005). Fluorescent staining (FITC) for chlamydial antigens supported an impairment

of chlamydial growth and inclusion body expansion after 405 nm exposure (Figure 2D-F, 20 J/cm2) compared to C. trachomatis infected cells alone (Figure 2A-C). We also analyzed the effect of irradiation application time post-infection to determine if it was growth phase specific. At 24 h post-infection, irradiation with 405 nm (20 J/cm2) LEDs still demonstrated a significant growth MK5108 chemical structure inhibition (Figure 1B, P < 0.005). C. trachomatis-infected cells treated with red 670 nm LEDs at similar energy densities (5-20 J/cm2) showed no significant effect on growth (data not shown). Figure 1 Effects of 405 nm irradiance on chlamydial growth in HeLa cells. Sotrastaurin supplier (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/cm2) either promptly after infection or 24 h post-infection (24 h Poziotinib supplier 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. Figure 2 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 Bortezomib chemical structure photodiodes. (D-F) Infected cells were exposed to 405 nm LEDs at 20 J/cm2 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. Considering many chronic chlamydial infections are

in a persistent stage of growth, we tested the effect of 405 nm on chlamydial growth after penicillin-induced persistence. As shown in Figure 1C, 405 nm retarded chlamydial growth during a persistent state (P < 0.05) at 20 J/cm2, though the result was not as pronounced as it was in the active state. Once again, no effect was seen with 670 nm treatment (data not shown). The effect of 405 nm irradiation on IL-6 production in C. trachomatis-infected HeLa cells Previous studies have identified IL-6 as a pro-inflammatory cytokine associated with immunopathologic effects in chronic C. trachomatis infections [12, 13]. In this study, we demonstrated elevated IL-6 levels post-chlamydial infection compared to uninfected cells (Figure 3A, P < 0.005). C.