title: HIV-1 penetrates coronary artery endothelial cells by transcytosis.
abstract: BACKGROUND
  The pathogenesis of HIV-1-related cardiomyopathy is poorly understood, but HIV-1 has been detected in cardiomyocytes. Whether HIV-1 penetrates into the myocardium by infection of coronary artery endothelial cells (CAEC) or using transcellular or paracellular routes across CAEC has not been resolved.
  
  
  METHODS
  A model of the CAEC barrier was constructed with primary CAEC (derived from human coronary vessels). Polymerase chain reaction (PCR) assay, infectious assay, and immunofluorescence were employed to show abortive nature of HIV-1 infection of CAEC. Tight junction (TJ) and cell adhesion proteins were visualized by immunofluorescence. The time course of HIV-1 invasion was measured by HIV-1 RNA assay. Inulin permeability assay determined paracellular leakage. Transmission electron microscopy demonstrated virus-induced endothelial vacuolization.
  
  
  RESULTS
  Despite a strong display on CAEC of CXCR4 and a lesser expression of CCR3 and CCR5, HIV-1 did not productively replicate in CAEC, as shown by infectious assay, immunofluorescence, and electron microscopy. HIV-1 infection of CAEC was abortive with minimal reverse transcription of strong stop DNA and pol but not full-length or two LTR DNA circles. Upon infection of the model with 1 million RNA copies of HIV-1JR-FL, virus penetration 2 hr postinfection (PI) was negligible but increased by 1,750% 24 hr PI. The paracellular permeability increased during this period by only 25%. Neither AOP-RANTES nor v-MIPII significantly reduced HIV-1JR-FL invasion. Virus infection did not alter the integral TJ protein occludin and the TJ-associated protein ZO-1. HIV-1 exposed CAEC and brain microvascular endothelial cells (BMVEC) developed extensive cytoplasmic vacuolization with retroviral-like particles in the vacuoles.
  
  
  CONCLUSIONS
  The endothelium is not an impenetrable barrier to HIV-1. The virus opens a transcellular route across coronary and brain endothelia in cytoplasmic vacuoles.
authors.affiliation: Department of Medicine, UCLA School of Medicine, Los Angeles, CA, USA.
authors.forename: C
authors.initials: C
authors.lastname: Gujuluva
authors.forename: A R
authors.initials: AR
authors.lastname: Burns
authors.forename: T
authors.initials: T
authors.lastname: Pushkarsky
authors.forename: W
authors.initials: W
authors.lastname: Popik
authors.forename: O
authors.initials: O
authors.lastname: Berger
authors.forename: M
authors.initials: M
authors.lastname: Bukrinsky
authors.forename: M C
authors.initials: MC
authors.lastname: Graves
authors.forename: M
authors.initials: M
authors.lastname: Fiala
mesh_terms: D001158:Arteries / Q000648:ultrastructure / Q000821:virology*; D001483:Base Sequence; D002478:Cells, Cultured; D018925:Chemokines / Q000502:physiology; D003331:Coronary Vessels / Q000648:ultrastructure / Q000821:virology*; D017931:DNA Primers; D004705:Endocytosis; D015684:Gene Products, pol / Q000235:genetics; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000502:physiology*; D008854:Microscopy, Electron; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014779:Virus Replication
chemical_list: D018925:Chemokines; D017931:DNA Primers; D015684:Gene Products, pol