h26  (Santa Cruz Biotechnology)

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: Total and specific IgG and IgA in BAL from SARS-CoV-2-infected and non-infected individuals. (A) Graphical representation showing the overall study design and the characteristics (number, age, body mass index (BMI), sex (F = female, M = male), diabetes, and fatality rates) of the individuals included in the study. Illustration with images from Servier Medical Art, licensed under the Creative Commons Attribution 3.0 Unported License. (B) Timeline of the course of disease for enrolled patients infected with SARS-CoV-2. (C) SARS-CoV-2 containing (SARS-CoV-2+) samples correspond to the early phase, whereas those lacking virus (SARS-CoV-2−) correspond to a late phase of the disease. Violin plots of time from onset of symptoms to sampling for each sample in SARS-CoV-2+ and SARS-CoV-2− BAL. p -Values were calculated by using Mann–Whitney test. (D) Comparison between values of total IgG and IgA (μg/ml) in BAL from SARS-CoV-2-infected individuals (SARS-CoV-2+ and SARS-CoV-2− BAL) and COVID-19 non-infected individuals. p -Values were calculated by using Wilcoxon test: *, p < 0.05; **, p < 0.01; ****, p < 0.0001. Dashed line: cutoff value for antibody detection. Negative values are not shown. BAL for bronchoalveolar lavage.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Infection, Sampling, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: S1-, RBD-, S2-, and NP-specific IgG and IgA in SARS CoV-2+ vs SARS-CoV-2− BAL. (A) S1-, S2-, RBD-, and NP-specific IgG and IgA responses in SARS-CoV-2+ BAL (B) S1-, S2-, RBD-, and NP-specific IgG and IgA responses in SARS-CoV-2− BAL. (A, B) Proportion of specific IgG or IgA over total IgG or IgA measured by ELISA. Specific (OD450)/total IgA or G (μg/ml) are shown. p -Values were calculated by using Mann–Whitney test: *, p < 0.05; **, p < 0.01; ***, p < 0.005. (C) Correlations between specific S1-, RBD-, S2-, and NP-specific IgG antibodies in SARS-CoV-2+ BAL (red dots) and SARS-CoV-2− BAL (gray dots) and onset of symptom to sampling date (days). (D) Correlations between S1-, RBD-, S2-, and NP-specific IgA antibodies SARS-CoV-2+ BAL (red dots) and SARS-CoV-2− BAL (gray dots) and onset of symptom to sampling date (days). (E) Correlation between specific S1, RBD, S2, and NP IgA and IgG antibodies in SARS-CoV-2+ BAL individuals. (F) Correlation between S1-, RBD-, S2-, and NP-specific IgA and IgG in SARS-CoV-2− BAL individuals. All correlations were calculated using Spearman’s test. RBD, receptor-binding domain; NP, nucleocapsid protein.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Sampling, Binding Assay

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: BAL from SARS-CoV-2-infected individuals contain IgG- and IgA-spike immune complexes (ICs). (A) Anti-spike IgG ICs were detected in 32 of the 48 samples analyzed by ELISA. Indicated with a red asterisk are individuals who had undetectable levels of IgG anti-spike/NP antibodies. (B) Anti-spike IgA ICs were detected in 25 of the 48 samples analyzed by ELISA. Indicated with red asterisk are individuals who had undetectable levels of IgA anti-spike/NP antibodies. (C, D) Comparison of presence of ICs made of spike with IgG (c) or IgA (D) in SARS-CoV-2+ BAL individuals and SARS-CoV-2− BAL subjects. p -Values were calculated by using Mann–Whitney test: *, p < 0.05. (E, F) Comparison of presence of ICs made of spike with IgG (E) or IgA (F) in survivors vs non-survivors. (G, H) Correlation between specific S1 and levels of ICs made of spike with IgG (G) or IgA (H) . Correlations were calculated using Spearman’s test. BAL, bronchoalveolar lavage; NP, nucleocapsid protein.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: IgG and IgA antibodies from BAL from SARS-CoV-2-infected individuals against RBD protein from SARS-CoV-2 Alpha (B.1.1.7), Beta (B.1.351), Gamma (P.1), and Delta (B.1.617) variants. (A) Specific IgG responses against Alpha, Beta, Gamma, and Delta RBD in BAL from SARS-CoV-2-infected individuals. (B) Specific IgA responses against Alpha, Beta, Gamma, and Delta RBD in BAL from SARS-CoV-2-infected individuals. (A, B) Proportion of specific IgG or IgA over total IgG or IgA measured by ELISA (specific (OD450)/total IgA or G (μg/ml)) are shown. (C) Pie charts showing the percentages of the different responses of IgG (left) and IgA (right) to Wuhan RBD and the different variants. (D–H) Comparison between specific IgG and IgA responses detected against the RBD from ancestral Wuhan strain (D) , Alpha (E) , Beta (F) , and Gamma (G) and Delta (H) variants. Correlation between IgG (I) and IgA (J) specific to Wuhan RBD and IgG and IgA antibodies specific for Alpha, Beta and Gamma variants. Correlations were calculated using Spearman’s test. p -Values were calculated by using Wilcoxon test. *, p < 0.05; **, p < 0.01; ***, p < 0.005, ****, p < 0.0001. Dashed line: cutoff value for antibody detection. BAL, bronchoalveolar lavage; RBD, receptor-binding domain.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: IC50 neutralization titers in BAL from COVID-19 individuals. (A) Comparison of IC50 neutralization titers between SARS-CoV-2+ BAL and SARS-CoV-2− BAL samples. (B) Correlations between IC50 neutralization titers and the onset of symptoms to sampling date in SARS-CoV-2+ BAL (light blue squares) and SARS-CoV-2− BAL (purple dots) individuals. (C) Correlation between IC50 neutralization titers and spike-specific IgG in SARS-CoV-2+ and SARS-CoV-2− BAL. (D) Correlation between IC50 neutralization titers and spike-specific IgA in SARS-CoV-2+ and SARS-CoV-2− BAL. (E) Correlation between neutralization activity and hospitalization time in SARS-CoV-2+ and SARS-CoV-2− BAL. (F) IC50 neutralization titers of BAL from SARS-CoV-2+ individuals against ancestral Wuhan, and Alpha, Beta, and Gamma SARS-CoV-2 variants. A specific color is associated with each individual. All correlations were calculated using Spearman’s test. p -Values were calculated by using Mann–Whitney test. *, p < 0.05; **, p < 0.01. BAL, bronchoalveolar lavage.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Neutralization, Sampling, Activity Assay, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: Specific IgG and IgA responses in COVID-19+ survivors vs non-survivors. (A) Specific S1-, S2-, RBD-, and NP-specific IgG and IgA responses in survivors and non-survivors. p -Values were calculated by using Wilcoxon test (A, B) Dashed line: cutoff value for antibody detection. (C) Comparison of the kinetics from the cross-sectional SARS-CoV-2-specific IgG responses in survivors (S) versus non-survivors (NS). (D) Comparison of the kinetics from the cross-sectional SARS-CoV-2-specific IgA responses in survivors (S) versus non-survivors (NS). All correlations were calculated using Spearman’s test. *, p < 0.05; **, p < 0.01. RBD, receptor-binding domain; NP, nucleocapsid protein.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Binding Assay

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: Neutralization activities in COVID-19+ survivors vs non-survivors. (A) Comparison between IC50 neutralization titers between survivors and non-survivors. (B) Cross-sectional representation of neutralizing antibodies in survivors vs non-survivors, shown as correlation between IC50 neutralization titters and time from symptom onset to sampling date in survivors and non-survivors. (C) Cross-sectional representation of neutralizing antibodies in non-survivors, shown as correlation between IC50 neutralization titters and time from symptom onset to sampling date, in SARS-CoV-2+ BAL and SARS-CoV-2− BAL individuals. All correlations were calculated using Spearman’s test. p -Values were calculated by using Wilcoxon test. BAL: bronchoalveolar lavage.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Neutralization, Sampling

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: Analysis of B-cell phenotype in BAL supernatant from SARS-CoV-2-infected individuals. From total B cells, five B-cell populations were defined including Naïve B cells, activated memory B cells, plasma B cells, resting memory B cells, and tissue memory B cells according to a gating strategy shown in Supplementary Figure 1 , which were further labeled for IgG (IgG+) and IgA (IgA+). (A) Frequencies of different IgG+ B-cell populations between COVID-19+ and COVID-19− individuals in the total B-cell population shown as violin plots. (B) Frequencies of different IgA+ B-cell populations between COVID-19+ and COVID-19− individuals in the total IgA+ B-cell population shown as violin plots. (C) Frequencies of different IgG+ B-cell populations between SARS CoV-2+ BAL and SARS CoV-2− BAL individuals in the total IgG+ B-cell population shown as violin plots. (D) Frequencies of different IgA+ B-cell populations between SARS CoV-2+ BAL and SARS CoV-2− BAL individuals in the total IgA+ B-cell population shown as violin plots. (E) Frequencies of different IgG+ B-cell populations between survivors and non-survivors in the total IgG+ B-cell population shown as violin plots. (F) Comparison of different IgA+ B-cell populations between survivors and non-survivors in the total IgA+ B-cell population shown as violin plots. p-Values were calculated by using Mann–Whitney test *, p < 0.05; **, p < 0.01.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Infection, Labeling, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Persistent but dysfunctional mucosal SARS-CoV-2-specific IgA and low lung IL-1β associate with COVID-19 fatal outcome: A cross-sectional analysis

doi: 10.3389/fimmu.2022.842468

Figure Lengend Snippet: Analysis of cytokines in BAL fluid. Quantification of the following cytokines in BAL from SARS-CoV-2-infected individuals: MIP-1α, G-CSF, IL-1β (IL-1β), IL-8, S100A8, TNF-a, MCP-1, CXCL10, IL-1α (IL-1A), IL-6, M-CSF, and S100B. (A) Mean amounts of cytokines (pg/ml) evaluated between COVID-19+ and COVID-19− individuals. (B) Mean amounts of cytokines (pg/ml) evaluated between BAL SARS CoV-2+ and BAL SARS CoV-2− individuals. (C) Cross-sectional concentration (pg/ml) of IL-8, IL-1β, and IL-1A as function of time from onset of symptoms to sampling date. (D) Correlation between RBD-specific IgA (shown as proportion of specific IgG or IgA over total IgG or IgA measured by ELISA (specific (OD450)/total IgA or G (μg/ml)) and IL-1β and IL-8 concentration (pg/ml). (E) Correlation between S2-specific IgA (shown as proportion of specific IgG or IgA over total IgG or IgA measured by ELISA (specific (OD450)/total IgA or G (μg/ml)) and S100A8 and IL-6 concentration. (F) Comparison of the levels of IL-1β (pg/ml) between survivors and non-survivors in SARS-CoV-2+ BAL individuals. (G) Comparison of the levels of IL-1β (pg/ml) between survivors and non-survivors in individuals BAL SARS-CoV-2−. p -Values were calculated by using Mann–Whitney test. *, p < 0.05, **, p < 0.01; ***, p < 0.005, ****, p < 0.0001. BAL, bronchoalveolar lavage; G-CSF, granulocyte colony-stimulating factor; RBD, receptor-binding domain.

Article Snippet: For IgG and IgA anti-S2 and anti-RBD quantification, 96-well, flat-bottomed plates (Nunc-Immun Microwell, Thermo Fisher Scientific, Odense C, Denmark) were coated overnight at 4°C with 1 ng/well, or 100 ng/well, of SARS-CoV-2 spike S2 protein and SARS-CoV-2 spike RBD Wuhan protein (LifeTein, Somerset, NJ, USA) and recombinant human SARS-CoV-2 spike RBD variants Alpha (B.1.1.7), Beta (B.1.351), P.1 (Gamma), and Delta (B.1.617) (Diaclone, Besançon, France).

Techniques: Infection, Concentration Assay, Sampling, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Binding Assay

Journal: Cell Discovery

Article Title: A trifunctional peptide broadly inhibits SARS-CoV-2 Delta and Omicron variants in hamsters

doi: 10.1038/s41421-022-00428-9

Figure Lengend Snippet: a Peptide sequences of H23, H26, H30, 2H30, 4H30, and reference HBD2. Peptide 2H30 and 4H30 were 2-branched and 4-branched H30 cross-linked by lysine at the C terminal of H30. b The antiviral activity of these peptides in the high salt concentration (150 mM, PBS) was determined by plaque reduction assay ( n = 3). c The antiviral activity of these peptides against SARS-CoV-2 in the low salt concentration (30 mM, PBS/5) was determined by plaque reduction assay ( n = 3). P values were generated by comparison with an untreated virus (0). d The antiviral activity of 2-branched H30 (2H30) and 4-branched H30 (4H30) against SARS-CoV-2 (HKU001a) in PBS was determined by plaque reduction assay ( n = 4). e The cytotoxicity of 4H30 in VeroE6 and Calu-3 cells was measured by MTT assay ( n = 3). f The antiviral activity of 4H30 was determined by anti-nucleocapsid (NP) immunofluorescent staining. SARS-CoV-2 with or without 4H30 treatment was added to cells for infection. Representative images were taken at 18 h post-infection (hpi) in VeroE6 cells. Scale bar, 20 μm. g The post-infection antiviral activity of 4H30 against SARS-CoV-2 (B.1.1.63, D614G) in VeroE6 and Calu-3 cells ( n = 6). At 6 hpi, 4H30 (50 μg/mL) was added to infected cells and viral titers in cell supernatants were measured at 24 hpi (for VeroE6 cells) or 30 hpi (for Calu-3 cells). h The broad-spectrum antiviral activities of 4H30 against SARS-CoV-2 variants and MERS-CoV (MERS) in VeroE6 cells were determined by plaque reduction assay ( n = 4). * P < 0.05 and ** P < 0.01 when compared with DMEM. P values were calculated by the two-tailed Student’s t -test. Data were presented as means ± SD of indicated biological samples with more than two independent experiments.

Article Snippet: H30, 2H30, 4H30, H26, and H23 shown in Fig. were synthesized by ChinaPeptide (Shanghai, China).

Techniques: Activity Assay, Concentration Assay, Generated, Comparison, Virus, MTT Assay, Staining, Infection, Two Tailed Test