Prevalence of Human Papillomavirus (HPV) Genotypes in HIV-1 Infected Women in Seattle, WA and Nairobi, Kenya Results from the Women HIV Interdisciplinary.

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Presentation transcript:

Prevalence of Human Papillomavirus (HPV) Genotypes in HIV-1 Infected Women in Seattle, WA and Nairobi, Kenya Results from the Women HIV Interdisciplinary Network (WHIN) HPV is considered the most common known sexually transmitted agent worldwide. At a any given point in time 10% of women are positive for cervical HPV DNA this represents approximately 291 million women globally. Cervical cancer is the number one cancer cause of year of life lost in the developing world. African women have the highest prevalence of HPV. The currently available HPV vaccine effectively protect against the most common oncogenic types of HPV ( 16 and 18). Given the type specific immunity of these vaccines and the potential for geographic variation on prevalence of HPV types it is important to understand the distribution of HPV genotypes in particular among HIV infected women Certain HPV types are associated with squamous intraepithelial lesions. Specifically HPV 16, 18, 45 and 31 cause over half of the low grade lesions and about 65% of the high grade lesions. Amneris E. Luque*, Jane Hitti, Christina Mwachari, Christopher Lane, Susan Messing, Susan E. Cohn, Robert Rose and Robert Coombs

Background-HPV Prevalence HPV is the most common known sexually transmitted agent worldwide At any given point in time 10% of women are positive for cervical HPV DNA ( 291 million women) Certain HPV types are associated with squamous intraepithelial lesions. HPV 16 and 18 cause about 65% of the high grade lesions

Cervical Cancer in the Developing World 83% of cases in developing world #1 cancer cause of YLL in developing world Source: NEJM 2005;353:2101-4

Background-HIV and HPV When compared with HIV-uninfected women, HIV Infected women have: higher rates of HPV infection higher prevalence of infection with multiple types of HPV higher prevalence of oncogenic (high-risk) HPV higher prevalence of cervical dysplasia A different distribution of HPV genotypes There appears to be a geographic variation in the distribution of HPV genotypes, but this has not been widely studied in HIV-infected women. HIV infection has a significant impact on HPV infection and the progression of HPV-related cervical disease. HIV infected women not only have a high prevalence of HPV infection but also are more likely to be infected with multiple HPV genotypes concurrently. They are also more likely to be infected with HPV genotypes that are considered high risk for progressing to cervical cancer.

Study Aim To assess the prevalence of HPV genotypes in two distinct populations of HIV-1 infected women To assess the prevalence of HPV genotypes in two geographically distinct populations, we compared the HPV genotype distribution between HIV-1 infected women from Seattle and Nairobi

Methods HIV-1 infected women were enrolled in Seattle, WA and had evaluations at set intervals, over a 3 year period (2003-2006) HIV-1 infected women with CD4+ cell count > 350 cells/mm3 were enrolled from Nairobi, Kenya and had similar evaluations as the Seattle participants at a single point in time

Methods (cont 1.) Informed consent Standardized history and physical examination Questionnaire Sample collection Cervical Cytology Cervico-vaginal lavage samples (CVL)

Methods (cont 2.) Total DNA extraction in CVL specimens was followed by PCR amplification and genotype analysis by reverse line blot Reverse line blot used a set of 27 immobilized oligonucleotide probes, each corresponding to an individual HPV genotype.

Reverse Line Blot Assay Type Specific Probe Application Rotate 90° The reverse line blot assay util;izes an immobilized probe array in which oligonucleotide probes are deposited in a linear array

Reverse Line Blot Assay Biotinylated PCR Product Application The PCR product is generated using biotinylated primers. The presence of biotinylated primers bound to specific probe is detected using streavidin-conjugated with alkaline phosphatase and a chromogenic soluble substrate to produce a color line at the position of the positive probe Detect using StrepAvidin-conjugated with Alkaline Phosphatase

On the right the different 27specific probes are listed with the Beta globin controls at the bottom. The numbers on the top of the columns represent consecutive patients 1,2,3,4,5,6,7,8,9,10 etc… ( only a number every 5 is written to prevent crowding.

Statistical Analyses Demographics and cross sectional data is presented by site. A logistic regression analysis was used to assess the relationship between grouped HPV types and cytological results. All statistical tests are two-sided with alpha <.05

Baseline demographic information, stratified by site Seattle n=38 Nairobi n= 50 Age (years) mean + sd 38.3 + 6.6 32.7 + 5.0 Parity 2.5 + 2.1 2.6 + 2.3 Race/ethnicity: n (%) Caucasian 12 (32) -- African American 20 (53) Asian/Pacific islander 2 (5) Hispanic Native American 1 (3) Other Kikuyo 24 (48) Luo 8 (16) Luhya 6 (12) Kamba 3 (6) Other/unknown 9 (18)

Baseline demographic information, stratified by site Seattle n=38 Nairobi n= 50 HIV Risk Factor: n (%) IVDU 9 (24) Heterosexual 25 (66) 50 (100) Blood transfusion 2 (5) Other/unknown Lifetime sexual partners Median ( IQR) 11(5-20) 3 (2-5)* Oral contraceptive use: n (%) Ever 29 (76) 30 (60) Current 2 (4) On ARVT: n (%) 20 (53) 0 (0) CD4 cell count: cells/mm3 346 538 ** HIV RNA: copies/ml 1,036 13,942 *P = .0001 Wilcoxon rank-sum test **P = .008

Results-Prevalence of HPV risk types and number of lifetime sexual partners No. lifetime sexual partners ( entire group) No HPV Only low risk HPV HPV 16/18 Other high-risk HPV <5 29 (85%) 1 (3%) 3 (9%) ≥5 17 (68%) 2 (8%) 1 (4%) 5 (20%)

Results-Prevalence of HPV risk types and number of lifetime sexual partners in the Nairobi group No. lifetime sexual partners No HPV High or Low Risk HPV <5 27 (84%) 5 (16%) ≥5 6 (50%) 6(50%) P =0.04 Fisher’s exact test

Results-HPV genotypes by site Cohort # of CVL samples Samples Positive for beta globin Samples Positive for HPV DNA HPV Infections HPV types (number of occurences) Seattle (n=37) 63 38 (60%) 15 (40%) Triple:1 Dual:5 Single:5 Non-typeable:4 56 (5), 66 (4), 6 (3), MM8(2), 16 (1), 33 (1), 53 (1), MM7 (1) Nairobi (n=50) 49 37 (76%) 11 (30%) Dual:1 Single: 9 53 (3), 6 (3), 33 (2), 58 (2) 16 (1), 18 (1), 62 (1), MM8 (1) Numbers in bold denote high-risk HPV types

Results-Cervical Cytology by Site Seattle n=64 Nairobi n=50 Total Normal 48 39 87 Abnormal 15 7 22 ASCUS 8 2 10 ASCUS-D LGSIL 4 6 HGSIL 1 3 Other Inadequate Missing

HPV detected in CVL specimen Results-Cervical cytology, according to HPV genotypes detected in CVL specimens Cytological results HPV detected in CVL specimen 56 66 6 53 33 MM8 16 18 58 other ASCUS 2 LGSIL 1 HGSIL Inadequate Other WNL 4 3 Total 5 6

Phylogenetic Tree of 118 HPV Types Virology, 324, 17-27 (2004)

Conclusions Participants from Nairobi had significantly fewer sexual partners and had more commonly a single HPV type detected in CVL than those from Seattle. HIV-1 infected women in Seattle and Nairobi had geographically distinct set of HPV genotypes Patients in this study had a higher prevalence of high-risk HPV types other than 16 and 18.

Conclusions Women with more than one HPV type detected in CVL were 6.25 more likely to have an abnormal Pap smear than those with only one or no HPV detected Women were 7.52 times more likely to have an abnormal pap smear when they had high risk HPV present vs. none. Currently available HPV vaccines may not protect against other prevalent HPV types in these women.

Acknowledgements Kenya Research Institute Christina Mwachari University of Washington in Seattle Bob Coombs Jane Hitti Katie Paul University of Rochester Richard C Reichman Robert Rose Susan Messing Chris Lane Susan Cohn Women in Nairobi and Seattle who participated in the study