Journal: International journal of molecular sciences

Article Title: Role of Citrullinated Collagen in Autoimmune Arthritis.

doi: 10.3390/ijms23179833

Figure Lengend Snippet: Figure 4. Phosphorylation of human LAIR1 by citrullinated collagen. Jurkat cells that overexpress the human LAIR1 (hLAIR1) were stimulated with PBS, collagen (200 µg/mL), or citrullinated collagen (200 µg/mL) for 5 min. Human LAIR1 was immunoprecipitated from 500 µg of total proteins by anti-human LAIR1 antibody (α-hLAIR1 (NKTA255), Thermo Fisher) using protein A/G bead. Phosphorylation of human LAIR1 was examined with the anti-phosphotyrosine antibody (α -pTyr 4G10, Millipore Human LAIR1 was used as control. The left panel represents type I collagen, and the right panel represents type II collagen. Data are representative of three separate analyses.

Article Snippet: In western blot studies, the anti-human LAIR1 and the anti-phospho-tyrosine specific antibodies were purchased from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Phospho-proteomics, Immunoprecipitation, Control

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: Relationship between LAIR1 expression and intraleukocytic haemozoin. (a) Pairwise comparisons of LAIR1 transcript levels between children with and without pigment-containing monocytes (PCM) and pigment-containing neutrophils (PCN) in the non-SMA and SMA groups were determined using Mann-Whitney U test. Data are presented as box-plots, where the box represents the interquartile range, the line through the box is the median, and whiskers show the 10th and 90th percentiles. LAIR1 transcript levels were lower in children with PCM compared to those without, while LAIR1 transcript levels were comparable between absence/presence of PCN. (b) Temporal kinetics of LAIR1 transcript levels in response to Pf Hz treatment of PBMC from malaria-naïve donors ( n = 6, measured in triplicate). Pairwise comparisons were determined using Student t -test. Significant ( P < .05) differences in transcript levels between no treatment and Pf Hz treatment (10 μg/mL) groups represented by *at 0.5, 2, and 4 h time points. Data represent average of individuals ( n = 3) with each condition performed in triplicate (error bars represent SEM). (c) Immunoblot analysis of non-treated (baseline) and Pf Hz-treated (10 μg/mL) PBMC lysates for 12, 24, and 48 h. (d) Densitometric analysis of normalised cellular LAIR1 protein production presented as mean ± SEM. Across group comparisons analysed using ANOVA. Pairwise comparisons analysed using Student t-test. Data represent average of individuals ( n = 6) with each condition performed in triplicate (error bars represent SEM). LAIR1 protein levels were lower in Pf Hz-treated PBMC lysates compared to no treatment at 12, 24, and 24 h.

Article Snippet: Following electrophoresis, proteins were electrotransferred to iBlot 2 nitrocellulose Mini Stacks (Life Technologies, Carlsbad, CA), and subsequently blocked with 5% blotting grade nonfat dry milk (Bio-Rad Laboratories) in PBS and 0.01% Tween 20 (PBST) at RT for 1 h. Membranes were then probed with anti-LAIR1 antibody (R&D Systems, Inc. Minneapolis, MN) dissolved in fresh blocking buffer.

Techniques: Expressing, MANN-WHITNEY, Western Blot

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: Effect of intraleukocytic Pf Hz on LAIR1 and SHP-1 phosphorylation. Temporal kinetics of the signalling pathway in response to the different treatment conditions was determined in PBMCs from malaria-naïve donors (n = 6, measured in triplicate). (a) Human Phospho-immunoreceptor array results showing LAIR1 and SHP-1 phosphorylation upon no treatment (baseline), C1q (25 μg/mL), Pf Hz (10 μg/mL), and Pf Hz + C1q. Phosphorylation (spots) in the different conditions with control represented by blue boxes, LAIR1 by red boxes, and SHP-1 by green boxes. (b) Densitometric analysis of human phosphor-immunoreceptor array data. Data presented as (mean ± SEM). Across group comparisons analysed using ANOVA. *indicates significant differences ( P < .05) determined by Student t-test in pLAIR1 and pSHP-1 compared to C1q treatment. Phagocytosis of Pf Hz resulted in a reduction of pLAIR1 relative to C1q (alone) treatment. Similarly, ingestion of Pf Hz also caused a marked decrease in pSHP-1 levels relative to C1q treatment. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Following electrophoresis, proteins were electrotransferred to iBlot 2 nitrocellulose Mini Stacks (Life Technologies, Carlsbad, CA), and subsequently blocked with 5% blotting grade nonfat dry milk (Bio-Rad Laboratories) in PBS and 0.01% Tween 20 (PBST) at RT for 1 h. Membranes were then probed with anti-LAIR1 antibody (R&D Systems, Inc. Minneapolis, MN) dissolved in fresh blocking buffer.

Techniques: Phospho-proteomics, Control

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: LAIR1 signalling pathway in malaria. The LAIR1 pathway is activated by attachment of collagen and collagenous ligands (C1q) to LAIR1 extracellular surface. This results in phosphorylation of intracellular LAIR1 ITIM tyrosine residues by Src family kinases. Phosphorylated ITIMs serve as docking sites for recruitment of SHP-1 and SHP-2 phosphatases. SHP-1 and SHP-2 become localized to phosphorylated ITIMs through their regulatory SH2 domains, subsequently inducing their phosphatase activity. Activated SHP-1 has been shown to block activation and nuclear translocation of nuclear factor nuclear factor-kappa beta (NF-κB) through de-phosphorylation of inhibitor of kappa-beta kinase complex (IKK). SHP-1 phosphatase activity also inhibits activation and translocation of IRFs from the cytoplasm to the nucleus by preventing TANK-binding kinase 1 (TANK-1) phosphorylation of interferon regulatory factors (IRFs). These events subsequently block transcription of inflammatory mediator encoding genes with response elements for IRFs and NF-κB. SHP-2 inhibits activation of IRF 8 and blocks expression of phagocyte NADPH oxidase (gp91 PHOX ). LAIR1 inhibitory signalling is regulated by soluble LAIR1 and LAIR2 (soluble homolog of LAIR1) through competition for collagenous ligands. Children with SMA had increased circulating levels of sLAIR1 and sLAIR2 indicative of enhanced receptor shedding. C1q was reduced in children with SMA, thereby, limiting ligand availability. Phagocytosis of Pf Hz antagonizes LAIR1 signalling through down-regulation of LAIR1 ITIM and SHP-1 phosphorylation, but does not alter SHP-2 phosphorylation. Leukocytic ingestion of Pf Hz also decreases LAIR1 transcripts and protein. These events result in NF-κB activation and the consequent production of pro-inflammatory mediators that enhance the pathogenesis of SMA. Red arrows represent ligand-receptor interaction, black solid arrows represent activation, and dashed black arrows represent blockade. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Following electrophoresis, proteins were electrotransferred to iBlot 2 nitrocellulose Mini Stacks (Life Technologies, Carlsbad, CA), and subsequently blocked with 5% blotting grade nonfat dry milk (Bio-Rad Laboratories) in PBS and 0.01% Tween 20 (PBST) at RT for 1 h. Membranes were then probed with anti-LAIR1 antibody (R&D Systems, Inc. Minneapolis, MN) dissolved in fresh blocking buffer.

Techniques: Phospho-proteomics, Activity Assay, Blocking Assay, Activation Assay, Translocation Assay, De-Phosphorylation Assay, Binding Assay, Expressing

Journal: Placenta

Article Title: Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1) is reduced with preeclampsia and small for gestational aged fetuses.

doi: 10.1016/j.placenta.2024.08.018

Figure Lengend Snippet: Fig. 1. LAIR1 mRNA and protein levels are reduced in placentas from women with early onset preeclampsia or small for gestational age. (A) LAIR1 mRNA expression was significantly reduced in placentas from women with early onset preeclampsia (blue triangles, n = 78), compared to gestation matched controls (black dots, n = 30). (B) LAIR1 mRNA expression was significantly reduced in placentas from women who delivered an infant small for gestational age (SGA; <10th birthweight centile, red diamonds, n = 32) compared to gestation matched controls (black dots, n = 30). (C) LAIR1 protein expression was significantly reduced in placentas from women with early onset preeclampsia (blue triangles, n = 43), compared to gestation matched controls (black dots, n = 21). (D) LAIR1 protein expression was significantly reduced in placentas from women who delivered an SGA infant (red diamonds, n = 10) compared to gestation matched controls (black dots, n = 21). Data points represent individual patients. Data are presented as box and whisker plot. **P < 0.01, ****P < 0.0001.

Article Snippet: Levels of LAIR1 in plasma and protein lysates were quantified using the Human LAIR1 DuoSet ELISA kit (DY2664–05, R&D System, Minnesota, USA) according to the manufacturer’s instructions.

Techniques: Expressing, Whisker Assay

Journal: Placenta

Article Title: Leukocyte-associated immunoglobulin-like receptor 1 (LAIR1) is reduced with preeclampsia and small for gestational aged fetuses.

doi: 10.1016/j.placenta.2024.08.018

Figure Lengend Snippet: Fig. 2. LAIR1 mRNA is reduced in placental explants exposed to hypoxia, but not inflammatory cytokines, IL6 or TNFα. Following IL6 treatment (10 ng/mL), LAIR1 mRNA expression (A) and protein expression in placental lysates (B), and media (C) of placental explants were not altered (red squares) compared to controls (0 ng/mL; black dots). Following TNFα treatment (10 ng/mL), LAIR1 mRNA expression (D) and protein expression in placental lysates (E), and media (F) of placental explants are not altered (blue squares) when compared to controls (black dots, 0 ng/mL). Following exposure to hypoxia (1 % O2), LAIR1 mRNA expression (G) is significantly reduced, while protein expression in placental lysates (H), and media (I) of placental explants were not significantly altered (green squares) compared to controls (black dots, 8 % O2). All experiments were done in triplicate, n = 5 patients. Data are presented as mean ± SEM. **P < 0.01.

Article Snippet: Levels of LAIR1 in plasma and protein lysates were quantified using the Human LAIR1 DuoSet ELISA kit (DY2664–05, R&D System, Minnesota, USA) according to the manufacturer’s instructions.

Techniques: Expressing

Journal: Cell Reports Medicine

Article Title: Surface CD52, CD84, and PTGER2 mark mature PMN-MDSCs from cancer patients and G-CSF-treated donors

doi: 10.1016/j.xcrm.2023.101380

Figure Lengend Snippet:

Article Snippet: anti-human LAIR-1-VioBright-FITC (clone REA447) – Lab 1 , Miltenyi Biotec , Cat# 130-106-852; RRID: AB_2657221.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Staining, Cell Isolation, Amplification, Multiplexing, Expressing, Software

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: Relationship between LAIR1 expression and intraleukocytic haemozoin. (a) Pairwise comparisons of LAIR1 transcript levels between children with and without pigment-containing monocytes (PCM) and pigment-containing neutrophils (PCN) in the non-SMA and SMA groups were determined using Mann-Whitney U test. Data are presented as box-plots, where the box represents the interquartile range, the line through the box is the median, and whiskers show the 10th and 90th percentiles. LAIR1 transcript levels were lower in children with PCM compared to those without, while LAIR1 transcript levels were comparable between absence/presence of PCN. (b) Temporal kinetics of LAIR1 transcript levels in response to Pf Hz treatment of PBMC from malaria-naïve donors ( n = 6, measured in triplicate). Pairwise comparisons were determined using Student t -test. Significant ( P < .05) differences in transcript levels between no treatment and Pf Hz treatment (10 μg/mL) groups represented by *at 0.5, 2, and 4 h time points. Data represent average of individuals ( n = 3) with each condition performed in triplicate (error bars represent SEM). (c) Immunoblot analysis of non-treated (baseline) and Pf Hz-treated (10 μg/mL) PBMC lysates for 12, 24, and 48 h. (d) Densitometric analysis of normalised cellular LAIR1 protein production presented as mean ± SEM. Across group comparisons analysed using ANOVA. Pairwise comparisons analysed using Student t-test. Data represent average of individuals ( n = 6) with each condition performed in triplicate (error bars represent SEM). LAIR1 protein levels were lower in Pf Hz-treated PBMC lysates compared to no treatment at 12, 24, and 24 h.

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques: Expressing, MANN-WHITNEY, Western Blot

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: Effect of intraleukocytic Pf Hz on LAIR1 and SHP-1 phosphorylation. Temporal kinetics of the signalling pathway in response to the different treatment conditions was determined in PBMCs from malaria-naïve donors (n = 6, measured in triplicate). (a) Human Phospho-immunoreceptor array results showing LAIR1 and SHP-1 phosphorylation upon no treatment (baseline), C1q (25 μg/mL), Pf Hz (10 μg/mL), and Pf Hz + C1q. Phosphorylation (spots) in the different conditions with control represented by blue boxes, LAIR1 by red boxes, and SHP-1 by green boxes. (b) Densitometric analysis of human phosphor-immunoreceptor array data. Data presented as (mean ± SEM). Across group comparisons analysed using ANOVA. *indicates significant differences ( P < .05) determined by Student t-test in pLAIR1 and pSHP-1 compared to C1q treatment. Phagocytosis of Pf Hz resulted in a reduction of pLAIR1 relative to C1q (alone) treatment. Similarly, ingestion of Pf Hz also caused a marked decrease in pSHP-1 levels relative to C1q treatment. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques:

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: LAIR1 signalling pathway in malaria. The LAIR1 pathway is activated by attachment of collagen and collagenous ligands (C1q) to LAIR1 extracellular surface. This results in phosphorylation of intracellular LAIR1 ITIM tyrosine residues by Src family kinases. Phosphorylated ITIMs serve as docking sites for recruitment of SHP-1 and SHP-2 phosphatases. SHP-1 and SHP-2 become localized to phosphorylated ITIMs through their regulatory SH2 domains, subsequently inducing their phosphatase activity. Activated SHP-1 has been shown to block activation and nuclear translocation of nuclear factor nuclear factor-kappa beta (NF-κB) through de-phosphorylation of inhibitor of kappa-beta kinase complex (IKK). SHP-1 phosphatase activity also inhibits activation and translocation of IRFs from the cytoplasm to the nucleus by preventing TANK-binding kinase 1 (TANK-1) phosphorylation of interferon regulatory factors (IRFs). These events subsequently block transcription of inflammatory mediator encoding genes with response elements for IRFs and NF-κB. SHP-2 inhibits activation of IRF 8 and blocks expression of phagocyte NADPH oxidase (gp91 PHOX ). LAIR1 inhibitory signalling is regulated by soluble LAIR1 and LAIR2 (soluble homolog of LAIR1) through competition for collagenous ligands. Children with SMA had increased circulating levels of sLAIR1 and sLAIR2 indicative of enhanced receptor shedding. C1q was reduced in children with SMA, thereby, limiting ligand availability. Phagocytosis of Pf Hz antagonizes LAIR1 signalling through down-regulation of LAIR1 ITIM and SHP-1 phosphorylation, but does not alter SHP-2 phosphorylation. Leukocytic ingestion of Pf Hz also decreases LAIR1 transcripts and protein. These events result in NF-κB activation and the consequent production of pro-inflammatory mediators that enhance the pathogenesis of SMA. Red arrows represent ligand-receptor interaction, black solid arrows represent activation, and dashed black arrows represent blockade. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques: Activity Assay, Blocking Assay, Activation Assay, Translocation Assay, De-Phosphorylation Assay, Binding Assay, Expressing

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: Relationship between LAIR1 expression and intraleukocytic haemozoin. (a) Pairwise comparisons of LAIR1 transcript levels between children with and without pigment-containing monocytes (PCM) and pigment-containing neutrophils (PCN) in the non-SMA and SMA groups were determined using Mann-Whitney U test. Data are presented as box-plots, where the box represents the interquartile range, the line through the box is the median, and whiskers show the 10th and 90th percentiles. LAIR1 transcript levels were lower in children with PCM compared to those without, while LAIR1 transcript levels were comparable between absence/presence of PCN. (b) Temporal kinetics of LAIR1 transcript levels in response to Pf Hz treatment of PBMC from malaria-naïve donors ( n = 6, measured in triplicate). Pairwise comparisons were determined using Student t -test. Significant ( P < .05) differences in transcript levels between no treatment and Pf Hz treatment (10 μg/mL) groups represented by *at 0.5, 2, and 4 h time points. Data represent average of individuals ( n = 3) with each condition performed in triplicate (error bars represent SEM). (c) Immunoblot analysis of non-treated (baseline) and Pf Hz-treated (10 μg/mL) PBMC lysates for 12, 24, and 48 h. (d) Densitometric analysis of normalised cellular LAIR1 protein production presented as mean ± SEM. Across group comparisons analysed using ANOVA. Pairwise comparisons analysed using Student t-test. Data represent average of individuals ( n = 6) with each condition performed in triplicate (error bars represent SEM). LAIR1 protein levels were lower in Pf Hz-treated PBMC lysates compared to no treatment at 12, 24, and 24 h.

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques: Expressing, MANN-WHITNEY, Western Blot

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: Effect of intraleukocytic Pf Hz on LAIR1 and SHP-1 phosphorylation. Temporal kinetics of the signalling pathway in response to the different treatment conditions was determined in PBMCs from malaria-naïve donors (n = 6, measured in triplicate). (a) Human Phospho-immunoreceptor array results showing LAIR1 and SHP-1 phosphorylation upon no treatment (baseline), C1q (25 μg/mL), Pf Hz (10 μg/mL), and Pf Hz + C1q. Phosphorylation (spots) in the different conditions with control represented by blue boxes, LAIR1 by red boxes, and SHP-1 by green boxes. (b) Densitometric analysis of human phosphor-immunoreceptor array data. Data presented as (mean ± SEM). Across group comparisons analysed using ANOVA. *indicates significant differences ( P < .05) determined by Student t-test in pLAIR1 and pSHP-1 compared to C1q treatment. Phagocytosis of Pf Hz resulted in a reduction of pLAIR1 relative to C1q (alone) treatment. Similarly, ingestion of Pf Hz also caused a marked decrease in pSHP-1 levels relative to C1q treatment. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques:

Journal: EBioMedicine

Article Title: Molecular basis of reduced LAIR1 expression in childhood severe malarial anaemia: Implications for leukocyte inhibitory signalling

doi: 10.1016/j.ebiom.2019.06.040

Figure Lengend Snippet: LAIR1 signalling pathway in malaria. The LAIR1 pathway is activated by attachment of collagen and collagenous ligands (C1q) to LAIR1 extracellular surface. This results in phosphorylation of intracellular LAIR1 ITIM tyrosine residues by Src family kinases. Phosphorylated ITIMs serve as docking sites for recruitment of SHP-1 and SHP-2 phosphatases. SHP-1 and SHP-2 become localized to phosphorylated ITIMs through their regulatory SH2 domains, subsequently inducing their phosphatase activity. Activated SHP-1 has been shown to block activation and nuclear translocation of nuclear factor nuclear factor-kappa beta (NF-κB) through de-phosphorylation of inhibitor of kappa-beta kinase complex (IKK). SHP-1 phosphatase activity also inhibits activation and translocation of IRFs from the cytoplasm to the nucleus by preventing TANK-binding kinase 1 (TANK-1) phosphorylation of interferon regulatory factors (IRFs). These events subsequently block transcription of inflammatory mediator encoding genes with response elements for IRFs and NF-κB. SHP-2 inhibits activation of IRF 8 and blocks expression of phagocyte NADPH oxidase (gp91 PHOX ). LAIR1 inhibitory signalling is regulated by soluble LAIR1 and LAIR2 (soluble homolog of LAIR1) through competition for collagenous ligands. Children with SMA had increased circulating levels of sLAIR1 and sLAIR2 indicative of enhanced receptor shedding. C1q was reduced in children with SMA, thereby, limiting ligand availability. Phagocytosis of Pf Hz antagonizes LAIR1 signalling through down-regulation of LAIR1 ITIM and SHP-1 phosphorylation, but does not alter SHP-2 phosphorylation. Leukocytic ingestion of Pf Hz also decreases LAIR1 transcripts and protein. These events result in NF-κB activation and the consequent production of pro-inflammatory mediators that enhance the pathogenesis of SMA. Red arrows represent ligand-receptor interaction, black solid arrows represent activation, and dashed black arrows represent blockade. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Soluble LAIR1 was measured in serum of study participants using the human LAIR1 ELISA matched antibody pair, and recombinant human LAIR1 (Creative Diagnostics, 45–16 Ramsey Rd, Shirley, NY 11967; Cat No. ABPR-0519).

Techniques: Activity Assay, Blocking Assay, Activation Assay, Translocation Assay, De-Phosphorylation Assay, Binding Assay, Expressing

Journal: bioRxiv

Article Title: IL-21-STAT3 axis negatively regulates LAIR1 expression in B cells

doi: 10.1101/2025.01.14.632971

Figure Lengend Snippet: (A) LAIR1 expression in total B cells (left) and the compositions of subsets in LAIR+ and LAIR1-B cells (right) (N=10). (B-D) LAIR1 expression in major B cell subsets (B; N=10), the different class of surface Ig (C; N=5) and major B cell subsets (D; N=9). (E) heatmap and volcano plot generated from RNA-seq data between LAIR1+ and LAIR1- SWM B cells (N=4). Differential gene expression was identified with Log 2 FC >0.5 and adjusted p value <0.05. Upregulated genes in LAIR1- are 278 genes and down-regulated genes in LAIR1- are 218 genes. (F) the potential of PC differentiation induced by TLR7 or TLR9 signaling. (G) the percentages of ANA+ autoreactive B cells in LAIR1+ and LAIR1- memory fraction (N=7). Data are shown as mean ± SEM with each symbol representing an individual subjects. P values were calculated with the Wilcoxon signed rank tests (F and G) and further adjusted for multiple comparisons using the Benjamini-Hochberg method (B, C and D). Asterisks indicate significant differences (* P < 0.05, ** P < 0.01).

Article Snippet: B cells (2.0×10 6 cells) isolated with EasySep human B cell isolation kit (Stem cell technologies) were rested in X-vivo medium (Lonza) at 37°C for 30 min and subsequently stimulated with biotinylated anti-LAIR1 polyclonal antibody (1.0 μg/mL, R&D Systems) or biotinylated isotype antibody (1.0 μg/mL, R&D Systems) in the presence of streptavidin (10 μg/mL, R&D Systems) for 20 min.

Techniques: Expressing, Generated, RNA Sequencing, Gene Expression

Journal: bioRxiv

Article Title: IL-21-STAT3 axis negatively regulates LAIR1 expression in B cells

doi: 10.1101/2025.01.14.632971

Figure Lengend Snippet: (A) Phosphorylated LAIR1 induced by its crosslinking with biotinylated anti-LAIR1 antibody and streptavidin (N=5). (B-D) The effects of co-crosslinking with BCR and LAIR1 in naïve B cells (B; N=5), IgM+ USWM B cells (C; N=5) and IgG+ SWM B cells (D; N=5). Data are shown as mean ± SEM with each symbol representing an individual subjects. P values were calculated with the Wilcoxon signed rank tests. Asterisks indicate significant differences (* P < 0.05).

Article Snippet: B cells (2.0×10 6 cells) isolated with EasySep human B cell isolation kit (Stem cell technologies) were rested in X-vivo medium (Lonza) at 37°C for 30 min and subsequently stimulated with biotinylated anti-LAIR1 polyclonal antibody (1.0 μg/mL, R&D Systems) or biotinylated isotype antibody (1.0 μg/mL, R&D Systems) in the presence of streptavidin (10 μg/mL, R&D Systems) for 20 min.

Techniques:

Journal: bioRxiv

Article Title: IL-21-STAT3 axis negatively regulates LAIR1 expression in B cells

doi: 10.1101/2025.01.14.632971

Figure Lengend Snippet: (A-B) Representative examples of LAIR1 expression in 7 subsets of B cells (A) and activated naïve B cells and DN2 B cells (B) from HD and SLE patients (each N=9). (C) The positivity of LAIR1 in ANA+ and ANA- B cells. (D) The expression level of LAIR1 in LAIR1 expressing ANA+ autoreactive B cells between HD (N=9) and SLE patients (N=8). Data are shown as mean ± SEM with each symbol representing an individual subjects. P values were calculated with the Mann-Whitney U test (A, B and D) or the Wilcoxon signed rank tests (C). Asterisks indicate significant differences (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: B cells (2.0×10 6 cells) isolated with EasySep human B cell isolation kit (Stem cell technologies) were rested in X-vivo medium (Lonza) at 37°C for 30 min and subsequently stimulated with biotinylated anti-LAIR1 polyclonal antibody (1.0 μg/mL, R&D Systems) or biotinylated isotype antibody (1.0 μg/mL, R&D Systems) in the presence of streptavidin (10 μg/mL, R&D Systems) for 20 min.

Techniques: Expressing, MANN-WHITNEY

Journal: bioRxiv

Article Title: IL-21-STAT3 axis negatively regulates LAIR1 expression in B cells

doi: 10.1101/2025.01.14.632971

Figure Lengend Snippet: (A) LAIR1 expression on B cells after 5 days co-culture with cTfh cells activated with anti-CD3/28 Beads in the presence of IL-21R-Fc protein (20 μg/mL) or isotype control (N=5). (B) The effects of IL-21 on LAIR1 expression in naïve B cells co-stimulated with CD40 or CD40/BCR in 3 days culture (N=12). (C) The effects of IL-21 on LAIR1 expression in SWM B cells co-stimulated with CD40 in 5 days culture (N=5). Data are shown as mean ± SEM with each symbol representing an individual subjects. P values were calculated with the Wilcoxon signed rank tests (C) and further adjusted for multiple comparisons using the Benjamini-Hochberg method (A and B). Asterisks indicate significant differences (* P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: B cells (2.0×10 6 cells) isolated with EasySep human B cell isolation kit (Stem cell technologies) were rested in X-vivo medium (Lonza) at 37°C for 30 min and subsequently stimulated with biotinylated anti-LAIR1 polyclonal antibody (1.0 μg/mL, R&D Systems) or biotinylated isotype antibody (1.0 μg/mL, R&D Systems) in the presence of streptavidin (10 μg/mL, R&D Systems) for 20 min.

Techniques: Expressing, Co-Culture Assay, Control

Journal: bioRxiv

Article Title: IL-21-STAT3 axis negatively regulates LAIR1 expression in B cells

doi: 10.1101/2025.01.14.632971

Figure Lengend Snippet: (A) The effects of IL-21 on LAIR1 expression in naïve B cells co-stimulated with TLR9 or TLR9/BCR in 3 days culture (N=7) (B) The effects of IL-21 on LAIR1 expression in ABCs differentiation from naïve B cells in 3 days culture (N=7). (C) The effects of IL-21 on LAIR1 expression in SWM B cells co-stimulated with TLR9 in 5 days culture (N=7). Data are shown as mean ± SEM with each symbol representing an individual subjects. P values were calculated with the Wilcoxon signed rank tests (B) and further adjusted for multiple comparisons using the Benjamini–Hochberg method (A and C). Asterisks indicate significant differences (* P < 0.05).

Article Snippet: B cells (2.0×10 6 cells) isolated with EasySep human B cell isolation kit (Stem cell technologies) were rested in X-vivo medium (Lonza) at 37°C for 30 min and subsequently stimulated with biotinylated anti-LAIR1 polyclonal antibody (1.0 μg/mL, R&D Systems) or biotinylated isotype antibody (1.0 μg/mL, R&D Systems) in the presence of streptavidin (10 μg/mL, R&D Systems) for 20 min.

Techniques: Expressing

Journal: bioRxiv

Article Title: IL-21-STAT3 axis negatively regulates LAIR1 expression in B cells

doi: 10.1101/2025.01.14.632971

Figure Lengend Snippet: (A) The effects of IL-21 on LAIR1 mRNA expression in naïve B cells co-stimulated with TLR9 or CD40/BCR in 2 days culture (N=6) (B) The effects of IL-21 on LAIR1 mRNA expression in naïve B cells in 2-24 hours culture (N=7). (C) IL-21-induced phosphorylation (left) and nuclear translocation (right) of STAT1/3/5 (N=3). (D) Amplification of IL-21-induced pSTAT3 level by TLR9 or CD40/BCR in the naïve B cell culture (N=4-5). (E-G) the effect of SD36 (1 μM) against STAT family proteins for 3.5 hours (E; N=3), LAIR1 mRNA expression change induced by IL-21 (F; N=6), pSTAT3 and the protein levels of STAT3 (G; N=6) and LAIR1 (H; N=11). Data are shown as mean ± SEM with each symbol representing an individual subjects. P values were calculated with the Wilcoxon signed rank tests (A, B, D, F and G). Asterisks indicate significant differences (* P < 0.05, ** P < 0.01).

Article Snippet: B cells (2.0×10 6 cells) isolated with EasySep human B cell isolation kit (Stem cell technologies) were rested in X-vivo medium (Lonza) at 37°C for 30 min and subsequently stimulated with biotinylated anti-LAIR1 polyclonal antibody (1.0 μg/mL, R&D Systems) or biotinylated isotype antibody (1.0 μg/mL, R&D Systems) in the presence of streptavidin (10 μg/mL, R&D Systems) for 20 min.

Techniques: Expressing, Phospho-proteomics, Translocation Assay, Amplification, Cell Culture

Journal: bioRxiv

Article Title: IL-21-STAT3 axis negatively regulates LAIR1 expression in B cells

doi: 10.1101/2025.01.14.632971

Figure Lengend Snippet: (A) STAT3 putative binding site in LAIR1 promoter region and its sequence (B) the binding of STAT3 of nuclear extract from IL-21 treated naïve B cells to LAIR1 promoter region (sequence #3) (N=3). P values were calculated with the Paired T test (C). Asterisk indicates significant difference (* P < 0.05).

Article Snippet: B cells (2.0×10 6 cells) isolated with EasySep human B cell isolation kit (Stem cell technologies) were rested in X-vivo medium (Lonza) at 37°C for 30 min and subsequently stimulated with biotinylated anti-LAIR1 polyclonal antibody (1.0 μg/mL, R&D Systems) or biotinylated isotype antibody (1.0 μg/mL, R&D Systems) in the presence of streptavidin (10 μg/mL, R&D Systems) for 20 min.

Techniques: Binding Assay, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Role of Citrullinated Collagen in Autoimmune Arthritis

doi: 10.3390/ijms23179833

Figure Lengend Snippet: LAIR-1 mediates suppression of CIA. Two groups ( n = 10 per group) of DR1/LAIR-1 +/+ mice and two groups of littermate DR1/LAIR-1 − /− mice were immunized with type II collagen/CFA. Then, each mouse was administered intraperitoneally a dose of either 100µg CI or PBS weekly for four weeks beginning one week after immunization. Each mouse was scored three times weekly beginning 2 weeks post immunization. The severity of the arthritis was statistically significant comparing CI treatment with PBS in LAIR-1 +/+ mice beginning on day 44 ( p ≤ 0.05) post immunization and continuing until the end of the experiment on day 56. ( p ≤ 0.001).

Article Snippet: The plates were treated with either an HRP-conjugated monoclonal antibody against murine LAIR-1 (Novus #NBP1-43324H) Novus Biologicals, (Centennial CO, USA) or a monoclonal antibody against human LAIR-1 (R&D #2664).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Role of Citrullinated Collagen in Autoimmune Arthritis

doi: 10.3390/ijms23179833

Figure Lengend Snippet: Cit-Collagen binds to LAIR with similar affinity as native Collagen. Wells were coated with CI, cit-CI, CII, or cit-CII before adding recombinant LAIR-1 (murine LAIR-1 in the ( upper panel ) and human LAIR-1 in the ( lower panel )) at the indicated concentrations. Binding was detected using an HRP conjugated α -murine LAIR-1 ab, or an α-human LAIR-1 ab followed by a biotinylated α IgG and streptavidin -HRP with reading of absorbance at 450. The control protein, ovalbulmin did not bind the collagens murine LAIR-1 or human LAIR-1.

Article Snippet: The plates were treated with either an HRP-conjugated monoclonal antibody against murine LAIR-1 (Novus #NBP1-43324H) Novus Biologicals, (Centennial CO, USA) or a monoclonal antibody against human LAIR-1 (R&D #2664).

Techniques: Recombinant, Binding Assay