Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Clinico-histopathological characteristics of the 136 cutaneous melanomas according to CD33, VISTA and PD-1 expression.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Correlations among CD33, VISTA, and PD-1 expression.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Expression of CD33 and VISTA in cutaneous melanoma. ( A ) High intratumoral expression (immunohistochemical score of 3, red color, cytoplasmic) of CD33 in acral melanoma (× 100). ( B ) High intratumoral expression (score 3, red color, cytoplasmic) of VISTA in nodular melanoma (× 100). ( C ) High peritumoral expression (score 3, red color, cytoplasmic) of CD33 in acral melanoma (× 100). ( D ) High peritumoral expression (score 3, red color, cytoplasmic) of VISTA in nodular melanoma (× 200). ( E ) Dual immunofluorescence staining of ( A ) CD33 and VISTA (CD33, red, cytoplasmic; VISTA, green, cytoplasmic) and ( F ) PD-1 and VISTA (PD-1, red, cytoplasmic; VISTA, green, cytoplasmic). Double-positive cells are shown in yellow (white arrow). VISTA, V-domain Ig suppressor of T-cell activation; PD-1, programmed cell death protein-1.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing, Immunohistochemical staining, Immunofluorescence, Staining, Activation Assay

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Patient survival according to CD33 and VISTA expression. ( A ) OS according to CD33 expression. ( B ) OS according to VISTA expression. ( C ) Worse OS in patients with high expression of both CD33 and VISTA. ( D ) OS according to the degree of CD33 and VISTA expression ( E ) Worse OS in cases with dual expression of VISTA and PD-1 among patients with high expression of PD-1. VISTA, V-domain Ig suppressor of T-cell activation; PD-1, programmed cell death protein-1.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing, Activation Assay

Journal: Scientific Reports

Article Title: The prognostic significance of VISTA and CD33-positive myeloid cells in cutaneous melanoma and their relationship with PD-1 expression

doi: 10.1038/s41598-020-71216-2

Figure Lengend Snippet: Univariate and multivariate analyses of OS and PFS.

Article Snippet: Paraffin-embedded sections were immunostained with anti-VISTA (1:200, ProSci Incorporation, CA, USA), anti-CD33 (1:100, Leica Biosystems, Newcastle, UK), or anti-PD-1 (1:100, Ventana, Tucson, AZ, USA) antibodies.

Techniques: Expressing

Journal: Parasites & Vectors

Article Title: Effect of B7-H4 downregulation induced by Toxoplasma gondii infection on dysfunction of decidual macrophages contributes to adverse pregnancy outcomes

doi: 10.1186/s13071-022-05560-9

Figure Lengend Snippet: Changes in B7-H4 expression on decidual macrophages (dMφ) and the effect of these changes on phagocytic activities during Toxoplasma gondii infection. a B7-H4 expression levels on human CD14 + dMφ in uninfected, infected and B7-H4 neutralized infected groups detected by flow cytometry. b B7-H4 expression on mouse F4/80 + dMφ detected in uninfected, infected and B7-H4 -/- infected mice by flow cytometry. c Fluorescence microscopy images of the number of phagocytic dMφ in the uninfected, infected and B7-H4 neutralized infected groups. Scale bar: 50 µm. d Flow cytometry results showing the phagocytotic capacity of dMφ in the uninfected, infected and B7-H4 neutralized infected groups. Data are presented as mean ± SD, with at least 6 pregnant mice or human samples in each group assayed individually by the unpaired t-test. Asterisks indicated significant differences between groups at * P < 0.05, ** P < 0.01. IgG-FITC, immunoglobulin G-fluorescein isothiocyanate complex

Article Snippet: Purified human primary CD14 + dMφ were equally divided into five groups: (i) uninfected; (ii) infected; (iii) STAT1-inhibitor infected; (iv) B7-H4-neutralized infected; and (v) B7-H4-neutralized STAT1-inhibitor infected group. dMφ of the STAT1-inhibitor infected group and B7-H4-neutralized STAT1-inhibitor infected group were preincubated with the STAT1 inhibitor fuldarabine (#HY-B0069; MedChemExpress) for 2 h, followed by addition of B7-H4 mAb.

Techniques: Expressing, Infection, Flow Cytometry, Fluorescence, Microscopy

Journal: Parasites & Vectors

Article Title: Effect of B7-H4 downregulation induced by Toxoplasma gondii infection on dysfunction of decidual macrophages contributes to adverse pregnancy outcomes

doi: 10.1186/s13071-022-05560-9

Figure Lengend Snippet: Reduction of B7-H4 expression on dMφ by T. gondii infection affects the expression of M1- and M2-type membrane functional molecules. a-e Flow cytometry analysis of CD80, CD86, CD206 and CD163 levels on human dMφ in uninfected, infected and B7-H4 neutralized infected groups. f-i Flow cytometry analysis of B7-H4, CD80, CD86 and CD206 expressions on dMφ in uninfected, infected, and B7-H4 -/- infected mice. Data are presented as the mean ± SD, with at least 6 pregnant mice or human samples in each group assayed individually by unpaired the t-test. Asterisks indicate significant differences between groups at * P < 0.05, ** P < 0.01. PE, Phycoerythrin; SSC, side scatter

Article Snippet: Purified human primary CD14 + dMφ were equally divided into five groups: (i) uninfected; (ii) infected; (iii) STAT1-inhibitor infected; (iv) B7-H4-neutralized infected; and (v) B7-H4-neutralized STAT1-inhibitor infected group. dMφ of the STAT1-inhibitor infected group and B7-H4-neutralized STAT1-inhibitor infected group were preincubated with the STAT1 inhibitor fuldarabine (#HY-B0069; MedChemExpress) for 2 h, followed by addition of B7-H4 mAb.

Techniques: Expressing, Infection, Membrane, Functional Assay, Flow Cytometry

Journal: Parasites & Vectors

Article Title: Effect of B7-H4 downregulation induced by Toxoplasma gondii infection on dysfunction of decidual macrophages contributes to adverse pregnancy outcomes

doi: 10.1186/s13071-022-05560-9

Figure Lengend Snippet: Reduction of B7-H4 expression on dMφ with T. gondii infection resulted in changes in the expression iNOS and Arg-1. a Levels of Arg-1 produced by dMφ in uninfected, infected and B7-H4 -/- infected mice analyzed by flow cytometry. b Flow cytometry analysis of iNOS produced by dMφ in uninfected, infected, and B7-H4 -/- infected mice. c Western blotting assay of B7-H4, iNOS and Arg-1 protein levels in uninfected, infected and B7-H4 neutralized infected groups of purified human macrophages. Data are presented as the mean ± SD, with at least 6 pregnant mice or human samples in each group assayed individually by the unpaired t-test. Asterisks indicate significant differences between groups at * P < 0.05, ** P < 0.01. Arg-1, Arginase-1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; iNOS, inducible nitric oxide synthase

Article Snippet: Purified human primary CD14 + dMφ were equally divided into five groups: (i) uninfected; (ii) infected; (iii) STAT1-inhibitor infected; (iv) B7-H4-neutralized infected; and (v) B7-H4-neutralized STAT1-inhibitor infected group. dMφ of the STAT1-inhibitor infected group and B7-H4-neutralized STAT1-inhibitor infected group were preincubated with the STAT1 inhibitor fuldarabine (#HY-B0069; MedChemExpress) for 2 h, followed by addition of B7-H4 mAb.

Techniques: Expressing, Infection, Produced, Flow Cytometry, Western Blot, Purification

Journal: Parasites & Vectors

Article Title: Effect of B7-H4 downregulation induced by Toxoplasma gondii infection on dysfunction of decidual macrophages contributes to adverse pregnancy outcomes

doi: 10.1186/s13071-022-05560-9

Figure Lengend Snippet: Reduction of B7-H4 expression on dMφ with T. gondii infection resulted in changes in the expression of TNF-α and IL-10. a-c Expression level of IL-10 and TNF-α in F4/80 + dMφ in uninfected, infected and B7-H4 -/- infected mice detected by flow cytometry. d , e Western blotting assay of TNF-α and IL-10 levels in CD14 + dMφ of the uninfected, infected and B7-H4 neutralized infected groups. f , g Representative immunofluorescent photographs of B7-H4, TNF-α and IL-10 of CD14 + dMφ in the uninfected, infected and B7-H4 neutralized infected groups. Scale bar: 50 µm. Data are presented as the mean ± SD, with at least 6 pregnant mice or human samples in each group assayed individually by the unpaired t-test. Asterisks indicate significant differences between groups at * P < 0.05, ** P < 0.01. DAPI, 4′,6-Diamidino-2-phenylindole; IL, interleukin; TNF, tumor necrosis factor

Article Snippet: Purified human primary CD14 + dMφ were equally divided into five groups: (i) uninfected; (ii) infected; (iii) STAT1-inhibitor infected; (iv) B7-H4-neutralized infected; and (v) B7-H4-neutralized STAT1-inhibitor infected group. dMφ of the STAT1-inhibitor infected group and B7-H4-neutralized STAT1-inhibitor infected group were preincubated with the STAT1 inhibitor fuldarabine (#HY-B0069; MedChemExpress) for 2 h, followed by addition of B7-H4 mAb.

Techniques: Expressing, Infection, Flow Cytometry, Western Blot

Journal: Parasites & Vectors

Article Title: Effect of B7-H4 downregulation induced by Toxoplasma gondii infection on dysfunction of decidual macrophages contributes to adverse pregnancy outcomes

doi: 10.1186/s13071-022-05560-9

Figure Lengend Snippet: Downregulation of B7-H4 by T. gondii infection resulted in changes in dMφ function by affecting the JAK2/STAT1 signaling pathways. a , b Representative western blot and histogram analysis of B7-H4, JAK2, p-JAK2, STAT1 and p-STAT1 levels of dMφ in uninfected, infected and B7-H4 neutralized infected groups. c , d Representative western blot and histograms analysis of B7-H4, STAT1, p-STAT1, iNOS and TNF-α of dMφ in the uninfected, infected, infected + STAT1 inhibitor (Infected + Flu), B7-H4 neutralized infected and B7-H4 neutralized infected + STAT1 inhibitor (Neutralized infected + Flu) groups. The data in all panels are representative of at least three independent experiments. Data are presented as the means ± SD, with the unpaired t-test. Asterisks indicate significant differences between groups at * P < 0.05, ** P < 0.01. Flu, Fludarabine; JAK2, Janus kinase 2; p-JAK2, phosphorylated JAK2; p-STAT1, phosphorylated signal transducer and activator of transcription 1; STAT1, signal transducer and activator of transcription 1

Article Snippet: Purified human primary CD14 + dMφ were equally divided into five groups: (i) uninfected; (ii) infected; (iii) STAT1-inhibitor infected; (iv) B7-H4-neutralized infected; and (v) B7-H4-neutralized STAT1-inhibitor infected group. dMφ of the STAT1-inhibitor infected group and B7-H4-neutralized STAT1-inhibitor infected group were preincubated with the STAT1 inhibitor fuldarabine (#HY-B0069; MedChemExpress) for 2 h, followed by addition of B7-H4 mAb.

Techniques: Infection, Protein-Protein interactions, Western Blot