Journal: Frontiers in Immunology

Article Title: Allergic T H 2 Response Governed by B-Cell Lymphoma 6 Function in Naturally Occurring Memory Phenotype CD4 + T Cells

doi: 10.3389/fimmu.2018.00750

Figure Lengend Snippet: Role of Bcl6-mediated MPT cell functions in NAT H 2 differentiation in an allergic murine model. [ (A) top] Mixture of purified KJ1-26 − MPT cells ( Bcl6 -WT or Bcl6 -KO) and KJ1-26 + WT naïve CD4 + T cells were transferred into BALB/c nu/nu mice intravenously (day 0). These mice were immunized with alum-conjugated OVA and then intratracheally challenged with OVA. [ (A) bottom] Absolute cell numbers of Neu, Eos, AM, and Lym in BALF, (B) hematoxylin and eosin-stained, formalin-fixed lung sections (magnification: 200×), and (C) T H 2 cytokine levels in the BALF of recipient mice 48 h after the last OVA challenge. (D) Relative Il4, Il5 , and Il13 expression mRNA in splenic KJ1-26 + T cells restimulated with anti-CD3 monoclonal antibodies 5 days after the last challenge. (E) OVA-specific IgE antibody titers in sera from each recipient of Bcl6 -WT NAT H 2 cells, plus MPT H 2 cells transferred from Bcl6 -TG, Bcl6 -WT, or Bcl6 -KO mice 2 days after the last challenge. All results are representative of four independent experiments with similar outcomes. Data are presented as the mean ± SEM ( n = 5–7). * P < 0.05, ** P < 0.01, comparison between two groups is indicated. AM, alveolar macrophages; BALF, bronchoalveolar lavage fluid; Bcl6, B-cell lymphoma 6; Eos, eosinophils; KJ + , KJ1-26-positive; KO, knockout; Lym, lymphocytes; MPT cell, memory phenotype CD4 + T cell; MPT H 2 cell, MPT cell-derived T H 2 cell; NAT H 2 cell; naïve CD4 + T cell-derived T H 2 cell; Neu, neutrophils; NS, not significant; OVA, ovalbumin; TG, transgenic; WT, wild-type.

Article Snippet: IgE anti-OVA Abs were detected using a mouse anti-OVA IgE Antibody Assay Kit (Chondrex, Redmond, WA, USA).

Techniques: Purification, Staining, Expressing, Knock-Out, Derivative Assay, Transgenic Assay

Journal: MethodsX

Article Title: Development and evaluation of mouse anti-Ara h 1 and Ara h 3 IgE monoclonal antibodies for advancing peanut allergy research

doi: 10.1016/j.mex.2023.102470

Figure Lengend Snippet: Dose dependency of each IgE monoclonal antibody against CPE in a sandwich ELISA.

Article Snippet: All plates were screened for anti-CPE IgE antibody-producing hybridomas by ELISA kits (Cat# 3063, Chondrex, Inc. WA, USA), according to the manufacturer's instructions.

Techniques: Sandwich ELISA

Journal: MethodsX

Article Title: Development and evaluation of mouse anti-Ara h 1 and Ara h 3 IgE monoclonal antibodies for advancing peanut allergy research

doi: 10.1016/j.mex.2023.102470

Figure Lengend Snippet: Dose dependency of each IgE monoclonal antibody against CPE in an indirect ELISA.

Article Snippet: All plates were screened for anti-CPE IgE antibody-producing hybridomas by ELISA kits (Cat# 3063, Chondrex, Inc. WA, USA), according to the manufacturer's instructions.

Techniques: Indirect ELISA

Journal: MethodsX

Article Title: Development and evaluation of mouse anti-Ara h 1 and Ara h 3 IgE monoclonal antibodies for advancing peanut allergy research

doi: 10.1016/j.mex.2023.102470

Figure Lengend Snippet: Summary of the Application of Anti-CPE IgE Monoclonal Antibodies.

Article Snippet: All plates were screened for anti-CPE IgE antibody-producing hybridomas by ELISA kits (Cat# 3063, Chondrex, Inc. WA, USA), according to the manufacturer's instructions.

Techniques: Indirect ELISA, Sandwich ELISA

Journal: ACS Central Science

Article Title: Injectable Hydrogels for Sustained Codelivery of Subunit Vaccines Enhance Humoral Immunity

doi: 10.1021/acscentsci.0c00732

Figure Lengend Snippet: Antibody concentration and affinity following immunization. (a) Timeline of the experimental setup shows subcutaneous (SC) injection of a model vaccine containing OVA and Poly(I:C) in a gel or bolus formulation at day 0, antibody analysis over time following a single administration, boost with a bolus vaccine formulation at day 90, and analysis of the immune response 15 days after the boost. (b) Serum anti-OVA IgG1 concentrations from day 0 to day 90 after a single injection of vaccines ( n = 5 to 19; one to four independent experiments; mean ± s.e.m.). *** p < 0.001 and **** p < 0.0001 compared to bolus, # p < 0.05, ## p < 0.005 compared to 1:5, determined by mixed-effects analysis with Tukey’s post hoc test. Serum anti-OVA (c) IgG end point titer, (d) IgG1 concentration, (e) IgG2b concentration, and (f) IgG2c concentration 15 days after bolus boost on day 90 for animals receiving either bolus, 1:5 gel, or 2:10 gel vaccines ( n = 4 to 5; mean ± s.d.). Reported p values determined by one-way ANOVA with Tukey’s post hoc test. (g) Model comparing competitive binding data with K D ranging from 1 to 10 4 nM. (h) Representative competitive binding curves for bolus, 1:5 gel, and 2:10 gel vaccine groups after the day 90 boost compared to an mAb reference competing with the same mAb. (i) Calculated K D values from fitted binding curves for bolus, 1:5 gel, and 2:10 gel vaccine groups ( n = 4; mean ± s.d.). p values determined by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: Anti-OVA IgG2b and IgG2c antibody concentrations were measured using mouse anti-OVA antibody assay kits for IgG2b (Chondrex, 3016) and IgG2c (Chondrex, 3029).

Techniques: Concentration Assay, Injection, Formulation, Vaccines, Binding Assay

Journal: ACS Central Science

Article Title: Injectable Hydrogels for Sustained Codelivery of Subunit Vaccines Enhance Humoral Immunity

doi: 10.1021/acscentsci.0c00732

Figure Lengend Snippet: Germinal center response to single vaccine administration. (a) Immunohistochemistry (IHC) of explanted inguinal lymph node 15 days after OVA + Poly(I:C) vaccine administration in 2:10 and bolus groups to visualize germinal centers (GC) (red) and naïve B cells (green). (b,c) The frequency of germinal center B cells (GCBCs) within total B cells at day 15 (b) and day 30 (c) after prime and (d,e) frequency of IgG1 + GCBCs within total GCBCs at day 15 (d) and day 30 (e) after prime in the inguinal lymph nodes were measured by flow cytometry ( n = 5 to 10). (f) Schematic of the GC response. (g) The percent of T follicular helper cells (Tfh) out of the CD4 + cell population and (h) and the ratio of lights zone (LZ) to dark zone (DZ) GCBCs in the inguinal lymph nodes at day 15 after vaccinatation ( n = 5 to 10). For (b,d,h), data come from two independent experiments, and all other graphs represent one independent experiment. All error bars are mean ± s.d.; p values are determined by one-way ANOVA with Tukey’s post hoc test.

Article Snippet: Anti-OVA IgG2b and IgG2c antibody concentrations were measured using mouse anti-OVA antibody assay kits for IgG2b (Chondrex, 3016) and IgG2c (Chondrex, 3029).

Techniques: Immunohistochemistry, Flow Cytometry