Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effects of 405 nm irradiance on chlamydial growth in HeLa cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5. ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (24 h post). Treatments are grouped based on post-hoc comparisons for convenience. The effect of 405 nm on chlamydial growth was assessed during active and persistent stages induced with penicillin ( B and C ). Growth was determined using quantitative real-time PCR to determine the ratio of chlamydial and eukaryotic housekeeping genes (16S: GAPDH respectively) 48 h post-infection on cDNA reverse transcribed from RNA. Mean ± standard deviation are plotted for the two replicated experiments. Statistical significance was determined post-hoc using a Bonferonni adjustment comparing all groups against C. trachomatis -infected HeLa cells alone (CTE); * P < 0.05, ** P < 0.005.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Real-time Polymerase Chain Reaction, Reverse Transcription, Standard Deviation

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Anti-chlamydial properties of 405 nm irradiance. ( A - C ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 without exposure to photodiodes. ( D - F ) Infected cells were exposed to 405 nm LEDs at 20 J/cm 2 promptly after infection to evaluate anti-chlamydial effects during an acute chlamydial infection. Cells were fixed and stained with dapi (blue) ( B and E ) and anti-chlamydial (green) ( C and F ) antibody 48 hours post-infection. Bar = 10μm.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Staining

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effect of 405 nm on IL-6 production in C. trachomatis -infected epithelial cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 (CTE5). ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (post-24 h). The effect of 405 nm on IL-6 production was assessed during active ( A and B ) and penicillin-induced persistent stages ( C ). Supernatants were collected and measured for IL-6 production using an ELISA. Treatments are grouped based on post-hoc comparisons for convenience. Mean ± SEM are plotted for the two replicated experiments. Statistical differences were determined post-hoc using a Bonferonni adjustment comparing all groups to C. trachomatis infected cells (CTE); *, P < 0.05; ** P < 0.005.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effect of 405 nm on CCL2 production in C. trachomatis -infected epithelial cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 (CTE5). ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (post-24 h). The effect of 405 nm on CCL2 was assessed during active ( B ) and persistent stages induced with penicillin ( C ). Supernatants were collected and measured for CCL2 production using an ELISA. Treatments are grouped based on post-hoc comparisons for convenience. Mean ± SEM are plotted for the two replicated experiments. Statistical differences were determined post-hoc using a Bonferonni adjustment comparing all groups to C. trachomatis infected cells (CTE); *, P < 0.001.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Enzyme-linked Immunosorbent Assay