anti-human igm antibody  (Advisains)

Journal: JCI Insight

Article Title: Balance between maternal antiviral response and placental transfer of protection in gestational SARS-CoV-2 infection

doi: 10.1172/jci.insight.167140

Figure Lengend Snippet: ( A ) Outline of participant recruitment, divided into pregnant women negative for SARS-CoV-2 (CoV-2 – ), recovered from infection in second (2R) or third (3R) trimesters, with ongoing infection (3O), or receiving COVID-19 vaccine (Vac). ( B ) Anti-spike and anti-RBD IgG endpoint titers ( n = 50 dyads). ( C ) Anti-spike and anti-RBD IgG transfer ratios ( n = 39). ( D ) Correlation between anti-spike and anti-RBD IgG transfer ratio and elapsed time between diagnosis and delivery ( n = 39). ( E ) Total IgG transfer ratios ( n = 50). ( F ) Anti-spike, anti-RBD, and total IgG transfer ratios ( n = 24). ( G ) Correlation between total IgM and anti-RBD IgM antibody levels ( n = 58). ( H ) Correlation between total IgG and anti-RBD IgG antibody levels ( n = 58). ( I ) Correlation between total IgA and anti-RBD IgA antibody levels ( n = 58). Data represent mean ± SD for parametric tests, or median ± IQR for nonparametric tests. NS, not significant. Significance determined by parametric paired, 2-tailed t test ( B ), nonparametric paired Wilcoxon’s test ( B ), ordinary ANOVA with post hoc Holm-Šidák ( C and F ), Kruskal-Wallis with post hoc Dunn’s ( E ), Pearson’s correlation ( D ), and Spearman’s correlation ( G – I ). Effect sizes were determined by Cohen’s d ( B ), correlation coefficient r ( B ), and η 2 ( C , E , and F ).

Article Snippet: Plates were washed and incubated for 30 minutes at room temperature with HRP-conjugated anti–human IgA, IgG, and IgM antibodies (Abcam; ab97225, ab97215, and ab97205) diluted 1:25,000 in 1% BSA/0.05% Tween–PBS.

Techniques: Infection

Journal: JCI Insight

Article Title: Balance between maternal antiviral response and placental transfer of protection in gestational SARS-CoV-2 infection

doi: 10.1172/jci.insight.167140

Figure Lengend Snippet: ( A ) NT 50 values for maternal and cord blood paired samples from SARS-CoV-2–infected (CoV-2 + ; n = 50 dyads) and vaccinated mothers (Vac, n = 6 dyads). ( B ) NT 50 for CoV-2 + maternal samples segregated by gestational age of infection ( n = 50). ( C ) Anti-RBD IgG transfer ratio in the presence (Sym) or absence (Asym) of symptoms in ongoing maternal infections (3O, n = 24). ( D ) Total IgG transfer ratio in the presence (Sym) or absence (Asym) of symptoms in ongoing maternal infections (3O, n = 35). ( E ) Correlation between maternal anti-spike IgA, IgG, and IgM and NT 50 in ongoing infections (3O, n = 35). ( F ) Correlation between maternal anti-spike IgA, IgG, and IgM and NT 50 in recovered infections (2R + 3R, n = 15). ( G ) Correlation between maternal anti-spike IgA, IgG, and IgM and NT 50 in vaccinated participants (Vac, n = 6). ( H ) Ratios of NT 50 values obtained from purified IgM and IgG fractions in maternal infections ( n = 33). ( I ) Ratios of NT 50 values obtained from purified IgA and IgG fractions in maternal infections ( n = 33). Data represent mean ± SD for parametric tests, or median ± IQR for nonparametric tests. nd, not detectable. NS, not significant. Significance determined by parametric paired, 2-tailed t test ( A ), unpaired, 2-tailed t test ( C ), nonparametric paired Wilcoxon’s test ( A ), Mann-Whitney test ( A , D , H , and I ), Kruskal-Wallis with post hoc Dunn’s ( B ), Spearman’s correlation ( E – G ), and Pearson’s correlation ( F and G ). Effect sizes were determined by Cohen’s d ( A and C ), correlation coefficient r ( A , D , H , and I ), and η 2 ( B ).

Article Snippet: Plates were washed and incubated for 30 minutes at room temperature with HRP-conjugated anti–human IgA, IgG, and IgM antibodies (Abcam; ab97225, ab97215, and ab97205) diluted 1:25,000 in 1% BSA/0.05% Tween–PBS.

Techniques: Infection, Purification, MANN-WHITNEY

Journal: JCI Insight

Article Title: Balance between maternal antiviral response and placental transfer of protection in gestational SARS-CoV-2 infection

doi: 10.1172/jci.insight.167140

Figure Lengend Snippet: ( A ) Maternal B cell frequency in ongoing SARS-CoV-2 (3O) infection in function of the presence (Sym) or absence (Asym) of symptoms ( n = 23). ( B ) Frequency of IgD – B cells as in A ( n = 23). ( C ) Frequency of CD4 + T cells as in A ( n = 22). ( D ) Frequency of CD69 + CD4 + T cells as in A ( n = 22). ( E ) Frequency of CCR6 + CD4 + T cell as in A ( n = 22). ( F ) Frequency of CXCR5 + CD4 + T cell as in A ( n = 16). ( G ) Maternal NK cell frequency in participants diagnosed for SARS-CoV-2 within 7 days of delivery in asymptomatic (Asym) and symptomatic (Sym) individuals ( n = 17). ( H ) Maternal anti-spike IgM levels as in G . ( I ) Cord blood NK cell frequency from participants diagnosed for SARS-CoV-2 within 7 days of delivery, segregated by the presence (Sym) or absence (Asym) of maternal symptoms ( n = 17). ( J ) Cord blood NK cell frequency upon either immediate or delayed umbilical cord clamping, in ongoing maternal infections (3O, n = 21). ( K ) Cord blood B cell frequency as in J ( n = 20). ( L ) Cord blood CD4 + T cell frequency as in J ( n = 22). ( M ) Cord blood NK cell frequency upon either immediate or delayed umbilical cord clamping, in second and third trimester recovered (2R + 3R) infection ( n = 10). Data represent mean ± SD for parametric tests, or median ± IQR for nonparametric tests. NS, not significant. Significance determined by unpaired, 2-tailed t test ( A – C , F , and I – L ) and Mann-Whitney test ( D , E , G , H , and M ). Effect sizes were determined by Cohen’s d ( A – C , F , and I – L ) and correlation coefficient r ( D , E , G , H , and M ).

Article Snippet: Plates were washed and incubated for 30 minutes at room temperature with HRP-conjugated anti–human IgA, IgG, and IgM antibodies (Abcam; ab97225, ab97215, and ab97205) diluted 1:25,000 in 1% BSA/0.05% Tween–PBS.

Techniques: Infection, MANN-WHITNEY

Journal: JCI Insight

Article Title: Balance between maternal antiviral response and placental transfer of protection in gestational SARS-CoV-2 infection

doi: 10.1172/jci.insight.167140

Figure Lengend Snippet: ( A ) Correlation between anti-RBD IgG transfer ratio and IL-6 concentration in CoV-2 + maternal plasma ( n = 39). ( B ) Correlation between anti-RBD IgG transfer ratio and IL-10 concentration in maternal plasma in ongoing (3O) infection ( n = 24). ( C ) As in B for IL-23. ( D ) Correlation between anti-RBD IgG levels and IL-6 concentration in CoV-2 + maternal plasma ( n = 48). ( E ) Correlation between anti-RBD IgG levels and IL-10 concentration in maternal plasma in ongoing (3O) infection ( n = 28). ( F ) As in E for IL-23. ( G ) Correlation between NT 50 and IL-6 concentration in maternal plasma in ongoing (3O) infection ( n = 23). ( H ) As in G for IL-10. ( I ) As in G for IL-23. ( J ) Correlation between anti-spike IgM levels and IL-10 concentration in CoV-2 + maternal plasma within 7 days between diagnosis and delivery ( n = 17). ( K ) Correlation between IL-10 concentration and NK cell frequency in ongoing (3O) infection ( n = 20). NS, not significant. Significance determined by Pearson’s correlation ( A – C , E , and F ) and Spearman’s correlation ( D and G – K ).

Article Snippet: Plates were washed and incubated for 30 minutes at room temperature with HRP-conjugated anti–human IgA, IgG, and IgM antibodies (Abcam; ab97225, ab97215, and ab97205) diluted 1:25,000 in 1% BSA/0.05% Tween–PBS.

Techniques: Concentration Assay, Infection

Journal: JCI Insight

Article Title: Balance between maternal antiviral response and placental transfer of protection in gestational SARS-CoV-2 infection

doi: 10.1172/jci.insight.167140

Figure Lengend Snippet: ( A and B ) Paired mother (M)–cord blood (C) dyad analysis of anti-spike IgA levels ( A ) and endpoint titers ( B ), with dyad 26 highlighted in pink and maroon ( n = 50). ( C and D ) As in A and B but for IgM. ( E ) Frequency of IgD – B cells in paired mother–cord blood dyads in recovered second (2R) and third (3R) trimester, or in ongoing (3O) infection, with dyad 26 highlighted in pink and in maroon ( n = 31). ( F ) Cytokine ratio between cord and maternal blood for dyad 26. Dashed line indicates assay cutoff; nd, not detectable. NS, not significant. Significance determined by nonparametric paired Wilcoxon’s test ( A , C , and E ) and parametric paired, 2-tailed t test ( E ). Effect sizes were determined by correlation coefficient r ( A , C , and E ) and Cohen’s d ( E ).

Article Snippet: Plates were washed and incubated for 30 minutes at room temperature with HRP-conjugated anti–human IgA, IgG, and IgM antibodies (Abcam; ab97225, ab97215, and ab97205) diluted 1:25,000 in 1% BSA/0.05% Tween–PBS.

Techniques: Infection

Journal: medRxiv

Article Title: Increased circulating fibronectin, depletion of natural IgM and heightened EBV, HSV-1 reactivation in ME/CFS and long COVID

doi: 10.1101/2023.06.23.23291827

Figure Lengend Snippet: a. Confocal images show mitochondrial architecture in primary HUVEC cells exposed to 1μg of purified IgG from patient sera. Two different representative images for each condition is shown. b. Average mitochondrial surface area in primary HUVEC cells exposed to 1μg of purified IgG from patient sera is quantified and compared between healthy controls, mild/moderate, severe ME/CFS. Data from three independent experiments from each serum sample is shown as a violin plot. n=3. Two-tailed non-parametric test. Healthy control vs mild/moderate ME/CFS, *P = 0.0329. Healthy control vs severe ME/CFS, **P = 0.0046. Mild/moderate vs severe ME/CFS, ****P < 0.0001. c. Immunoblot analysis shows decrease in mitofusin1 (Mfn1) and PLD6 protein levels in HUVEC cells exposed to 1μg of purified IgG from patient sera for 12 h. Actin staining was used as a loading control. Fold change values were derived from densitometric analysis of bands after normalization with the same for actin. n=2. HD, healthy donors; CFS, severe CFS patients. d . Heat map of log2 fold LFQ intensities of proteins detected within purified immune complexes from patient sera. Three proteins that showed differential protein levels between healthy controls and ME/CFS patients are shown. e . Multivariate analysis of clusters based on distance metrics derived from IgM antibody levels for a panel of autoantigens. Log-transformed scaled data showing relative differences between different variable in both healthy controls and patients. f . The Variables Factor map for the Principal Components (for Patients and Healthy Controls combined data) shows the projection of the top 10 Autoantigen variables projected onto the plane spanned by the first two Principal Components. g . Serum fibronectin (FN1) levels in patient sera. Log2 values of FN1 are presented as a violin plot. Two-tailed parametric t-test. Healthy control (HC) vs ME/CFS, **P = 0.005. h. Kernel density plot showing the bivariate serum FN1 distributions among healthy controls and ME/CFS patients. FN1 concentrations on both X- and Y-axis are presented as μg/ml. i. Circulating fibronectin (FN1) levels correlates with ME/CFS severity associated Bell score. Log2 fold FN1 vales are presented as a violin plot. Two-tailed parametric t-test. HC vs Bell 0-20, ****P < 0.0001. Bell 0-20 vs Bell 30-50, **P = 0.0032. j. AUROC analysis for circulating FN1 levels in healthy controls (HC) vs severe ME/CFS patients. k. Serum fibronectin (FN1) levels in different patient groups post SARS-CoV-2 infection. Log2 values of FN1 are presented as a violin plot. Two-tailed parametric t-test. Healthy control (HC) vs mild LC, **P = 0.0032. HC vs severe LC, *P = 0.0488. ns, not significant.

Article Snippet: For IgG and IgM standard wells, 100 μl 25.60 ∼ 0 pg/μl series dilutions of native human IgM protein (Abcam, ab90348) or human IgG isotype control (Thermo Fisher Scientific, 31154) was coated onto the plates in carbonate coating buffer (Thermo Fisher Scientific, CB01100) overnight at 4°C.

Techniques: Purification, Two Tailed Test, Western Blot, Staining, Derivative Assay, Transformation Assay, Infection

Journal: medRxiv

Article Title: Increased circulating fibronectin, depletion of natural IgM and heightened EBV, HSV-1 reactivation in ME/CFS and long COVID

doi: 10.1101/2023.06.23.23291827

Figure Lengend Snippet: a. IgM levels against fibronectin (IgM-FN1) in patient sera. Log2 fold IgM-FN1 amounts are presented as a violin plot. Two-tailed parametric t-test. Healthy control (HC) vs SARS CoV-2 positive but without long COVID (No LC), HC vs mild LC, HC vs severe LC, ****P < 0.00001. No LC vs severe LC, *P = 0.0376. ns, not significant. b. IgG levels against fibronectin (IgM-FN1) in patient sera. Log2 fold IgG-FN1 amounts are presented as a violin plot. Two-tailed parametric t-test. Healthy control (HC) vs SARS CoV-2 positive but without long COVID (No LC), HC vs mild LC, HC vs severe LC, ****P < 0.00001. ns, not significant. c. IgM-fibronectin (FN1) levels correlates with ME/CFS severity associated Bell scores. Log2 fold IgM-FN1 vales are presented as a violin plot. Two-tailed parametric t-test. HC vs Bell 0-20, **P = 0.0046. Bell 0-20 vs Bell 30-50, ***P = 0.0002. d. IgG-fibronectin (FN1) levels does not correlate with ME/CFS severity associated Bell scores. Log2 fold IgG-FN1 vales are presented as a violin plot. Two-tailed parametric t-test. Ns, not significant. e. IgM levels against phosphorylcholine (IgM-PC) in patient sera. Log2 fold IgM-PC amounts are presented as a violin plot. Two-tailed parametric t-test. Healthy control (HC) vs mild LC, ***P = 0.0002. HC vs severe LC, ***P = 0.0006. No LC vs severe LC, *P = 0.0216. No LC vs mild LC, *P = 0.0111. ns, not significant.\ f. IgM levels against malondialdehyde (IgM-MDA) in patient sera. Log2 fold IgM-MDA amounts are presented as a violin plot. Two-tailed parametric t-test. Healthy control (HC) vs mild LC, *P = 0.0175. HC vs severe LC, **P = 0.0068. No LC vs severe LC, *P = 0.0209. No LC vs mild LC, *P = 0.0525. ns, not significant. g. AUROC analysis for IgM-FN1 levels in healthy controls (HC) vs SARS CoV-2 positive but without long COVID (No LC) patients. h. AUROC analysis for IgM-FN1 levels in healthy controls (HC) vs mild LC patients. i. AUROC analysis for IgM-FN1 levels in healthy controls (HC) vs severe LC patients.

Article Snippet: For IgG and IgM standard wells, 100 μl 25.60 ∼ 0 pg/μl series dilutions of native human IgM protein (Abcam, ab90348) or human IgG isotype control (Thermo Fisher Scientific, 31154) was coated onto the plates in carbonate coating buffer (Thermo Fisher Scientific, CB01100) overnight at 4°C.

Techniques: Two Tailed Test

Journal: medRxiv

Article Title: Increased circulating fibronectin, depletion of natural IgM and heightened EBV, HSV-1 reactivation in ME/CFS and long COVID

doi: 10.1101/2023.06.23.23291827

Figure Lengend Snippet: Distribution of serum IgM-FN1 concentrations in different groups of patients separated by gender. Two-tailed parametric t-test. ME/CFS male vs female, *P = 0.0398. b. Distribution of serum IgG-FN1 concentrations in different groups of patients separated by gender. c. Comparison of gender-specific IgM-FN1 concentrations (log2 values) among different patient groups. Two-tailed parametric t-test. Healthy control (HC) male vs No LC male, **P = 0.003; HC male vs mild LC male, **P = 0.0018; HC male vs severe LC male, ***P = 0.0007; HC female vs no LC female, HC female vs mild LC female, HC female vs severe LC female, ****P < 0.0001.

Article Snippet: For IgG and IgM standard wells, 100 μl 25.60 ∼ 0 pg/μl series dilutions of native human IgM protein (Abcam, ab90348) or human IgG isotype control (Thermo Fisher Scientific, 31154) was coated onto the plates in carbonate coating buffer (Thermo Fisher Scientific, CB01100) overnight at 4°C.

Techniques: Two Tailed Test