alair1  (R&D Systems)

Journal: JCI Insight

Article Title: LAIR1 prevents excess inflammatory tissue damage in Staphylococcus aureus skin infection and cutaneous T cell lymphoma

doi: 10.1172/jci.insight.183935

Figure Lengend Snippet: ( A – C ) CD4 + cells were isolated from CTCL patient peripheral blood and analyzed by single cell RNA-Seq. UMAP projections show 92,496 mononuclear cells from 16 peripheral blood samples from 6 patients with CTCL across multiple time points. ( A ) Cell type assignment by highest spearman rho of purified immune populations in the Human Primary Cell Atlas. ( B and C ) UMAP plots show expression of LAIR1 ( B ) and LAIR2 ( C ). ( D ) Illustration depicts hypothesis that LAIR2 overexpression reduces LAIR1 signaling in CTCL. ( E ) Illustration depicts mouse model for loss of LAIR1 signaling via KO of Lair1 .

Article Snippet: WT C57BL/6J and Lair1 –/– (B6.Cg-Lair1tm1.1Jco/J, JAX Strain #:032788) mice ( ) were purchased from The Jackson Laboratory and housed in a pathogen-free environment in the WUSM animal facility.

Techniques: Isolation, Single Cell, RNA Sequencing, Purification, Expressing, Over Expression

Journal: JCI Insight

Article Title: LAIR1 prevents excess inflammatory tissue damage in Staphylococcus aureus skin infection and cutaneous T cell lymphoma

doi: 10.1172/jci.insight.183935

Figure Lengend Snippet: WT and Lair1- KO mice were infected s.c. with 1 × 10 7 CFU S . aureus USA300 LAC. ( A and B ) Lesions were monitored over 14 days and quantified as abscess ( A ) and dermonecrosis ( B ) area (noninvasive measurements, area calculated: A = ([pi/2] × l × w). Statistical analysis by repeated measures 2-way ANOVA with Bonferroni post hoc tests. Results are representative of 5 independent experiments ( n = 40 WT, n = 40 Lair1 KO). ( C ) Representative gross images of S . aureus skin lesions in WT (left) and Lair1 -KO (right) mice on 4dpi. Abscess boundaries are indicated with the dashed lines. Dermonecrosis is the central erythematous lesion. ( D ) Representative H&E stains of lesions at 2 dpi. Scale bars: 1 mm. Asterisks indicate abscesses, and solid lines indicate areas of dermonecrosis, which are larger in Lair1 KO.

Article Snippet: WT C57BL/6J and Lair1 –/– (B6.Cg-Lair1tm1.1Jco/J, JAX Strain #:032788) mice ( ) were purchased from The Jackson Laboratory and housed in a pathogen-free environment in the WUSM animal facility.

Techniques: Infection

Journal: JCI Insight

Article Title: LAIR1 prevents excess inflammatory tissue damage in Staphylococcus aureus skin infection and cutaneous T cell lymphoma

doi: 10.1172/jci.insight.183935

Figure Lengend Snippet: ( A – F ) Skin lesions were collected at 18 hours after infection, homogenized, and cytokine production measured by multiplex cytokine array. ( A ) Heatmap shows average WT and Lair1 KO z -scored cytokine concentrations. ( B – F ) Bar graphs show all cytokines with significantly different expression between WT and Lair1 KO. ( G – L ) BMDMs were generated in vitro and treated with PBS or S . aureus at a multiplicity of infection (MOI) of 10:1 for 18 hours. Cell supernatant was analyzed by multiplex cytokine array. ( G ) Heatmap shows average WT and Lair1- KO cytokine concentrations in PBS and S . aureus groups. ( H – L ) Bar graphs show all cytokines with significantly different expression between WT and Lair1 KO ( n = 2 independent infections, 2–6 mice per group). ( B – F and G – L ) Two-way ANOVA with Fisher’s LSD for multiple comparisons. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: WT C57BL/6J and Lair1 –/– (B6.Cg-Lair1tm1.1Jco/J, JAX Strain #:032788) mice ( ) were purchased from The Jackson Laboratory and housed in a pathogen-free environment in the WUSM animal facility.

Techniques: Infection, Multiplex Assay, Expressing, Generated, In Vitro

Journal: JCI Insight

Article Title: LAIR1 prevents excess inflammatory tissue damage in Staphylococcus aureus skin infection and cutaneous T cell lymphoma

doi: 10.1172/jci.insight.183935

Figure Lengend Snippet: ( A and C ) BMDM from WT and Lair1-KO mice were treated with PBS or PAM3CSK4 for 6 hours and were then subjected to RNA-Seq. ( B and D ) Skin punch biopsies from WT and Lair1 -KO mouse S . aureus skin infections were taken at 2 dpi and were then subjected to RNA-Seq. GSEA was used to define the top 20 mouse Reactome pathways with enrichment scores and nominal P value in genes upregulated in Lair1 KO in BMDM treated with PAM3CSK4 ( A ) and 2 dpi SSTI ( B ); collagen-related pathways common to both analyses are bolded in teal. Genes from the 4 pathways, many of which overlapped, were classified into 3 general categories: collagen genes, collagen synthesis genes, and extracellular matrix (ECM) remodeling genes. Heatmaps show z -scored expression for these genes for BMDM ( C ) and S . aureus SSTI ( D ). ( E and F ) BMDM from WT and Lair1 -KO mice were treated with PBS or PAM3CSK4 for 24 hours and were then fixed and stained for Serpin H1. Representative images from 3 independent experiments are shown ( E ). Scale bars: 50 μm. Images were analyzed in CellProfiler and mean Serpin H1 cytoplasmic fluorescence intensity per cell was calculated ( F ). Mean ± SEM are indicated by black bars. Statistical analysis was done by 2-way ANOVA with multiple comparisons.

Article Snippet: WT C57BL/6J and Lair1 –/– (B6.Cg-Lair1tm1.1Jco/J, JAX Strain #:032788) mice ( ) were purchased from The Jackson Laboratory and housed in a pathogen-free environment in the WUSM animal facility.

Techniques: RNA Sequencing, Expressing, Staining, Fluorescence

Journal: JCI Insight

Article Title: LAIR1 prevents excess inflammatory tissue damage in Staphylococcus aureus skin infection and cutaneous T cell lymphoma

doi: 10.1172/jci.insight.183935

Figure Lengend Snippet: WT and Lair1- KO neutrophils were isolated from mouse bone marrow. ( A ) Amplex red fluorescence measured by plate reader indicates hydrogen peroxide (H 2 O 2 ) production in neutrophils treated with PBS or S . aureus at MOI 50:1 for 30 minutes. ( B ) Ratio of WT/KO lysosome probe MFI measured by flow cytometry indicates lysosome acidification in neutrophils treated with PBS or S . aureus at MOI 100:1 for 30 minutes. ( C ) Neutrophils incubated for 30 minutes with PBS or S . aureus bioparticles (pHrodo) that fluoresce when acidified in the lysosome were subjected to flow cytometry. ( D ) Neutrophils incubated for 30 minutes with PBS or mCherry-expressing S . aureus at MOI 50:1 were subjected to flow cytometry. mCherry MFI is shown as a ratio of KO to WT. Statistical analysis for all panels was done by 2-way ANOVA with Fisher’s LSD. ** P < 0.01.

Article Snippet: WT C57BL/6J and Lair1 –/– (B6.Cg-Lair1tm1.1Jco/J, JAX Strain #:032788) mice ( ) were purchased from The Jackson Laboratory and housed in a pathogen-free environment in the WUSM animal facility.

Techniques: Isolation, Fluorescence, Flow Cytometry, Incubation, 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: After a washing step, the sorted cells were rested in X-vivo medium at 37°C for 30 min and 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 absence or presence of streptavidin (5 μg/mL, Sigma-Aldrich) 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: After a washing step, the sorted cells were rested in X-vivo medium at 37°C for 30 min and 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 absence or presence of streptavidin (5 μg/mL, Sigma-Aldrich) 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: After a washing step, the sorted cells were rested in X-vivo medium at 37°C for 30 min and 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 absence or presence of streptavidin (5 μg/mL, Sigma-Aldrich) 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: After a washing step, the sorted cells were rested in X-vivo medium at 37°C for 30 min and 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 absence or presence of streptavidin (5 μg/mL, Sigma-Aldrich) 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: After a washing step, the sorted cells were rested in X-vivo medium at 37°C for 30 min and 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 absence or presence of streptavidin (5 μg/mL, Sigma-Aldrich) 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: After a washing step, the sorted cells were rested in X-vivo medium at 37°C for 30 min and 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 absence or presence of streptavidin (5 μg/mL, Sigma-Aldrich) 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: After a washing step, the sorted cells were rested in X-vivo medium at 37°C for 30 min and 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 absence or presence of streptavidin (5 μg/mL, Sigma-Aldrich) for 20 min.

Techniques: Binding Assay, Sequencing