Fig. 6

Blockage of ICAM-1 significantly attenuates meningitic E. coli-caused neuroinflammatory responses and extends the survival. A Schematics briefly showing the design of two gRNAs (gRNA1 and gRNA2) in the ICAM-1 Exon2 and the identification of the ICAM-1 knockout (KO) through PCR amplification, sequencing, and western blot analysis. Specific primers (P1 and P2) were used to analyze the ICAM-1 sequence in the genome. A total of 30 bp was deleted in ICAM-1 KO cells, in which the ICAM-1 protein was not expressed. B The induction of ICAM-1 by meningitic E. coli was completely abolished in hBMECs accompanying with the knockout of ICAM-1. C Meningitic E. coli-mediated increased monocytes (THP-1) adhesion to hBMECs was significantly blocked when ICAM-1 was knocked out. All data were presented as mean ± SD for three individual experiments. D MSD analysis of the IL-1β and TNF-α amount in serum and brain lysates from control mice, meningitic E. coli-challenged mice with or without ICAM-1-neutralizing antibody pretreatment. Data were expressed as mean ± SD (n = 5). E ICAM-1-neutralizing antibody pretreatment significantly prolongs mice survival time during bacterial challenge. Survival of mice in each group was monitored for 20 h after tail vein injection of meningitic E. coli. Data were collected and shown as Kaplan-Meier survival curves (n = 6)