Journal: ImmunoHorizons

Article Title: ICAM-1 autoantibodies detected in healthy individuals and cross-react with functional epitopes

doi: 10.1093/immhor/vlaf025

Figure Lengend Snippet: ICAM-1 IgG autoantibody and sICAM-1 levels across healthy individuals and those with severe COVID-19, high BMI, and autoimmune conditions. (A) Dot plot graph of AUC on the log axis of the optical density at 450 nm measurement by ELISA for determining the levels of IgG antibodies targeting ICAM-1 within plasma from healthy individuals (n = 40), individuals with severe COVID-19 (n = 40), individuals with a high BMI over 30 kg/m 2 (n = 50), and individuals with select autoimmune conditions, SLE or RA (n = 39). (B) Dot plot graph of concentration (pg/mL) on the log axis measured by ELISA for determining the levels of sICAM-1 within plasma from healthy individuals (n = 40), individuals with severe COVID-19 (n = 40), individuals with a high BMI over 30, (n = 50), and individuals with autoimmune conditions SLE or RA (n = 39). Statistical significance was determined using the Mann-Whitney test. P value results are shown.

Article Snippet: Secondary antibodies were purchased from the following: Goat anti-human IgG (109-036-098, Lot# 149163; Jackson ImmunoResearch), Goat Anti-Human IgA alpha chain (ab97215, Lot# GR3373878-8; Abcam), Goat Anti-Human IgM mu chain (ab97205, Lot# GR3396429-1; Abcam), Mouse Anti-Human IgG1 (9054-05, Lot#B2023-NM34C; Southern Biotech), Mouse Anti-Human IgG2 (9060-05, Lot#K3220-YF80; Southern Biotech), Mouse Anti-Human IgG3 (9210-05, Lot#DO720-QB4017; Southern Biotech), and Mouse Anti-Human IgG4 (9200-05, Lot#E1723-RB54B; Southern Biotech).

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Concentration Assay, MANN-WHITNEY

Journal: ImmunoHorizons

Article Title: ICAM-1 autoantibodies detected in healthy individuals and cross-react with functional epitopes

doi: 10.1093/immhor/vlaf025

Figure Lengend Snippet: Correlation of anti-ICAM-1 IgG autoantibody levels with age. (A) Dot plot graph of the AUC of the optical density at 450 nm on the log y-axis of IgG antibodies targeting ICAM-1 within plasma compared with age (x-axis) from healthy pediatric individuals ranging in age from 6 mo to 15 yr old (n = 37). (B) Dot plot graph of the AUC of the optical density at 450 nm on the log y-axis of IgG antibodies targeting ICAM-1 within plasma compared with age (x-axis) from healthy adult individuals ranging in age from 22 to 75 yr old (n = 40). Statistical significance was determined using the Pearson correlation test. P value results are shown.

Article Snippet: Secondary antibodies were purchased from the following: Goat anti-human IgG (109-036-098, Lot# 149163; Jackson ImmunoResearch), Goat Anti-Human IgA alpha chain (ab97215, Lot# GR3373878-8; Abcam), Goat Anti-Human IgM mu chain (ab97205, Lot# GR3396429-1; Abcam), Mouse Anti-Human IgG1 (9054-05, Lot#B2023-NM34C; Southern Biotech), Mouse Anti-Human IgG2 (9060-05, Lot#K3220-YF80; Southern Biotech), Mouse Anti-Human IgG3 (9210-05, Lot#DO720-QB4017; Southern Biotech), and Mouse Anti-Human IgG4 (9200-05, Lot#E1723-RB54B; Southern Biotech).

Techniques: Clinical Proteomics

Journal: ImmunoHorizons

Article Title: ICAM-1 autoantibodies detected in healthy individuals and cross-react with functional epitopes

doi: 10.1093/immhor/vlaf025

Figure Lengend Snippet: High-resolution IgG epitope mapping of ICAM-1 autoantibodies using ICAM-1 peptide microarrays. (A) Heatmap of z scores for ICAM-1 epitope binding for individual samples (n = 20). ICAM-1 domains 1 through 5 are annotated above the heatmap. (B) Line graphs of median group z scores of ICAM-1 autoantibody binding to individual ICAM-1 peptides in the peptide array. Epitopes of recurrent high binding were qualified as median z scores of ≥0.95 to represent 1 SD ±0.05 above the median, points in red. (C) Table of recurrent high binding peptides, with associated z score and sequence. (D) Table of high binding clusters with associated peptide IDs, z score range, and sequence. Overlapping sequence motifs are bolded, and the common leucine threonine epitope is underlined.

Article Snippet: Secondary antibodies were purchased from the following: Goat anti-human IgG (109-036-098, Lot# 149163; Jackson ImmunoResearch), Goat Anti-Human IgA alpha chain (ab97215, Lot# GR3373878-8; Abcam), Goat Anti-Human IgM mu chain (ab97205, Lot# GR3396429-1; Abcam), Mouse Anti-Human IgG1 (9054-05, Lot#B2023-NM34C; Southern Biotech), Mouse Anti-Human IgG2 (9060-05, Lot#K3220-YF80; Southern Biotech), Mouse Anti-Human IgG3 (9210-05, Lot#DO720-QB4017; Southern Biotech), and Mouse Anti-Human IgG4 (9200-05, Lot#E1723-RB54B; Southern Biotech).

Techniques: Binding Assay, Peptide Microarray, Sequencing

Journal: ImmunoHorizons

Article Title: ICAM-1 autoantibodies detected in healthy individuals and cross-react with functional epitopes

doi: 10.1093/immhor/vlaf025

Figure Lengend Snippet: Immunoglobulin isotype and subclass determination of anti-ICAM-1 antibodies. (A) Dot plot graph of concentration (µg/mL) on the log y-axis obtained by ELISA for determining the levels of immunoglobulin isotype class within high-autoantibody-presenting plasma, defined as the top 16 positive samples by qualitative ELISA regardless of disease group (n = 16). (B) Dot plot graph of concentration (µg/mL) obtained by ELISA for determining the levels of IgG subclass within high-autoantibody-presenting plasma, defined as the top 16 positive samples by qualitative ELISA regardless of disease group (n = 16). Statistical significance was determined using the Mann-Whitney test. P value results are shown.

Article Snippet: Secondary antibodies were purchased from the following: Goat anti-human IgG (109-036-098, Lot# 149163; Jackson ImmunoResearch), Goat Anti-Human IgA alpha chain (ab97215, Lot# GR3373878-8; Abcam), Goat Anti-Human IgM mu chain (ab97205, Lot# GR3396429-1; Abcam), Mouse Anti-Human IgG1 (9054-05, Lot#B2023-NM34C; Southern Biotech), Mouse Anti-Human IgG2 (9060-05, Lot#K3220-YF80; Southern Biotech), Mouse Anti-Human IgG3 (9210-05, Lot#DO720-QB4017; Southern Biotech), and Mouse Anti-Human IgG4 (9200-05, Lot#E1723-RB54B; Southern Biotech).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, MANN-WHITNEY

Journal: ImmunoHorizons

Article Title: ICAM-1 autoantibodies detected in healthy individuals and cross-react with functional epitopes

doi: 10.1093/immhor/vlaf025

Figure Lengend Snippet: Anti-ICAM-1 IgG autoantibodies interact with protein glycans. (A) SDS-PAGE separation of ICAM-1 under nonreducing conditions. Lane 1 shows ICAM-1 protein, lane 2 shows ICAM-1 in reaction buffer without PNGase F, and lane 3 shows ICAM-1 with PNGase F. Molecular masses are displayed on the left margin in kDa. (B) Dot plot graph of the AUC of the optical density at 450 nm on the log y-axis obtained by ELISA for determining the levels of ICAM-1 IgG autoantibodies targeting ICAM-1 with glycans (control) compared with enzymatically deglycosylated ICAM-1 (PNGASE) within plasma with high levels of detected ICAM-1 autoantibodies, defined as the top 8 positive samples by qualitative ELISA regardless of disease group (n = 8). Statistical significance was determined using the Wilcoxon matched pairs signed rank test. P value results are shown.

Article Snippet: Secondary antibodies were purchased from the following: Goat anti-human IgG (109-036-098, Lot# 149163; Jackson ImmunoResearch), Goat Anti-Human IgA alpha chain (ab97215, Lot# GR3373878-8; Abcam), Goat Anti-Human IgM mu chain (ab97205, Lot# GR3396429-1; Abcam), Mouse Anti-Human IgG1 (9054-05, Lot#B2023-NM34C; Southern Biotech), Mouse Anti-Human IgG2 (9060-05, Lot#K3220-YF80; Southern Biotech), Mouse Anti-Human IgG3 (9210-05, Lot#DO720-QB4017; Southern Biotech), and Mouse Anti-Human IgG4 (9200-05, Lot#E1723-RB54B; Southern Biotech).

Techniques: SDS Page, Enzyme-linked Immunosorbent Assay, Control, Clinical Proteomics