title: APOBEC3G expression is restricted to epithelial cells of the proximal convoluted tubules and is not expressed in the glomeruli of macaques.
abstract: The Vif protein of human immunodeficiency virus-1 (HIV-1) interacts with members of the APOBEC family of cytidine deaminases. In this study, we isolated RNA from renal cortex as well as from isolated glomeruli and tubulointerstitial fractions from two pigtailed macaques that were exsanguinated and perfused with saline. RT-PCR results indicate that APOBEC3G was detected in the tubule fractions but not in the glomerular fractions. Immunoblot analysis using lysates prepared from these same fractions and a monoclonal antibody to APOBEC3G confirmed the RT-PCR findings. To determine which cell types express APOBEC3G, immunohistochemical studies were performed using this monoclonal antibody on renal cortical sections. Our results clearly show that the glomeruli do not express APOBEC3G but that select tubules within the cortex express APOBEC3G at high levels. To further differentiate the distribution of APOBEC3G expression, serial sections were stained with the lectins Dolichos biflorus agglutinin (DBA) and Phaseolus vulgaris erythroagglutinin (PHA-E), which differentially bind to epithelial cells of the tubules and glomeruli. Our results indicate that APOBEC3G expression was restricted to PHA-E-staining tubules and not DBA-staining tubules, suggesting that APOBEC3G expression was restricted to proximal convoluted tubules. These findings suggest that infection of epithelial cells of proximal renal tubules could suppress Vif-defective HIV-1 replication, whereas infection of cells of the glomeruli, a major target of HIV-associated nephropathy, could act as a reservoir for the replication of Vif-defective HIV-1.
authors.affiliation: Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
authors.forename: M Sarah
authors.initials: MS
authors.lastname: Hill
authors.forename: Autumn
authors.initials: A
authors.lastname: Ruiz
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Gomez
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Miller
authors.forename: Nancy E J
authors.initials: NE
authors.lastname: Berman
authors.forename: Edward B
authors.initials: EB
authors.lastname: Stephens
mesh_terms: D000818:Animals; D003564:Cytidine Deaminase / Q000096:biosynthesis* / Q000235:genetics; D004847:Epithelial Cells / Q000201:enzymology*; D015151:Immunoblotting; D007150:Immunohistochemistry; D007678:Kidney Glomerulus / Q000201:enzymology*; D007684:Kidney Tubules / Q000166:cytology / Q000201:enzymology*; D008254:Macaca nemestrina; D010835:Phytohemagglutinins; D037121:Plant Lectins; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D010835:Phytohemagglutinins; D037121:Plant Lectins; C036554:dolichos biflorus agglutinin; C067494:erythroagglutinating phytohemagglutinin; D003564:Cytidine Deaminase