Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Bacterial strains used in this study .

Article Snippet: Urogenital tract (ME180) , Ng , Neisseria gonorrhoeae MS11 (ATCC BAA1833) , Lc , Lactobacillus crispatus MV24-1a.

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Bacteria mediated EGR1 induction in host epithelial cells. (A–D) EGR1 expression was evaluated using qPCR at 1, 2, 4, and 6 h after the addition of bacteria at a multiplicity of infection (MOI) = 100. The data is represented as fold change relative to uninfected. (E–H) EGR1 expression evaluated using Western blotting. The blots represent the 4 h time point during the course of infection. β-actin expression was used as a loading control. (I–L) Bacterial attachment to cells at different time points after bacterial inoculation (MOI = 100). Bacteria were diluted and plated for viable counts to determine colony forming units (CFU)/ml. (A,E,I) Pharyngeal FaDu cells were inoculated with different strains of N. meningitidis (Nm-A, Nm-B, Nm-C, Nm-W), P. aeruginosa (Pa), N. subflava (Ns), and N. lactamica . (B,F,J) FaDu cells were inoculated with S. pyogenes (Sp-M1, Sp-M3, Sp-M5, Sp-M6), S. aureus (Sa), L. reuteri (Lr), and L. salivarius (C, G, K) Gastric AGS cells were inoculated with H. pylori (Hp-J99, Hp-6721) and L. rhamnosus (Lrh). Cervical ME-180 cells were infected with N. gonorrhoeae (Ng) and L. crispatus (Lc). (D,H,L) Intestinal Caco-2 cells were inoculated with E. coli (Ec-B09, Ec-O11, Ec-DH5α), L. rhamnosus GG (Lrh-GG), Salmonella enterica serovar Enteritidis (SE-3934) and S. enterica serovar Typhimurium (STM-42).

Article Snippet: Urogenital tract (ME180) , Ng , Neisseria gonorrhoeae MS11 (ATCC BAA1833) , Lc , Lactobacillus crispatus MV24-1a.

Techniques: Bacteria, Expressing, Infection, Western Blot, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Cell type specificity in the bacteria-mediated induction of EGR1. (A–E) Cell line specific induction of EGR1 was investigated during bacterial infection of epithelial cell lines of a pharyngeal (FaDu), gastric (AGS), intestinal (Caco-2), and cervical origin (ME180). Expression of EGR1 was monitored using qPCR at 1, 2, 4, and 6 h post infection. (F–J) Bacterial attachment levels to epithelial cells at different time points. Bacteria were diluted and plated for viable counts to determine CFU/ml. Bacteria were added at a MOI of 100 for all experiments. Cells were infected with N. meningitidis (Nm-C), S. pyogenes (Sp-M6), H. pylori (Hp-J99), N. gonorrhoeae (Ng) or S. Enteritidis (SE-3934).

Article Snippet: Urogenital tract (ME180) , Ng , Neisseria gonorrhoeae MS11 (ATCC BAA1833) , Lc , Lactobacillus crispatus MV24-1a.

Techniques: Bacteria, Infection, Expressing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Bacterial viability and direct contact between bacteria and the host cells affect upregulation of EGR1 . Induction of EGR1 was monitored by qPCR after infection of the host epithelial cells with live or dead bacteria. Dead bacteria were obtained by heat treatment at 95°C for 10 min. Viable counts were used to ensure complete killing of the bacteria. The role of bacterial contact with the epithelial cells in the EGR1 induction was studied using a 0.4 μm Millicell filter, which helps to physically separate the bacteria and host cells, but still allows diffusion of secreted factors in the cell growth media (Sup.). (A) Pharyngeal FaDu cells infected with N. meningitidis (Nm-C). (B) Pharyngeal FaDu cells infected with S. pyogenes (Sp-M6). (C) Gastric AGS cells infected with H. pylori (Hp-J99). (D) Cervical ME180 cells infected with N. gonorrhoeae (Ng). (E) Intestinal Caco-2 cells infected with S. Enteritidis (SE3439). Bacteria were added to a MOI of 100 in all experiments. The data was analyzed at 2 h post infection for Nm-C, Sp-M6, SE3439, and Hp-J99. The data was analyzed at 4 h post infection of Ng.

Article Snippet: Urogenital tract (ME180) , Ng , Neisseria gonorrhoeae MS11 (ATCC BAA1833) , Lc , Lactobacillus crispatus MV24-1a.

Techniques: Bacteria, Infection, Diffusion-based Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: EGR1 is primarily activated by the EGFR–ERK1/2 pathway upon bacterial infection . Host epithelial cells were pretreated with PD153035, PD184352, SP600125, P38-IV and PKA-14:22 (inhibiting EGFR, ERK1/2, JNK, p38 and PKA, respectively) 1 h prior to infection. Bacterial infection of the host epithelial cells was carried out by co-incubation with the inhibitors for 2 h, except infection with N. gonorrhoeae that continued for 4 h. The expression of EGR1 was analyzed by qPCR. Expression of cells treated with DMSO was set to 1. (A) FaDu infected with N. meningitidis serogroup C (Nm-C). (B) FaDu infected with S. pyogenes serogroup M6 (Sp-M6). (C) AGS infected with H. pylori J99 (Hp-J99). (D) ME180 infected with N. gonorrhoea MS11 (Ng). (E) Caco-2 infected with S. Enteritidis (SE-3934). Bacteria were added to a MOI of 100 in all experiments. The white bars represent uninfected controls. The colored bars represent the infected samples. The significant difference between the infected control (DMSO) and the infected samples is marked with asterisk.

Article Snippet: Urogenital tract (ME180) , Ng , Neisseria gonorrhoeae MS11 (ATCC BAA1833) , Lc , Lactobacillus crispatus MV24-1a.

Techniques: Infection, Incubation, Expressing, Bacteria, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Integrin mediated signaling in bacteria induced EGR1 expression . The host epithelial cells were transfected with control siRNA (si-NT), siRNA targeted against β1-integrin (si-Integrin) or directed against integrin-linked kinase (si-ILK) for 60–68 h. The cells were then infected with bacteria with a MOI of 100 for 2 h, except the infection with N. gonorrhoeae that continued for 4 h. EGR1 expression was analyzed by qPCR. The graphs represent fold difference in EGR1 expression between infected and uninfected epithelial cells. (A) FaDu infected with N. meningitidis serogroup C (Nm-C). (B) FaDu infected with S. pyogenes serogroup M6 (Sp-M6). (C) MKN45 infected with H. pylori J99 (Hp-J99). (D) ME180 infected with N. gonorrhoea MS11 (Ng). (E) Caco-2 infected with S. Enteritidis (SE-3934).

Article Snippet: Urogenital tract (ME180) , Ng , Neisseria gonorrhoeae MS11 (ATCC BAA1833) , Lc , Lactobacillus crispatus MV24-1a.

Techniques: Bacteria, Expressing, Transfection, Control, Infection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Host Cell Transcription Factor EGR1 Is Induced by Bacteria through the EGFR–ERK1/2 Pathway

doi: 10.3389/fcimb.2017.00016

Figure Lengend Snippet: Hypothetical model of bacteria-mediated EGR1 induction in host epithelial cells . At the host cell surface, the bacteria may stimulate β1-integrin and/or EGFR. All strains tested exhibit absolute requirement for β1-integrin expression to trigger EGR1 induction (shown in Figure ). Inhibition of EGFR by PD153035 blocked induction of EGR1 by all strains (shown in Figure ). Thereby, suggesting that activation of EGFR through β1-integrin as a possible mechanism for bacteria mediated induction of EGR1. Bacterial contact with the host cells was required for the induction of EGR1 response (shown in Figure ). Integrins and EGFR can activate several signaling molecules inside the host cell that can in turn lead to induction of EGR1. Signaling through ERK1/2 was critical for all strains, since the induction of EGR1 was blocked by ERK1/2 inhibitor (Figure ). Also other signaling factors are partially involved in a species-dependent manner. Data suggest that H. pylori, N. gonorrhoeae and S. Enteritidis can signal through JNK to upregulate EGR1, whereas PKA is utilized only by S. Enteritidis .

Article Snippet: Urogenital tract (ME180) , Ng , Neisseria gonorrhoeae MS11 (ATCC BAA1833) , Lc , Lactobacillus crispatus MV24-1a.

Techniques: Bacteria, Expressing, Inhibition, Activation Assay