cmp neu5ac  (MedChemExpress)

Journal: Cell host & microbe

Article Title: Interferon-epsilon, an estrogen-induced type I interferon, is uniquely exploited by Neisseria gonorrhoeae via effects on sialic acid metabolism.

doi: 10.1016/j.chom.2025.05.015

Figure Lengend Snippet: Figure 6. Estrogen upregulates CMAS mRNA in the genital tract via IFN-ε Estrogen (E2) upregulates CMP-sialic acid synthetase (CMAS) mRNA in an IFN-ε-dependent fashion but does not upregulate the cathelicidin mCRAMP. Genital tract tissue harvested from WT and Ifnε−/−female mice at 24 h post-implantation of 17β-estradiol (E2) pellets or sham treatment (n = 5–6/group). mRNA isolated from Trizol extracts was analyzed by qPCR, and mRNA levels of (A) IFN-ε, (B) CMAS, and (C) mCRAMP were determined relative to TATA-binding protein (TBP), a housekeeping gene. (D–I) CMP-Neu5Ac, the product of CMAS, enhances gonococcal burden in C57BL/6 J WT and Ifnε−/−mice (D–F) and BALB/c mice treated with anti-IFNAR mAb (G–I) when administered intravaginally. (D–F) Estradiol pellet-treated WT C57BL/6 mice (n = 10/group) and Ifnε−/−mice (n = 8–9/group) were colonized with N. gonorrhoeae FA1090 (3.5 × 107 CFU) on day 0 and treated daily intravaginally (starting on day 0 at the time of infection and on subsequent days after swabs for CFU enumeration were obtained) with either CMP-Neu5Ac (25 μg) or saline. Vaginas were swabbed daily to enumerate CFUs. (G–I) Estradiol pellet-treated WT BALB/c mice (n = 9–10/group) were either given function-blocking anti-IFNAR mAb (MAR1-5A3, i.p., 1 mg, 500, 250, and 250 μg on days −2, 0, 3, and 5, respectively) or saline, were inoculated with N. gonorrhoeae FA1090 (3.1 × 107 CFU) intravaginally on day 0, and were treated daily intravaginally (starting on day

Article Snippet: Additional cohorts of mice were treated intravaginally with 25 μg CMP-Neu5AC (MedChem Express, Cat. HY-112942A) or saline administered daily starting on day 0 at the time of infection, and on subsequent days after swabs.

Techniques: Isolation, Binding Assay, Infection, Saline, Blocking Assay

Journal: Cell host & microbe

Article Title: Interferon-epsilon, an estrogen-induced type I interferon, is uniquely exploited by Neisseria gonorrhoeae via effects on sialic acid metabolism.

doi: 10.1016/j.chom.2025.05.015

Figure Lengend Snippet: Figure 7. Two different strains of gonococci are equally susceptible to complement killing in the absence of IFN signaling (A–F) Anti-IFNAR treatment of Cmah−/−mice (lacking the enzyme CMP-Neu5Ac hydroxylase and, therefore, expressing a human-like sialome) displays reduced colonization by N. gonorrhoeae FA1090 (A–D) and MS11 (E–H) when compared with control PBS-treated Cmah−/−mice (n = 8 mice/group). (A and E) Kaplan- Meier curves showing time to clearance (comparisons were made by Mantel-Cox test). (B and F) Log10 CFU vs. time (mean [SEM]). Comparisons across groups were made by two-way ANOVA. *p < 0.05, **p < 0.01, ****p < 0.0001. (C and G) AUC analysis. The mean (95% CI) is shown. Comparisons across groups are by Mann-Whitney’s non-parametric test. (D and H) Gonococci recovered directly (i.e., without subpassage on artificial media) from the vaginas of Cmah−/−mice treated with PBS are more resistant to killing by normal human serum than anti-IFNAR-treated Cmah−/−mice. N. gonorrhoeae FA1090 and MS11 were recovered from vaginal secretions from the experiments shown above (D and H), respectively. Secretions from 4 mice in each group were pooled to yield two pools from each group and tested for resistance to human complement. Serial dilutions of secretions were incubated with 10% normal human serum, and duplicate aliquots were plated at 0 and 30 min. Percent survival at 30 min (t30) relative to 0 min (t0) is shown on the y axis. Reaction mixtures that yielded 50 to 300 CFU at 0 min were used to calculate survival.

Article Snippet: Additional cohorts of mice were treated intravaginally with 25 μg CMP-Neu5AC (MedChem Express, Cat. HY-112942A) or saline administered daily starting on day 0 at the time of infection, and on subsequent days after swabs.

Techniques: Expressing, Control, Incubation