Journal: Journal of Human Immunity

Article Title: Oncostatin M receptor deficiency as a novel candidate genetic cause of autosomal recessive hyper-IgE syndrome

doi: 10.70962/jhi.20250119

Figure Lengend Snippet: B cell−extrinsic elevation of circulating plasmablasts and increase in IgE isotype switching in the patient. (A) Flow plots of main B cell subsets (top row) and antigen-experienced B cell subsets (bottom row) in one healthy control (left panels) and the patient (right panels). Representative of two independent analyses. (Data shown in rows 1 and 2, column 1 are identical to data in , row 1, column 4 and 5.) (B) Gate frequencies for antigen-experienced B cell subsets of five healthy controls (HCs) and the patient. Representative of two independent analyses. (C) Fraction of IgE class-switched cells among antigen-experienced B cell subsets of three healthy controls and the patient. The individual points for healthy control and patient represent one each of three sampling methods (conventional Ficoll prep, separator tubes, and whole-blood RBC lysis) from one experiment. (D) Schematic overview of iGB culture setup (created with BioRender). (E) Cell counts for one representative of two similar experiments. (F) IgM, IgG1, IgG3, and IgE secretion in cultures, normalized to cell count. Full gating strategies are shown in . Statistics were calculated using the unpaired t test. * = P < 0.05. aMBC_PC, aMBC/plasma cell; DN, double-negative; MBC_only, conventional memory B cell; SWmem, switched memory; T1, T2, T3, transitional 1, 2, 3 subsets.

Article Snippet: Wells of FluoroNunc MaxiSorp 96-well plates were coated with 100 μl capture antibody diluted in PBS, either 0.5 μg/ml goat α-human IgM-Hc (2023-01; Southern Biotech), 5 μg/ml mouse α-human IgG1-Fc (9054-01; Southern Biotech), 1 μg/ml mouse α-human IgG3-hinge (9210-01; Southern Biotech), or 2.5 μg/ml mouse α-human IgE-Fc (9240-01; Southern Biotech).

Techniques: Control, Sampling, Red Blood Cell Lysis, Cell Characterization, Clinical Proteomics

Journal: Vaccines

Article Title: Fine-Mapping and Protective Analysis of Immunodominant Linear B-Cell Epitopes of FimA Antigen of Klebsiella Pneumoniae

doi: 10.3390/vaccines14040347

Figure Lengend Snippet: Analysis of specific antibody and subtype levels induced by recombinant FimA protein immunization. Serum samples were collected from immunized BALB/c mice seven days after the final immunization. An enzyme-linked immunosorbent assay (ELISA) was performed to measure the potency of FimA-specific immunoglobulin G (IgG) and its subtypes. ( A ) Reciprocal of serum dilution in all immunized mice. ( B ) Antibody titer of IgG in all immunized mice. ( C ) The OD450 of each subtype of antibodies in the serum of mice immunized with FimA plus Al(OH) 3 . ( D ) The titers of each subtype of antibodies in the serum of mice immunized with FimA plus Al(OH) 3 (*, p < 0.05; ***, p < 0.001; ****, p < 0.001; ns, not significant).

Article Snippet: Add horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG1, IgG2a, IgG2b, IgG3, IgM, and IgA antibodies (Alpha Diagnostic) at a dilution of 1:10,000 to the wells and incubate at 37 °C for 1 h. Wash the wells three times with PBST to remove unbound secondary antibodies.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Vaccines

Article Title: Fine-Mapping and Protective Analysis of Immunodominant Linear B-Cell Epitopes of FimA Antigen of Klebsiella Pneumoniae

doi: 10.3390/vaccines14040347

Figure Lengend Snippet: Analysis of immunodominant peptides of FimA antigen in clinical human convalescent sera after K. pneumoniae infection. In this experiment, serum samples from patients infected with K. pneumoniae were collected. ( A ) Reciprocal of FimA-specific antisera dilution of the human convalescent sera from K. pneumoniae infection. OVA 192–201 peptide-specific antisera were used as a negative control. ( B ) The mean titer of FimA-specific IgG antibodies in human sera was 1:14,720, which was significantly different from that of the OVA 192–201 peptide group (****, p < 0.0001). ( C ) The levels of each subtype of antibodies in the antisera of mice immunized with the single peptide-KLH or mixed peptides-KLH. ( D ) Fine mapping of B-cell immunodominant epitopes of FimA non-human convalescent sera after K. pneumoniae infection. The four novel immunodominant epitopes, including FimA 97–114 (*, p = 0.0257), FimA 103–120 (**, p = 0.0092), FimA 109–126 (**, p = 0.0084), and FimA 145–160 (****, p < 0.0001), identified in the FimA immunized mice were also found to be immunodominant in the human convalescent sera. ( E ) All overlapping peptides were used to map the linear B-cell epitopes recognized by human serum samples. In the antibody of human convalescent sera, there were also three other positive immunodominant epitopes, namely FimA 1–18 (**, p = 0.0087), FimA 49–60 (***, p = 0.0009), and FimA 55–72 (***, p = 0.0009).

Article Snippet: Add horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG1, IgG2a, IgG2b, IgG3, IgM, and IgA antibodies (Alpha Diagnostic) at a dilution of 1:10,000 to the wells and incubate at 37 °C for 1 h. Wash the wells three times with PBST to remove unbound secondary antibodies.

Techniques: Infection, Negative Control