Journal: bioRxiv

Article Title: Immune receptor LILRB1 mediates cis -signalling which is targeted by RIFINs of the malaria parasite

doi: 10.64898/2026.04.16.718981

Figure Lengend Snippet: a) Gating strategy to assess LILRB1 expression on NK92 cells. b) Expression of LILRB1 on NK92 cells through either anti-LILRB1 staining (left) or GFP expression (right). c) Gating strategy to assess expression of HLA-A/B/C on HEK293 cells. d) Expression of HLA-A/B/C on HEK293 WT or HEK293 β2M −/- cells.

Article Snippet: Primary antibodies were anti-HLA-A,B,C Alexa Fluor 647 (Biolegend, 311414) and anti-LILRB1 (R&D systems, AF2017).

Techniques: Expressing, Staining

Journal: Cellular immunology

Article Title: In vitro ovarian tumor-conditioned CD163+ human macrophages retain phagocytic response to CD47 blockade.

doi: 10.1016/j.cellimm.2025.104932

Figure Lengend Snippet: Fig. 2. Tumor conditioning induces macrophage CD163 and IL-10 gene expression. Tumor-conditioned human monocyte-derived macrophages (M(A2780)) were generated and their gene expression was compared to control macrophages by RT-qPCR. A-F: Presents gene expression of polarization markers (CD163, IL-10, and TNF-α) and phagocytosis checkpoints (SIRPα, LILRB1, and Siglec-10) in tumor-associated macrophages compared to controls. Statistics: comparisons were made against controls using paired t-tests and a significance level of p = 0.05. When indicated by a logarithmic y-axis, comparisons were made on log-transformed data. Data are presented as individual data points with mean ± SD. N = 4.

Article Snippet: To measure sLILRB1, microtitre plates were coated with polyclonal anti-human LILRB1 antibody (R&D systems, AF2017) and incubated overnight.

Techniques: Gene Expression, Derivative Assay, Generated, Control, Quantitative RT-PCR, Transformation Assay

Journal: Cellular immunology

Article Title: In vitro ovarian tumor-conditioned CD163+ human macrophages retain phagocytic response to CD47 blockade.

doi: 10.1016/j.cellimm.2025.104932

Figure Lengend Snippet: Fig. 3. Tumor cell conditioning induces CD163, CD206, CD80 and LILRB1 on the membrane protein level. Tumor-conditioned human monocyte-derived macrophages (M(A2780)) were generated and their membrane protein expression compared to control macrophages by flow cytometry. The gating strategy is presented in Supplementary fig. 1. A-G: Presents median fluorescence intensities (MFI) of polarization markers (CD163, CD206, and CD80) and MFI values and percent macrophages positive for phagocytosis checkpoints (LILRB1, and Siglec-10). Statistics: comparisons were made against controls using paired t-tests and a significance level of p = 0.05. When indicated by a logarithmic y-axis, comparisons were made on log-transformed data. Data are presented as individual data points with mean ± SD. N = 9.

Article Snippet: To measure sLILRB1, microtitre plates were coated with polyclonal anti-human LILRB1 antibody (R&D systems, AF2017) and incubated overnight.

Techniques: Membrane, Derivative Assay, Generated, Expressing, Control, Flow Cytometry, Fluorescence, Transformation Assay

Journal: Cellular immunology

Article Title: In vitro ovarian tumor-conditioned CD163+ human macrophages retain phagocytic response to CD47 blockade.

doi: 10.1016/j.cellimm.2025.104932

Figure Lengend Snippet: Fig. 2. Tumor conditioning induces macrophage CD163 and IL-10 gene expression. Tumor-conditioned human monocyte-derived macrophages (M(A2780)) were generated and their gene expression was compared to control macrophages by RT-qPCR. A-F: Presents gene expression of polarization markers (CD163, IL-10, and TNF-α) and phagocytosis checkpoints (SIRPα, LILRB1, and Siglec-10) in tumor-associated macrophages compared to controls. Statistics: comparisons were made against controls using paired t-tests and a significance level of p = 0.05. When indicated by a logarithmic y-axis, comparisons were made on log-transformed data. Data are presented as individual data points with mean ± SD. N = 4.

Article Snippet: After block and wash, culture supernatants (diluted 1:2), controls and standards were applied and incubated for 1 h. Subsequently, monoclonal anti-human LILRB1 antibody (R&D systems, MAB2017) was added and incubated for 1 h. After washing, polyclonal anti-mouse horseradish peroxidase-conjugated antibodies (Dako, P0447) were added and incubated for 1 h. After wash, TMB One (Kementec, 4380 L) was added, and the plates were incubated in the dark and stopped by 1 M phosphoric acid.

Techniques: Gene Expression, Derivative Assay, Generated, Control, Quantitative RT-PCR, Transformation Assay

Journal: Cellular immunology

Article Title: In vitro ovarian tumor-conditioned CD163+ human macrophages retain phagocytic response to CD47 blockade.

doi: 10.1016/j.cellimm.2025.104932

Figure Lengend Snippet: Fig. 3. Tumor cell conditioning induces CD163, CD206, CD80 and LILRB1 on the membrane protein level. Tumor-conditioned human monocyte-derived macrophages (M(A2780)) were generated and their membrane protein expression compared to control macrophages by flow cytometry. The gating strategy is presented in Supplementary fig. 1. A-G: Presents median fluorescence intensities (MFI) of polarization markers (CD163, CD206, and CD80) and MFI values and percent macrophages positive for phagocytosis checkpoints (LILRB1, and Siglec-10). Statistics: comparisons were made against controls using paired t-tests and a significance level of p = 0.05. When indicated by a logarithmic y-axis, comparisons were made on log-transformed data. Data are presented as individual data points with mean ± SD. N = 9.

Article Snippet: After block and wash, culture supernatants (diluted 1:2), controls and standards were applied and incubated for 1 h. Subsequently, monoclonal anti-human LILRB1 antibody (R&D systems, MAB2017) was added and incubated for 1 h. After washing, polyclonal anti-mouse horseradish peroxidase-conjugated antibodies (Dako, P0447) were added and incubated for 1 h. After wash, TMB One (Kementec, 4380 L) was added, and the plates were incubated in the dark and stopped by 1 M phosphoric acid.

Techniques: Membrane, Derivative Assay, Generated, Expressing, Control, Flow Cytometry, Fluorescence, Transformation Assay

Journal: bioRxiv

Article Title: Natural killer cell function is regulated by TGF-β signaling in pregnancy and tumor progression

doi: 10.1101/2024.09.18.613652

Figure Lengend Snippet: a, Heatmap showing gene-gene correlation of top variable genes within NK metacells with gene modules from three distinct NK subsets. b , Barplot showing the top 30 genes that are most correlated with GZMA across NK metacells. This set of 30 cytotoxic genes is subsequently used to calculate cytotoxic scores. Scatterplots depicting the cytotoxic score per metacell versus log enrichment of a selected set of chemotactic genes. c , Similar to panel b but showing the top 30 genes most correlated with XCL1 , defining the chemotactic score, and scatterplots depicting per metacell the chemotactic score versus log enrichment of several cytotoxic genes. d , Barplots showing expression fold change (log2 scale) of selected genes in human cytotoxic and chemotactic decidual NK cells when compared with circulating blood NK cells. Genes are grouped into gene families. e , Scatterplot depicting cytotoxic score versus chemotactic score in all NK metacells. f , Heatmap showing expression levels from Mass Cytometry time-of-flight (CyTOF) assay of selected proteins (NCAM1 (CD56), FCGR3A (CD16) and ENTPD1 (CD39)) on a tSNE map of all analyzed CD56+, CD3-cells from healthy donor peripheral blood and representative decidua sample. g , Paired point plot showing intracellular protein levels measured by flow cytometry in cytotoxic-dNKs (LILRB1 + ) and chemotactic-dNKs (LILRB1 - ).

Article Snippet: Human samples were stained using the following antibodies: CD45 ((PerCP-Cy5.5, Biolegend, clone EO1, cat. no. 368504), CD56 (Cytognos, clone: C5.9, cat. no. CYT-56PE), CD16 (PE/cy7, Biolegend, clone: 3gG8, cat. no. 302015), CD16 (BV510, Biolegend, clone: 3gG8, cat. No. 302047), CD3 (FITC, Beckman Coulter, cat. No. A07746), CD19 (Beckman Coulter, clone: J3.119, cat. no. IM3628U), CD8 (APC-H7, BD Bioscience, clone: SK1, cat. No. 560179), CD14 (APC-Cy7, BD Bioscience, clone: MOPg, cat. No. 557831), LILRB1 (FITC, R&D systems, clone:292305, cat. No. FAB-20171F), CD96 (PE/cy7, Biolegend, clone: NK92.39 cat. no. 338415), CD7 (APC, Biolegend, clone: CD7-6B7, cat. No.343107), CD44 (APC/cy7, Biolegend, clone: IM7, cat. No. 103027), XCL1 (APC, LSbio, cat. no. LS-C716131-100), PRF1 (APC, Biolegend, clone: gD9, cat. No. 308112), GZMB (AF647, Biolegend, clone: GB11, cat. No. 515406), GZMA (APC, LSbio, cat. no. LS-C715841-100), GNLY (AF647, Biolegend, clone: DH2, cat. no. 348006).

Techniques: Expressing, Mass Cytometry, Flow Cytometry

Journal: bioRxiv

Article Title: Natural killer cell function is regulated by TGF-β signaling in pregnancy and tumor progression

doi: 10.1101/2024.09.18.613652

Figure Lengend Snippet: a , Volcano plots showing the differentially expressed genes between human cytotoxic decidual NK cells with human CD56 dim blood NK cells, human chemotactic decidual NK cells with human CD56 dim blood NK cells, and human cytotoxic decidual NK cells with human chemotactic decidual NK cells, with the highlights of selected important genes. Mann-Whitney test with Bonferroni adjustment was used to calculate the FDR for the comparison. b , Barplot showing the top 30 genes that are most correlated with FGFBP2 across NK metacells. This set of 30 genes is subsequently used to calculate blood NK cell scores. Scatterplots depicting the blood NK cell score per metacell versus log enrichment of a selected set of blood NK cell genes. c-d , Mass Cytometry (CyTOF) analysis of CD56 + , CD3 - cells in peripheral blood of healthy donors ( c ) and 3 decidual samples ( d ). e , Normalized expression levels of gene LILRB1 across the entire metacell model, color coded by cell type, with the highlights of metacells for cytotoxic decidual NK cells and chemotactic decidual NK cells. f. dNK cytotoxic assay: x axis denotes effector to target cell ratio; y axis denotes mean percentage lysis of target cells by dNK cells; Stars denote p value (*p < 0.05; **p < 0.01) of a student t-test comparing assays using LILRB1 + /LILRB1 - sorted dNK cell subsets.

Article Snippet: Human samples were stained using the following antibodies: CD45 ((PerCP-Cy5.5, Biolegend, clone EO1, cat. no. 368504), CD56 (Cytognos, clone: C5.9, cat. no. CYT-56PE), CD16 (PE/cy7, Biolegend, clone: 3gG8, cat. no. 302015), CD16 (BV510, Biolegend, clone: 3gG8, cat. No. 302047), CD3 (FITC, Beckman Coulter, cat. No. A07746), CD19 (Beckman Coulter, clone: J3.119, cat. no. IM3628U), CD8 (APC-H7, BD Bioscience, clone: SK1, cat. No. 560179), CD14 (APC-Cy7, BD Bioscience, clone: MOPg, cat. No. 557831), LILRB1 (FITC, R&D systems, clone:292305, cat. No. FAB-20171F), CD96 (PE/cy7, Biolegend, clone: NK92.39 cat. no. 338415), CD7 (APC, Biolegend, clone: CD7-6B7, cat. No.343107), CD44 (APC/cy7, Biolegend, clone: IM7, cat. No. 103027), XCL1 (APC, LSbio, cat. no. LS-C716131-100), PRF1 (APC, Biolegend, clone: gD9, cat. No. 308112), GZMB (AF647, Biolegend, clone: GB11, cat. No. 515406), GZMA (APC, LSbio, cat. no. LS-C715841-100), GNLY (AF647, Biolegend, clone: DH2, cat. no. 348006).

Techniques: MANN-WHITNEY, Comparison, Mass Cytometry, Expressing, Lysis