title: Molecular characterization of hypoxia-hypothermia-conditioned human endothelial cells and their interaction with human monocytes.
abstract: This in vitro study was designed to characterize the molecular profiling of human endothelial cells (ECs) during the early phase of hypoxia-hypothermia (HH) conditioning and to evaluate their interactions with allogeneic monocytes. The HH-conditioned ECs were analyzed using real-time quantitative polymerase chain reaction (RT-PCR). A cell adhesion assay was performed to assess adhesion of purified allogeneic monocytes as well as CD4- and CD8-positive T cells to HH-conditioned ECs with or without blocking antibodies specific for CD15s and CD162. Uptake of EC membrane by monocytes with or without scavenger receptor blockade was examined using fluorescence-activated cell scanning. The RT-PCR revealed up-regulation of gene transcripts for inflammatory cytokines, monocyte-associated growth factors, costimulatory, and apoptosis-related molecules in HH-conditioned ECs. Analysis using fluorescence-activated cell scanning showed minimal CD54 up-regulation in HH-conditioned ECs. We noted low-level adhesion of CD4- or CD8-positive cells to resting and HH-conditioned ECs. High-level adhesion of monocytes to HH-conditioned ECs was observed when compared with resting ECs. Blockade of CD15s and CD162 dramatically reduced monocyte adhesion to normal and HH-conditioned ECs. Monocytes but not T cells showed uptake of EC membranes during their interactions with HH-conditioned ECs, which was inhibited by scavenger receptor blockade. These data characterized the molecular features of ECs during early HH-conditioning. The EC transcripts related to monocyte recruitment and interaction between monocytes and HH-conditioned ECs dominated the early post-HH condition. Blockade of CD15s and CD162 prevented monocyte adhesion to ECs. These findings suggest that the initial interaction between monocytes and HH-conditioned ECs has a central role during the early phase of reperfusion injury.
authors.affiliation: Department of Transplantation, Jinan City Central Hospital, Jinan, China.
authors.forename: X
authors.initials: X
authors.lastname: Wang
authors.forename: Z
authors.initials: Z
authors.lastname: Liu
authors.forename: B
authors.initials: B
authors.lastname: Zhu
authors.forename: P
authors.initials: P
authors.lastname: Wang
authors.forename: C
authors.initials: C
authors.lastname: Wu
authors.forename: H
authors.initials: H
authors.lastname: Xu
mesh_terms: D015703:Antigens, CD / Q000097:blood; D001011:Aorta; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005434:Flow Cytometry; D006684:HLA-DR Antigens / Q000097:blood; D006801:Humans; D007035:Hypothermia / Q000503:physiopathology*; D000860:Hypoxia / Q000503:physiopathology*; D007962:Leukocytes / Q000166:cytology / Q000502:physiology; D009000:Monocytes / Q000166:cytology / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014184:Transplantation, Homologous; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D015703:Antigens, CD; D017077:Culture Media, Conditioned; D006684:HLA-DR Antigens; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha