Journal: Infection and Immunity

Article Title: Eosinophils Affect Antibody Isotype Switching and May Partially Contribute to Early Vaccine-Induced Immunity against Coxiella burnetii

doi: 10.1128/IAI.00376-19

Figure Lengend Snippet: SSChigh granulocytes express CD11b and the interleukin-5 receptor. BALB/c mice were vaccinated s.c. with 10 μg of PIV or PIIV, and spleens were processed for flow cytometry as previously described. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. Naïve mice were used to establish baseline values. (A) Gating strategy. FSC and SSC were used to remove cell debris from the analysis. CD45+ leukocytes were then identified and further subgated based on SSC and expression of CD11b. CD11b+ SSChigh granulocytes were then further subgated based on expression of the interleukin-5 receptor alpha subunit (IL-5Rα, CD125). (B and C) Population frequencies and absolute cell counts were determined at 7 and 14 dpv. Each experimental group includes 4 to 6 mice, with error bars representing the standard deviation from the mean. *, P < 0.05; **, P < 0.01, as determined by one-way ANOVA with Tukey’s multiple-comparison test.

Article Snippet: Recombinant anti-mouse CD125 (IL-5Ra, clone REA343; conjugated to phycoerythrin [PE]), Siglec-F (clone REA798; PE-Vio770), and CD45R (B220, clone REA755; fluorescein isothiocyanate [FITC]) as well as rat anti-mouse CD19 (clone 6D5; FITC) were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany).

Techniques: Flow Cytometry, Adjuvant, Expressing, Standard Deviation, Comparison

Journal: Infection and Immunity

Article Title: Eosinophils Affect Antibody Isotype Switching and May Partially Contribute to Early Vaccine-Induced Immunity against Coxiella burnetii

doi: 10.1128/IAI.00376-19

Figure Lengend Snippet: SSChigh CD11b+ CD125+ cells are Siglec-F+ eosinophils. BALB/c mice were vaccinated s.c. with 10 μg of PIV or PIIV, and spleens were processed for flow cytometry. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. Naïve mice were used to establish baseline values. (A) Gating strategy. SSChigh CD11b+ CD125+ cells were examined for expression of Siglec-F (eosinophils) and CD19/B220 (B cells). (B and C) Population frequencies and absolute cell counts were determined at 7 and 14 dpv. Each experimental group includes 4 to 6 mice, with error bars representing the standard deviations from the means. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, as determined by one-way ANOVA with Tukey’s multiple-comparison test.

Article Snippet: Recombinant anti-mouse CD125 (IL-5Ra, clone REA343; conjugated to phycoerythrin [PE]), Siglec-F (clone REA798; PE-Vio770), and CD45R (B220, clone REA755; fluorescein isothiocyanate [FITC]) as well as rat anti-mouse CD19 (clone 6D5; FITC) were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany).

Techniques: Flow Cytometry, Adjuvant, Expressing, Comparison

Journal: Infection and Immunity

Article Title: Eosinophils Affect Antibody Isotype Switching and May Partially Contribute to Early Vaccine-Induced Immunity against Coxiella burnetii

doi: 10.1128/IAI.00376-19

Figure Lengend Snippet: Eosinophil accumulation in PIV-vaccinated mice is CD4+ T cell dependent. WT C57BL/6 and CD4-deficient mice were vaccinated s.c. with 10 μg of PIV, and 7 days later spleens were harvested for flow cytometry or ex vivo restimulation studies. Mice receiving Alhydrogel adjuvant alone served as unvaccinated controls. Naïve mice were used to establish baseline values. (A) SSChigh CD11b+ CD125+ Siglec-F+ eosinophils were enumerated using the gating strategy described in the legends for Fig. 4 and ​and5.5. (B) Total splenocytes were isolated and restimulated with medium or with 1 × 107 C. burnetii NMI bacteria for 3 and 6 days, and cell culture supernatants were evaluated for IFN-γ production by cytokine-specific ELISA. Each experimental group includes 5 mice, with error bars representing the standard deviations from the means. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, as determined by one-way ANOVA with Tukey’s multiple-comparison test. Cb, C. burnetii; KO, knockout.

Article Snippet: Recombinant anti-mouse CD125 (IL-5Ra, clone REA343; conjugated to phycoerythrin [PE]), Siglec-F (clone REA798; PE-Vio770), and CD45R (B220, clone REA755; fluorescein isothiocyanate [FITC]) as well as rat anti-mouse CD19 (clone 6D5; FITC) were purchased from Miltenyi Biotec (Bergisch Gladbach, Germany).

Techniques: Flow Cytometry, Ex Vivo, Adjuvant, Isolation, Bacteria, Cell Culture, Enzyme-linked Immunosorbent Assay, Comparison, Knock-Out

Journal: Frontiers in Immunology

Article Title: Local Innate Markers and Vaginal Microbiota Composition Are Influenced by Hormonal Cycle Phases

doi: 10.3389/fimmu.2022.841723

Figure Lengend Snippet: Antibodies used to characterize the neutrophil subpopulations.

Article Snippet: CDw125 , REA705 , PE , 2.5 μL , 130-110-544 , Milteny.

Techniques: Clone Assay, Concentration Assay