human cd155 pvr transcript variant 1 gene cdna  (Sino Biological)


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    Name:
    CD155 PVR cDNA ORF Clone Human C GFPSpark tag
    Description:
    Full length Clone DNA of Human poliovirus receptor PVR transcript variant 1 with C terminal GFPSpark tag
    Catalog Number:
    HG10109-ACG
    Price:
    225.0
    Category:
    cDNA Clone
    Applications:
    Stable or Transient mammalian expression
    Size:
    1Unit
    Product Aliases:
    CD155 cDNA ORF Clone Human, HVED cDNA ORF Clone Human, Necl-5 cDNA ORF Clone Human, NECL5 cDNA ORF Clone Human, PVS cDNA ORF Clone Human, TAGE4 cDNA ORF Clone Human
    Molecule Name:
    PVR,Tage4,CD155
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    Structured Review

    Sino Biological human cd155 pvr transcript variant 1 gene cdna
    CD155 PVR cDNA ORF Clone Human C GFPSpark tag
    Full length Clone DNA of Human poliovirus receptor PVR transcript variant 1 with C terminal GFPSpark tag
    https://www.bioz.com/result/human cd155 pvr transcript variant 1 gene cdna/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human cd155 pvr transcript variant 1 gene cdna - by Bioz Stars, 2021-05
    93/100 stars

    Images

    1) Product Images from "IL-15 stimulation with TIGIT blockade reverses CD155-mediated NK cell dysfunction in melanoma"

    Article Title: IL-15 stimulation with TIGIT blockade reverses CD155-mediated NK cell dysfunction in melanoma

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    doi: 10.1158/1078-0432.CCR-20-0575

    Membrane-bound CD155 induces CD226 degradation and NK cell dysfunction. ( A-C ). cNKs isolated from MP were incubated with L cells or L-CD155 for 16 h +/− concanamycin-A (Conca). ( A ) Representative histograms of flow cytometry showing the effects of Conca on cell surface and total CD226 expression (MFI) by NK cells in the presence of L-CD155 cells or L cells as compared with DMSO control. ( B and C) Cell surface and total CD226 ( B ), TIGIT ( C ), and surface CD45 (as control) expressions by cNKs (normalized MFI expression as compared to cNKs + L cells). ( D and E ) cNKs were incubated 48 h with either sCD155, IgG- or CD155-Fc-beads +/− blocking antibodies and washed before functional assay in presence of FO-I. ( D ) CD107a expression (%) by cNKs pre-incubated or not with CD155-Fc beads ( n =11) ( E ) CD107a expression (fold change) by cNKs pre-incubated with either CD155-Fc-beads ( n =6), IgG-beads ( n =3), or sCD155 ( n =3) +/− aCD226 or aTIGIT blocking mAbs as compared with IgG-beads (dotted line). Horizontal bars depict means ± SEM ( B , C , and E ) and P values were obtained by one-way ANOVA tests followed by Tukey’s ( B and C ) or Dunnett’s ( E ) multiple comparisons test or by paired t -tests ( D ) with ns (non-significant), P > 0.05; *, P
    Figure Legend Snippet: Membrane-bound CD155 induces CD226 degradation and NK cell dysfunction. ( A-C ). cNKs isolated from MP were incubated with L cells or L-CD155 for 16 h +/− concanamycin-A (Conca). ( A ) Representative histograms of flow cytometry showing the effects of Conca on cell surface and total CD226 expression (MFI) by NK cells in the presence of L-CD155 cells or L cells as compared with DMSO control. ( B and C) Cell surface and total CD226 ( B ), TIGIT ( C ), and surface CD45 (as control) expressions by cNKs (normalized MFI expression as compared to cNKs + L cells). ( D and E ) cNKs were incubated 48 h with either sCD155, IgG- or CD155-Fc-beads +/− blocking antibodies and washed before functional assay in presence of FO-I. ( D ) CD107a expression (%) by cNKs pre-incubated or not with CD155-Fc beads ( n =11) ( E ) CD107a expression (fold change) by cNKs pre-incubated with either CD155-Fc-beads ( n =6), IgG-beads ( n =3), or sCD155 ( n =3) +/− aCD226 or aTIGIT blocking mAbs as compared with IgG-beads (dotted line). Horizontal bars depict means ± SEM ( B , C , and E ) and P values were obtained by one-way ANOVA tests followed by Tukey’s ( B and C ) or Dunnett’s ( E ) multiple comparisons test or by paired t -tests ( D ) with ns (non-significant), P > 0.05; *, P

    Techniques Used: Isolation, Incubation, Flow Cytometry, Expressing, Blocking Assay, Functional Assay

    IL-15 together with TIGIT blockade reverses CD155-induced NK cell dysfunction. ( A ) Pooled data showing the effect of STAT3 and STAT5 inhibitors (Stat3i and Stat5i, respectively) on CD226 and TIGIT expression (MFI) by cNKs after 6 d incubation +/− IL-15 ( n =6). ( B ) Pooled data of from three independent experiments showing CD226 and TIGIT mRNA relative expression by cNKs from MPs after 48 h incubation with indicated cell lines +/− IL-15 ( n =4 and n =5, respectively). ( C ) Surface and total CD226 expression by cNKs after 48 h incubation with L-CD155 or FO-I +/− IL-15 (normalized MFI ratio as compared with cNKs in medium alone). (D ) Pool of 30 randomly selected cNKs from one representative ImageStream analysis ( n =3), showing CD226 internalization ratio after 1 h incubation with indicated cell lines. ( E ) CD226 and TIGIT expression (MFI and fold change) by TiNKs, ex-vivo and after 6 d incubation with IL-15 ( n= 13). ( F ) Pooled data comparing CD107a expression by cNKs treated 48 h with indicated beads (frequency and fold change), and by TiNKs (frequency) after 16 h or 6 d IL-15 stimulation ( n =7 and n =10, respectively), in response to FO-I. ( G) Fold change of CD107a + TiNKs after 16 h or 6 d IL-15 stimulation prior to incubation with FO-I +/− aTIGIT mAbs ( n =16) as compared with 16 h IL-15 + IgG control mAb (dotted line). Horizontal bars depict means ± SD ( A and B ) or ± SEM ( C ). P values were obtained by paired t -tests ( E and F ), ordinary ( D ) or repeated-measures one-way ANOVA tests followed by Tukey’s ( B, and G ) or Dunnett’s ( C ) multiple comparisons test, with *, P
    Figure Legend Snippet: IL-15 together with TIGIT blockade reverses CD155-induced NK cell dysfunction. ( A ) Pooled data showing the effect of STAT3 and STAT5 inhibitors (Stat3i and Stat5i, respectively) on CD226 and TIGIT expression (MFI) by cNKs after 6 d incubation +/− IL-15 ( n =6). ( B ) Pooled data of from three independent experiments showing CD226 and TIGIT mRNA relative expression by cNKs from MPs after 48 h incubation with indicated cell lines +/− IL-15 ( n =4 and n =5, respectively). ( C ) Surface and total CD226 expression by cNKs after 48 h incubation with L-CD155 or FO-I +/− IL-15 (normalized MFI ratio as compared with cNKs in medium alone). (D ) Pool of 30 randomly selected cNKs from one representative ImageStream analysis ( n =3), showing CD226 internalization ratio after 1 h incubation with indicated cell lines. ( E ) CD226 and TIGIT expression (MFI and fold change) by TiNKs, ex-vivo and after 6 d incubation with IL-15 ( n= 13). ( F ) Pooled data comparing CD107a expression by cNKs treated 48 h with indicated beads (frequency and fold change), and by TiNKs (frequency) after 16 h or 6 d IL-15 stimulation ( n =7 and n =10, respectively), in response to FO-I. ( G) Fold change of CD107a + TiNKs after 16 h or 6 d IL-15 stimulation prior to incubation with FO-I +/− aTIGIT mAbs ( n =16) as compared with 16 h IL-15 + IgG control mAb (dotted line). Horizontal bars depict means ± SD ( A and B ) or ± SEM ( C ). P values were obtained by paired t -tests ( E and F ), ordinary ( D ) or repeated-measures one-way ANOVA tests followed by Tukey’s ( B, and G ) or Dunnett’s ( C ) multiple comparisons test, with *, P

    Techniques Used: Expressing, Incubation, Ex Vivo

    Membrane-bound CD155 induces TIGIT and CD226 internalization in NK cells. (A and B) CD226 ( A ) and TIGIT ( B ) expression by cNKs (fold change of MFI) after 48 h incubation with indicated targets +/− aCD155 blocking mAb, as compared with control (no target, n =15, or IgG-coated beads, n =8). ( C ) Fold change of CD226 expression in MFI by cNKs incubated 48 h with sCD155 or the supernatants of melanoma cell lines ( n =7). (D ) CD226 mRNA relative expression by cNKs either alone, co-cultured for 48 h with L-cells, or with L-CD155 cells. Dots are means of triplicates ( n =7). ( E and F ) Representative pictures from ImageStream analysis of cNKs alone or with indicated cell lines (1 h) depicting CD226 ( E ) and TIGIT ( F ) membrane (yellow) and intracellular (violet) expression, as well as surface CD45 expression (turquoise). ( G ) Representative histograms gated on total live cNKs (left panels) and statistical analysis from a pool of 30 randomly selected cNKs (right panels) showing the ratio of CD226 and TIGIT internalization by cNKs in the presence of L-CD155 and FO-I as compared with L cells or cNKs alone. Cell surface CD45 expression was used to calculate the internalization ratio of CD226 and TIGIT intracellular staining using IDEAS software. P values were obtained from one-way ANOVA tests ( A and B ) or repeated-measure ANOVA tests ( D and G ) followed by Dunnett’s multiple comparisons test with *, P
    Figure Legend Snippet: Membrane-bound CD155 induces TIGIT and CD226 internalization in NK cells. (A and B) CD226 ( A ) and TIGIT ( B ) expression by cNKs (fold change of MFI) after 48 h incubation with indicated targets +/− aCD155 blocking mAb, as compared with control (no target, n =15, or IgG-coated beads, n =8). ( C ) Fold change of CD226 expression in MFI by cNKs incubated 48 h with sCD155 or the supernatants of melanoma cell lines ( n =7). (D ) CD226 mRNA relative expression by cNKs either alone, co-cultured for 48 h with L-cells, or with L-CD155 cells. Dots are means of triplicates ( n =7). ( E and F ) Representative pictures from ImageStream analysis of cNKs alone or with indicated cell lines (1 h) depicting CD226 ( E ) and TIGIT ( F ) membrane (yellow) and intracellular (violet) expression, as well as surface CD45 expression (turquoise). ( G ) Representative histograms gated on total live cNKs (left panels) and statistical analysis from a pool of 30 randomly selected cNKs (right panels) showing the ratio of CD226 and TIGIT internalization by cNKs in the presence of L-CD155 and FO-I as compared with L cells or cNKs alone. Cell surface CD45 expression was used to calculate the internalization ratio of CD226 and TIGIT intracellular staining using IDEAS software. P values were obtained from one-way ANOVA tests ( A and B ) or repeated-measure ANOVA tests ( D and G ) followed by Dunnett’s multiple comparisons test with *, P

    Techniques Used: Expressing, Incubation, Blocking Assay, Cell Culture, Staining, Software

    Related Articles

    Variant Assay:

    Article Title: IL15 Stimulation with TIGIT Blockade Reverses CD155-mediated NK-Cell Dysfunction in Melanoma.
    Article Snippet: Natural killer (NK) cells play a critical role in tumor immunosurveillance. .. Natural killer (NK) cells play a critical role in tumor immunosurveillance. ..

    Article Title: IL-15 stimulation with TIGIT blockade reverses CD155-mediated NK cell dysfunction in melanoma
    Article Snippet: K562 and L cells were purchased (ATCC). .. Human CD155/PVR transcript variant 1 Gene cDNA Clone (full-length ORF Clone, Sino Biological Inc.) was transfected into L cells using lipofectamine 3000 (Invitrogen) according to manufacturer’s instructions. ..

    Transfection:

    Article Title: IL15 Stimulation with TIGIT Blockade Reverses CD155-mediated NK-Cell Dysfunction in Melanoma.
    Article Snippet: Natural killer (NK) cells play a critical role in tumor immunosurveillance. .. Natural killer (NK) cells play a critical role in tumor immunosurveillance. ..

    Article Title: IL-15 stimulation with TIGIT blockade reverses CD155-mediated NK cell dysfunction in melanoma
    Article Snippet: K562 and L cells were purchased (ATCC). .. Human CD155/PVR transcript variant 1 Gene cDNA Clone (full-length ORF Clone, Sino Biological Inc.) was transfected into L cells using lipofectamine 3000 (Invitrogen) according to manufacturer’s instructions. ..

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  • 93
    Sino Biological human cd155 pvr transcript variant 1 gene cdna
    Membrane-bound <t>CD155</t> induces CD226 degradation and NK cell dysfunction. ( A-C ). cNKs isolated from MP were incubated with L cells or L-CD155 for 16 h +/− concanamycin-A (Conca). ( A ) Representative histograms of flow cytometry showing the effects of Conca on cell surface and total CD226 expression (MFI) by NK cells in the presence of L-CD155 cells or L cells as compared with DMSO control. ( B and C) Cell surface and total CD226 ( B ), TIGIT ( C ), and surface CD45 (as control) expressions by cNKs (normalized MFI expression as compared to cNKs + L cells). ( D and E ) cNKs were incubated 48 h with either sCD155, IgG- or CD155-Fc-beads +/− blocking antibodies and washed before functional assay in presence of FO-I. ( D ) CD107a expression (%) by cNKs pre-incubated or not with CD155-Fc beads ( n =11) ( E ) CD107a expression (fold change) by cNKs pre-incubated with either CD155-Fc-beads ( n =6), IgG-beads ( n =3), or sCD155 ( n =3) +/− aCD226 or aTIGIT blocking mAbs as compared with IgG-beads (dotted line). Horizontal bars depict means ± SEM ( B , C , and E ) and P values were obtained by one-way ANOVA tests followed by Tukey’s ( B and C ) or Dunnett’s ( E ) multiple comparisons test or by paired t -tests ( D ) with ns (non-significant), P > 0.05; *, P
    Human Cd155 Pvr Transcript Variant 1 Gene Cdna, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human cd155 pvr transcript variant 1 gene cdna/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    human cd155 pvr transcript variant 1 gene cdna - by Bioz Stars, 2021-05
    93/100 stars
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    92
    Sino Biological plasmid pcmv3 c ha cd155
    Membrane-bound <t>CD155</t> induces CD226 degradation and NK cell dysfunction. ( A-C ). cNKs isolated from MP were incubated with L cells or L-CD155 for 16 h +/− concanamycin-A (Conca). ( A ) Representative histograms of flow cytometry showing the effects of Conca on cell surface and total CD226 expression (MFI) by NK cells in the presence of L-CD155 cells or L cells as compared with DMSO control. ( B and C) Cell surface and total CD226 ( B ), TIGIT ( C ), and surface CD45 (as control) expressions by cNKs (normalized MFI expression as compared to cNKs + L cells). ( D and E ) cNKs were incubated 48 h with either sCD155, IgG- or CD155-Fc-beads +/− blocking antibodies and washed before functional assay in presence of FO-I. ( D ) CD107a expression (%) by cNKs pre-incubated or not with CD155-Fc beads ( n =11) ( E ) CD107a expression (fold change) by cNKs pre-incubated with either CD155-Fc-beads ( n =6), IgG-beads ( n =3), or sCD155 ( n =3) +/− aCD226 or aTIGIT blocking mAbs as compared with IgG-beads (dotted line). Horizontal bars depict means ± SEM ( B , C , and E ) and P values were obtained by one-way ANOVA tests followed by Tukey’s ( B and C ) or Dunnett’s ( E ) multiple comparisons test or by paired t -tests ( D ) with ns (non-significant), P > 0.05; *, P
    Plasmid Pcmv3 C Ha Cd155, supplied by Sino Biological, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid pcmv3 c ha cd155/product/Sino Biological
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plasmid pcmv3 c ha cd155 - by Bioz Stars, 2021-05
    92/100 stars
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    93
    Sino Biological full length orf
    Membrane-bound <t>CD155</t> induces CD226 degradation and NK cell dysfunction. ( A-C ). cNKs isolated from MP were incubated with L cells or L-CD155 for 16 h +/− concanamycin-A (Conca). ( A ) Representative histograms of flow cytometry showing the effects of Conca on cell surface and total CD226 expression (MFI) by NK cells in the presence of L-CD155 cells or L cells as compared with DMSO control. ( B and C) Cell surface and total CD226 ( B ), TIGIT ( C ), and surface CD45 (as control) expressions by cNKs (normalized MFI expression as compared to cNKs + L cells). ( D and E ) cNKs were incubated 48 h with either sCD155, IgG- or CD155-Fc-beads +/− blocking antibodies and washed before functional assay in presence of FO-I. ( D ) CD107a expression (%) by cNKs pre-incubated or not with CD155-Fc beads ( n =11) ( E ) CD107a expression (fold change) by cNKs pre-incubated with either CD155-Fc-beads ( n =6), IgG-beads ( n =3), or sCD155 ( n =3) +/− aCD226 or aTIGIT blocking mAbs as compared with IgG-beads (dotted line). Horizontal bars depict means ± SEM ( B , C , and E ) and P values were obtained by one-way ANOVA tests followed by Tukey’s ( B and C ) or Dunnett’s ( E ) multiple comparisons test or by paired t -tests ( D ) with ns (non-significant), P > 0.05; *, P
    Full Length Orf, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/full length orf/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    full length orf - by Bioz Stars, 2021-05
    93/100 stars
      Buy from Supplier

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    Membrane-bound CD155 induces CD226 degradation and NK cell dysfunction. ( A-C ). cNKs isolated from MP were incubated with L cells or L-CD155 for 16 h +/− concanamycin-A (Conca). ( A ) Representative histograms of flow cytometry showing the effects of Conca on cell surface and total CD226 expression (MFI) by NK cells in the presence of L-CD155 cells or L cells as compared with DMSO control. ( B and C) Cell surface and total CD226 ( B ), TIGIT ( C ), and surface CD45 (as control) expressions by cNKs (normalized MFI expression as compared to cNKs + L cells). ( D and E ) cNKs were incubated 48 h with either sCD155, IgG- or CD155-Fc-beads +/− blocking antibodies and washed before functional assay in presence of FO-I. ( D ) CD107a expression (%) by cNKs pre-incubated or not with CD155-Fc beads ( n =11) ( E ) CD107a expression (fold change) by cNKs pre-incubated with either CD155-Fc-beads ( n =6), IgG-beads ( n =3), or sCD155 ( n =3) +/− aCD226 or aTIGIT blocking mAbs as compared with IgG-beads (dotted line). Horizontal bars depict means ± SEM ( B , C , and E ) and P values were obtained by one-way ANOVA tests followed by Tukey’s ( B and C ) or Dunnett’s ( E ) multiple comparisons test or by paired t -tests ( D ) with ns (non-significant), P > 0.05; *, P

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: IL-15 stimulation with TIGIT blockade reverses CD155-mediated NK cell dysfunction in melanoma

    doi: 10.1158/1078-0432.CCR-20-0575

    Figure Lengend Snippet: Membrane-bound CD155 induces CD226 degradation and NK cell dysfunction. ( A-C ). cNKs isolated from MP were incubated with L cells or L-CD155 for 16 h +/− concanamycin-A (Conca). ( A ) Representative histograms of flow cytometry showing the effects of Conca on cell surface and total CD226 expression (MFI) by NK cells in the presence of L-CD155 cells or L cells as compared with DMSO control. ( B and C) Cell surface and total CD226 ( B ), TIGIT ( C ), and surface CD45 (as control) expressions by cNKs (normalized MFI expression as compared to cNKs + L cells). ( D and E ) cNKs were incubated 48 h with either sCD155, IgG- or CD155-Fc-beads +/− blocking antibodies and washed before functional assay in presence of FO-I. ( D ) CD107a expression (%) by cNKs pre-incubated or not with CD155-Fc beads ( n =11) ( E ) CD107a expression (fold change) by cNKs pre-incubated with either CD155-Fc-beads ( n =6), IgG-beads ( n =3), or sCD155 ( n =3) +/− aCD226 or aTIGIT blocking mAbs as compared with IgG-beads (dotted line). Horizontal bars depict means ± SEM ( B , C , and E ) and P values were obtained by one-way ANOVA tests followed by Tukey’s ( B and C ) or Dunnett’s ( E ) multiple comparisons test or by paired t -tests ( D ) with ns (non-significant), P > 0.05; *, P

    Article Snippet: Human CD155/PVR transcript variant 1 Gene cDNA Clone (full-length ORF Clone, Sino Biological Inc.) was transfected into L cells using lipofectamine 3000 (Invitrogen) according to manufacturer’s instructions.

    Techniques: Isolation, Incubation, Flow Cytometry, Expressing, Blocking Assay, Functional Assay

    IL-15 together with TIGIT blockade reverses CD155-induced NK cell dysfunction. ( A ) Pooled data showing the effect of STAT3 and STAT5 inhibitors (Stat3i and Stat5i, respectively) on CD226 and TIGIT expression (MFI) by cNKs after 6 d incubation +/− IL-15 ( n =6). ( B ) Pooled data of from three independent experiments showing CD226 and TIGIT mRNA relative expression by cNKs from MPs after 48 h incubation with indicated cell lines +/− IL-15 ( n =4 and n =5, respectively). ( C ) Surface and total CD226 expression by cNKs after 48 h incubation with L-CD155 or FO-I +/− IL-15 (normalized MFI ratio as compared with cNKs in medium alone). (D ) Pool of 30 randomly selected cNKs from one representative ImageStream analysis ( n =3), showing CD226 internalization ratio after 1 h incubation with indicated cell lines. ( E ) CD226 and TIGIT expression (MFI and fold change) by TiNKs, ex-vivo and after 6 d incubation with IL-15 ( n= 13). ( F ) Pooled data comparing CD107a expression by cNKs treated 48 h with indicated beads (frequency and fold change), and by TiNKs (frequency) after 16 h or 6 d IL-15 stimulation ( n =7 and n =10, respectively), in response to FO-I. ( G) Fold change of CD107a + TiNKs after 16 h or 6 d IL-15 stimulation prior to incubation with FO-I +/− aTIGIT mAbs ( n =16) as compared with 16 h IL-15 + IgG control mAb (dotted line). Horizontal bars depict means ± SD ( A and B ) or ± SEM ( C ). P values were obtained by paired t -tests ( E and F ), ordinary ( D ) or repeated-measures one-way ANOVA tests followed by Tukey’s ( B, and G ) or Dunnett’s ( C ) multiple comparisons test, with *, P

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: IL-15 stimulation with TIGIT blockade reverses CD155-mediated NK cell dysfunction in melanoma

    doi: 10.1158/1078-0432.CCR-20-0575

    Figure Lengend Snippet: IL-15 together with TIGIT blockade reverses CD155-induced NK cell dysfunction. ( A ) Pooled data showing the effect of STAT3 and STAT5 inhibitors (Stat3i and Stat5i, respectively) on CD226 and TIGIT expression (MFI) by cNKs after 6 d incubation +/− IL-15 ( n =6). ( B ) Pooled data of from three independent experiments showing CD226 and TIGIT mRNA relative expression by cNKs from MPs after 48 h incubation with indicated cell lines +/− IL-15 ( n =4 and n =5, respectively). ( C ) Surface and total CD226 expression by cNKs after 48 h incubation with L-CD155 or FO-I +/− IL-15 (normalized MFI ratio as compared with cNKs in medium alone). (D ) Pool of 30 randomly selected cNKs from one representative ImageStream analysis ( n =3), showing CD226 internalization ratio after 1 h incubation with indicated cell lines. ( E ) CD226 and TIGIT expression (MFI and fold change) by TiNKs, ex-vivo and after 6 d incubation with IL-15 ( n= 13). ( F ) Pooled data comparing CD107a expression by cNKs treated 48 h with indicated beads (frequency and fold change), and by TiNKs (frequency) after 16 h or 6 d IL-15 stimulation ( n =7 and n =10, respectively), in response to FO-I. ( G) Fold change of CD107a + TiNKs after 16 h or 6 d IL-15 stimulation prior to incubation with FO-I +/− aTIGIT mAbs ( n =16) as compared with 16 h IL-15 + IgG control mAb (dotted line). Horizontal bars depict means ± SD ( A and B ) or ± SEM ( C ). P values were obtained by paired t -tests ( E and F ), ordinary ( D ) or repeated-measures one-way ANOVA tests followed by Tukey’s ( B, and G ) or Dunnett’s ( C ) multiple comparisons test, with *, P

    Article Snippet: Human CD155/PVR transcript variant 1 Gene cDNA Clone (full-length ORF Clone, Sino Biological Inc.) was transfected into L cells using lipofectamine 3000 (Invitrogen) according to manufacturer’s instructions.

    Techniques: Expressing, Incubation, Ex Vivo

    Membrane-bound CD155 induces TIGIT and CD226 internalization in NK cells. (A and B) CD226 ( A ) and TIGIT ( B ) expression by cNKs (fold change of MFI) after 48 h incubation with indicated targets +/− aCD155 blocking mAb, as compared with control (no target, n =15, or IgG-coated beads, n =8). ( C ) Fold change of CD226 expression in MFI by cNKs incubated 48 h with sCD155 or the supernatants of melanoma cell lines ( n =7). (D ) CD226 mRNA relative expression by cNKs either alone, co-cultured for 48 h with L-cells, or with L-CD155 cells. Dots are means of triplicates ( n =7). ( E and F ) Representative pictures from ImageStream analysis of cNKs alone or with indicated cell lines (1 h) depicting CD226 ( E ) and TIGIT ( F ) membrane (yellow) and intracellular (violet) expression, as well as surface CD45 expression (turquoise). ( G ) Representative histograms gated on total live cNKs (left panels) and statistical analysis from a pool of 30 randomly selected cNKs (right panels) showing the ratio of CD226 and TIGIT internalization by cNKs in the presence of L-CD155 and FO-I as compared with L cells or cNKs alone. Cell surface CD45 expression was used to calculate the internalization ratio of CD226 and TIGIT intracellular staining using IDEAS software. P values were obtained from one-way ANOVA tests ( A and B ) or repeated-measure ANOVA tests ( D and G ) followed by Dunnett’s multiple comparisons test with *, P

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    Article Title: IL-15 stimulation with TIGIT blockade reverses CD155-mediated NK cell dysfunction in melanoma

    doi: 10.1158/1078-0432.CCR-20-0575

    Figure Lengend Snippet: Membrane-bound CD155 induces TIGIT and CD226 internalization in NK cells. (A and B) CD226 ( A ) and TIGIT ( B ) expression by cNKs (fold change of MFI) after 48 h incubation with indicated targets +/− aCD155 blocking mAb, as compared with control (no target, n =15, or IgG-coated beads, n =8). ( C ) Fold change of CD226 expression in MFI by cNKs incubated 48 h with sCD155 or the supernatants of melanoma cell lines ( n =7). (D ) CD226 mRNA relative expression by cNKs either alone, co-cultured for 48 h with L-cells, or with L-CD155 cells. Dots are means of triplicates ( n =7). ( E and F ) Representative pictures from ImageStream analysis of cNKs alone or with indicated cell lines (1 h) depicting CD226 ( E ) and TIGIT ( F ) membrane (yellow) and intracellular (violet) expression, as well as surface CD45 expression (turquoise). ( G ) Representative histograms gated on total live cNKs (left panels) and statistical analysis from a pool of 30 randomly selected cNKs (right panels) showing the ratio of CD226 and TIGIT internalization by cNKs in the presence of L-CD155 and FO-I as compared with L cells or cNKs alone. Cell surface CD45 expression was used to calculate the internalization ratio of CD226 and TIGIT intracellular staining using IDEAS software. P values were obtained from one-way ANOVA tests ( A and B ) or repeated-measure ANOVA tests ( D and G ) followed by Dunnett’s multiple comparisons test with *, P

    Article Snippet: Human CD155/PVR transcript variant 1 Gene cDNA Clone (full-length ORF Clone, Sino Biological Inc.) was transfected into L cells using lipofectamine 3000 (Invitrogen) according to manufacturer’s instructions.

    Techniques: Expressing, Incubation, Blocking Assay, Cell Culture, Staining, Software