fannyhessea vaginae  (DSMZ)

Journal: bioRxiv

Article Title: Genetic transformation of Gardnerella species and characterization of vaginolysin and sialidase mutants

doi: 10.1101/2025.05.09.653190

Figure Lengend Snippet: Regions of homology with the G. vaginalis ATCC 14018 chromosome are shown in gray. pAKK186 contains the rpsB promoter from ATCC 14018 for constitutive expression of a gene cloned into the multiple cloning site (MCS). Plasmid integrants will be Erm R Spc R , while double recombinants will be Erm R Spc S .

Article Snippet: Gardnerella vaginalis ATCC 14018 was used to develop a method for electrotransformation starting with the methods for transformation with oligos described by Garcia ( ).

Techniques: Expressing, Clone Assay, Cloning, Plasmid Preparation

Journal: bioRxiv

Article Title: Vaginal bacteria-derived extracellular vesicles diffuse through human cervicovaginal mucus to enable bacterial signaling to upper female reproductive tract tissues

doi: 10.1101/2025.04.28.651063

Figure Lengend Snippet: Analysis was completed via nanoparticle tracking analysis. ( A ) Isolated bEVs were confirmed to be membrane-bound particles via TEM. Scale bars denote 200 nm. ( B, C ) G. vaginalis -derived bEVs were significantly lower in concentration (1.30 ± 0.089 × 10 9 ) compared to isolated bEVs from L. crispatus (2.05 ± 0.16 × 10 9 particles/mL, p = 0.0156) and M. mulieris cultures (2.14 ± 0.24 × 10 9 particles/mL, p = 0.0110). ( D ) L. crispatus cultures (7.15 ± 1.35 × 10 9 bEVs/CFU) had significantly higher bEV production compared to G. vaginalis cultures (1.67 ± 0.36 × 10 9 bEVs/CFU; p = 0.0048) and M. mulieris cultures (0.24 ± 0.05 × 10 9 bEVs/CFU, p = 0.0015). L. iners cultures (5.49 ± 1.42 × 10 9 bEVs/CFU) had higher bEVs production compared to M. mulieris cultures ( p = 0.0145). ( E ) No differences in bEV protein content were observed. ( F ) G. vaginalis bEVs (134.0 ± 2.293 nm) and M. mulieris -derived bEVs (131.7 ± 2.809 nm) were smaller than L. crispatus - and L. iners -derived bEVs (143.6 ± 1.327 nm, p ≤ 0.0182; 154.5 ± 1.785 nm, p < 0.0001, respectively). ( G ) G. vaginalis -derived bEVs had a more neutral ζ-potential (−26.5 ± 1.221 mV) compared to L. crispatus -derived bEVs (−30.85 ± 0.537 mV, p = 0.0317), L. iners -derived bEVs (−31.90 ± 1.070 mV, p = 0.0072), and M. mulieris -derived bEVs (−31.30 ± 1.107 mV, p = 0.0257). Values are reported as mean ± SEM. Statistical significance ( p ≤ 0.05) is represented by a letter corresponding to the species of comparison (a, L. crispatus; b, L. iners ; c, G. vaginalis ; d, M. mulieris ). (n = 6 per group, 2 outliers removed from the M. mulieris group in accordance with Grubb’s outlier test due to size and concentration).

Article Snippet: Human-derived strains of Lactobacillus crispatus (Brygoo and Aladame, 33820), Lactobacillus iners (BAA-3226), Gardnerella vaginalis (Gardner and Dukes, 14018), and Mobiluncus mulieris (43064), as well as VK2/E6E7 (CRL-2616), and Kaighn’s Modification of Ham’s F-12 media, were sourced from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Isolation, Membrane, Derivative Assay, Concentration Assay, Comparison

Journal: bioRxiv

Article Title: Vaginal bacteria-derived extracellular vesicles diffuse through human cervicovaginal mucus to enable bacterial signaling to upper female reproductive tract tissues

doi: 10.1101/2025.04.28.651063

Figure Lengend Snippet: ( A ) Geometric means of the average mean squared displacement at 1 second for all samples. bEVs demonstrate increased mobility compared to whole bacteria ( B ) All CVM samples showed limited mobility of L. crispatus whole bacteria, compared to bEVs ( p ≤ 0.01). ( C ) 9/10 samples showed limited mobility of L. iners whole bacteria compared to bEVs ( p ≤ 0.0001). ( D ) 6/10 samples showed increased mobility of G. vaginalis bEVs compared to whole bacteria ( p ≤ 0.01). ( E ) 9/10 samples demonstrated limited mobility of whole bacteria compared to bEVs (p ≤ 0.0001). Box and Whiskers are shown as 5-95% confidence intervals. Two-tailed Mann-Whitney non-parametric tests were performed to compare whole bacteria and bEV mobility in each individual participant. n = 10 samples.

Article Snippet: Human-derived strains of Lactobacillus crispatus (Brygoo and Aladame, 33820), Lactobacillus iners (BAA-3226), Gardnerella vaginalis (Gardner and Dukes, 14018), and Mobiluncus mulieris (43064), as well as VK2/E6E7 (CRL-2616), and Kaighn’s Modification of Ham’s F-12 media, were sourced from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Bacteria, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Vaginal bacteria-derived extracellular vesicles diffuse through human cervicovaginal mucus to enable bacterial signaling to upper female reproductive tract tissues

doi: 10.1101/2025.04.28.651063

Figure Lengend Snippet: ( A ) Confocal images reveal uptake of bEVs to the cytosol. Scale bars denote 20 µm. ( B ) Based on 2-way ANOVAs, uptake assays determined that there were significant differences in the uptake based on bEV type. ( C ) Vaginal epithelial cells internalized M. mulieris -derived bEVs (23.88 ± 2.43%) significantly more than L. crispatus -derived bEVs (13.30 ± 1.99%, p = 0.0001), L. iners -derived bEVs (15.52 ± 2.13%, p = 0.0074), or G. vaginalis -derived bEVs (13.86 ± 2.03%, p = 0.0005) at 24 h (n=8 per dosage per timepoint. Replicates below the standard curve were assumed to be 0%.) ( D ) Vaginal epithelial cells did not exhibit a decrease in viability after 24 h incubation with bEVs of any dosage or species. (n=8 for treatment group, n=16 for vehicle control). ( E ) VK2/E6E7 cells exhibit a pro-inflammatory response after treatment with G. vaginalis- and M. mulieris-derived bEVs (n=8). Statistics for uptake and viability were performed using 2-way ANOVA. Statistics for cytokine production were performed using 1-way ANOVA for each cytokine using raw cytokine concentrations.

Article Snippet: Human-derived strains of Lactobacillus crispatus (Brygoo and Aladame, 33820), Lactobacillus iners (BAA-3226), Gardnerella vaginalis (Gardner and Dukes, 14018), and Mobiluncus mulieris (43064), as well as VK2/E6E7 (CRL-2616), and Kaighn’s Modification of Ham’s F-12 media, were sourced from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Derivative Assay, Incubation, Control

Journal: bioRxiv

Article Title: Vaginal bacteria-derived extracellular vesicles diffuse through human cervicovaginal mucus to enable bacterial signaling to upper female reproductive tract tissues

doi: 10.1101/2025.04.28.651063

Figure Lengend Snippet: ( A ) Confocal images reveal uptake of bEVs to the cytosol. Scale bars denote 20 µm. ( B ) Based on 2-way ANOVAs, uptake assays determined that there were significant differences in the uptake based on bEV type. ( C ) L. crispatus-derived bEVs (14.06 ± 0.91%) saw the highest uptake at 24 h compared to L. iners-derived bEVs (9.97 ± 0.67%, p < 0.0001), G. vaginalis-derived bEVs (1.468 ± 0.52%, p < 0.0001), and M. mulieris-derived bEVs (7.09 ± 0.73%, p < 0.0001) (n=8 per dosage per timepoint. Replicates below the standard curve were assumed to be 0%.) ( D ) Endometrial cells did not exhibit a decrease in viability after 24 h incubation with bEVs of any dosage or species. (n=8) ( E ) Endometrial cells exhibit changes in IL-8 production after dosage with G. vaginalis- and M. mulieris-derived bEVs (n=9). Treatment with G. vaginalis-derived bEVs resulted in an increase in log 2 fold change (1.27 ± 0.10, p < 0.0001). Statistics for uptake and viability were performed using 2-way ANOVA. Statistics for cytokine production were performed using 1-way ANOVA for each cytokine using the raw concentrations.

Article Snippet: Human-derived strains of Lactobacillus crispatus (Brygoo and Aladame, 33820), Lactobacillus iners (BAA-3226), Gardnerella vaginalis (Gardner and Dukes, 14018), and Mobiluncus mulieris (43064), as well as VK2/E6E7 (CRL-2616), and Kaighn’s Modification of Ham’s F-12 media, were sourced from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Derivative Assay, Incubation

Journal: bioRxiv

Article Title: Vaginal bacteria-derived extracellular vesicles diffuse through human cervicovaginal mucus to enable bacterial signaling to upper female reproductive tract tissues

doi: 10.1101/2025.04.28.651063

Figure Lengend Snippet: ( A ) Confocal images show uptake of bEVs to the cytosol. Scale bars denote 20 µm. ( B ) Based on 2-way ANOVAs, uptake assays determined that there were significant differences in the uptake based on bEV type. ( C ) L. crispatus -derived bEVs were most internalized after 24 h (41.58 ± 2.502%). G. vaginalis -derived bEVs exhibited the second highest uptake, reaching 22.21 ± 0.988%. L. iners - and M. mulieris -derived bEVs saw lower uptake, at 13.890 ± 1.244% and 5.097 ± 0.544% respectively (n=8 per dosage per timepoint. Replicates below the standard curve were assumed to be 0%.) ( D ) BeWo-b30 cells did decrease in viability after 24 h incubation with bEVs of any dosage or species. (n=8 for treatment group, n=16 for vehicle control). ( E ) Multiplex assays studies revealed significant increases of the log 2 fold change in IL-6 (0.43 ± 0.06, p = 0.0010) and TNFα (1.91 ± 0.014, p < 0.0001 ) after dosage with G. vaginalis -derived bEVs. Statistics for uptake and viability were performed using 2-way ANOVA. Statistics were performed using 1-way ANOVA for raw concentrations of each cytokine.

Article Snippet: Human-derived strains of Lactobacillus crispatus (Brygoo and Aladame, 33820), Lactobacillus iners (BAA-3226), Gardnerella vaginalis (Gardner and Dukes, 14018), and Mobiluncus mulieris (43064), as well as VK2/E6E7 (CRL-2616), and Kaighn’s Modification of Ham’s F-12 media, were sourced from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Derivative Assay, Incubation, Control, Multiplex Assay