Characterization of influenza virus-induced leukocyte adherence to human umbilical vein endothelial cell monolayers.
The adherence of undifferentiated 51Cr-labeled HL-60 (0.5 x 10(6) HL-60 cells/well) cells was monitored on influenza virus-infected HUVEC monolayers. Whereas only 3.0 +/- 1.6% (n = 36) of HL-60 cells adhered to uninfected HUVEC, adherence was increased to 41.7 +/- 2.2% (n = 6), 79.7 +/- 1.2% (n = 6), 83.9 +/- 0.7% (n = 6), and 84.4 +/- 0.5% (n = 6) on HUVEC infected for 7 h at a MOI of 1, 3, 6, and 9, respectively. In comparison, HL-60 cell adherence increased to 35% when HUVEC monolayers were stimulated with LPS (0.2-20 micrograms) for 4 h. Increased adherence to infected HUVEC occurred at 5 h postinfection, peaked at 7 h, and was maintained at 24 h postinfection. Active virus and metabolically active endothelial cells were required to mediate the virus-induced adherence. E-selectin and ICAM-1 Ag were upregulated 78.3- and 4.1-fold, respectively, by LPS (0.02-20 micrograms, 4 h) whereas virus infection (7 h) only increased these proteins 2.6- and 1.4-fold with a MOI > or = 16. Although the time courses of expression for both adhesion molecules after LPS treatment of virus infection were similar, the difference in the magnitude of upregulation suggests that virus-induced adherence is not a result of upregulation of E-selectin and ICAM-1. In contrast, surface expression of HA is involved in HL-60 cell adherence to virus-infected HUVEC because (1) the time course and magnitude of HA AG expression paralleled the time course and magnitude of HL-60 cell adherence after virus infection of HUVEC; (2) HL-60 cell aggregates were absent on infected HUVEC monolayers in the presence of anti-HA; (3) HL-60 cells competed with RBC for infected endothelial cells stained for cellular HA Ag and (4) anti-HA abolished the virus-induced adherence. Furthermore, it appears that HL-60 cells are binding directly to HA because HL-60 cell adherence to a cell-free surface was increased if virus was prebound and neuraminidase treatment of HL-60 cells prevented the HL-60 cell adherence to influenza virus-infected endothelial monolayers.
Colden-Stanfield
M
M
Department of Physiology, Armed Forces Radiobiology Research Institute, Bethesda, MD 20889-5603.
Ratcliffe
D
D
Cramer
E B
EB
Gallin
E K
EK
D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism; D019040:E-Selectin; D004730:Endothelium, Vascular / Q000166:cytology*; D006389:Hemagglutinins, Viral / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D009980:Influenza A virus; D007251:Influenza, Human / Q000276:immunology* / Q000473:pathology; D018799:Intercellular Adhesion Molecule-1; D007962:Leukocytes / Q000166:cytology*; D009439:Neuraminidase / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014471:Umbilical Veins / Q000166:cytology
D015815:Cell Adhesion Molecules; D019040:E-Selectin; D006389:Hemagglutinins, Viral; D018799:Intercellular Adhesion Molecule-1; D009439:Neuraminidase