Journal: Frontiers in Immunology

Article Title: Intrahepatic activated leukocyte cell adhesion molecule induces CD6 high CD4 + T cell infiltration in autoimmune hepatitis

doi: 10.3389/fimmu.2022.967944

Figure Lengend Snippet: Elevated hepatic and serum ALCAM were observed in patients with AIH. (A) Representative immunohistochemistry images of ALCAM in liver biopsies from HC (n=3) and patients with AIH (n=6). (B) Representative immunofluorescence staining for CD4, CD6 and ALCAM in interface hepatitis lesion of liver sections from patients with AIH (n=3). (C) Concentration of serum ALCAM in HC (n=28) and AIH (n=86) was measured by ELISA assay. Individual correlation between clinical indicators and serum ALCAM was calculated in patients with AIH (n=86). (D) Correlation between the number of hepatic CD6 + cells and paired serum ALCAM concentration was calculated (n=27). ***p < 0.001.

Article Snippet: Recombinant human ALCAM Fc chimera (R&D Systems, Minneapolis, MN, USA 7187-AL, referred as rhALCAM) was added when required.

Techniques: Immunohistochemistry, Immunofluorescence, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Intrahepatic activated leukocyte cell adhesion molecule induces CD6 high CD4 + T cell infiltration in autoimmune hepatitis

doi: 10.3389/fimmu.2022.967944

Figure Lengend Snippet: ALCAM promoted CD6 high CD4 + T cells trans-endothelial migration in vitro . (A, B) Human CD4 + T cells were magnetically isolated from PBMC of healthy donors and stimulated with αCD3/28 for 3 days in a flat-bottom 96-well plate, the expression of CD6 and cell proliferation was detected with flow cytometry. (C) After stimulation, the expression of cytokines, surface markers and transcription factors was compared between the CD6 high and CD6 low subsets. (D) Pre-activated CD4 + T cells were placed on a transwell chamber with 5μm pore for 24 hours in the presence of rhALCAM (3ug/ml) or vehicle (PBS). The expression of CD6 and CD69 was measured by flow cytometry. Experiments were repeated at least three times. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: Recombinant human ALCAM Fc chimera (R&D Systems, Minneapolis, MN, USA 7187-AL, referred as rhALCAM) was added when required.

Techniques: Migration, In Vitro, Isolation, Expressing, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Intrahepatic activated leukocyte cell adhesion molecule induces CD6 high CD4 + T cell infiltration in autoimmune hepatitis

doi: 10.3389/fimmu.2022.967944

Figure Lengend Snippet: Schematic diagram for this study. Upregulated ALCAM on hepatocytes promoted the trans-endothelial migration of pathogenic CD6 high CD4 + T cells, which further aggravated hepatic inflammation of patients with AIH. This study revealed a putative therapeutic approach for patients with AIH.

Article Snippet: Recombinant human ALCAM Fc chimera (R&D Systems, Minneapolis, MN, USA 7187-AL, referred as rhALCAM) was added when required.

Techniques: Migration

Journal: Nature Communications

Article Title: Identification of a common mesenchymal stromal progenitor for the adult haematopoietic niche

doi: 10.1038/ncomms13095

Figure Lengend Snippet: ( a ) Representative FACS profiles of stromal cells (CD45 − Ter119 − CD31 − ) near the endosteum (bone-disassociated). They were separated based on their CD166, CD146 and Sca1 expression profiles. Numbers shown in the gate represent the percentage of total live cells. ( b ) Single-cell colony forming efficiency of BM stromal progenitors (*: P ≤0.05, three repeats of 60 cells each, student's t -test). ( c ) Sections of femur from Flk1-GFP mouse stained with antibodies against Sca1 and scanned with an iCys Research imaging cytometer. Nuclei were stained with Dapi. ( d ) Confocal maximum intensity projections of sections of femurs from Flk1-GFP mice stained with antibodies against Sca1 or CD146; scanned near the epiphyseal plate. ( e ) Confocal maximum intensity projections of sections of femur stained with antibodies against Sca1 or CD146 and laminin; scanned near epiphyseal plate. White arrows point to mesenchymal progenitors yellow arrows point to endothelial cells.

Article Snippet: Sections were blocked with 1% BSA, then probed with primary antibody at 4 °C overnight; purified rat-anti-Sca1 (1:50 BioLegend 108102), purified rat-anti-CD146 (1:50, BioLegend 134702), CD166-Fitc (1:25 R&D Systems FAB1172F), CD45-Alexa647 (1:50 BioLegend 103124) and/or rabbit-anti-laminin (1:200 Sigma-Aldrich L9393-.2ML).

Techniques: Expressing, Staining, Imaging, Cytometry

Journal: Nature Communications

Article Title: Identification of a common mesenchymal stromal progenitor for the adult haematopoietic niche

doi: 10.1038/ncomms13095

Figure Lengend Snippet: ( a – b ) FACS analysis of bone-disassociated stromal cells (CD45 − TER119 − CD31 − ) from ( a ) Col2.3-GFP or ( b ) Osx1-Cre:GFP mice. The fequency of the parent gate is displayed. ( c – d ) The frequency of Sca1+, CD146+, CD166+ and Sca1- in the GFP+ population for (mean±s.d.) ( c ) Col2.3-GFP and ( d ) Osx1-Cre:GFP obtained from FACS analysis ( n =3). ( e – f ) Sections of femurs from ( e ) Col2.3-GFP or ( f ) Osx1-Cre:GFP mice stained with antidodies to Sca1 or CD146.

Article Snippet: Sections were blocked with 1% BSA, then probed with primary antibody at 4 °C overnight; purified rat-anti-Sca1 (1:50 BioLegend 108102), purified rat-anti-CD146 (1:50, BioLegend 134702), CD166-Fitc (1:25 R&D Systems FAB1172F), CD45-Alexa647 (1:50 BioLegend 103124) and/or rabbit-anti-laminin (1:200 Sigma-Aldrich L9393-.2ML).

Techniques: Staining

Journal: Nature Communications

Article Title: Identification of a common mesenchymal stromal progenitor for the adult haematopoietic niche

doi: 10.1038/ncomms13095

Figure Lengend Snippet: ( a ) Representative qRT-PCR of 18 single cells from CD146 + , CD166 + or Sca1 + stromal populations either near the endosteum (bone-disassociated) or in the marrow (Marrow) using the Fluidigm 48.48 dynamic array. Each column within a given population represents expression in an individual cell. ( b ) Comparison of the first and second prinicpal components from a principal component analysis of single-cell PCR gene expression data from all 47 genes. ( c – e ) Expression patterns of genes that are significantly overexpressed in mean±s.d. ( c ) Sca1 + cells ( d ) Sca1 + cells and/or CD146 + cells, or ( e ) CD166 + cells. Statistical significance was calculated using one-way ANOVA analysis * P ≤0.05, ** P ≤0.005, *** P ≤0.00; n =72. ANOVA, analysis of variance.

Article Snippet: Sections were blocked with 1% BSA, then probed with primary antibody at 4 °C overnight; purified rat-anti-Sca1 (1:50 BioLegend 108102), purified rat-anti-CD146 (1:50, BioLegend 134702), CD166-Fitc (1:25 R&D Systems FAB1172F), CD45-Alexa647 (1:50 BioLegend 103124) and/or rabbit-anti-laminin (1:200 Sigma-Aldrich L9393-.2ML).

Techniques: Quantitative RT-PCR, Expressing, Comparison, Gene Expression

Journal: Nature Communications

Article Title: Identification of a common mesenchymal stromal progenitor for the adult haematopoietic niche

doi: 10.1038/ncomms13095

Figure Lengend Snippet: ( a ) Sca1 + progenitors were transduced with lentivirus as indicated and 1,000 sorted GFP + Sca1 + progenitors were co-transplanted with unmarked fetal skeletal progenitors under the kidney capsule. ( b ) Representative cross sections of the engrafted ectopic bone stained with H&E (right) or GFP (left) to identify the donor origin, harvested 1 month after transplant. ( c ) Representative FACS profiles of LSK frequencies that were pre-gated for live, CD45 + lineage − cells. ( d ) Cellularity of the ectopic niche marrow (* P <0.05, n =5, student's t -test). ( e ) Rescue of lethally irradiated mice with 2 × 10 5 marrow cells from engrafted ectopic niche generated with either mixed Sca1 + /shKITL progenitors (red) or with Sca1 + /Scrambled shRNA progenitors (yellow) or control bone marrow from host tibia (black) ( n =8). ( f ) The schematic summaries our findings such that Sca1+ cells are the most primative giving rise to CXCL12 producing stromal cells, CD146+ cells and CD166+ cells. CD146+ and CD166+ cells are osteo-progenitors. It is possible that CD146+ is an intermidiary cell fate between Sca1+ cells and CD166+ cells.

Article Snippet: Sections were blocked with 1% BSA, then probed with primary antibody at 4 °C overnight; purified rat-anti-Sca1 (1:50 BioLegend 108102), purified rat-anti-CD146 (1:50, BioLegend 134702), CD166-Fitc (1:25 R&D Systems FAB1172F), CD45-Alexa647 (1:50 BioLegend 103124) and/or rabbit-anti-laminin (1:200 Sigma-Aldrich L9393-.2ML).

Techniques: Transduction, Staining, Irradiation, Generated, shRNA, Control

Journal: Frontiers in Immunology

Article Title: Longitudinal assessment of urinary ALCAM, HPX, and PRDX6 in Korean patients with systemic lupus erythematosus: implications for disease activity monitoring and treatment response

doi: 10.3389/fimmu.2024.1369385

Figure Lengend Snippet: Comparison of urine biomarkers among patients with SLE, LN, and healthy controls. (A) Urine level of ALCAM. (B) Urine level of HPX. (C) Urine level of PRDX6. Statistical analyses were conducted using the Mann–Whitney U test. SLE, systemic lupus erythematosus; LN, lupus nephritis; ALCAM, activated leukocyte cell adhesion molecule; HPX, hemopexin; PRDX, peroxiredoxin.

Article Snippet: The concentration of biomarker levels in mouse urine was analyzed using enzyme-linked immunosorbent assay (ELISA) kits specific for ALCAM (Catalog # DY656; R&D Systems, Minneapolis, MN, USA), HPX (Catalog # ab108860; Abcam Inc, Toronto, ON, Canada) and PRDX 6 (Catalog # MBS067069; Mybiosource, San Diego, CA, USA), according to the manufacturer’s protocols.

Techniques: Comparison, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Longitudinal assessment of urinary ALCAM, HPX, and PRDX6 in Korean patients with systemic lupus erythematosus: implications for disease activity monitoring and treatment response

doi: 10.3389/fimmu.2024.1369385

Figure Lengend Snippet: ROC curves for urine ALCAM, HPX, and PRDX in the diagnosis of SLE and LN compared to healthy controls. (A) For SLE (non-renal + renal) diagnosis, the AUC was 0.801 for the urine ALCAM (95% CI, 0.734 – 0.868), 0.707 for the urine hemopexin (95% CI, 0.622 – 0.792), and 0.697 for the urine PRDX (95% CI, 0.612 – 0.782). (B) For LN diagnosis, the AUC was 0.850 for the urine ALCAM (95% CI, 0.778 – 0.921), 0.781 for the urine hemopexin (95% CI, 0.695 – 0.867), and 0.714 for the urine PRDX (95% CI, 0.617 – 0.812). ROC, Receiver operating characteristic; SLE, systemic lupus erythematosus; LN, lupus nephritis; ALCAM, activated leukocyte cell adhesion molecule; HPX, hemopexin; PRDX, peroxiredoxin; AUC, area under the receiver operating characteristic curve; CI, confidence interval.

Article Snippet: The concentration of biomarker levels in mouse urine was analyzed using enzyme-linked immunosorbent assay (ELISA) kits specific for ALCAM (Catalog # DY656; R&D Systems, Minneapolis, MN, USA), HPX (Catalog # ab108860; Abcam Inc, Toronto, ON, Canada) and PRDX 6 (Catalog # MBS067069; Mybiosource, San Diego, CA, USA), according to the manufacturer’s protocols.

Techniques: Biomarker Discovery

Journal: Frontiers in Immunology

Article Title: Longitudinal assessment of urinary ALCAM, HPX, and PRDX6 in Korean patients with systemic lupus erythematosus: implications for disease activity monitoring and treatment response

doi: 10.3389/fimmu.2024.1369385

Figure Lengend Snippet: Correlation between urine biomarkers and disease activity markers in patients with SLE.

Article Snippet: The concentration of biomarker levels in mouse urine was analyzed using enzyme-linked immunosorbent assay (ELISA) kits specific for ALCAM (Catalog # DY656; R&D Systems, Minneapolis, MN, USA), HPX (Catalog # ab108860; Abcam Inc, Toronto, ON, Canada) and PRDX 6 (Catalog # MBS067069; Mybiosource, San Diego, CA, USA), according to the manufacturer’s protocols.

Techniques: Activity Assay

Journal: Frontiers in Immunology

Article Title: Longitudinal assessment of urinary ALCAM, HPX, and PRDX6 in Korean patients with systemic lupus erythematosus: implications for disease activity monitoring and treatment response

doi: 10.3389/fimmu.2024.1369385

Figure Lengend Snippet: Comparison of ALCAM, HPX, and PRDX according to clinical manifestations in patients with SLE.

Article Snippet: The concentration of biomarker levels in mouse urine was analyzed using enzyme-linked immunosorbent assay (ELISA) kits specific for ALCAM (Catalog # DY656; R&D Systems, Minneapolis, MN, USA), HPX (Catalog # ab108860; Abcam Inc, Toronto, ON, Canada) and PRDX 6 (Catalog # MBS067069; Mybiosource, San Diego, CA, USA), according to the manufacturer’s protocols.

Techniques: Comparison

Journal: Frontiers in Immunology

Article Title: Longitudinal assessment of urinary ALCAM, HPX, and PRDX6 in Korean patients with systemic lupus erythematosus: implications for disease activity monitoring and treatment response

doi: 10.3389/fimmu.2024.1369385

Figure Lengend Snippet: Changes in urine biomarker levels in follow-up observations of patients with SLE without LN and patients with LN. (A) Urine level of ALCAM. (B) Urine level of HPX. (C) Urine level of PRDX6. Statistical analyses were conducted using the Mann–Whitney U test. SLE, systemic lupus erythematosus; LN, lupus nephritis; ALCAM, activated leukocyte cell adhesion molecule; HPX, hemopexin; PRDX, peroxiredoxin.

Article Snippet: The concentration of biomarker levels in mouse urine was analyzed using enzyme-linked immunosorbent assay (ELISA) kits specific for ALCAM (Catalog # DY656; R&D Systems, Minneapolis, MN, USA), HPX (Catalog # ab108860; Abcam Inc, Toronto, ON, Canada) and PRDX 6 (Catalog # MBS067069; Mybiosource, San Diego, CA, USA), according to the manufacturer’s protocols.

Techniques: Biomarker Discovery, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Longitudinal assessment of urinary ALCAM, HPX, and PRDX6 in Korean patients with systemic lupus erythematosus: implications for disease activity monitoring and treatment response

doi: 10.3389/fimmu.2024.1369385

Figure Lengend Snippet: Changes in urine biomarker Levels of patients with SLE without LN and patients with LN only follow-up. (A) Urine level of ALCAM. (B) Urine level of HPX. (C) Urine level of PRDX6. Statistical analyses were conducted using the Mann–Whitney U test. SLE, systemic lupus erythematosus; LN, lupus nephritis; ALCAM, activated leukocyte cell adhesion molecule; HPX, hemopexin; PRDX, peroxiredoxin.

Article Snippet: The concentration of biomarker levels in mouse urine was analyzed using enzyme-linked immunosorbent assay (ELISA) kits specific for ALCAM (Catalog # DY656; R&D Systems, Minneapolis, MN, USA), HPX (Catalog # ab108860; Abcam Inc, Toronto, ON, Canada) and PRDX 6 (Catalog # MBS067069; Mybiosource, San Diego, CA, USA), according to the manufacturer’s protocols.

Techniques: Biomarker Discovery, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Longitudinal assessment of urinary ALCAM, HPX, and PRDX6 in Korean patients with systemic lupus erythematosus: implications for disease activity monitoring and treatment response

doi: 10.3389/fimmu.2024.1369385

Figure Lengend Snippet: Association between changes in biomarker levels and disease activity during follow-up examinations. (A) Correlation between ΔALCAM and ΔSLEDAI-2k. (B) Correlation between HPX and SLEDAI-2k. (C) Correlation between Δ PRDX6 and ΔSLEDAI-2k. SLEDAI, SLE disease activity index 2000; ALCAM, activated leukocyte cell adhesion molecule; HPX, hemopexin; PRDX, peroxiredoxin.

Article Snippet: The concentration of biomarker levels in mouse urine was analyzed using enzyme-linked immunosorbent assay (ELISA) kits specific for ALCAM (Catalog # DY656; R&D Systems, Minneapolis, MN, USA), HPX (Catalog # ab108860; Abcam Inc, Toronto, ON, Canada) and PRDX 6 (Catalog # MBS067069; Mybiosource, San Diego, CA, USA), according to the manufacturer’s protocols.

Techniques: Biomarker Discovery, Activity Assay