d562 human pharyngeal epithelial cells  (ATCC)

Journal: mSphere

Article Title: Heme sequestration by hemophilin from Haemophilus haemolyticus reduces respiratory tract colonization and infection with non-typeable Haemophilus influenzae

doi: 10.1128/msphere.00006-24

Figure Lengend Snippet: H. haemolyticus reduces NTHi growth and adherence to human airway epithelial cells in an Hpl-dependent manner. ( A ) Growth rate of NTHi strain H632 represented as the log change in bacterial density during exponential phase per hour, α·hr −1 , in cultures co-inoculated with H. haemolyticus strain WT BW1 or Δhpl BW1. Cultures were performed under minimal heme conditions with or without supplementation as indicated ( n = 3 independent experiments). ( B ) Attachment and invasion of NTHi strain H632 to human respiratory tract epithelial cell lines, A549 cells and D562 cells, with or without 4-hour pre-exposure to H. haemolyticus strain WT BW1 or Δhpl BW1 ( n = 3 independent experiments). ( C ) Growth rate of NTHi strain H632 as in ( A ), in cultures co-inoculated with H. haemolyticus strains RHH (RHH122) or NF5 ( n = 3 independent experiments). ( D ) Attachment and invasion of NTHi strain H632 to A549 cells and D562 cells as in ( B ) with or without 4-hour pre-exposure to H. haemolyticus strains RHH (RHH122) or NF5 ( n = 3 independent experiments). Data are displayed as the mean of two technical replicates per condition for each independent experiment (biological replicate) ( A, C ) or median of three technical replicates per condition for each independent experiment (biological replicate) ( B, D ) ±SEM. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001, two-way ANOVA with Dunnett’s post hoc analysis for multiple comparisons ( A, C ), or Kruskal–Wallis test with Dunn’s post hoc analysis for multiple comparisons ( B, D ).

Article Snippet: A549 (ATCC #CCL-185) human lung carcinoma epithelial cells and Detroit 562 (D562) human pharyngeal carcinoma epithelial cells (ATCC #CCL-138) were maintained in Ham’s F-12 (Kaighn’s) medium (ThermoFisher Scientific) supplemented with 10% fetal bovine serum (CPS Serum) and 10,000 U/mL penicillin–streptomycin (ThermoFisher Scientific).

Techniques:

Journal: mSphere

Article Title: Heme sequestration by hemophilin from Haemophilus haemolyticus reduces respiratory tract colonization and infection with non-typeable Haemophilus influenzae

doi: 10.1128/msphere.00006-24

Figure Lengend Snippet: Recombinant Hpl is sufficient to reduce NTHi burdens in vivo and decrease adherence to human respiratory tract epithelial cells. ( A ) rHpl exposure and NTHi strain H632 challenge experimental model. Mice were treated with three doses of rHpl at 24-hour intervals prior to NTHi infection. ( B–C ) Burdens of NTHi detected in the nasopharyngeal lavage ( B ) and lower airway BAL and lung tissue ( C ) at 24 hours post-infection with 10 8 CFU/mouse i.n. with rHpl pre-exposures at the indicated doses ( n = 11–15 mice/group). ( D ) H. haemolyticus strain Δhpl BW1 pre-exposure with or without rHpl prior to NTHi strain H632 challenge experimental model. ( E–F ) Burdens of NTHi detected in the nasopharyngeal lavage ( E ) and lower airway BAL and lung tissue ( F ) at 24 hours post-infection with H. haemolyticus strain Δhpl BW1 and rHpl pre-exposures as indicated ( n = 14 mice/group). ( G ) Percent adherence of NTHi to human respiratory tract epithelial cell lines, A549 cells and D562 cells, with or without 4-hour pre-exposure to rHpl as indicated ( n = 3 independent experiments, conducted with three technical replicates per condition). Data are pooled from three to four independent experiments and are displayed as mean ± SEM. * P < 0.05, ** P < 0.01, Kruskal–Wallis test with Dunn’s post hoc analysis for multiple comparisons ( B–C ), Mann-Whitney U test ( E, F ), and one-way ANOVA with Dunnett’s post hoc analysis for multiple comparisons ( G ).

Article Snippet: A549 (ATCC #CCL-185) human lung carcinoma epithelial cells and Detroit 562 (D562) human pharyngeal carcinoma epithelial cells (ATCC #CCL-138) were maintained in Ham’s F-12 (Kaighn’s) medium (ThermoFisher Scientific) supplemented with 10% fetal bovine serum (CPS Serum) and 10,000 U/mL penicillin–streptomycin (ThermoFisher Scientific).

Techniques: Recombinant, In Vivo, Infection, MANN-WHITNEY