Human papillomavirus proteins are found in peripheral blood and semen Cd20+ and Cd56+ cells during Hpv-16 semen infection

BMC Infectious Diseases

Luca De Toni²  Andrea Lenzi¹  Silvia Mason²  Damiano Pizzol²  Andrea Garolla²  Alessandro Bertoldo²  Carlo Foresta² 

[1] Department of Medical Physiopathology, 1st University of Rome “La Sapienza”, Rome, Italy;Department of Molecular Medicine and Centre for Human Reproduction Pathology, University of Padova, Via Gabelli 63, 35121, Padova, Italy

关键词: Hpv-related disease;    Peripheral blood mononuclear cells;    Fluorescence in situ hybridization;    Semen infection;    Human papillomavirus;   

Others  :  1137804
DOI  :  10.1186/1471-2334-13-593

 received in 2013-10-09, accepted in 2013-12-05,  发布年份 2013

【 摘 要 】

Background

Human papillomavirus (HPV) currently represents an important risk factor for cancer development and infertility in humans. Whilst binding of HPV to spermatozoa has been associated with male infertility, an investigation about the presence of HPV-DNA in non-spermatozoal semen cells is lacking. Previous findings documented the presence of HPV in peripheral blood leukocytes. The aim of this study was to investigate the expression of HPV markers in semen and blood leukocytes during HPV-16 infection.

Methods

A total of 32 subjects, 16 patients affected by HPV-16 semen infection and 16 controls, were evaluated in our andrological centre and enrolled in the study. Semen non-spermatozoal cells from all subjects were isolated and evaluated for the expression of HPV-16 markers (DNA and L1, E6 proteins) and further characterized for their molecular phenotype. Analogue determination was performed on peripheral blood mononuclear cells.

Results

The presence of HPV-DNA by FISH analysis in a round cell population from semen, confirmed to be CD45+ leukocytes, was observed. These HPV-DNA containing-cells also displayed HPV-16-E6 and HPV-16-L1 viral proteins and, upon further investigation, were found to be CD20+ and CD56+, likely phenotypes of B cells and natural killer cells (NK) respectively. In 25% of the patient group, a very small population of peripheral blood mononuclear cells was found to be positive for HPV-DNA via FISH. These cells displayed the CD20+ and CD56+ phenotype alike. None of the control subjects displayed HPV-DNA in either semen or peripheral blood.

Conclusion

Considering the role of CD20+ and CD56+ cell populations in the antiviral immune response, the detection of HPV markers on leukocytes may reflect the presence of virus particles within the endosomal compartment. However, the presence of HPV markers in circulating mononuclear cells raise concerns about the risk of developing cancers to distal organs.

【 授权许可】

   
2013 Foresta et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Schiller JT, Lowy DR: Understanding and learning from the success of prophylactic human papillomavirus vaccines. Nat Rev Microbiol 2012, 10:681-692.
• [2]Miralles-Guri C, Bruni L, Cubilla AL, Castellsagué X, Bosch FX, de Sanjosé S: Human papillomavirus prevalence and type distribution in penile carcinoma. J Clin Pathol 2009, 62:870-878.
• [3]Rintala MA, Grénman SE, Pöllänen PP, Suominen JJ, Syrjänen SM: Detection of high-risk HPV DNA in semen and its association with the quality of semen. Int J STD AIDS 2004, 15:740-743.
• [4]Lai YM, Lee JF, Huang HY, Soong YK, Yang FP, Pao CC: The effect of human papillomavirus infection on sperm cell motility. Fertil Steril 1997, 67:1152-1155.
• [5]Foresta C, Pizzol D, Moretti A, Barzon L, Palù G, Garolla A: Clinical and prognostic significance of human papillomavirus DNA in the sperm or exfoliated cells of infertile patients and subjects with risk factors. Fertil Steril 2010, 94:1723-1727.
• [6]Pérez-Andino J, Buck CB, Ribbeck K: Adsorption of human papillomavirus 16 to live human sperm. PLoS One 2009, 4:e5847.
• [7]Foresta C, Patassini C, Bertoldo A, Menegazzo M, Francavilla F, Barzon L, Ferlin A: Mechanism of human papillomavirus binding to human spermatozoa and fertilizing ability of infected spermatozoa. PLoS One 2011, 6:e15036.
• [8]Williams MA, Smith DC: RNA and protein synthesis in the nonspermatozoal cells of normal human semen. J Anat 1996, 188:137-147.
• [9]Keck C, Gerber-Schäfer C, Clad A, Wilhelm C, Breckwoldt M: Seminal tract infections: impact on male fertility and treatment options. Hum Reprod Update 1998, 4:891-903.
• [10]Kay P, Allan B, Denny L, Hoffman M, Williamson AL: Detection of HPV 16 and HPV 18 DNA in the blood of patients with cervical cancer. J Med Virol 2005, 75:435-439.
• [11]Chen AC, Keleher A, Kedda MA, Spurdle AB, McMillan NA, Antonsson A: Human papillomavirus DNA detected in peripheral blood samples from healthy Australian male blood donors. J Med Virol 2009, 81:1792-1796.
• [12]Roperto S, Comazzi S, Ciusani E, Paolini F, Borzacchiello G, Esposito I, Lucà R, Russo V, Urraro C, Venuti A, Roperto F: PBMCs are additional sites of productive infection of bovine papillomavirus type 2. J Gen Virol 2011, 92:1787-1794.
• [13]Hartl B, Hainisch EK, Shafti-Keramat S, Kirnbauer R, Corteggio A, Borzacchiello G, Tober R, Kainzbauer C, Pratscher B, Brandt S: Inoculation of young horses with bovine papillomavirus type 1 virions leads to early infection of PBMCs prior to pseudo-sarcoid formation. J Gen Virol 2011, 92:2437-2445.
• [14]Bodaghi S, Wood LV, Roby G, Ryder C, Steinberg SM, Zheng ZM: Could human papillomaviruses be spread through blood? J Clin Microbiol 2005, 43:5428-5434.
• [15]Rombaldi RL, Serafini EP, Mandelli J, Zimmermann E, Losquiavo KP: Perinatal transmission of human papilomavirus DNA. Virol J 2009, 6:83. BioMed Central Full Text
• [16]Barzon L, Militello V, Pagni S, Palù G: Comparison of INNO-LiPA genotyping extra and hybrid capture 2 assays for detection of carcinogenic human papillomavirus genotypes. J Clin Virol 2012, 55:256-261.
• [17]World Health Organization: WHO Laboratory Manual for the Examination and Processing of Human Semen. 5th edition. Geneva; 2010.
• [18]Garolla A, Pizzol D, Bertoldo A, De Toni L, Barzon L, Foresta C: Association, prevalence, and clearance of human papillomavirus and antisperm antibodies in infected semen samples from infertile patients. Fertil Steril 2013, 99:125-131.
• [19]Saunders AE, Johnson P: Modulation of immune cell signalling by the leukocyte common tyrosine phosphatase, CD45. Cell Signal 2010, 22:339-348.
• [20]Shafti-Keramat S, Handisurya A, Kriehuber E, Meneguzzi G, Slupetzky K, Kirnbauer R: Different heparan sulfate proteoglycans serve as cellular receptors for human papillomaviruses. J Virol 2003, 77:13125-13135.
• [21]Pao CC, Lin SS, Lin CY, Maa JS, Lai CH, Hsieh TT: Identification of human papillomavirus DNA sequences in peripheral blood mononuclear cells. Am J Clin Pathol 1991, 95:540-546.
• [22]Carter JJ, Koutsky LA, Hughes JP, Lee SK, Kuypers J, Kiviat N, Galloway DA: Comparison of human papillomavirus types 16, 18, and 6 capsid antibody responses following incident infection. J Infect Dis 2000, 181:1911-1919.
• [23]Amador-Molina A, Hernández-Valencia JF, Lamoyi E, Contreras-Paredes A, Lizano M: Role of Innate Immunity against Human Papillomavirus (HPV) Infections and Effect of Adjuvants in Promoting Specific Immune Response. Viruses 2013, 5:2624-2642.
• [24]Li H, Ou X, Xiong J, Wang T: HPV16E7 mediates HADC chromatin repression and downregulation of MHC class I genes in HPV16 tumorigenic cells through interaction with an MHC class I promoter. Biochem Biophys Res Commun 2006, 349:1315-21.
• [25]Ashrafi GH, Brown DR, Fife KH, Campo MS: Down-regulation of MHC class I is a property common to papillomavirus E5 proteins. Virus Res 2006, 120:208-211.
• [26]Foster CA, Yokozeki H, Rappersberger K, Koning F, Volc-Platzer B, Rieger A, Coligan JE, Wolff K, Stingl G: Human epidermal T cells predominantly belong to the lineage expressing alpha/beta T cell receptor. J Exp Med 1990, 171:997-1013.
• [27]Zentz C, Wiesner M, Man S, Frankenberger B, Wollenberg B, Hillemanns P, Zeidler R, Hammerschmidt W, Moosmann A: Activated B cells mediate efficient expansion of rare antigen-specific T cells. Hum Immunol 2007, 68:75-85.
• [28]Da Silva DM, Fausch SC, Verbeek JS, Kast WM: Uptake of human papillomavirus virus-like particles by dendritic cells is mediated by Fcgamma receptors and contributes to acquisition of T cell immunity. J Immunol 2007, 178:7587-7597.
• [29]Nicholls PK, Moore PF, Anderson DM, Moore RA, Parry NR: Regression of canine oral papillomas is associated with infiltration of CD4+ and CD8+ lymphocytes. Virology 2001, 283:31-39.
• [30]Wilgenburg BJ, Budgeon LR, Lang CM, Griffith JW, Christensen ND: Characterization of immune responses during regression of rabbit oral papillomavirus infections. Comp Med 2005, 55:431-439.
• [31]Lenz P, Thompson CD, Day PM, Bacot SM, Lowy DR, Schiller JT: Interaction of papillomavirus virus-like particles with human myeloid antigen-presenting cells. Clin Immunol 2003, 106:231-237.
• [32]Renoux VM, Bisig B, Langers I, Dortu E, Clémenceau B, Thiry M, Deroanne C, Colige A, Boniver J, Delvenne P, Jacobs N: Human papillomavirus entry into NK cells requires CD16 expression and triggers cytotoxic activity and cytokine secretion. Eur J Immunol 2011, 41:3240-3252.
• [33]Reijmers RM, Spaargaren M, Pals ST: Heparan sulfate proteoglycans in the control of B cell development and the pathogenesis of multiple myeloma. FEBS J 2013, 280:2180-2193.
• [34]Evander M, Frazer IH, Payne E, Qi YM, Hengst K, McMillan NA: Identification of alpha6 integrin as a candidate receptor for papillomaviruses. J Virol 1997, 71:2449-2456.
• [35]Woodham AW, Da Silva DM, Skeate JG, Raff AB, Ambroso MR, Brand HE, Isas JM, Langen R, Kast WM: The S100A10 subunit of the annexin A2 heterotetramer facilitates L2-mediated human papillomavirus infection. PLoS One 2012, 7:e43519.
• [36]Syrjanen S: Current concepts on human papillomavirus infections in children. APMIS 2010, 118:494-450.
• [37]Armbruster-Moraes E, Ioshimoto LM, Leão E, Zugaib M: Presence of human papillomavirus DNA in amniotic fluids of pregnant women with cervical lesions. Gynecol Oncol 1994, 54:152-158.
• [38]Righini A, Parazzini C, Doneda C, Avagliano L, Arrigoni F, Rustico M, Consonni D, Re TJ, Bulfamante G, Triulzi F: Early formative stage of human focal cortical gyration anomalies: fetal MRI. AJR Am J Roentgenol 2012, 198:439-447.
• [39]Chen J, Tsai V, Parker WE, Aronica E, Baybis M, Crino PB: Detection of human papillomavirus in human focal cortical dysplasia type IIB. Ann Neurol 2012, 72:881-892.
• [40]Crino PB: mTOR: a pathogenic signaling pathway in developmental brain malformations. Trends Mol Med 2011, 17:734-742.