mouse isolate e coli a3  (ATCC)

Journal: Frontiers in Immunology

Article Title: Removal of natural anti-αGal antibodies elicits protective immunity against Gram-negative bacterial infections

doi: 10.3389/fimmu.2023.1232924

Figure Lengend Snippet: Treatment with GAS914 impacts GalT-KO mice survival after CLP. (A) DNA stratification by pulsed-field gel electrophoresis (PFGE) of E coli isolated from mouse blood after 12 h – 24 h of CLP in GalT-KO mice treated with GAS914 (n = 8) or PBS (n = 8). (B) Influence of GAS914 or PBS treatment on GalT-KO mouse survival after CLP in animals treated before and after the procedure (left, n = 17) or beginning the treatment 12 h after CLP (right, n = 14). Data are represented in Kaplan-Maier curves and were compared using the long-rank (Mantel-Cox) test, **P < 0.01.

Article Snippet: We also measured in vitro GalT-KO mice serum bactericidal activity, and the antibody and complement deposition against the pathogenic mouse isolate E. coli A3 and E. coli O86:B7 (ATCC ® 12701 TM ), a bacterium with high α-galactosyl content ( , ).

Techniques: Pulsed-Field Gel, Electrophoresis, Isolation

Journal: Frontiers in Immunology

Article Title: Removal of natural anti-αGal antibodies elicits protective immunity against Gram-negative bacterial infections

doi: 10.3389/fimmu.2023.1232924

Figure Lengend Snippet: Treatment with GAS914 increases GalT-KO mice serum bactericidal capacity against E coli A3 isolated after CLP. (A) Effect of GAS914 and PBS in GalT-KO bactericidal activity against E coli A3. The bactericidal activity was calculated as the percentage of bacteria surviving in reaction mixtures containing the tested serum compared to the control (growth). Growth: control bacterial growth, ACP, alternative complement pathway; SMS, standard mouse serum. Individual data of PBS and GAS914 represent the mean of three experiments. (B) Median fluorescence intensity of IgG, IgM, C3, C4, and IgG subclasses on the surface of E coli A3 (n = 6). Individual data represents the mean of three experiments. Unpaired non-parametric Mann-Whitney test was used to compare PBS and GAS914 groups, ** P < 0.01.

Article Snippet: We also measured in vitro GalT-KO mice serum bactericidal activity, and the antibody and complement deposition against the pathogenic mouse isolate E. coli A3 and E. coli O86:B7 (ATCC ® 12701 TM ), a bacterium with high α-galactosyl content ( , ).

Techniques: Isolation, Activity Assay, Bacteria, Control, Fluorescence, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Removal of natural anti-αGal antibodies elicits protective immunity against Gram-negative bacterial infections

doi: 10.3389/fimmu.2023.1232924

Figure Lengend Snippet: Neutralizing anti-αGal antibodies from human sera with GAS914 increases serum bactericidal killing and decreases IgG2 binding to Gram-negative bacteria. (A) Bactericidal killing after exposing the human sera to PBS or GAS914 against E coli O86:B7, P. aeruginosa 21565, and K pneumonia 35204 (n = 8). The bactericidal killing was calculated as the percentage of the number of bacteria surviving in reaction mixtures containing the tested serum compared to the control (growth). Growth: control bacterial growth, ACP, alternative complement pathway. Individual data of PBS and GAS914 represent the mean of three experiments. (B) Median fluorescence intensity of IgG, IgM, IgA, IgG subclasses, and complement C3 and C4 deposition on the surface of E coli O86:B7 (n = 8). Individual data represents the mean of three experiments. Comparisons were analyzed by paired t -tests, * P < 0.05, ** P < 0.01.

Article Snippet: We also measured in vitro GalT-KO mice serum bactericidal activity, and the antibody and complement deposition against the pathogenic mouse isolate E. coli A3 and E. coli O86:B7 (ATCC ® 12701 TM ), a bacterium with high α-galactosyl content ( , ).

Techniques: Binding Assay, Bacteria, Control, Fluorescence

Journal: Frontiers in Immunology

Article Title: Removal of natural anti-αGal antibodies elicits protective immunity against Gram-negative bacterial infections

doi: 10.3389/fimmu.2023.1232924

Figure Lengend Snippet: Length and expression level of lipopolysaccharide (LPS) O-antigen chains and αGal in Gram-negative bacteria. (A) Separation by SDS-PAGE (13%) of repeating oligosaccharide units of LPS extracted from Escherichia coli O86:B7 (lane 1), Pseudomonas aeruginosa 21565 (lane 2), Klebsiella pneumoniae 35204 (lane 3) and Escherichia coli A3 (lane 4). Standard LPS from E coli O111:B4 (lane 5, Sigma-Aldrich, St. Louis, MO, USA) was used as a reference. (B) Median fluorescence intensity of α-galactosyl expression with isolectin IB4 and αGal expression with anti-αGal IgM and IgG monoclonal antibodies in E coli A3, K pneumoniae 35204, P. aeruginosa 21565, and E coli O86:B7. Data are representative of three independent experiments.

Article Snippet: We also measured in vitro GalT-KO mice serum bactericidal activity, and the antibody and complement deposition against the pathogenic mouse isolate E. coli A3 and E. coli O86:B7 (ATCC ® 12701 TM ), a bacterium with high α-galactosyl content ( , ).

Techniques: Expressing, Bacteria, SDS Page, Fluorescence, Bioprocessing