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igg4 hp6025 antibodies conjugated  (SouthernBiotech)


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    SouthernBiotech igg4 hp6025 antibodies conjugated
    Igg4 Hp6025 Antibodies Conjugated, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 94/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/mouse+anti+human+igg4+fc+ap/Mouse+Anti-Human+IgG4+Fc-AP/pm40796093-75-23-30
    Average 94 stars, based on 97 article reviews
    igg4 hp6025 antibodies conjugated - by Bioz Stars, 2026-10
    94/100 stars

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    Modification:

    Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection.
    Article Snippet: Plates were incubated for 2 h at 37 °C with 300 rpm shaking and following washing with PBS/T samples were further incubated with mouse anti-human IgG1 hinge-AP, mouse anti-human IgG3 hinge-AP, goat anti-human IgA-AP and goat anti-human IgM-AP (Southern Biotech) for 1 h at 37 °C with 300 rpm used for detection and the optical density at 405 nm was measured until the internal control reached an OD405 of 1. .. For detection of antispike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405. ..

    Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection
    Article Snippet: Plates were incubated for 2 h at 37 °C with 300 rpm shaking and following washing with PBS/T samples were further incubated with mouse anti-human IgG1 hinge-AP, mouse anti-human IgG3 hinge-AP, goat anti-human IgA-AP and goat anti-human IgM-AP (Southern Biotech) for 1 h at 37 °C with 300 rpm used for detection and the optical density at 405 nm was measured until the internal control reached an OD 405 of 1. .. For detection of anti-spike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405. ..

    Control:

    Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection.
    Article Snippet: Plates were incubated for 2 h at 37 °C with 300 rpm shaking and following washing with PBS/T samples were further incubated with mouse anti-human IgG1 hinge-AP, mouse anti-human IgG3 hinge-AP, goat anti-human IgA-AP and goat anti-human IgM-AP (Southern Biotech) for 1 h at 37 °C with 300 rpm used for detection and the optical density at 405 nm was measured until the internal control reached an OD405 of 1. .. For detection of antispike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405. ..

    Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection
    Article Snippet: Plates were incubated for 2 h at 37 °C with 300 rpm shaking and following washing with PBS/T samples were further incubated with mouse anti-human IgG1 hinge-AP, mouse anti-human IgG3 hinge-AP, goat anti-human IgA-AP and goat anti-human IgM-AP (Southern Biotech) for 1 h at 37 °C with 300 rpm used for detection and the optical density at 405 nm was measured until the internal control reached an OD 405 of 1. .. For detection of anti-spike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405. ..

    Recombinant:

    Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection.
    Article Snippet: Plates were incubated for 2 h at 37 °C with 300 rpm shaking and following washing with PBS/T samples were further incubated with mouse anti-human IgG1 hinge-AP, mouse anti-human IgG3 hinge-AP, goat anti-human IgA-AP and goat anti-human IgM-AP (Southern Biotech) for 1 h at 37 °C with 300 rpm used for detection and the optical density at 405 nm was measured until the internal control reached an OD405 of 1. .. For detection of antispike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405. ..

    Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection
    Article Snippet: Plates were incubated for 2 h at 37 °C with 300 rpm shaking and following washing with PBS/T samples were further incubated with mouse anti-human IgG1 hinge-AP, mouse anti-human IgG3 hinge-AP, goat anti-human IgA-AP and goat anti-human IgM-AP (Southern Biotech) for 1 h at 37 °C with 300 rpm used for detection and the optical density at 405 nm was measured until the internal control reached an OD 405 of 1. .. For detection of anti-spike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405. ..

    Microplate Reader Absorbance Measurement:

    Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection.
    Article Snippet: Plates were incubated for 2 h at 37 °C with 300 rpm shaking and following washing with PBS/T samples were further incubated with mouse anti-human IgG1 hinge-AP, mouse anti-human IgG3 hinge-AP, goat anti-human IgA-AP and goat anti-human IgM-AP (Southern Biotech) for 1 h at 37 °C with 300 rpm used for detection and the optical density at 405 nm was measured until the internal control reached an OD405 of 1. .. For detection of antispike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405. ..

    Article Title: Divergent trajectories of antiviral memory after SARS-CoV-2 infection
    Article Snippet: Plates were incubated for 2 h at 37 °C with 300 rpm shaking and following washing with PBS/T samples were further incubated with mouse anti-human IgG1 hinge-AP, mouse anti-human IgG3 hinge-AP, goat anti-human IgA-AP and goat anti-human IgM-AP (Southern Biotech) for 1 h at 37 °C with 300 rpm used for detection and the optical density at 405 nm was measured until the internal control reached an OD 405 of 1. .. For detection of anti-spike IgG2 and IgG4 steps modified as follows: (1) Plates were additionally coated with commercially available human immunoglobulin control (recombinant human IgG2 lambda or recombinant human IgG4 lambda (Bio-Rad)) to serve as internal controls, (2) Mouse anti-human IgG2 Fd-AP or mouse anti-human IgG4 Fc-AP (Southern Biotech) were used, and (3) Optical density at 405 nm was measured using an ELx808 absorbance reader (BioTek) until the immunoglobulin control reached a specified OD405. ..

    other:

    Article Title: Development of antibody-dependent cellular cytotoxicity in response to recombinant and live-attenuated herpes zoster vaccines.
    Article Snippet: Supernatants were collected and assayed for the presence of TNFα using a commercial ELISA kit as per manufacturer’s instructions (R&D Systems, Minneapolis, MN).

    Enzyme-linked Immunosorbent Assay:

    Article Title: Phase 1/2 trial of SARS-CoV-2 vaccine ChAdOx1 nCoV-19 with a booster dose induces multifunctional antibody responses.
    Article Snippet: .. This antibody is routinely tested in ELISA (Source: https://www.southernbiotech.com/? catno=9210-04&type=Monoclonal#&panel2-1) • Mouse Anti-Human IgG4 Fc-AP, Southern Biotech, 9200-04, HP6025, B3317-M788G. ..



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    FIGURE 1 Vaccine-specific plasma cell and memory <t>IgG+</t> 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.
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    Image Search Results


    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.

    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

    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.

    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

    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.

    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

    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.

    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

    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.

    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

    (A) Heatmap indicating antibody functions (antibody-dependent complement deposition, ADCD; cellular phagocytosis, ADCP; neutrophil phagocytosis, ADNP; NK activation with production of CD107a, IFNγ and MIP1β), capacity to bind Fcγ receptors (FcγR 2A, 2B, 3A, 3B) and total IgG of different SARS-CoV-2 specific antigens (Spike, S1, S2, RBD and NC) in individuals who did not develop (no PASC) or developed neurological PASC (neuroPASC). Each row corresponds to a single individual. Z-scores, positive (i.e. higher than the mean) in purple, negative (i.e. lower than the mean) in blue. NC= nucleocapsid, RBD=receptor binding domain (B) Bar dot plots show the level of IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA, FcγR binding capacity ( , , , ) and functions (ADCD, ADNP, ADNKA with production of CD107a) of Spike-specific antibodies in individuals who did not develop (no PASC, in blue) or developed neurological PASC (neuroPASC, in purple). MFI=Median Fluorescence Intensity. Mann-Whitney, *p<0.05, **p<0.01, ***p<0.001 ****p<0.0001. (C) Radial plots indicate the levels of IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM, IgA1, IgA2, binding capacity to FcγR ( , , , ), FcαR, and functions (ADCD, ADCP, ADNP, ADNKA with production of CD107a, IFNγ and MIP1β) of Spike-specific antibodies in individuals who did not develop (noPASC) or developed neurological PASC (neuroPASC). Each sector represents a z-scored antibody feature. (D) Heatmap indicating the average difference (%) of Spike-specific functions (ADCD, ADCP, ADNP, ADNKA with production of CD107a, IFNγ and MIP1β), binding capacity to FcγR (2A, 2B, 3A, 3B), FcαR, IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA between individuals with no PASC and with neuroPASC. Each column represents an antibody feature. Positive values indicate features enriched in no PASC, null values indicate features equally present in both no PASC and neuroPASC. None of the features were enriched in neuroPASC.

    Journal: medRxiv

    Article Title: Neurologic sequalae of COVID-19 are determined by immunologic imprinting from previous Coronaviruses

    doi: 10.1101/2022.11.07.22282030

    Figure Lengend Snippet: (A) Heatmap indicating antibody functions (antibody-dependent complement deposition, ADCD; cellular phagocytosis, ADCP; neutrophil phagocytosis, ADNP; NK activation with production of CD107a, IFNγ and MIP1β), capacity to bind Fcγ receptors (FcγR 2A, 2B, 3A, 3B) and total IgG of different SARS-CoV-2 specific antigens (Spike, S1, S2, RBD and NC) in individuals who did not develop (no PASC) or developed neurological PASC (neuroPASC). Each row corresponds to a single individual. Z-scores, positive (i.e. higher than the mean) in purple, negative (i.e. lower than the mean) in blue. NC= nucleocapsid, RBD=receptor binding domain (B) Bar dot plots show the level of IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA, FcγR binding capacity ( , , , ) and functions (ADCD, ADNP, ADNKA with production of CD107a) of Spike-specific antibodies in individuals who did not develop (no PASC, in blue) or developed neurological PASC (neuroPASC, in purple). MFI=Median Fluorescence Intensity. Mann-Whitney, *p<0.05, **p<0.01, ***p<0.001 ****p<0.0001. (C) Radial plots indicate the levels of IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM, IgA1, IgA2, binding capacity to FcγR ( , , , ), FcαR, and functions (ADCD, ADCP, ADNP, ADNKA with production of CD107a, IFNγ and MIP1β) of Spike-specific antibodies in individuals who did not develop (noPASC) or developed neurological PASC (neuroPASC). Each sector represents a z-scored antibody feature. (D) Heatmap indicating the average difference (%) of Spike-specific functions (ADCD, ADCP, ADNP, ADNKA with production of CD107a, IFNγ and MIP1β), binding capacity to FcγR (2A, 2B, 3A, 3B), FcαR, IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA between individuals with no PASC and with neuroPASC. Each column represents an antibody feature. Positive values indicate features enriched in no PASC, null values indicate features equally present in both no PASC and neuroPASC. None of the features were enriched in neuroPASC.

    Article Snippet: After a 2 hour incubation at room temperature, immune complexes were washed, and subclasses and isotypes were detected using phycoerythrin (PE)-conjugated mouse anti-human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM (Southern Biotech) at 1.3 μg/ml.

    Techniques: Activation Assay, Binding Assay, Fluorescence, MANN-WHITNEY

    (A) Bar dot plots show the level of IgG1 antibodies against non-coronaviruses (EBV, Flu, HSV1) in individuals who did not develop (no PASC, in blue) or developed neurological PASC (neuroPASC, in purple). MFI=Median Fluorescence Intensity. Mann-Whitney, ns= statistically not significant. (B) Heatmap indicates the average difference (%) of SARS-CoV-2 -specific (Spike, S1, S2, RBD and NC) and common Coronaviruses-specific (229E, OC43, NL63, HKU1) IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA1, IgA2, binding capacity to FcγRs (2A, 2B, 3A, 3B), FcαR, between individuals with no PASC and with neuroPASC. Each column represents an antibody feature. Positive values (purple) indicate features enriched in no PASC, negative values (blue) indicate features enriched in neuroPASC, null values (white) indicate features equally present in both no PASC and neuroPASC. Common CoVs = other non-SARS-CoV-2 common Coronaviruses; NC= SARS-CoV-2 nucleocapsid; RBD= SARS-CoV-2 receptor binding domain. Mann-Whitney was used to define statistically significant differences at a univariate level between no PASC and neuroPASC, *p<0.01 or less. ( C-D ) Multivariate analysis of antibody signatures in individuals with neuroPASC and no PASC. Partial least square discriminant analysis (PLSDA) on LASSO-selected features was used to resolve antibody profiles in neuroPASC and no PASC. Dots represent individual samples (no PASC, blue; neuroPASC, red) across SARS-CoV-2 specific (Spike, S1, S2, RBD, NC), and common Coronaviruses (229E, OC43, NL63, HKU1) specific antibody features. Bar graph shows LV1 loadings of LASSO-selected features ranked by their Variable Importance in Projection (VIP). Features enriched in no PASC are in blue, features enriched in neuroPASC are in red. Ten-fold cross validation was performed, resulting in 84% cross validation accuracy (p<0.01). (E) Chord plots indicate the Spearman correlation coefficients between the LASSO-selected features enriched in no PASC (left) and in neuroPASC (right), and the non-LASSO selected antibody features across SARS-CoV-2 antigens (Spike, S1, S2, RBD, NC), and other common Coronaviruses antigens (229E, OC43, NL63, HKU1). Only coefficients >0.5 are plotted, p<0.01 after Benjamini-Hochberg correction for multiple comparisons. Positive values represent direct correlations, negative values represent inverse correlations (none of the features were inversely correlated with any other feature).

    Journal: medRxiv

    Article Title: Neurologic sequalae of COVID-19 are determined by immunologic imprinting from previous Coronaviruses

    doi: 10.1101/2022.11.07.22282030

    Figure Lengend Snippet: (A) Bar dot plots show the level of IgG1 antibodies against non-coronaviruses (EBV, Flu, HSV1) in individuals who did not develop (no PASC, in blue) or developed neurological PASC (neuroPASC, in purple). MFI=Median Fluorescence Intensity. Mann-Whitney, ns= statistically not significant. (B) Heatmap indicates the average difference (%) of SARS-CoV-2 -specific (Spike, S1, S2, RBD and NC) and common Coronaviruses-specific (229E, OC43, NL63, HKU1) IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA1, IgA2, binding capacity to FcγRs (2A, 2B, 3A, 3B), FcαR, between individuals with no PASC and with neuroPASC. Each column represents an antibody feature. Positive values (purple) indicate features enriched in no PASC, negative values (blue) indicate features enriched in neuroPASC, null values (white) indicate features equally present in both no PASC and neuroPASC. Common CoVs = other non-SARS-CoV-2 common Coronaviruses; NC= SARS-CoV-2 nucleocapsid; RBD= SARS-CoV-2 receptor binding domain. Mann-Whitney was used to define statistically significant differences at a univariate level between no PASC and neuroPASC, *p<0.01 or less. ( C-D ) Multivariate analysis of antibody signatures in individuals with neuroPASC and no PASC. Partial least square discriminant analysis (PLSDA) on LASSO-selected features was used to resolve antibody profiles in neuroPASC and no PASC. Dots represent individual samples (no PASC, blue; neuroPASC, red) across SARS-CoV-2 specific (Spike, S1, S2, RBD, NC), and common Coronaviruses (229E, OC43, NL63, HKU1) specific antibody features. Bar graph shows LV1 loadings of LASSO-selected features ranked by their Variable Importance in Projection (VIP). Features enriched in no PASC are in blue, features enriched in neuroPASC are in red. Ten-fold cross validation was performed, resulting in 84% cross validation accuracy (p<0.01). (E) Chord plots indicate the Spearman correlation coefficients between the LASSO-selected features enriched in no PASC (left) and in neuroPASC (right), and the non-LASSO selected antibody features across SARS-CoV-2 antigens (Spike, S1, S2, RBD, NC), and other common Coronaviruses antigens (229E, OC43, NL63, HKU1). Only coefficients >0.5 are plotted, p<0.01 after Benjamini-Hochberg correction for multiple comparisons. Positive values represent direct correlations, negative values represent inverse correlations (none of the features were inversely correlated with any other feature).

    Article Snippet: After a 2 hour incubation at room temperature, immune complexes were washed, and subclasses and isotypes were detected using phycoerythrin (PE)-conjugated mouse anti-human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM (Southern Biotech) at 1.3 μg/ml.

    Techniques: Fluorescence, MANN-WHITNEY, Binding Assay, Biomarker Discovery

    (A) The heatmap indicates functions (ADCD, ADCP, ADNP, ADNKA with production of CD107a, IFNγ and MIP1β), IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM, IgA, capacity to bind to Fcγ receptors (FcγR2A, 2B, 3A, 3B) and FcαR of antibodies to different SARS-CoV-2 specific antigens (Spike, S1, S2, RBD and NC) in serum and CSF from individuals with neuroPASC. Each row corresponds to a single sample. Z-scores, positive (i.e. higher than the mean) in purple, negative (i.e. lower than the mean) in blue. NC= nucleocapsid, RBD=receptor binding domain (B) Radial plots indicate levels of IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA1, IgA2, binding capacity to FcγR ( , , , ), FcαR, and functions (ADCD, ADCP, ADNP, ADNKA with production of CD107a, IFNγ and MIP1β) of Spike-specific antibodies in serum and CSF from individuals with neuroPASC. Each sector represents a z-scored antibody feature. (C) Bar dot plots show the level of IgG subclasses (IgG1, IgG2, IgG3), IgM and IgA, binding capacity to FcγR ( , , , ) and FcαR, and functions (ADCD, ADNP, ADNKA) of Spike-specific antibodies in serum (purple) and CSF (blue) from individuals with neuroPASC. MFI=Median Fluorescence Intensity. Mann-Whitney, *p<0.05, **p<0.01, ***p<0.001 ****p<0.0001. (D) The bar plots indicate the percentage of neuroPASC subjects/samples with positive Spike-specific antibody responses of each IgG subclass in serum and CSF. Positive responses were considered if MFI of each sample was 2-fold higher than the median MFI of PBS level (background). (E) Heatmap indicating the average difference (%) of Spike-specific IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA1, IgA2, binding capacity to FcγR (2A, 2B, 3A, 3B) and FcαR, between CSF and serum from individuals with neuroPASC. Each column represents an antibody feature. Positive values (purple) indicate features enriched in serum, negative values (blue) indicate features enriched in CSF, null values (white) indicate features equally present in serum and CSF. No features were enriched in CSF.

    Journal: medRxiv

    Article Title: Neurologic sequalae of COVID-19 are determined by immunologic imprinting from previous Coronaviruses

    doi: 10.1101/2022.11.07.22282030

    Figure Lengend Snippet: (A) The heatmap indicates functions (ADCD, ADCP, ADNP, ADNKA with production of CD107a, IFNγ and MIP1β), IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM, IgA, capacity to bind to Fcγ receptors (FcγR2A, 2B, 3A, 3B) and FcαR of antibodies to different SARS-CoV-2 specific antigens (Spike, S1, S2, RBD and NC) in serum and CSF from individuals with neuroPASC. Each row corresponds to a single sample. Z-scores, positive (i.e. higher than the mean) in purple, negative (i.e. lower than the mean) in blue. NC= nucleocapsid, RBD=receptor binding domain (B) Radial plots indicate levels of IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA1, IgA2, binding capacity to FcγR ( , , , ), FcαR, and functions (ADCD, ADCP, ADNP, ADNKA with production of CD107a, IFNγ and MIP1β) of Spike-specific antibodies in serum and CSF from individuals with neuroPASC. Each sector represents a z-scored antibody feature. (C) Bar dot plots show the level of IgG subclasses (IgG1, IgG2, IgG3), IgM and IgA, binding capacity to FcγR ( , , , ) and FcαR, and functions (ADCD, ADNP, ADNKA) of Spike-specific antibodies in serum (purple) and CSF (blue) from individuals with neuroPASC. MFI=Median Fluorescence Intensity. Mann-Whitney, *p<0.05, **p<0.01, ***p<0.001 ****p<0.0001. (D) The bar plots indicate the percentage of neuroPASC subjects/samples with positive Spike-specific antibody responses of each IgG subclass in serum and CSF. Positive responses were considered if MFI of each sample was 2-fold higher than the median MFI of PBS level (background). (E) Heatmap indicating the average difference (%) of Spike-specific IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA1, IgA2, binding capacity to FcγR (2A, 2B, 3A, 3B) and FcαR, between CSF and serum from individuals with neuroPASC. Each column represents an antibody feature. Positive values (purple) indicate features enriched in serum, negative values (blue) indicate features enriched in CSF, null values (white) indicate features equally present in serum and CSF. No features were enriched in CSF.

    Article Snippet: After a 2 hour incubation at room temperature, immune complexes were washed, and subclasses and isotypes were detected using phycoerythrin (PE)-conjugated mouse anti-human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM (Southern Biotech) at 1.3 μg/ml.

    Techniques: Binding Assay, Fluorescence, MANN-WHITNEY

    (A) Heatmap indicating the average difference (%) of antibody responses to SARS-CoV-2 (Spike, S1, S2, RBD and NC), other common Coronaviruses (229E, OC43, NL63, HKU1) and non-coronaviruses (EBV, Flu, HHV1), including IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA, binding capacity to FcγR (2A, 2B, 3A, 3B) and FcαR, between serum and CSF from individuals with neuroPASC. Positive values (purple) indicate features enriched in serum, negative values (blue) indicate features enriched in CSF, null values (white) indicate features equally present in both serum and CSF. There were no features enriched in CSF. Common CoVs = other non-SARS-CoV-2 common Coronaviruses (229E, OC43, NL63, HKU1); NC= nucleocapsid; RBD= receptor binding domain (B-C) Multivariate analyses of antibody signatures in serum and CSF from individuals with neuroPASC. Multilevel partial least square discriminant analysis (M-PLSDA) on LASSO-selected features was used to resolve antibody profiles in serum:CSF pairs. Dots represent individual samples (serum, blue; CSF, purple) across SARS-CoV-2 antigens (Spike, S1, S2, RBD, NC), and common Coronaviruses (229E, OC43, NL63, HKU1). Bar graph shows LV1 loadings of LASSO-selected features ranked by their Variable Importance in Projection (VIP). Features enriched in serum are in purple, no features were enriched in CSF. Ten-fold cross validation was performed, resulting in 100% cross validation accuracy (p<0.01). (D-E) Bar dot plots indicate CSF to serum ratios of IgG1, FcγR (2A, 2B, 3A) and FcαR binding capacity for SARS-CoV-2 (Spike, S1, S2, RBD, NC antigens combined) and other common Coronaviruses (229E, OC43, NL63, HKU1 combined) antibodies in individuals with neuroPASC. Mann-Whitney, *p<0.05, ***p<0.001 ****p<0.0001. CoV=common Coronaviruses; NC= nucleocapsid; RBD= receptor binding domain (F) Bar dot plots show the binding capacity of antibodies to the neonatal Fc receptor (FcRn) of serum and CSF SARS-CoV-2 antibodies targeting different antigens (Spike, S1, S2, RDB, NC) in individuals with neuroPASC. Mann-Whitney, *p<0.05, **p<0.01, ***p<0.001 ****p<0.0001. MFI=Median Fluorescence Intensity; NC=nucleocapsid; RBD=receptor binding domain (G) Bar dot plots indicate the FcRn binding capacity of serum and CSF SARS-CoV-2 antibodies (Spike, S1, S2, RBD, NC, combined), compared to common Coronaviruses (229E, OC43, NL63, HKU1 combined) and non-coronaviruses (EBV, Flu, HSV1 combined). Kruskal Wallis test, Dunn’s correction for multiple comparisons. **p<0.01, ****p<0.0001. MFI=Median Fluorescence Intensity; NC= nucleocapsid; RBD= receptor binding domain

    Journal: medRxiv

    Article Title: Neurologic sequalae of COVID-19 are determined by immunologic imprinting from previous Coronaviruses

    doi: 10.1101/2022.11.07.22282030

    Figure Lengend Snippet: (A) Heatmap indicating the average difference (%) of antibody responses to SARS-CoV-2 (Spike, S1, S2, RBD and NC), other common Coronaviruses (229E, OC43, NL63, HKU1) and non-coronaviruses (EBV, Flu, HHV1), including IgG subclasses (IgG1, IgG2, IgG3, IgG4), IgM and IgA, binding capacity to FcγR (2A, 2B, 3A, 3B) and FcαR, between serum and CSF from individuals with neuroPASC. Positive values (purple) indicate features enriched in serum, negative values (blue) indicate features enriched in CSF, null values (white) indicate features equally present in both serum and CSF. There were no features enriched in CSF. Common CoVs = other non-SARS-CoV-2 common Coronaviruses (229E, OC43, NL63, HKU1); NC= nucleocapsid; RBD= receptor binding domain (B-C) Multivariate analyses of antibody signatures in serum and CSF from individuals with neuroPASC. Multilevel partial least square discriminant analysis (M-PLSDA) on LASSO-selected features was used to resolve antibody profiles in serum:CSF pairs. Dots represent individual samples (serum, blue; CSF, purple) across SARS-CoV-2 antigens (Spike, S1, S2, RBD, NC), and common Coronaviruses (229E, OC43, NL63, HKU1). Bar graph shows LV1 loadings of LASSO-selected features ranked by their Variable Importance in Projection (VIP). Features enriched in serum are in purple, no features were enriched in CSF. Ten-fold cross validation was performed, resulting in 100% cross validation accuracy (p<0.01). (D-E) Bar dot plots indicate CSF to serum ratios of IgG1, FcγR (2A, 2B, 3A) and FcαR binding capacity for SARS-CoV-2 (Spike, S1, S2, RBD, NC antigens combined) and other common Coronaviruses (229E, OC43, NL63, HKU1 combined) antibodies in individuals with neuroPASC. Mann-Whitney, *p<0.05, ***p<0.001 ****p<0.0001. CoV=common Coronaviruses; NC= nucleocapsid; RBD= receptor binding domain (F) Bar dot plots show the binding capacity of antibodies to the neonatal Fc receptor (FcRn) of serum and CSF SARS-CoV-2 antibodies targeting different antigens (Spike, S1, S2, RDB, NC) in individuals with neuroPASC. Mann-Whitney, *p<0.05, **p<0.01, ***p<0.001 ****p<0.0001. MFI=Median Fluorescence Intensity; NC=nucleocapsid; RBD=receptor binding domain (G) Bar dot plots indicate the FcRn binding capacity of serum and CSF SARS-CoV-2 antibodies (Spike, S1, S2, RBD, NC, combined), compared to common Coronaviruses (229E, OC43, NL63, HKU1 combined) and non-coronaviruses (EBV, Flu, HSV1 combined). Kruskal Wallis test, Dunn’s correction for multiple comparisons. **p<0.01, ****p<0.0001. MFI=Median Fluorescence Intensity; NC= nucleocapsid; RBD= receptor binding domain

    Article Snippet: After a 2 hour incubation at room temperature, immune complexes were washed, and subclasses and isotypes were detected using phycoerythrin (PE)-conjugated mouse anti-human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM (Southern Biotech) at 1.3 μg/ml.

    Techniques: Binding Assay, Biomarker Discovery, MANN-WHITNEY, Fluorescence

    (A) The radial plots indicate the levels of IgG1 and FcγR2A binding capacity of SARS-CoV-2 antibodies (Spike, S1, S2, RBD, upper half circle) and common Coronaviruses (229E, OC43, NL63, lower half circle) in individuals with neuroPASC, divided into those with good versus poor outcome. Each sector represents a z-scored antibody feature. RBD=receptor binding domain (B-C) Volcano plots showing the correlation of each SARS-CoV-2 and common Coronaviruses (229E, OC43, NL63, HKU1)-antibody feature with modified Rankin Scale (mRS). Spearman correlation coefficients are indicated in the x-axis (positive correlation on the right, red; negative correlation on the left, blue), and the statistical significance is indicated in the y-axis (-log10[p-values]). Values above black dashed line indicate statistically significant correlations (p adjusted value<0.01, Benjamini-Hochberg correction for multiple comparisons). Negative correlations, indicating good outcome, were identified with serum antibodies (left graph, in blue), whereas negative correlations, indicating poor outcome, were identified with CSF antibody profiles (right graph, in red).

    Journal: medRxiv

    Article Title: Neurologic sequalae of COVID-19 are determined by immunologic imprinting from previous Coronaviruses

    doi: 10.1101/2022.11.07.22282030

    Figure Lengend Snippet: (A) The radial plots indicate the levels of IgG1 and FcγR2A binding capacity of SARS-CoV-2 antibodies (Spike, S1, S2, RBD, upper half circle) and common Coronaviruses (229E, OC43, NL63, lower half circle) in individuals with neuroPASC, divided into those with good versus poor outcome. Each sector represents a z-scored antibody feature. RBD=receptor binding domain (B-C) Volcano plots showing the correlation of each SARS-CoV-2 and common Coronaviruses (229E, OC43, NL63, HKU1)-antibody feature with modified Rankin Scale (mRS). Spearman correlation coefficients are indicated in the x-axis (positive correlation on the right, red; negative correlation on the left, blue), and the statistical significance is indicated in the y-axis (-log10[p-values]). Values above black dashed line indicate statistically significant correlations (p adjusted value<0.01, Benjamini-Hochberg correction for multiple comparisons). Negative correlations, indicating good outcome, were identified with serum antibodies (left graph, in blue), whereas negative correlations, indicating poor outcome, were identified with CSF antibody profiles (right graph, in red).

    Article Snippet: After a 2 hour incubation at room temperature, immune complexes were washed, and subclasses and isotypes were detected using phycoerythrin (PE)-conjugated mouse anti-human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM (Southern Biotech) at 1.3 μg/ml.

    Techniques: Binding Assay, Modification