1. Human Papilloma Virus (HPV) in potentially malignant and malignant oral mucosal disease Faden A, 1 Teo CG, 2 Moles DR, 3 Porter SR 1 1 Oral Medicine and 3 Health Services Research, Division of Microbial Diseases, Eastman Dental Institute, UCL; 3 Health Protection Agency, Colindale, London, UK

2. Human papilloma virus and oral squamous cell carcinoma HPV not always detected OR of HPV for OSCC much lower than the cervical malignancy (~74) Geographic variation in HPV detection in OSCC No increase OSCC in HIV disease without HAART (Reviewed by Scully, 2002; Ha and Califano, 2004; Frezzini et al, 2005)

3. AIMS 1.Establish the frequency of carriage of HPV DNA within a) OSCC from two different locales (UK and KSA) b) Potentially malignant oral mucosal disease (OED, PVL, OLP) c)Normal oral mucosa 2.Establish the frequency of carriage of EBV, CMV and HHV-8 within similar disease 3.Determine the OR of HPV for OSCC, OED and related disease

4. Samples/No.Gender Median AgeAge Range M:F (Year) (Year) Normal (26) 10:16 63 21- 97 LP-UK (30) 16:14 58 40-79 LP-KSA (32) 13:19 43.5 23-65 Dysplasia(45) 23:22 60 29-77 PVL (12) 6:6 74.5 48-85 SCC-UK (34) 22:12 70 31-96 SCC-KSA (32) 23:7 60 30-81 HIV-UK (25) 22:3 43 31-53 Summary of patients

5. Materials and Methods Extraction of DNA from Paraffin Embedded Tissues (PET). (Howe et al., 1997; Coombs & Primrose., 1999; Akula et al., 1999) βeta-globin gene detection assessment for DNA integrity (Fouret et al., 1995) HPV PCR with “degenerate” consensus primers (MY09/MY11), followed by nested PCR with “general” primers (GP5+/GP6+) for L1 region. (Strauss et al., 2000; Husnjak et al., 2000) Reverse line blotting hybridization (RLB) for HPV genotyping. (Jordens et al., 2000; Van den Brule et al., 2002)

6. Materials and Methods Nested PCR detection of CMV DNA (gN region) (Beyari et al., 2003) Nested PCR detection of EBV DNA (BamHI K region) (Triantos et al.,1998) Nested PCR detection of HHV-8 (ORF K1 and ORF 26 regions) (Cooke et al., 2002)

7. Descriptive – Frequencies Analytical – Logistic regression -Unadjusted -Adjusted Statistical Methods

8. HPV DNA in potentially malignant and malignant disease Sample No Country HPV Normal 26 UK 7.7% Lichen Planus 32 KSA 28% Lichen Planus 30 UK 30% Dysplasia 45 UK 42% PVL 12 UK 58% SCC 30 KSA 57% SCC 34 UK 75%

9. HPV DNA in potentially malignant and malignant diseases OutcomeCountryUnadjusted Odds Ratio for HPV 95%CIP-value DysplasiaUK1.00.5, 2.10.889 KSAn/a SCC or DysplasiaUK3.31.8, 5.9<0.001 KSAn/a SCCUK7.73.2, 18.8<0.001 PVL KSA UK 3.7 2.0 1.4, 9.3 0.6, 6.8 0.006 0.225 Logistic regression (unadjusted)

10. HPV DNA in OSCC Logistic regression (adjusted for smoking habit, gender and ethnicity) OutcomeCountryAdjusted Odds Ratio for HPV 95%CIP-value SCCUK10.03.9, 25.3<0.001 KSA3.61.3, 10.30.017

11. HPV DNA in HIV, OLP and PVL Logistic regression (adjusted for smoking habit, gender and ethnicity) OutcomeCountryAdjusted Odds Ratio for HPV 95%CIP-value HIVUK0.80.3, 1.90.570 KSAn/a OLPUK0.40.2, 1.00.069 KSA0.60.1, 2.40.354 PVLUK1.80.5, 6.30.379 KSAn/a

12. Herpes Viruses Sample/ Country CMV % EBV % HHV8-K1 % HHV8-26 % Normal-UK (26) 0 0 4 0 OED-UK (45) 15.6 7 0 7 PVL-UK (12) 17 50 42 8 Lichen Planus-UK (30) 0 0 10 0 Lichen Planus-KSA (32) 0 0 38 3 SCC-UK (34) 0 3 10 0 SCC-KSA (30) 0 0 23 7 HIV-UK (25) 0 0 36 24

13. Conclusions and discussion HPV strongly associated with OSCC in the UK or KSA But HPV present in only 40% of OED lesions HPV 16 most common oral genotype Herpes viruses not strongly associated with such disease A causal role for HPV in OSCC would seem possible

14. Conclusions and discussion Implications of HPV within oral lichen planus are unclear Impact of immune restoration disease of HIV infection are concerning Clinical advice should include discussion of modification of lifestyle

15. Acknowledgements Histopathology support: Professor Paul Speight Dr William Barrett Mr Paul Darkins Advice: Professor Crispian Scully CBE Funding: Kingdom of Saudi Arabia (KSA)

16. Human Papilloma Virus (HPV) in potentially malignant and malignant oral mucosal disease Faden A, 1 Teo CG, 2 Moles D, 3 Porter SR 1 1 Oral Medicine and 3 Health Services Research, Division of Microbial Diseases, Eastman Dental Institute, UCL; 3 Health Protection Agency, Colindale, London, UK