most likely due to a rise in the incidence of HPV infections. 3 Lifetime risk to become infected with HPV in western societies is around 80% and $40% of all sexually active, female adolescents are at least once infected with hrHPV. [4][5][6][7] When hrHPV persists (in less than 10% of cases), premalignant disorders of the anogenital tract, such as uVIN, can develop. 8-10 uVIN has invasive potential (10% of untreated cases will progress in 1-4 years) and needs to be treated proactively. 11,12 The host immune response is of critical importance in determining clearance or persistence of an HPV-infection. In natural life, during the early stages of a viral infection, CpGrich regions in the viral DNA are recognized by toll-like receptor (TLR) 7 and 9 on the cell surface of immature dendritic cells (DCs). Upon binding, TLRs activate kinase cascades eventually activating NFjB, which will result in the production of cytokines, adhesions molecules and other effectors of the innate immune response. 13 Other receptor and adhesion molecules at the DC surface can bind viral antigen and will internalize and digest the antigen, which is followed by expression on the cell membrane in major histocompatibility complex...

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Section: Patientsmentioning

confidence: 99%

Imiquimod‐induced clearance of HPV is associated with normalization of immune cell counts in usual type vulvar intraepithelial neoplasia

Terlou

Seters

KleinJan

et al. 2010

Intl Journal of Cancer

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“…18 HPV genotypes were identified by the INNO-LiPA line probe assay (Innogenetics NV, Ghent, Belgium). 19,20 Immunohistochemistry Antigen retrieval using citrate buffer was performed on dewaxed sections prior to blocking with swine serum. Primary antibodies to Ki67 (MIB-1; Dako Cytomation, Glostrup, Denmark; M 7240, dilution 1 : 50) and p16 INK4A (Ab-7; Neomarkers, Fremont, CA, U.S.A.; MS-1064-P0, dilution 1 : 100) were applied for 1 h and detected using an Envision Kit (Dako Cytomation; K4006).…”

Section: Patient Samples Dna Preparation and Human Papillomavirus Gementioning

confidence: 99%

Human papillomavirus-associated increase in p16INK4A expression in penile lichen sclerosus and squamous cell carcinoma

Prowse

Ktori

Chandrasekaran

et al. 2007

British Journal of Dermatology

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Background Human papillomaviruses (HPVs) are sexually transmitted human carcinogens that may play a role in the oncogenesis of penile cancer. Objectives To investigate the role of HPV infection and expression of the tumour suppressor protein p16 INK4A in the pathogenesis of penile cancer. Methods By means of polymerase chain reaction amplification and reverse hybridization line probe assay to detect HPV infection, and immunohistochemical staining for p16 INK4A and Ki67, we analysed 26 penile squamous cell carcinomas (SCCs) and 20 independent penile lichen sclerosus (LS) lesions from 46 patients. Results HPV DNA was found in 54% of penile SCCs and 33% of penile LS cases in single and multiple infections. High-risk HPV 16 was the predominant HPV type detected. No relationship between Ki67 expression and HPV infection was observed. Strong immunostaining for p16 INK4A correlated with HPV 16/18 infection in both penile LS and penile SCC. In our penile SCC series the cancer margins were also associated with penile LS in 13 of 26 lesions, and HPV was detected in seven of the 13 SCC cases associated with LS and in six of the 11 SCC lesions not involving LS. Conclusions Our study shows a high prevalence of HPV 16 and p16 INK4A expression in penile lesions, consistent with an active role for HPV in interfering with the retinoblastoma pathway. High-risk HPV infection could be involved in the tumorigenic process in 50% of penile cancers, and the use of prophylactic HPV vaccines has the potential to prevent these cancers.

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“…12 HPV DNA obtained from plucked eyebrow hair follicles and whole tissue sections (WTS) obtained from formalin-fixed paraffin embedded blocks of the 11 removed lesions were typed for a (genital-and skin-tropic types), b, l and m genera, as previously described. 13,14 The HPV genotypes detected in plucked hair follicles were HPV28 (cutaneous a genus, species 2) and HPV8, HPV9, HPV14, HPV20 and HPV24 from the b genus. All the skin lesions contained one or both of the b-HPV types 20 and 24, and eight lesions were also positive for HPV8.…”

Section: Case Reportmentioning

confidence: 99%

Characterization of skin lesions induced by skin-tropic α- and β-papillomaviruses in a patient with epidermodysplasia verruciformis

et al. 2014

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Epidermodysplasia verruciformis (EV) is a rare, lifelong, autosomal recessive skin disease associated with an unusual susceptibility to infections with ubiquitous β-human papillomaviruses (β-HPVs), and in some cases also skin-tropic α genotypes. In this case report, HPV infection patterns were correlated with pathology and clinical manifestations of skin lesions from a patient with EV, without loss-of-function mutations in the EVER genes. HPV infection was investigated by both polymerase chain reaction (PCR) and laser capture microdissection (LCM) PCR, alongside immunofluorescence for the viral proteins E4 and L1. Analysis of eyebrow hair bulbs revealed multiple β-genus HPV infections, including HPV20 and HPV24, which were consistently found in all 11 skin lesions on the patient. Six lesions were also positive for the skin tropic α-genotype, HPV27. Clear-cut differences between two wart-like lesions, one caused by a skin-tropic α-genotype and the other by β-genotypes (as detected by LCM PCR) are shown, including the high cellular proliferation rate in β-HPV-induced lesions. Widespread expression of the early protein E4 was also evident in skin lesions positive for HPV20 by LCM PCR in both tumours and nearby intraepidermal proliferative areas. L1 expression was restricted to areas of intraepidermal proliferation showing productive infection. The patient's inability to control HPV infections is conclusive to the uncontrolled replication of few genotypes from both α and β genera, which cause proliferative lesions with clear-cut clinical and histological features.

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Development and Clinical Evaluation of a Highly Sensitive PCR-Reverse Hybridization Line Probe Assay for Detection and Identification of Anogenital Human Papillomavirus

Cited by 601 publications

References 39 publications

Imiquimod‐induced clearance of HPV is associated with normalization of immune cell counts in usual type vulvar intraepithelial neoplasia

Imiquimod‐induced clearance of HPV is associated with normalization of immune cell counts in usual type vulvar intraepithelial neoplasia

Human papillomavirus-associated increase in p16INK4A expression in penile lichen sclerosus and squamous cell carcinoma

Characterization of skin lesions induced by skin-tropic α- and β-papillomaviruses in a patient with epidermodysplasia verruciformis

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