hmgb1 blocking antibody  (Santa Cruz Biotechnology)

Journal: PLoS Medicine

Article Title: Role of heme in lung bacterial infection after trauma hemorrhage and stored red blood cell transfusion: A preclinical experimental study

doi: 10.1371/journal.pmed.1002522

Figure Lengend Snippet: (A) C57BL/6 wild-type mice underwent trauma hemorrhage (TH) and were randomly resuscitated with either fresh ( n = 3) or stored ( n = 3) red blood cells (RBCs) with plasma (1:1). Blood was drawn 6 h after resuscitation, and high mobility group box 1 (HMGB1) levels were measured by ELISA. Data are mean ± SEM; each symbol represents an individual mouse. * p = 0.018 compared to mice resuscitated with fresh blood by unpaired t test. (B) Rat microvascular endothelial cell (RMVEC) monolayers were exposed to vehicle (0.01 M NaOH, n = 5) or hemin (10 μM, n = 5) for 6 h, and supernatants were analyzed by ELISA for extracellular HMGB1. Data show mean ± SEM. * p = 0.003 compared to vehicle by unpaired t test. (C) For phagocytosis assays, MH-S cells were pretreated with anti-HMGB1 blocking antibody or isotype control antibody prior to exposure to hemin and P . aeruginosa K-strain (PAK). Colonies were counted, and phagocytosis expressed as percent of control, with 100 percent being represented by colony number in plates in vehicle-treated cells. Data were collected from 3 independent experiments with 3 replicates in each experiment. Data are mean ± SEM; each symbol represents an individual experimental mean. * p = 0.023 relative to control and ** p = 0.045 relative to heme + isotype antibody by 1-way repeated measures ANOVA with Tukey post-test. (D) C57BL/6 wild-type mice underwent TH and resuscitation with stored RBCs with plasma (1:1). Forty-eight hours later, these mice were instilled with PAK, and survival monitored. Before instillation of PAK, mice were injected peritoneally with either anti-HMGB1 blocking antibody ( n = 6) or isotype control antibody ( n = 4). * p = 0.001 compared to isotype control by log-rank test.

Article Snippet: Anti-HMGB1 blocking antibody was a kind gift from Kevin Tracey (Feinstein Institute for Medical Research, New York, NY).

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Blocking Assay, Control, Injection

Journal: PLoS Medicine

Article Title: Role of heme in lung bacterial infection after trauma hemorrhage and stored red blood cell transfusion: A preclinical experimental study

doi: 10.1371/journal.pmed.1002522

Figure Lengend Snippet: We hypothesize that when resuscitation with stored red blood cells (RBCs) leads to increased circulating heme that is in excess of hemopexin, 2 parallel pathways are activated involving (i) release of HMGB1 and subsequent decreased immune-cell-dependent bacterial clearance and (ii) increased endothelial permeability. These combine to promote acute lung injury and increase sensitivity to lung infection. We also underscore the potential that heme-dependent effects synergize with other components of stored RBCs, including hemoglobin, non-transferrin-bound iron (NTBI), and microvesicles, to mediate adverse outcomes.

Article Snippet: Anti-HMGB1 blocking antibody was a kind gift from Kevin Tracey (Feinstein Institute for Medical Research, New York, NY).

Techniques: Permeability, Infection