Epstein‐Barr virus and human papillomavirus serum antibodies define the viral status of nasopharyngeal carcinoma in a low endemic country

Abstract

Epstein-Barr virus (EBV) causes nasopharyngeal carcinoma (NPC) in endemic regions,where almost every tumor is EBV-positive. In Western populations, NPC is rare, and human papillomavirus infection (HPV) has been suggested as another viral cause. We validated multiplex serology with molecular tumor markers, to define EBV-positive, HPV-positive and EBV-/HPV-negative NPCs in the United Kingdom, and analyzed survival differences between those groups. Sera from NPC cases (n = 98) and ageand sex-matched controls (n = 142) from the Head and Neck 5000 clinical cohort study were analyzed. IgA and IgG serum antibodies against 13 EBV antigens were measured and compared with EBER in situ hybridization (EBER-ISH) data of 41 NPC tumors (29 EBER-ISH positive, 12 negative). IgG antibodies to EBV LF2 correctly diagnosed EBV-positive NPCs in 28 of 29 cases, while all EBER-ISH negative NPCs were seronegative to LF2 IgG (specificity = 100%, sensitivity = 97%). HPV early antigen serology was compared to HPV molecular markers (p16 expression, HPV DNA and RNA) available for 41 NPCs (13 positive, 28 negative). Serology matched molecular HPV markers in all but one case (specificity = 100%, sensitivity = 92%). EBV and HPV infections were mutually exclusive. Overall, 67% of the analyzed NPCs were defined as EBV-positive, 18% as HPV-positive and 14% as EBV/HPV-negative. There was no statistical evidence of a difference in survival between the three groups.These data provide evidence that both, EBV-positive and HPV-positive NPCs are present in a low incidence country, and that EBV and HPV serum antibodies correlate with the viral status of the tumor. K E Y W O R D S nasopharyngeal carcinoma, multiplex serology, Epstein-Barr virus, human papillomavirus Abbreviations: CI, confidence interval; EA-D, early antigen diffuse; EBER-1, Epstein-Barr virus small RNA; EBNA1, EBV nuclear antigen 1; EBV, Epstein-Barr virus; HPV, human papillomavirus; HPyV, human polyomavirus; IHC, immunohistochemistry; IMD, index of multiple deprivation; ISH, in situ hybridization; LSCC, laryngeal squamous cell carcinoma; MFI, median fluorescence intensity; MPG, multiplex papillomavirus genotyping; NCT,

HPV Positive Status Is a Favorable Prognostic Factor in Non-Nasopharyngeal Head and Neck Squamous Cell Carcinoma Patients: A Retrospective Study From the Surveillance, Epidemiology, and End Results Database

“…Previous studies had showed that HPV infection in NPC was observed but it was relatively rare compared to EBV infection and the prognostic role of HPV infection in NPC was controversial (33). There were studies suggesting that there was no statistical difference in the survival between HPV positive and EBV positive NPC patients (34). While, existing literature showed that HPV positive patients had worse outcomes compared to patients with EBV-positive NPC (35).…”

Development of High-Throughput Multiplex Serology to Detect Serum Antibodies against Coxiella burnetii

“…We demonstrated that multiplex serology is a valuable tool for large-scale studies addressing C. burnetii infections. Analyzing up to 2000 serum samples per day, this technique has been used in diverse seroepidemiological studies to detect antibodies against a variety of different pathogens, e.g., Helicobacter pylori, human papillomaviruses, and herpes viruses [25][26][27][28]. The inclusion of C. burnetii broadens the spectrum of measurable pathogens and enables powerful studies to investigate the association between C. burnetii infections and potential clinical outcomes.…”

“…This bead-based technique follows the principle of an antigen ELISA, while further enabling the simultaneous measurement of antibodies against multiple antigens in a single reaction. Up to 2000 samples per day can be processed cost effectively, as it has been demonstrated in multiple studies [25][26][27][28]. By including C. burnetii antigens, we enable powerful large-scale studies which will help to better understand the role of C. burnetii in potential clinical manifestations, e.g., by investigating respective case-control studies.…”

High-Risk Human Papillomavirus and Epstein–Barr Virus Coinfection: A Potential Role in Head and Neck Carcinogenesis

“…However, in non-endemic areas, HPV/EBV coinfection is less common. Indeed, some authors reported that HPV and EBV infections are mutually exclusive in populations with a low NPC incidence [ 86 , 90 ].…”