Identification of Human Papilloma Virus (HPV) in Saliva of Human Immunodeficiency Virus (HIV): Seropositive Adults Six Months Post HAART

Field: Medicine (Health Sciences)

INTRODUCTION:
The number of people living with HIV globally is 33.4 millions. Systemic and oral lesions in HIV infection reflect the immune status of the patients and these lesions are important not only for the morbidity that they cause, but also for their diagnostic value in monitoring the immune status of the patient. The occurrence of oral lesions indicate a great susceptibility for opportunistic infections and a great possibility for a rapid progression to AIDS.
The introduction of antiretroviral therapy (ART), especially combination therapy –has led to the reduction of opportunistic infections, mortality and morbidity in HIV infected patients. Highly active antiretroviral therapy (HAART) is defined as an antiretroviral regimen containing a minimum of 3 drugs, nucleotide reverse transcriptase inhibitor (NRTI), protease inhibitor and non nucleotide reverse transcriptase inhibitor (NNRTI).
HPV infection is recognized as the most common sexually transmitted infection. The association between HPV and cervical cancer has been recognized. In humans more than 70 types have been described. Mucosal and genital HPV consisting of about 30 types, are divided into low risk (HPV 6,11,42,43 and 44) and high risk (HPV 16,18,31,33,35,45,51,52 and 56), according to their presence in malignant lesions of the cervix. There are circumstantial evidence suggesting that high risk HPVs are involved in oral carcinogenesis.
Estimates of oral HPV prevalence is variable due to methodological or population differences and ranges from 9.2 – 18.8%. When HPV lesions are present in association with HIV infection, some patients experience a rather sudden occurrence and growth of new lesions in the mouth concurrently with deterioriation of the immune status. The oral warts caused by HPV are usually 1-3mm in diameter, and may be located anywhere in the mouth, but most often on the labial or buccal mucosa or the gingiva.
Prevalence of HPV associated oral condylomas have reportedly increased in HIV infected individuals since the introduction of HAART and oral warts due to HPV also have dramatically increased in the HAART era. Greenspan et al studied the effect of HAART on frequency of oral warts and found that HIV infected patients on HAART were six times more likely to develop warts than patients not on HAART. They also postulated that the functionally incomplete reconstitution of the immune system, observed in patients initiated on HAART could alter its effectiveness with regard to different potentially pathogenic microorgansisms and may probably lead to the development of oral warts in the context of an overall reduction of opportunistic infections.
AIM OF THE STUDY:
To identify Human Papilloma Virus in Saliva of Human Immunodeficiency Virus (HIV) - Seropositive adults 6 months post HAART.
OBJECTIVES:
To detect the presence of Human Papilloma Virus (HPV) in saliva of
• HIV seropositive adults on six months of highly active anti retroviral therapy (HAART)
• HIV seronegative adults.
STUDY DESIGN:
A prospective study was done to ascertain oral HPV status in the saliva by using PCR to amplify the late gene L1 of HPV genome.
STUDY SUBJECTS:
GROUP I: Ten HIV seropositive patients who were on HAART ≥ 6 months for whom HPV status was ascertained prior to initiation of HAART
GROUP II: Ten HIV seronegative individuals.
STUDY SETTING:
• Saliva samples were collected from patients attending the out patient wing of YRG CARE, VHS, Chennai.
• Study was conducted at Ragas Dental College and Hospital, Chennai.
STUDY PROCEDURE:
• Ethical clearance was obtained from the institutional review board of Ragas Dental College and from YRGCARE.
• Informed consent was obtained from the patient.
• Demographic details of the patient were recorded.
• A thorough oral examination was done and findings were recorded in prestructured case sheets.
SAMPLE COLLECTION:
1. The patient was asked to sit on a chair, and after spitting any residual saliva present in his mouth, he was asked to spit into a sterile container every minute for 10 minutes to collect unstimulated whole saliva.
2. 5ml of unstimulated whole saliva was collected in the morning after breakfast (9:00 am – 11:00 am) from the study subjects (to avoid diurnal variation).
3. Buccal mucosal scrapping was collected from 5 patients of group I selected randomly
4. The sample was immediately transported to the research laboratory at Ragas dental college and hospital and stored at -700C until DNA extraction.
CONCLUSION:
This study was done to detect HPV in the saliva of HIV seropositive patients on HAART therapy (for ateast six months)
1. Ten HIV seropositive patients were recruited for the study≥ six months of initiation of HAART (group I) and 10 HIV seronegative subjects were controls (Group II).
2. DNA was extracted from all the salivary samples. The DNA was quantified (≥25 μg/dl)
3. Extracted DNA was amplified for the identification of HPV using L1 consensus primers and primers to amplify the house keeping gene by multiplex PCR method.
4. All samples were positive for β globin (house keeping) gene and positive control (infected HeLa cell line DNA) was positive for HPV DNA.
5. We were unable to detect HPV DNA in our study subjects of both group.
We used the most sensitive method, PCR to detect HPV genome in our study subjects. To understand the biology of HPV infection in HIV seropositive patients, more longitudinal studies are necessary especially in the context of HAART.