chlamydia trachomatis c t serovar d svd  (ATCC)

Journal: NPJ Vaccines

Article Title: Post-exposure vaccine protection of CTH522/CAF ® 01 against reinfection with Chlamydia trachomatis requires Th1/Th17 but not Th2-immunity

doi: 10.1038/s41541-025-01117-w

Figure Lengend Snippet: Groups of female B6C3F1 mice were given a transcervical infection (infection 1, I1) 13 weeks after, the animals were given another infection (infection 2, I2). 7 days after the second infection the animals were taken out for analysis. a Bacterial burden in genital tract (GT) swabs is measured ( n = 12). Bacterial numbers are calculated as log10 of inclusion-forming units (IFU). Line represents median log 10 (IFU) levels with 25th and 75th percentiles. b Overview of the model using antibiotics: The animals were infected at week 0 (W0) with 1.5×10 3 IFU of Chlamydia trachomatis (C.t.) serovar D (SvD). 1, 2 or 3 weeks after the infection, the animals were treated with antibiotics. 13 weeks after the first infection, the mice were given a second infection with 1.5×10 3 IFU of C.t . SvD. c Bacterial levels measured at day 7 post infection no. 2 ( n = 4–12). d , e Total IgG in serum at day 7 post infection 2 shown as titration curve with calculated area under the curve (AUC). f , g , h The effect of week 1 antibiotic treatment on natural immunity 5 weeks post treatment ( n = 8). f Total UV-SvD-specific IgG in serum shown as titration curve g with calculated AUC. Points and error bars indicate means ± SD. h the frequency of cytokine-producing CD44 high(hi) CD4 T cells (cyt+, expressing combinations of TNFα, IL-2, IFNγ and/or IL-17) among all CD4 T cells in GT (D, n = 8) in response to in vitro UV-SvD stimulation (gating strategy Supplementary Fig. ). Bars indicate means ± SD. Statistical significance in ( a ) was evaluated Mann–Whitney test. Statistical significance in ( c ) and ( e ) was determined by Kruskal–Wallis test followed by Dunn’s multiple comparisons test. Unpaired t test was used in ( g , h ) for comparison among groups. * p < 0.05, *** p < 0.001, **** p < 0.0001, ns not significant.

Article Snippet: Chlamydia trachomatis(C.t) Serovar D (SvD) (ATCC) were grown in HeLa cells (ATCC) in RPMI 1640 media (Invitrogen) supplemented with 1%HEPES, 1% of Non-essential amino acids (NEAA) (MP Biomedicals), 1% L-Glutamin (Gibco) and 1% pyruvate (Gibco).

Techniques: Infection, Titration, Expressing, In Vitro, MANN-WHITNEY, Comparison

Journal: Human Vaccines & Immunotherapeutics

Article Title: Intranasal delivery of Salmonella OMVs decorated with Chlamydia trachomatis antigens induces specific local and systemic immune responses

doi: 10.1080/21645515.2024.2330768

Figure Lengend Snippet: Production of OMV-based C. trachomatis vaccines. (a-b) Schematic representation of the strategy for vaccine production. Soluble CTH522-SpT2 ( A ) or SpT2-HtrA (B) are conjugated to Salmonella OMVs displaying HbpD-SpC to yield the OMV-CTH522 and OMV-HtrA vaccine formulations. (c-d) Protein profiles of OMV-CTH522 ( C ) and OMV-HtrA ( D ) compared to non-conjugated control OMVs ( OMV-Ctrl ) as analyzed by SDS-PAGE and Coomassie staining. Adducts ( HbpD-SpC-SpT2-CTH522 , HbpD-SpC-SpT2-HtrA ), non-conjugated HbpD-SpC, non-conjugated antigens ( CTH522-SpT2 , SpT2-HtrA ) and the major outer membrane proteins ( OMPs ) as present in the OMV membrane are indicated. Molecular weight markers (kDa) are indicated at the left side of the panels.

Article Snippet: Initially, an E. coli optimized synthetic DNA fragment encoding C. trachomatis SvD HtrA was purchased from Geneart.

Techniques: Vaccines, Control, SDS Page, Staining, Membrane, Molecular Weight

Journal: Human Vaccines & Immunotherapeutics

Article Title: Intranasal delivery of Salmonella OMVs decorated with Chlamydia trachomatis antigens induces specific local and systemic immune responses

doi: 10.1080/21645515.2024.2330768

Figure Lengend Snippet: Intranasal immunization strategy and mouse tissue collection. B6C3F1 female mice were primed on day 0 and received two subsequent boosters on days 14 and 28, with each administration comprising 12 µg of either OMV-CTH522, OMV-HtrA, or OMV-Ctrl. Serum, vaginal fluid, draining cervical lymph nodes (cLns), and spleens of the vaccinated mice were collected 10 d after the third immunization.

Article Snippet: Initially, an E. coli optimized synthetic DNA fragment encoding C. trachomatis SvD HtrA was purchased from Geneart.

Techniques:

Journal: Human Vaccines & Immunotherapeutics

Article Title: Intranasal delivery of Salmonella OMVs decorated with Chlamydia trachomatis antigens induces specific local and systemic immune responses

doi: 10.1080/21645515.2024.2330768

Figure Lengend Snippet: Humoral immune responses to C. trachomatis antigens. ELISA was used to assess antibody responses. (a-b) Post-vaccination kinetics of total serum and vaginal IgG against CTH522. (c-d) Post-vaccination total serum and vaginal IgG against HtrA. (e-f) pattern of IgG1 and IgG2a antibody responses specific to CTH522 in serum post-vaccination. (g-h) Post-vaccination IgG1 and IgG2a antibody responses specific to HtrA in serum. The error bars represent the standard error of mean (SEM). GraphPad Prism version 8.0 was used to analyze the statistically significant difference between the OMV-CTH522/OMV-HtrA and OMV-Ctrl vaccinated groups using ANOVA and Sidak’s multiple comparisons test. * p < .05, ** p < .01, *** p < .001, **** p < .0001 and ns (not significant).

Article Snippet: Initially, an E. coli optimized synthetic DNA fragment encoding C. trachomatis SvD HtrA was purchased from Geneart.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Human Vaccines & Immunotherapeutics

Article Title: Intranasal delivery of Salmonella OMVs decorated with Chlamydia trachomatis antigens induces specific local and systemic immune responses

doi: 10.1080/21645515.2024.2330768

Figure Lengend Snippet: Antigen-specific cytokine released in the culture supernatants of cLN and splenic cells. Splenic and cLN cell suspensions (1 × 10 6 ) were restimulated in vitro for 72 h with purified CTH522, HtrA, or OVA and released cytokines in cell culture supernatants were measured using Luminex assay. Levels of IFN-γ and IL-17a cytokines secreting from cLN cells (a-b) and splenocytes (e-f) after CTH522 and OVA stimulation. Levels of IFN-γ and IL-17a cytokines secreting from cLN cells (e-f) and splenocytes (g-h) after restimulation with HtrA and OVA. The standard error of mean (SEM) was used to express the results. ANOVA and Sidak’s multiple comparisons tests were used in GraphPad prism version 8.0 to determine statistically significant differences between the OMV-CTH522/OMV-HtrA and OMV-Ctrl vaccinated groups. * p < .05, ** p < .01, *** p < .001, **** p < .0001 and ns (not significant).

Article Snippet: Initially, an E. coli optimized synthetic DNA fragment encoding C. trachomatis SvD HtrA was purchased from Geneart.

Techniques: In Vitro, Purification, Cell Culture, Luminex

Journal: Cytometry

Article Title: A flow cytometry‐based assay to determine the phagocytic activity of both clinical and nonclinical antibody samples against Chlamydia trachomatis

doi: 10.1002/cyto.a.23353

Figure Lengend Snippet: Gating strategies for CFSE‐labeled C. trachomatis bacteria and phagocytosis assay. All gatings were first set on singlet events (FSC‐A vs FSC‐H), followed by gating on target cell population (FSC‐A vs. SSC‐A). (A) Bacteria were then shown in APC‐A versus FITC‐A (stained either with mouse anti‐ C. trachomatis LPS monoclonal Ab and a goat anti‐mouse‐IgG‐Alexaflour™647 secondary Ab or labeled with CFSE). (B) PLB‐985 cells were stained with fixable viability dye eFluor ® 780 (FvD) and gated on living cells with FSC‐A versus APC‐Cy7‐A. (C) CFSE‐labeled SvD bacteria were preincubated with no serum, serum from naïve rabbits, or serum from rabbits vaccinated with Hirep1 for 40 min at 37°C and incubated for 4 h with DMF‐stimulated PLB‐985 cells. CFSE‐signal was measured by flow cytometry in the FITC channel. Cells were gated on CFSE‐positive (=phagocytosing) events. Pseudo‐color dot plots show PLB‐985 cells alone or after incubation with noncoated or coated bacteria.

Article Snippet: C. trachomatis SvD (UW‐3/Cx; ATCC VR‐885) were propagated in Hela 229 cells and purified as described elsewhere .

Techniques: Labeling, Bacteria, Phagocytosis Assay, Staining, Incubation, Flow Cytometry

Journal: Cytometry

Article Title: A flow cytometry‐based assay to determine the phagocytic activity of both clinical and nonclinical antibody samples against Chlamydia trachomatis

doi: 10.1002/cyto.a.23353

Figure Lengend Snippet: Phagocytosis assay using human and murine serum. CFSE‐labeled SvD bacteria at a MOI of 10 were incubated with (A) serum from infected/vaccinated mice or a monoclonal mouse‐a‐ C. trachomatis LPS Ab, or (B) serum from an infected human donor. Serum of naïve mice/humans were used as negative controls. Murine sera were titrated from 1:10 to 1:10000 ( C. trachomatis LPS mAb was pre‐diluted to 1 ng/μl) and human sera from 1:100 to 1:10000. Phagocytosis was measured by flow cytometry. (C) Representative pseudo‐color dot plots of the phagocytosis assay with all sera at a dilution of 1:10 (for human sera 1:100). Mean and SD are shown at a sample size of n = 3 (A and B).

Article Snippet: C. trachomatis SvD (UW‐3/Cx; ATCC VR‐885) were propagated in Hela 229 cells and purified as described elsewhere .

Techniques: Phagocytosis Assay, Labeling, Bacteria, Incubation, Infection, Flow Cytometry