HIV-1 penetrates coronary artery endothelial cells by transcytosis

Background: The pathogenesis of HIV-1-related cardiomyopathy is poorly unde rstood, but HIV-1 has been detected in cardiomyocytes. Whether HIV-1 penetr ates into the myocardium by infection of coronary artery endothelial cells (CAEC) or using transcellular or paracellular routes across CAEC has not be en resolved. Materials and Methods: A model of the CAEC barrier was constructed with pri mary 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 a dhesion proteins were visualized by immunofluorescence. The time course of HIV-1 invasion was measured by HIV-1 RNA assay. Inulin permeability assay d etermined paracellular leakage. Transmission electron microscopy demonstrat ed virus-induced endothelial vacuolization. Results: Despite a strong display on CAEC of CXCR4 and a lesser expression of CCR3 and CCR5, HIV-I did not productively replicate in CAEC, as shown by infectious assay, immunofluorescence, and electron microscopy. HIV-1 infec tion 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-1(JR-FL), virus penetration 2 hr postinfection (PI) was negligible but increased by 1750% 24 hr PI. The par acellular permeability increased during this period by only 25%. Neither AO P-RANTES nor v-MIPII significantly reduced HIV-1(JR-FL) invasion. Virus inf ection 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 v irus opens a transcellular route across coronary and brain endothelia in cy toplasmic vacuoles.