Umbilical Cord Blood Mononuclear Cell HIV-1 LTR Binding Activities
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Published on: May 7, 2012
P.S. Kedar1, K. Arden, M. Foyle
1Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital,Brisbane, Australia.
Insights
Researchers identified HIV-1 LTR binding factors in cord blood, revealing differences from adult cells that may influence pediatric HIV infection and disease. This study sheds light on host factors affecting fetal and newborn HIV replication.
Area of Science:
- Virology
- Immunology
- Pediatrics
Background:
- Vertical transmission of HIV is a major pediatric concern.
- Understanding host factors is crucial for characterizing HIV replication in fetuses and newborns.
Purpose of the Study:
- To identify HIV-1 Long Terminal Repeat (LTR) binding factors in cord blood mononuclear cells.
- To compare these factors with those found in adult peripheral blood mononuclear cells.
Main Methods:
- Electrophoretic mobility shift assays (EMSAs) were used to detect protein-DNA interactions.
- UV cross-linking studies and transient transfection experiments were performed.
- Nuclear extracts from cord blood and adult peripheral blood were analyzed.
Main Results:
- Cord blood extracts showed significant binding to multiple regions of the HIV-1 LTR, particularly NF-kB sites.
- Qualitative differences in binding activity were observed between cord blood and adult peripheral blood extracts.
- NF-kB and Sp1 sequences, along with upstream regions, were important for HIV-1 LTR expression in cord blood cells.
Conclusions:
- Cord blood cells contain factors that bind to key HIV-1 LTR regulatory sites.
- Distinct binding activities in cord blood compared to adult blood may play a role in the pathogenesis of neonatal HIV infection.
Abstract:
Vertically transmitted HIV disease constitutes a significant problem in pediatrics. In order to characterize some of the possible host factors involved in HIV replication in fetuses and newborns, we surveyed the HIV-1 LTR binding factors present in nuclear extracts from cord blood mononuclear cells. A series of electrophoretic mobility shift assays (EMSAs) showed that protein extracts from cord blood interacted with several regions of the HIV LTR. The most prominent binding activities involved the NF-kB sites, but other regions of the LTR also showed factor binding with the cord blood extracts. Some of these cord blood extract binding activities displayed qualitative differences when compared to adult peripheral blood mononuclear cell extracts in EMSA and UV cross-linking studies. Transient transfection experiments indicated that the NF-kB and Sp1 sequences were important for wild type levels of expression in cord blood cells, but that additional sequences 5' to the NF-kB sites also contributed activity. Thus, factors that interact with many of the well-known HIV LTR regulatory sites are present in cord blood cells. However, certain qualitative differences distinguished cord blood and adult peripheral blood binding activities and these may contribute to pathogenesis of HIV infection in neonates.
