human lair1  (R&D Systems)

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

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: 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