mouse anti human igg3 hinge ap hp6050  (SouthernBiotech)

Journal: The Journal of Infectious Diseases

Article Title: Serum IgG1 and IgG3 Antibodies to Chlamydia trachomatis Pgp3 and Hsp60 in Men of Subfertile Couples

doi: 10.1093/infdis/jiaf535

Figure Lengend Snippet: Overlap between Chlamydia trachomatis Pgp3 immunoglobulin G1 (IgG1), Pgp3 immunoglobulin G3 (IgG3), Hsp60 IgG1, and Hsp60 IgG3 seropositivity.

Article Snippet: To test for IgG3 antibodies, Mouse Anti-Human IgG3 Hinge-AP (HP6050) (Southern Biotech) was used.

Techniques:

Journal: The Journal of Infectious Diseases

Article Title: Serum IgG1 and IgG3 Antibody Responses to Chlamydia trachomatis Pgp3 and Hsp60 in Tubal Factor Infertility

doi: 10.1093/infdis/jiaf092

Figure Lengend Snippet: The absorbance values of Chlamydia trachomatis Pgp3 IgG1 ( A ) and IgG3 ( B ) antibody by the severity of tubal factor infertility (TFI; 0 = no occlusion, 1 = unilateral occlusion, 2 = bilateral occlusion).

Article Snippet: To test for IgG3 antibodies, mouse anti-human IgG3 hinge-AP (HP6050; Southern Biotech) was used.

Techniques:

Journal: The Journal of Infectious Diseases

Article Title: Serum IgG1 and IgG3 Antibody Responses to Chlamydia trachomatis Pgp3 and Hsp60 in Tubal Factor Infertility

doi: 10.1093/infdis/jiaf092

Figure Lengend Snippet: The absorbance values of Chlamydia trachomatis and Hsp60 IgG1 ( A ) and IgG3 ( B ) antibody according to the severity of tubal factor infertility (TFI; 0 = no occlusion, 1 = unilateral occlusion, 2 = bilateral occlusion.

Article Snippet: To test for IgG3 antibodies, mouse anti-human IgG3 hinge-AP (HP6050; Southern Biotech) was used.

Techniques:

Journal: Frontiers in immunology

Article Title: Analyses of human vaccine-specific circulating and bone marrow-resident B cell populations reveal benefit of delayed vaccine booster dosing with blood-stage malaria antigens.

doi: 10.3389/fimmu.2023.1193079

Figure Lengend Snippet: FIGURE 1 Vaccine-specific plasma cell and memory IgG+ B cell responses. PBMC from pre-vaccination (Day 0) and post-final vaccination (FV) time points were analysed for B cell responses by flow cytometry; gating strategies are as described in Methods and Supplementary Figures 1 and 3. Frequencies of DBPRII-specific B cells – identified by probe staining – were compared between vaccine platforms (A, B) or protein/adjuvant dosing regimens (C, D) within both plasma cell (A, C) or memory IgG+ B cell (B, D) populations. Similarly, frequencies of RH5-specific B cells were compared between protein/adjuvant dosing regimens within plasma cells (E) and memory IgG+ B cells (F). IgM+, IgA+, activated and resting memory B cell responses are shown in Supplementary Figures 2 and 4. VV = ChAd63-MVA viral vectors [monthly and delayed dosing]; PA = PvDBPII protein/adjuvant [PA-M and PA-D]; PA-M = PvDBPII protein/adjuvant monthly dosing; PA-D = PvDBPII protein/adjuvant delayed booster dosing; PA-DB = PvDBPII protein/ adjuvant delayed booster dosing with extra booster; M = RH5.1/adjuvant monthly dosing; D = RH5.1/adjuvant delayed booster dosing. Post- vaccination comparisons were performed between DBPRII platforms (A, B) or RH5 dosing regimens (E, F) with Mann-Whitney U tests or between PvDBPII protein/adjuvant dosing regimens by Kruskal Wallis test with Dunn’s correction for multiple comparisons (C, D). Sample sizes for all assays were based on sample availability; each circle represents a single sample. (A, B) VV/PA: Day 0 = 8/5-6, FV+7 = 8/12, FV+14 = 8/12, FV+28 = 8/10. (C, D) PA-M/PA-D/PA-DB: Day 0 = 3-4/2/na, FV+7 = 4/8/5, FV+14 = 4/8/4, FV+28 = 4/6/5. (E, F) M/D: Day 0 = 5/1-4, FV+7 = 4-5/3-4, FV+14 = 4/6, FV+28 = 4/4. PA-D vaccinees returning in the PA-DB group are connected by lines. Bars represent medians. * p < 0.05, ** p < 0.01.

Article Snippet: The antibody used was dependent on the isotype or subclass being assayed and were as follows: IgG-AP (A3187, Thermo Scientific), IgG1 Fc-AP (9054-04, Southern Biotech), IgG3 Hinge-AP (9210-04, Southern Biotech), IgG4 Fc-AP (9200-04, Southern Biotech), IgA-AP (2050-04, Southern Biotech), IgA1-AP (9130-04, Southern Biotech), and IgM-AP (2020-04, Southern Biotech).

Techniques: Clinical Proteomics, Cytometry, Staining, Adjuvant, MANN-WHITNEY

Journal: Frontiers in immunology

Article Title: Analyses of human vaccine-specific circulating and bone marrow-resident B cell populations reveal benefit of delayed vaccine booster dosing with blood-stage malaria antigens.

doi: 10.3389/fimmu.2023.1193079

Figure Lengend Snippet: FIGURE 2 Vaccine-specific responses within agnostically-defined B cell populations using CITRUS. CITRUS was run on single live (B cell-enriched) lymphocyte flow cytometry fcs files to agnostically define the main B cell populations within either DBPRII (A–C) or RH5 (D, E) trial samples. Clusters identified by CITRUS are visualised in dendrograms (A, D), colour-coded for example markers of interest [(A)- IgG, CD38; (D)- IgG]. Each node represents a cluster. Median marker expression within each cluster was used to define gating strategies for B cell populations in FlowJo, which were re-analysed for DBPRII- (B, C) or RH5-specific (E) responses through probe staining (gating shown in Supplementary Figure 5). See Table 2 and Supplementary Figures 6, 7 for a full list of populations identified via CITRUS clusters for further analysis. VV-M = ChAd63-MVA viral vector monthly dosing; VV-D ChAd63-MVA delayed booster dosing; PA-M = PvDBPII protein/adjuvant monthly dosing; PA-D = PvDBPII protein/adjuvant delayed booster dosing; PA-DB = PvDBPII protein/adjuvant delayed booster dosing with extra booster; M = RH5.1/adjuvant monthly dosing; D = RH5.1/adjuvant delayed booster dosing. FV = final vaccination. Post-vaccination comparisons were performed between PvDBPII protein/adjuvant dosing regimens by Kruskal Wallis test with Dunn’s correction for multiple comparisons (B, C) or RH5 dosing regimens (E) with Mann-Whitney U tests. Sample sizes for all assays were based on sample availability; each circle represents a single sample. (B, C) VV-M/VV-D/PA-M/PA-D/PA-DB: Day 0 = 6/2/4/2/na, FV+7 = 6/2/4/ 8/5, FV+14 = 6/2/4/8/4, FV+28 = 6/2/4/6/5. (E) M/D: Day 0 = 5/2, FV+7 = 5/4, FV+14 = 4/6, FV+28 = 4/4. PA-D vaccinees returning in the PA-DB group are connected by lines. Bars represent medians. * p < 0.05, ** p < 0.01.

Article Snippet: The antibody used was dependent on the isotype or subclass being assayed and were as follows: IgG-AP (A3187, Thermo Scientific), IgG1 Fc-AP (9054-04, Southern Biotech), IgG3 Hinge-AP (9210-04, Southern Biotech), IgG4 Fc-AP (9200-04, Southern Biotech), IgA-AP (2050-04, Southern Biotech), IgA1-AP (9130-04, Southern Biotech), and IgM-AP (2020-04, Southern Biotech).

Techniques: Cytometry, Marker, Expressing, Staining, Plasmid Preparation, Adjuvant, MANN-WHITNEY

Journal: Frontiers in immunology

Article Title: Analyses of human vaccine-specific circulating and bone marrow-resident B cell populations reveal benefit of delayed vaccine booster dosing with blood-stage malaria antigens.

doi: 10.3389/fimmu.2023.1193079

Figure Lengend Snippet: FIGURE 3 DBPRII-specific peak antibody responses and serum maintenance. Standardised ELISAs were developed to report anti-DBPRII specific antibody responses against the Sal I strain in pre-vaccination (Day 0) and post-final vaccination (FV) serum samples. Responses were compared between protein/adjuvant dosing regimens for IgG1 (A), IgG3 (B), IgG4 (C), IgA (D), IgA1 (E), and IgM (F). Fold change between C+96 and FV+14 was calculated for total IgG (G) and specific isotypes/subclasses (H) to compare monthly (M: VV-M, PA-M) and delayed (D: VV-D, PA-D) booster regimens. IgG4 and IgA1 were excluded from this analysis as ≥1 vaccinee had undetectable antibodies at both time points. Comparisons between vaccine platforms are shown in Supplementary Figure 8. VV-M = ChAd63-MVA viral vector monthly dosing; VV-D ChAd63-MVA delayed booster dosing; PA-M = protein/adjuvant monthly dosing; PA-D = protein/adjuvant delayed booster dosing; PA-DB = protein/adjuvant delayed booster dosing with extra booster. C+96 = 96 days after controlled human malaria infection (approximately 16 weeks after FV). Post-vaccination comparisons were performed between protein/adjuvant dosing regimens by Kruskal Wallis test with Dunn’s correction for multiple comparisons (A– F) or fold changes with Mann-Whitney U tests (G, H). Sample sizes for all assays were based on sample availability; each circle represents a single sample [triangles indicate viral vector samples in (G, H)]. (A–F) VV-M/VV-D/PA-M/PA-D/PA-DB: Day 0 = 6/2/4/8/na, FV+14 = 6/2/4/8/4, FV+28 = 6/ 2/4/6/5. (G, H) M = 9-10, D = 9. Bars represent medians. * p < 0.05, ** p < 0.01.

Article Snippet: The antibody used was dependent on the isotype or subclass being assayed and were as follows: IgG-AP (A3187, Thermo Scientific), IgG1 Fc-AP (9054-04, Southern Biotech), IgG3 Hinge-AP (9210-04, Southern Biotech), IgG4 Fc-AP (9200-04, Southern Biotech), IgA-AP (2050-04, Southern Biotech), IgA1-AP (9130-04, Southern Biotech), and IgM-AP (2020-04, Southern Biotech).

Techniques: Adjuvant, Plasmid Preparation, Infection, MANN-WHITNEY

Journal: Frontiers in immunology

Article Title: Analyses of human vaccine-specific circulating and bone marrow-resident B cell populations reveal benefit of delayed vaccine booster dosing with blood-stage malaria antigens.

doi: 10.3389/fimmu.2023.1193079

Figure Lengend Snippet: FIGURE 5 Correlations between circulating DBPRII-specific B cells and in vivo growth inhibition of P. vivax parasites or maintenance of serum antibody. In vivo growth inhibition (IVGI) of P. vivax parasites following post-vaccination controlled human malaria infection (CHMI) was calculated from qPCR data as described in the Methods. Spearman correlations were performed between IVGI and the peak frequency of DBPRII-specific memory IgG+ B cells at FV+14 (A) or plasma cells at FV+7 (B) as defined in Supplementary Figure 1 and reported in Figure 1. Spearman correlations were also performed between C+96/FV+14 fold change in total anti-DBPRII IgG (Sal I strain; see Figure 3) and memory IgG+ B cells at FV+14 (C) or plasma cell at FV+7 (D). VV-M = ChAd63-MVA viral vector monthly dosing; VV-D ChAd63-MVA delayed booster dosing; PA-M = protein/adjuvant monthly dosing; PA-D = protein/adjuvant delayed booster dosing. C+96 = 96 days after controlled human malaria infection (approximately 16 weeks after FV). Spearman rho, p-values, and sample sizes are annotated on individual graphs. Each circle represents a single sample.

Article Snippet: The antibody used was dependent on the isotype or subclass being assayed and were as follows: IgG-AP (A3187, Thermo Scientific), IgG1 Fc-AP (9054-04, Southern Biotech), IgG3 Hinge-AP (9210-04, Southern Biotech), IgG4 Fc-AP (9200-04, Southern Biotech), IgA-AP (2050-04, Southern Biotech), IgA1-AP (9130-04, Southern Biotech), and IgM-AP (2020-04, Southern Biotech).

Techniques: In Vivo, Inhibition, Infection, Clinical Proteomics, Plasmid Preparation, Adjuvant

Journal: Frontiers in immunology

Article Title: Analyses of human vaccine-specific circulating and bone marrow-resident B cell populations reveal benefit of delayed vaccine booster dosing with blood-stage malaria antigens.

doi: 10.3389/fimmu.2023.1193079

Figure Lengend Snippet: FIGURE 6 RH5-specific bone marrow plasma cell responses and correlations with serum antibody or circulating RH5-specific cells. RH5-specific bone marrow plasma cells were detected in B cells enriched from pre- and post-final vaccination (FV) bone marrow mononuclear cells and assayed by IgG antibody-secreting cell ELISPOT as described in the Methods. The frequency of RH5-specific IgG plasma cells (antibody-secreting cells) per million bone marrow B cells was compared between dosing regimens (A). Spearman correlation analyses were performed between RH5-specific bone marrow B cells and matched time point serum IgG (B), matched time point frequency of RH5-specific cells within CITRUS-guided “Population 12” [CD19+CD20+CD21+CD27+CD138-CD38+IgM-IgA-IgG+; see Table 2, Figure 2, Supplementary Figure 5] (C), and between RH5-specific bone marrow B cells at FV+28 and “Population 12” at FV+14 (D). M = RH5.1/adjuvant monthly dosing; D = RH5.1/adjuvant delayed booster dosing. Post- vaccination comparisons were performed between dosing regimens by Mann Whitney U test [(A); not significant]. Spearman rho, p-values, and sample sizes are annotated on individual graphs. Each circle represents a single sample.

Article Snippet: The antibody used was dependent on the isotype or subclass being assayed and were as follows: IgG-AP (A3187, Thermo Scientific), IgG1 Fc-AP (9054-04, Southern Biotech), IgG3 Hinge-AP (9210-04, Southern Biotech), IgG4 Fc-AP (9200-04, Southern Biotech), IgA-AP (2050-04, Southern Biotech), IgA1-AP (9130-04, Southern Biotech), and IgM-AP (2020-04, Southern Biotech).

Techniques: Clinical Proteomics, Enzyme-linked Immunospot, Adjuvant, MANN-WHITNEY