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The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Clinical & Epidemiology Abstract 53 KAPOSI’S SARCOMA-ASSOCIATED HERPESVIRUS SEROCONVERSION AND SEROREVERSION IN A MATERNAL-INFANT COHORT IN ZAMBIA Veenu Minhas, 1 Kay L. Crabtree, 1 Janet Wojcicki, Tendai J. M’soka, 2 Chipepo Kankasa, 2 Charles D. Mitchell, 3 and Charles Wood 1 1 Nebraska Center for Virology, School of Biological Sciences, University of Nebraska Lincoln, Lincoln, NE USA; 2 Department of Paediatrics and Child Health, University Teaching Hospital, Lusaka, Zambia; 3 Department of Pediatrics, University of Miami School of Medicine, Miami, FL USA. Abstract Zambia is currently experiencing an HIV-1 epidemic leading to an increase in the incidence of Kaposi’s sarcoma among adults and children. This is a prospective study designed to investigate Kaposi’s sarcoma-associated herpesvirus (KSHV) seroincidence and associated risk factors for KSHV seroconversion and seroreversion in a mother-infant cohort, by following them for 48 months after delivery. Our results show that overall KSHV seroincidence in KSHV negative Zambian women was 0.7 per 100 person-months which was lower as compared to children followed in the same cohort. We observed that up to 40% of Zambian children in this cohort may be infected by 48 months of age, indicating that primary infection occurs mostly in early childhood. HIV-1 infection was associated with higher risk for seroconversion in women and children. In adult women, KSHV seroconversion was not associated with most socio-economic factors, sexual habits or medical history. This indicates that sexual transmission during adulthood may not be an important route of transmission in this cohort. Our laboratory has earlier reported that KSHV antibodies may not persist over time in children and that seroreversion leading to partial or complete loss of KSHV antibodies is prevalent. We observed seroreversion in up to 55% of women and this was more common in single women and in women delivering for the first time. These results demonstrate that multiple longitudinal samples per patient may be required to accurately determine KSHV infection status. Presenting author Email: veenu@bigred.unl.edu 80
The 11 th International Workshop on KSHV & Related Agents, Birmingham, UK Clinical & Epidemiology Abstract 54 DISTRIBUTION OF KSHV MICRORNA POLYMORPHISMS IN AIDS-KS, CLASSICAL KS, AND MULTICENTRIC CASTLEMAN’S DISEASE Vickie Marshall, 1 Eliza Martró, 2 Elizabeth Brown, 3 Dian Wang, 1 Alex Ray, 1 Maria Nazzarena Labo, 1 The Classical Kaposi’s Sarcoma Working Group, Robert Yarchoan, 4 Jordi Casabona, 2 Rolfe Renne, 5 and Denise Whitby 1 1 NCI-Frederick, Frederick, MD, USA; 2 Hospital Universitari Germans Trias i Pujol, Badalona, Spain; 3 University of Alabama at Birmingham, Birmingham, AL, USA; 4 Division of Clinical Services, NCI, NIH, Bethesda, MD, USA; 5 University of Florida Shands Cancer Center, Gainsville, FL, USA Abstract MicroRNAs are short regulatory RNAs of approximately 19-24 nucleotides in length that bind specifically to mRNA in a process that interferes with translation or promotes degradation. KSHV encodes twelve microRNAs, all located within the latency associated transcript which is highly expressed in KSHV-associated malignancies. We amplified, cloned, and sequenced a 2,712 bp non-coding region containing a cluster of 10 microRNAs and a 646 bp fragment of the K12/T0.7 gene which contains two microRNAs. We successfully characterized KSHV microRNA sequences from lymphocytes collected as part of two Kaposi’s sarcoma case-control studies (one AIDS-KS, one Classic–KS) and MCD clinic attendees from the NCI. We have completely or partly characterized all twelve KSHV microRNAs from a total of 37 Kaposi sarcoma cases, 21 KSHV positive controls, and 8 MCD patients. Phylogenetic analysis of both regions demonstrates extreme conservation of microRNAs overall. However, several distinct polymorphisms were discovered in this study, some occurring within the mature microRNA transcript. Phylogenetic characterization of the KSHV K1 and viral microRNA sequences in this study indicated all were subtypes A/C. However, we did detect four microRNA polymorphisms we previously described as specific to KSHV viral subtypes A5/B. Some polymorphisms appear to be more common in Kaposi’s sarcoma and MCD cases than in KSHV positive controls. Our data demonstrate that most KSHV microRNAs are highly conserved suggesting that they contribute to the biological activity and possibly viral pathogenesis of KSHV. However, some KSHV microRNAs have distinct polymorphisms which may affect processing, function, and mRNA target recognition. Presenting author Email: marshall@ncifcrf.gov 81
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