Journal:

Article Title: Recombinant Urease and Urease DNA of Coccidioides immitis Elicit an Immunoprotective Response against Coccidioidomycosis in Mice

doi: 10.1128/IAI.69.5.2878-2887.2001

Figure Lengend Snippet: (A) Expression of Th1 and Th2 cytokines and HPRT mRNA in lung tissue of infected mice which either had not been immunized (lane 1) or had been immunized as described in Materials and Methods with one of the selected antigens plus CpG ODN adjuvant (i.e., rHSP60 [lanes 2 and 3], rURE [lanes 4 and 5], BSA [lane 6 and 7]). The RT-PCR and electrophoresis conditions were the same as those used for analysis of cytokine expression in Fig. ​Fig.2.2. HPRT was used as an internal control. The results are representative of three separate experiments. (B) Total Ig and IgG subclass responses of immunized and infected BALB/c mice to the indicated antigens. Mice were sacrificed and terminally bled as described in Materials and Methods; plasma samples were tested for antigen-specific total Ig, IgG2a, and IgG1 by ELISA as indicated. The results are expressed as reciprocal titers (1/dilution) and presented as means plus standard deviations for three test sera in each group of immunized, infected mice.

Article Snippet: Alkaline phosphatase conjugated with goat anti-mouse Ig (IgM, IgG, and IgA; heavy and light chains), goat anti-mouse IgG2a, or goat anti-mouse IgG1 (Southern Biotechnology Associates, Inc., Birmingham, Ala.) was added to the wells at a dilution of 1:1,000 in PBS to detect bound, primary antibody.

Techniques: Expressing, Infection, Reverse Transcription Polymerase Chain Reaction, Electrophoresis, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Diversity of immune responses in children highly exposed to SARS-CoV-2

doi: 10.3389/fimmu.2023.1105237

Figure Lengend Snippet: Flowchart of the immunoglobulin profile in children to evaluate their immunological status. Four groups were finally defined: IgG − IgM − , IgG − IgM low , IgG − IgM high , and IgG + . Sera were analyzed as described in the Material and methods . Results shown are the mean values of one duplicate experiment of the three independent experiments performed with similar qualitative results.

Article Snippet: Bound antibodies were detected by incubation with mouse anti‐human IgG1 or IgM secondary antibody coupled to horseradish peroxidase (HRP; Southern Biotech) diluted 1/6,000 in 1% BSA in PBS, which was then detected using an ABTS substrate solution (Invitrogen).

Techniques:

Journal: Frontiers in Immunology

Article Title: Diversity of immune responses in children highly exposed to SARS-CoV-2

doi: 10.3389/fimmu.2023.1105237

Figure Lengend Snippet: Levels of soluble ACE2 (A) and ANG Il (B) in serum of children according to their immunological status. Serum levels were analysed by ELISA as described in methods in the indicated four groups: IgG - IgM, IgG - IgM low , IgG - IgM high , IgG + . Results shown are the mean values of one duplicate experiment of the 2 independent performed with similar qualitative results. *p-adjusted<0,05.

Article Snippet: Bound antibodies were detected by incubation with mouse anti‐human IgG1 or IgM secondary antibody coupled to horseradish peroxidase (HRP; Southern Biotech) diluted 1/6,000 in 1% BSA in PBS, which was then detected using an ABTS substrate solution (Invitrogen).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Diversity of immune responses in children highly exposed to SARS-CoV-2

doi: 10.3389/fimmu.2023.1105237

Figure Lengend Snippet: Levels of cytokines in serum of children according to their immunological status. Serum levels were mesured by Multiplex Cytokine Assay as described in methods in the indicated four groups: IgG - IgM - , IgG - IgM low , IgG - IgM high , IgG + . The statistical differences between the IgG - IgM low , IgG - IgM high and IgG. groups are indicated. *p-adjusted<0,05; **p-adjusted<0,01; ***p-adjusted<0,005.

Article Snippet: Bound antibodies were detected by incubation with mouse anti‐human IgG1 or IgM secondary antibody coupled to horseradish peroxidase (HRP; Southern Biotech) diluted 1/6,000 in 1% BSA in PBS, which was then detected using an ABTS substrate solution (Invitrogen).

Techniques: Multiplex Assay, Cytokine Assay

Journal: Frontiers in Immunology

Article Title: Diversity of immune responses in children highly exposed to SARS-CoV-2

doi: 10.3389/fimmu.2023.1105237

Figure Lengend Snippet: Neutralization. (A) The neutralization assay was carried out with the pseudotyped virus as described in the Material and methods . (B) Group IgG − IgM low showed a significant correlation between the neutralizing activity and the levels of IgA anti-S in the serum.

Article Snippet: Bound antibodies were detected by incubation with mouse anti‐human IgG1 or IgM secondary antibody coupled to horseradish peroxidase (HRP; Southern Biotech) diluted 1/6,000 in 1% BSA in PBS, which was then detected using an ABTS substrate solution (Invitrogen).

Techniques: Neutralization, Virus, Activity Assay