Journal: International Journal of Molecular Sciences

Article Title: Polymerized Type I Collagen Downregulates STAT-1 Phosphorylation Through Engagement with LAIR-1 in Circulating Monocytes, Avoiding Long COVID

doi: 10.3390/ijms26031018

Figure Lengend Snippet: Binding of PTIC to LAIR1 and its effect on M1. ( A ) ELISA binding assay of PTIC or CI to LAIR; ( B ) SRP binding assay of PTIC or CI to LAIR1. M1 stimulated with anti-LAIR1 antibody (1:100) for 24 h to detect ( C ) M1 (CD16 + /CD36 + /CD86 + /IL-1β + ) and ( D ) M2 (CD14 + /CD16 hi /CD163 + /IL-10 + ). M1 stimulated with anti-LAIR1 antibody (1:100) + 10% PTIC for 24 h to detect ( E ) M1 and ( F ) M2; ( G ) M1 and M2 cell percentage. ( H ) Cytokine mRNA expression in M1 macrophages. They were treated with a constant stimulus of 20 ng/mL of IFN-γ (pink squares). M1 macrophages were treated with a constant stimulus of 20 ng/mL of IFN-γ and 10% PTIC (green circles), and M1 macrophages were cultured with 10% PTIC (blue triangles). All values were reported as ΔΔCT.

Article Snippet: Subsequently, a 1:500 dilution mouse anti-human LAIR1 (HycultBiotech # HM2364-100UG) was added overnight at 4 °C.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: Polymerized Type I Collagen Downregulates STAT-1 Phosphorylation Through Engagement with LAIR-1 in Circulating Monocytes, Avoiding Long COVID

doi: 10.3390/ijms26031018

Figure Lengend Snippet: The binding of PTIC to LAIR1 induces downregulation of STAT1 phosphorylation. ( A ) Western blotting of the relative expression of STAT1 and p STAT1. The bands were analyzed based on normalization to actin expression to eliminate inconsistencies caused by the amount of protein in the lane. ( B ) Relative expression of STAT1 and pSTAT1. Both were normalized for actin and were adjusted to 1 unit. ( C ) Effect of STAT1 phosphorylation by activating LAIR1 with anti-LAIR1 antibody in M1. The bands were analyzed based on normalization to actin expression to eliminate inconsistencies caused by the amount of protein in the lane. ( D ) Relative expression of phosphorylated STAT1.

Article Snippet: Subsequently, a 1:500 dilution mouse anti-human LAIR1 (HycultBiotech # HM2364-100UG) was added overnight at 4 °C.

Techniques: Binding Assay, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Polymerized Type I Collagen Downregulates STAT-1 Phosphorylation Through Engagement with LAIR-1 in Circulating Monocytes, Avoiding Long COVID

doi: 10.3390/ijms26031018

Figure Lengend Snippet: Representative flow plots of circulating Mo1 subset in SARS-CoV2-infected symptomatic outpatients at the baseline 8, 15, and 90 days post-treatment with PTIC ( n = 20) or a placebo ( n = 20). CD86 + /CD11c + /CD3 − /IP-10 + -expressing cells in ( A ) healthy donors and patients at ( B ) the baseline, ( C ) 8 days, ( D ) 15 days, and ( E ) 90 days post-treatment. ( F ) Flow plots at the baseline (red) and 90 days post-treatment with PTIC (blue). ( G ) CD86 + /CD11c + /CD3 − /IP-10 + -producing cells are expressed as a mean ± SEM. ( H ) CD14 + /CD11c + /CD86 + /LAIR1 + -producing cells are expressed as a mean ± SEM. Blue stars show the day the treatment reached p < 0.05 compared to the PTIC treatment baseline. Pink stars depict the day the therapy reached p < 0.05 compared to the baseline for the placebo. The numbers on the graph represent the statistical significance between the patients treated with PTIC vs. the placebo. ** p ≤ 0.001 depict statistically significant differences from the baseline (blue: PTIC, pink: placebo, green: healthy donors).

Article Snippet: Subsequently, a 1:500 dilution mouse anti-human LAIR1 (HycultBiotech # HM2364-100UG) was added overnight at 4 °C.

Techniques: Infection, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Polymerized Type I Collagen Downregulates STAT-1 Phosphorylation Through Engagement with LAIR-1 in Circulating Monocytes, Avoiding Long COVID

doi: 10.3390/ijms26031018

Figure Lengend Snippet: Representative flow plots of circulating Mo2 subset in SARS-CoV2-infected symptomatic outpatients at the baseline 8, 15, and 90 days post-treatment with PTIC ( n = 20) or a placebo ( n = 20). CD11b + /CD16 + /CD163 + /IDO + -expressing cells in ( A ) healthy donors and patients at ( B ) the baseline, ( C ) 8 days, ( D ) 15 days, and ( E ) 90 days post-treatment. ( F ) Flow plots at the baseline (red) and 90 days post-treatment with PTIC (blue). ( G ) CD16 + /CD11b + /CD163 + /IDO + -producing cells are expressed as a mean ± SEM. ( H ) CD16 + /CD11b + /CD163 + /LAIR1 + -producing cells are expressed as a mean ± SEM. Blue stars show the day the treatment reached p < 0.05 compared to the PTIC treatment the baseline. Pink stars depict the day the treatment reached p < 0.05 compared to the baseline for the placebo. The numbers on the graph represent the statistical significance between the patients treated with PTIC vs. the placebo. * p ≤ 0.05 and ** p ≤ 0.001 depict statistically significant differences from the baseline (blue: PTIC, pink: placebo, green: healthy donors).

Article Snippet: Subsequently, a 1:500 dilution mouse anti-human LAIR1 (HycultBiotech # HM2364-100UG) was added overnight at 4 °C.

Techniques: Infection, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Polymerized Type I Collagen Downregulates STAT-1 Phosphorylation Through Engagement with LAIR-1 in Circulating Monocytes, Avoiding Long COVID

doi: 10.3390/ijms26031018

Figure Lengend Snippet: PTIC regulates STAT1 phosphorylation through LAIR1 in M1 and favors polarization towards M2. ( A ) Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1 or CD305) is a type I transmembrane glycoprotein that contains one extracellular Ig-like domain and two immunoreceptor tyrosine-based inhibitory motifs (ITIMs) in its intracellular domain. LAIR1 is expressed in most hematopoietic lineages, including monocytes, macrophages, dendritic cells (DCs), natural killer (NK) cells, and many T and B cell populations. Its extracellular domain binds to glycine–proline–hydroxyproline collagen repeats, and its ITIMs recruit the phosphatases SHP1 and SHP2. Collagens, C1q, MBL, surface protein-D (SP-D), Rifins, and Colec12 have been reported as ligands for LAIR1. It downregulates T, B, and natural killer (NK) cell functions by the recruitment of the SHP1 and SHP2 phosphatases. ( B ) The pre-polarized (M0) macrophage subsets challenge LPS/IFN-γ and induce polarization to M1. The human monocytic cell line THP-1 expresses high levels of LAIR1. ( C ) LAIR1 binding type I collagen regulates the immune system balance and protects against tissue damage against a hyperactive immune response or autoimmune dysfunction through SHP1, SHP2, CSK, and pSTAT1 intracellular signaling. ( D ) Polymerized type I collagen induces the downregulation of the phosphorylation of STAT1 in M1 and promotes polarization to M2. The arrow pointing downwards means downregulation. The arrow pointing upwards means upregulation. The horizontal arrow means polarization.

Article Snippet: Subsequently, a 1:500 dilution mouse anti-human LAIR1 (HycultBiotech # HM2364-100UG) was added overnight at 4 °C.

Techniques: Binding Assay