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anti cd8a rabbit polyclonal antibody (Bioss)

Name: anti cd8a rabbit polyclonal antibody
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Rating: 95 (102 reviews)

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Bioss anti cd8a rabbit polyclonal antibody

( A ) Representative digital three-dimensional (3D) rendering from MRI images of 4-month-old STING-sufficient Nf1 f/f ; DhhCre and STING-deficient STING gt/gt ; Nf1 f/f ; DhhCre. ( B ) Quantification of PNF volume. ( C ) Representative gross dissections of mice with WT STING and mutant STING at 7 months old. ( D ) Quantification of the number of tumors. ( E ) Representative hematoxylin and eosin (H&E) stain, toluidine blue stain for mast cells, and Iba-1 for macrophages. ( F ) PNF flow cytometer of Nf1 f/f ; DhhCre or STING gt/gt Nf1 f/f ; DhhCre mice for MHCII + CD11c + CD172a − XCR-1 + cDC1 and MHCII + CD11c + CD172a + XCR-1 − cDC2 DCs, ( G ) TCRβ + T cells and ( H ) CD4 and <t>CD8</t> T cells with ( I ) represented analysis of activation markers (CD44 and CD69). Statistical significance ( P < 0.05). n.s., not significant. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, *** P < 0.003, and **** P < 0.0001.

Anti Cd8a Rabbit Polyclonal Antibody, supplied by Bioss, used in various techniques. Bioz Stars score: 95/100, based on 102 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti cd8a rabbit polyclonal antibody - by Bioz Stars, 2026-02

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1) Product Images from "Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance"

Article Title: Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance

Journal: Science Advances

doi: 10.1126/sciadv.ado6342

Figure Legend Snippet: ( A ) Representative digital three-dimensional (3D) rendering from MRI images of 4-month-old STING-sufficient Nf1 f/f ; DhhCre and STING-deficient STING gt/gt ; Nf1 f/f ; DhhCre. ( B ) Quantification of PNF volume. ( C ) Representative gross dissections of mice with WT STING and mutant STING at 7 months old. ( D ) Quantification of the number of tumors. ( E ) Representative hematoxylin and eosin (H&E) stain, toluidine blue stain for mast cells, and Iba-1 for macrophages. ( F ) PNF flow cytometer of Nf1 f/f ; DhhCre or STING gt/gt Nf1 f/f ; DhhCre mice for MHCII + CD11c + CD172a − XCR-1 + cDC1 and MHCII + CD11c + CD172a + XCR-1 − cDC2 DCs, ( G ) TCRβ + T cells and ( H ) CD4 and CD8 T cells with ( I ) represented analysis of activation markers (CD44 and CD69). Statistical significance ( P < 0.05). n.s., not significant. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, *** P < 0.003, and **** P < 0.0001.

Techniques Used: Mutagenesis, Staining, Flow Cytometry, Activation Assay

( A ) Schematic diagram of STING inhibitor (STINGi; H-151) treatment regimen. ( B ) Representative images of gross dissected paraspinal tumors from vehicle or H-151–treated mice and the corresponding graphical quantification of tumor numbers and size. ( C ) Representative images of IHC against p-STING, p-TBK1, and p-IRF3 and ( D ) quantified in vehicle versus H-151–treated mice. ( E ) Toluidine blue staining for mast cells, immunostaining of Iba-1 for detection of macrophages and CD3 for T cells in vehicle and H-151–treated groups and ( F ) quantified wherein each panel and each dot represents data from a single mouse, in which 5 HPF (high-power field) were analyzed per animal. ( G and H ) Flow cytometric quantification of activated (CD44hi CD69hi) CD8 T cells and cDC1 in vehicle or H-151–treated mice. Student’s t-test: * P < 0.05; ** P < 0.001.

Figure Legend Snippet: ( A ) Schematic diagram of STING inhibitor (STINGi; H-151) treatment regimen. ( B ) Representative images of gross dissected paraspinal tumors from vehicle or H-151–treated mice and the corresponding graphical quantification of tumor numbers and size. ( C ) Representative images of IHC against p-STING, p-TBK1, and p-IRF3 and ( D ) quantified in vehicle versus H-151–treated mice. ( E ) Toluidine blue staining for mast cells, immunostaining of Iba-1 for detection of macrophages and CD3 for T cells in vehicle and H-151–treated groups and ( F ) quantified wherein each panel and each dot represents data from a single mouse, in which 5 HPF (high-power field) were analyzed per animal. ( G and H ) Flow cytometric quantification of activated (CD44hi CD69hi) CD8 T cells and cDC1 in vehicle or H-151–treated mice. Student’s t-test: * P < 0.05; ** P < 0.001.

Techniques Used: Staining, Immunostaining

( A ) Representative IHC images of human nerve or neurofibroma biopsies for the detection of TCRβ T cells and ( B ) double labeling for TCRβ (brown) with CD4 or CD8 (red) T cells and CD11c (brown) and XCR-1 (red) for DCs. ( C ) Representative IHC images WT DRG or Nf1 f/f ; DhhCre neurofibroma. ( D ) Representative flow cytometric analysis of TCRβ + CD3 + T cells and quantification. ( E ) CD4 and CD8 T cells subtypes and in PNF tissue corresponding flow cytometric quantification. ( F ) Immunofluorescence showing CD4 and CD8 T cells subtypes in neurofibroma tissues. ( G ) Representative flow cytometric analysis of antigen primed and activation marker CD44 high CD69 high of CD4 or CD8 T cell subpopulation from a WT DRG or Nf1 f/f ; DhhCre neurofibromas and graphical quantification showing frequency of effector T cells indicated by CD44 + CD69 + expression of CD4 or CD8 T cells. ( H ) Flow cytometric analysis of conventional DCs gated from MHCII + and CD11c + cells in WT DRG or Nf1 f/f ; DhhCre neurofibromas with corresponding quantification of Xcr-1 + (cDC1) and Sirpα + (cDC2). ( I ) Flow cytometric quantification of B cells marked by CD19 + cells in Nf1 f/f ; DhhCre neurofibroma compared to those in WT DRG. Data represent means ± SD; t test, P < 0.05. DAPI, 4′,6-diamidino-2-phenylindole. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, and *** P < 0.003.

Figure Legend Snippet: ( A ) Representative IHC images of human nerve or neurofibroma biopsies for the detection of TCRβ T cells and ( B ) double labeling for TCRβ (brown) with CD4 or CD8 (red) T cells and CD11c (brown) and XCR-1 (red) for DCs. ( C ) Representative IHC images WT DRG or Nf1 f/f ; DhhCre neurofibroma. ( D ) Representative flow cytometric analysis of TCRβ + CD3 + T cells and quantification. ( E ) CD4 and CD8 T cells subtypes and in PNF tissue corresponding flow cytometric quantification. ( F ) Immunofluorescence showing CD4 and CD8 T cells subtypes in neurofibroma tissues. ( G ) Representative flow cytometric analysis of antigen primed and activation marker CD44 high CD69 high of CD4 or CD8 T cell subpopulation from a WT DRG or Nf1 f/f ; DhhCre neurofibromas and graphical quantification showing frequency of effector T cells indicated by CD44 + CD69 + expression of CD4 or CD8 T cells. ( H ) Flow cytometric analysis of conventional DCs gated from MHCII + and CD11c + cells in WT DRG or Nf1 f/f ; DhhCre neurofibromas with corresponding quantification of Xcr-1 + (cDC1) and Sirpα + (cDC2). ( I ) Flow cytometric quantification of B cells marked by CD19 + cells in Nf1 f/f ; DhhCre neurofibroma compared to those in WT DRG. Data represent means ± SD; t test, P < 0.05. DAPI, 4′,6-diamidino-2-phenylindole. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, and *** P < 0.003.

Techniques Used: Labeling, Immunofluorescence, Activation Assay, Marker, Expressing

( A ) Representative image of gross dissection of the spinal cord with attached DRG or tumors of Nf1 f/f ; DhhCre mice with heterozygous or homozygous loss of Batf3 transcription factor necessary for cDC1 development. ( B and C ) Quantification of the number of tumors and size in Nf1 f/f ; DhhCre mice with or without cDC1 subset deficiency. ( D ) Representative H&E and S100B staining of DRG/tumors from Nf1 f/f ; DhhCre or Batf3 −/− ; Nf1 f/f ; DhhCre mice. ( E ) Flow cytometric analysis of cDC1 (Xcr-1 + ) and cDC2 (SIRPα + ) population in Nf1 f/f ; DhhCre or Batf3 −/− ; Nf1 f/f ; DhhCre tumors. ( F and G ) Representative flow cytometry and quantification of activated (CD44 + CD69 + ) CD4 or CD8 T cells. ( H ) Electron micrographs of the saphenous nerve of 7-month-old WT control, Nf1 f/f ; DhhCre neurofibroma, and Batf3 −/− ; Nf1 f/f ; DhhCre mice and quantification of intact Remak bundle. Scale bar, 100 μm. Student’s t-test: *adj. * P < 0.05, *** P < 0.003, and **** P < 0.0001.

Figure Legend Snippet: ( A ) Representative image of gross dissection of the spinal cord with attached DRG or tumors of Nf1 f/f ; DhhCre mice with heterozygous or homozygous loss of Batf3 transcription factor necessary for cDC1 development. ( B and C ) Quantification of the number of tumors and size in Nf1 f/f ; DhhCre mice with or without cDC1 subset deficiency. ( D ) Representative H&E and S100B staining of DRG/tumors from Nf1 f/f ; DhhCre or Batf3 −/− ; Nf1 f/f ; DhhCre mice. ( E ) Flow cytometric analysis of cDC1 (Xcr-1 + ) and cDC2 (SIRPα + ) population in Nf1 f/f ; DhhCre or Batf3 −/− ; Nf1 f/f ; DhhCre tumors. ( F and G ) Representative flow cytometry and quantification of activated (CD44 + CD69 + ) CD4 or CD8 T cells. ( H ) Electron micrographs of the saphenous nerve of 7-month-old WT control, Nf1 f/f ; DhhCre neurofibroma, and Batf3 −/− ; Nf1 f/f ; DhhCre mice and quantification of intact Remak bundle. Scale bar, 100 μm. Student’s t-test: *adj. * P < 0.05, *** P < 0.003, and **** P < 0.0001.

Techniques Used: Dissection, Staining, Flow Cytometry, Control

( A ) Schematic diagram of the T cell proliferation assay: CD8 + T cells were purified by negative selection using a Miltenyi kit. T cells from tumor-bearing mice were labeled with CFSE and mixed with APCs from WT or tumor-bearing Nf1 f/f ; DhhCre mice. Four days later, cells were harvested and assessed for CFSE dilution. Graph shows the percent of CD8 + T cells that had proliferated (CFSE low). ( B ) Quantification on proliferating CD8 T cells from WT mice cocultured with APC from WT or Nf1 f/f ; DhhCre mice. ( C ) CD8 T cell proliferation from Nf1 f/f ; DhhCre mice cocultured with APC from WT or Nf1 f/f ; DhhCre mice. (C) Representative images of gross dissections of mouse spinal cord with the associated neurofibromas of the dorsal root in Nf1 f/f ; DhhCre mice not observed in Rag1 −/− ; Nf1 f/f ; DhhCre mice that lack adaptive immune cells at 7 months of age. ( D ) Quantification of the number of tumors observed per mouse and tumor diameters between Nf1 f/f ; DhhCre and Rag1 −/− ; Nf1 f/f ; DhhCre mice. ( E ) Representative tissue histochemistry of DRGs/PNF in Nf1 f/f ; DhhCre or Rag1 −/− ; Nf1 f/f ; DhhCre stained with H&E, S100β, and Ki67. ( F ) Immunofluorescence of Iba-1 + macrophages in DRG or neurofibromas, and the number of Iba-1 + per field of view (FOV). ( G ) Toluidine blue stain for detection of mast cells, and the number of toluidine blue–positive mast cells per field of view. ( H ) Electron micrographs of the saphenous nerve of 7-month-old WT control, Nf1 f/f ; DhhCre neurofibroma, and Rag1 −/− ; Nf1 f/f ; DhhCre mice. ( I ) Quantification of intact Remak bundle. * P < 0.05; Student’s t-test: **adj. ** P < 0.001; *** P < 0.003, and **** P < 0.0001.

Figure Legend Snippet: ( A ) Schematic diagram of the T cell proliferation assay: CD8 + T cells were purified by negative selection using a Miltenyi kit. T cells from tumor-bearing mice were labeled with CFSE and mixed with APCs from WT or tumor-bearing Nf1 f/f ; DhhCre mice. Four days later, cells were harvested and assessed for CFSE dilution. Graph shows the percent of CD8 + T cells that had proliferated (CFSE low). ( B ) Quantification on proliferating CD8 T cells from WT mice cocultured with APC from WT or Nf1 f/f ; DhhCre mice. ( C ) CD8 T cell proliferation from Nf1 f/f ; DhhCre mice cocultured with APC from WT or Nf1 f/f ; DhhCre mice. (C) Representative images of gross dissections of mouse spinal cord with the associated neurofibromas of the dorsal root in Nf1 f/f ; DhhCre mice not observed in Rag1 −/− ; Nf1 f/f ; DhhCre mice that lack adaptive immune cells at 7 months of age. ( D ) Quantification of the number of tumors observed per mouse and tumor diameters between Nf1 f/f ; DhhCre and Rag1 −/− ; Nf1 f/f ; DhhCre mice. ( E ) Representative tissue histochemistry of DRGs/PNF in Nf1 f/f ; DhhCre or Rag1 −/− ; Nf1 f/f ; DhhCre stained with H&E, S100β, and Ki67. ( F ) Immunofluorescence of Iba-1 + macrophages in DRG or neurofibromas, and the number of Iba-1 + per field of view (FOV). ( G ) Toluidine blue stain for detection of mast cells, and the number of toluidine blue–positive mast cells per field of view. ( H ) Electron micrographs of the saphenous nerve of 7-month-old WT control, Nf1 f/f ; DhhCre neurofibroma, and Rag1 −/− ; Nf1 f/f ; DhhCre mice. ( I ) Quantification of intact Remak bundle. * P < 0.05; Student’s t-test: **adj. ** P < 0.001; *** P < 0.003, and **** P < 0.0001.

Techniques Used: Proliferation Assay, Purification, Selection, Labeling, Staining, Immunofluorescence, Control

( A ) Schematic of CD8 T cell depletion. ( B ) Flow cytometric quantification in the proportion of CD4 or CD8 of the total blood CD3 T cells in Nf1 f/f ; DhhCre compared to that in Rag1 −/− ; Nf1 f/f ; DhhCre. ( C ) Percentage of CD3 T cells at 1 and 2 months (1Mo and 2Mo, respectively) after adoptive transfer. ( D ) Percentage of CD4 and CD8 T cells present in circulation after adoptive transfer. ( E ) Representative flow cytometric analysis of blood CD3 T cells for the markers CD44 and CD127 at 2 months old after total T cell AT. ( F ) Representative MRI image of Nf1 f/f ; DhhCre neurofibroma bearing mice and Rag1 −/− ; Nf1 f/f ; DhhCre that were adoptively transferred with T cells or PBS as control. ( G ) Representative images of gross dissection of spinal cord with the associated neurofibromas of the dorsal root in 7-month-old Rag1 −/− ; Nf1 f/f ; DhhCre after adoptive transfer of pan-T cells. ( H ) Representative flow cytometric analysis for CD44 and CD127 of blood from Rag1 −/− ; Nf1 f/f ; DhhCre mice at 2 months old after adoptive transfer (AT) of CD8 or CD4 T cells. ( I ) Representative MRI image of Rag1 −/− ; Nf1 f/f ; DhhCre that were adoptively transferred with CD4 T cells or CD8 T cells alone. ( J ) Quantification of T cell recipient Rag1 −/− ; Nf1 f/f ; DhhCre mice that developed neurofibromas after adoptively transferred with or without total T cells or CD4 or CD8 T cells. ( K ) Predicted gene expression of CCL5 and its receptor CCR5 in immune cells of PNF tissue. m.o., months old. ( L ) CCL5 protein expression determined by flow cytometry of PNF tissue. Student’s t-test: *adj. * P < 0.05, *** P < 0.003, and **** P < 0.0001..

Figure Legend Snippet: ( A ) Schematic of CD8 T cell depletion. ( B ) Flow cytometric quantification in the proportion of CD4 or CD8 of the total blood CD3 T cells in Nf1 f/f ; DhhCre compared to that in Rag1 −/− ; Nf1 f/f ; DhhCre. ( C ) Percentage of CD3 T cells at 1 and 2 months (1Mo and 2Mo, respectively) after adoptive transfer. ( D ) Percentage of CD4 and CD8 T cells present in circulation after adoptive transfer. ( E ) Representative flow cytometric analysis of blood CD3 T cells for the markers CD44 and CD127 at 2 months old after total T cell AT. ( F ) Representative MRI image of Nf1 f/f ; DhhCre neurofibroma bearing mice and Rag1 −/− ; Nf1 f/f ; DhhCre that were adoptively transferred with T cells or PBS as control. ( G ) Representative images of gross dissection of spinal cord with the associated neurofibromas of the dorsal root in 7-month-old Rag1 −/− ; Nf1 f/f ; DhhCre after adoptive transfer of pan-T cells. ( H ) Representative flow cytometric analysis for CD44 and CD127 of blood from Rag1 −/− ; Nf1 f/f ; DhhCre mice at 2 months old after adoptive transfer (AT) of CD8 or CD4 T cells. ( I ) Representative MRI image of Rag1 −/− ; Nf1 f/f ; DhhCre that were adoptively transferred with CD4 T cells or CD8 T cells alone. ( J ) Quantification of T cell recipient Rag1 −/− ; Nf1 f/f ; DhhCre mice that developed neurofibromas after adoptively transferred with or without total T cells or CD4 or CD8 T cells. ( K ) Predicted gene expression of CCL5 and its receptor CCR5 in immune cells of PNF tissue. m.o., months old. ( L ) CCL5 protein expression determined by flow cytometry of PNF tissue. Student’s t-test: *adj. * P < 0.05, *** P < 0.003, and **** P < 0.0001..

Techniques Used: Adoptive Transfer Assay, Control, Dissection, Expressing, Flow Cytometry

( A ) Experimental design of CD8 T cell depletion, in which, after adoptive transfer of CD8 T cells in Rag1 −/− ; Nf1 f/f DhhCre and confirming PNF development, CD8 antibody (Ab) or IgG control were administered once a week at 250 μg per mouse. ( B ) MRI images of Rag1 −/− ; Nf1 f/f ; DhhCre after CD8 T cell adoptive transfer confirming PNF development. ( C ) Flow cytometry of circulatory T cells after three treatment doses. ( D ) Gross dissection of mice after 2 months from treatment of CD8 antibody revealing reduced PNF formation. ( E ) Toluidine blue staining for mast cells of nerve or PNF sections. ( F ) Immunofluorescence of TCRβ for T cells and Iba-1 for macrophages in nerve or PNF. ( G ) Immunofluorescence of CCR5 (green) expressing F4/80 macrophages (red). ( H ) Flow cytometry of circulatory T cells after at the 6-month-old endpoint indicates reduced CD8 T cells mice treated with CD8 depletion antibody compared to that in IgG-treated control. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, and *** P < 0.003.

Figure Legend Snippet: ( A ) Experimental design of CD8 T cell depletion, in which, after adoptive transfer of CD8 T cells in Rag1 −/− ; Nf1 f/f DhhCre and confirming PNF development, CD8 antibody (Ab) or IgG control were administered once a week at 250 μg per mouse. ( B ) MRI images of Rag1 −/− ; Nf1 f/f ; DhhCre after CD8 T cell adoptive transfer confirming PNF development. ( C ) Flow cytometry of circulatory T cells after three treatment doses. ( D ) Gross dissection of mice after 2 months from treatment of CD8 antibody revealing reduced PNF formation. ( E ) Toluidine blue staining for mast cells of nerve or PNF sections. ( F ) Immunofluorescence of TCRβ for T cells and Iba-1 for macrophages in nerve or PNF. ( G ) Immunofluorescence of CCR5 (green) expressing F4/80 macrophages (red). ( H ) Flow cytometry of circulatory T cells after at the 6-month-old endpoint indicates reduced CD8 T cells mice treated with CD8 depletion antibody compared to that in IgG-treated control. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, and *** P < 0.003.

Techniques Used: Adoptive Transfer Assay, Control, Flow Cytometry, Dissection, Staining, Immunofluorescence, Expressing

Image Search Results

Journal: Science Advances

Article Title: Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance

doi: 10.1126/sciadv.ado6342

Figure Lengend Snippet: ( A ) Representative digital three-dimensional (3D) rendering from MRI images of 4-month-old STING-sufficient Nf1 f/f ; DhhCre and STING-deficient STING gt/gt ; Nf1 f/f ; DhhCre. ( B ) Quantification of PNF volume. ( C ) Representative gross dissections of mice with WT STING and mutant STING at 7 months old. ( D ) Quantification of the number of tumors. ( E ) Representative hematoxylin and eosin (H&E) stain, toluidine blue stain for mast cells, and Iba-1 for macrophages. ( F ) PNF flow cytometer of Nf1 f/f ; DhhCre or STING gt/gt Nf1 f/f ; DhhCre mice for MHCII + CD11c + CD172a − XCR-1 + cDC1 and MHCII + CD11c + CD172a + XCR-1 − cDC2 DCs, ( G ) TCRβ + T cells and ( H ) CD4 and CD8 T cells with ( I ) represented analysis of activation markers (CD44 and CD69). Statistical significance ( P < 0.05). n.s., not significant. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, *** P < 0.003, and **** P < 0.0001.

Article Snippet: The following antibodies were used: phospho-STING (Ser 366 ) (E9A9K) rabbit monoclonal antibody (mAb) (Cell Signaling Technology), phospho-STING (Ser 365 ) (D1C4T) rabbit mAb (no. 62912, Cell Signaling Technology), phospho-STING (Ser 366 ) (E9A9K) rabbit mAb (no. 50907, Cell Signaling Technology), phospho-IRF3 (Ser 396 ) (4D4G) rabbit mAb (Cell Signaling Technology), phospho-TBK1/NAK (Ser 172 ) (D52C2) XP rabbit mAb (Cell Signaling Technology), phospho-IRF3 (Ser 396 ) (D6O1M) rabbit mAb (no. 29047S), F4/80 (BM8.1) rat mAb (Cell Signaling Technology), mouse CCR5 antibody (MAB6138-SP, Thermo Fisher Scientific), CoraLite488 anti-mouse TCRβ (H57-597) (CL488-65106, Proteintech), anti-CD4 rabbit polyclonal antibody (BS-0647R, Bioss Antibodies), anti-CD8A rabbit polyclonal antibody (BS-0648R, Bioss Antibodies), XCR1 (D2F8T) rabbit mAb (no. 44665S, Cell Signaling Technology), CD103/INGAE (integrin alpha e) (EP206) rabbit mAb (no. 95835S, Cell Signaling Technology), anti-CD8 antibody, rabbit monoclonal (clone SP16) (SAB5500074, Sigma-Aldrich), CD3ε (E4T1B) XP Rabbit mAb (no. 78588, Cell Signaling Technology), and Iba-1 rabbit polyclonal (no. 019-19741, Wako).

Techniques: Mutagenesis, Staining, Flow Cytometry, Activation Assay

Journal: Science Advances

Article Title: Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance

doi: 10.1126/sciadv.ado6342

Figure Lengend Snippet: ( A ) Schematic diagram of STING inhibitor (STINGi; H-151) treatment regimen. ( B ) Representative images of gross dissected paraspinal tumors from vehicle or H-151–treated mice and the corresponding graphical quantification of tumor numbers and size. ( C ) Representative images of IHC against p-STING, p-TBK1, and p-IRF3 and ( D ) quantified in vehicle versus H-151–treated mice. ( E ) Toluidine blue staining for mast cells, immunostaining of Iba-1 for detection of macrophages and CD3 for T cells in vehicle and H-151–treated groups and ( F ) quantified wherein each panel and each dot represents data from a single mouse, in which 5 HPF (high-power field) were analyzed per animal. ( G and H ) Flow cytometric quantification of activated (CD44hi CD69hi) CD8 T cells and cDC1 in vehicle or H-151–treated mice. Student’s t-test: * P < 0.05; ** P < 0.001.

Techniques: Staining, Immunostaining

Journal: Science Advances

Article Title: Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance

doi: 10.1126/sciadv.ado6342

Figure Lengend Snippet: ( A ) Representative IHC images of human nerve or neurofibroma biopsies for the detection of TCRβ T cells and ( B ) double labeling for TCRβ (brown) with CD4 or CD8 (red) T cells and CD11c (brown) and XCR-1 (red) for DCs. ( C ) Representative IHC images WT DRG or Nf1 f/f ; DhhCre neurofibroma. ( D ) Representative flow cytometric analysis of TCRβ + CD3 + T cells and quantification. ( E ) CD4 and CD8 T cells subtypes and in PNF tissue corresponding flow cytometric quantification. ( F ) Immunofluorescence showing CD4 and CD8 T cells subtypes in neurofibroma tissues. ( G ) Representative flow cytometric analysis of antigen primed and activation marker CD44 high CD69 high of CD4 or CD8 T cell subpopulation from a WT DRG or Nf1 f/f ; DhhCre neurofibromas and graphical quantification showing frequency of effector T cells indicated by CD44 + CD69 + expression of CD4 or CD8 T cells. ( H ) Flow cytometric analysis of conventional DCs gated from MHCII + and CD11c + cells in WT DRG or Nf1 f/f ; DhhCre neurofibromas with corresponding quantification of Xcr-1 + (cDC1) and Sirpα + (cDC2). ( I ) Flow cytometric quantification of B cells marked by CD19 + cells in Nf1 f/f ; DhhCre neurofibroma compared to those in WT DRG. Data represent means ± SD; t test, P < 0.05. DAPI, 4′,6-diamidino-2-phenylindole. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, and *** P < 0.003.

Techniques: Labeling, Immunofluorescence, Activation Assay, Marker, Expressing

Journal: Science Advances

Article Title: Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance

doi: 10.1126/sciadv.ado6342

Figure Lengend Snippet: ( A ) Representative image of gross dissection of the spinal cord with attached DRG or tumors of Nf1 f/f ; DhhCre mice with heterozygous or homozygous loss of Batf3 transcription factor necessary for cDC1 development. ( B and C ) Quantification of the number of tumors and size in Nf1 f/f ; DhhCre mice with or without cDC1 subset deficiency. ( D ) Representative H&E and S100B staining of DRG/tumors from Nf1 f/f ; DhhCre or Batf3 −/− ; Nf1 f/f ; DhhCre mice. ( E ) Flow cytometric analysis of cDC1 (Xcr-1 + ) and cDC2 (SIRPα + ) population in Nf1 f/f ; DhhCre or Batf3 −/− ; Nf1 f/f ; DhhCre tumors. ( F and G ) Representative flow cytometry and quantification of activated (CD44 + CD69 + ) CD4 or CD8 T cells. ( H ) Electron micrographs of the saphenous nerve of 7-month-old WT control, Nf1 f/f ; DhhCre neurofibroma, and Batf3 −/− ; Nf1 f/f ; DhhCre mice and quantification of intact Remak bundle. Scale bar, 100 μm. Student’s t-test: *adj. * P < 0.05, *** P < 0.003, and **** P < 0.0001.

Techniques: Dissection, Staining, Flow Cytometry, Control

Journal: Science Advances

Article Title: Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance

doi: 10.1126/sciadv.ado6342

Figure Lengend Snippet: ( A ) Schematic diagram of the T cell proliferation assay: CD8 + T cells were purified by negative selection using a Miltenyi kit. T cells from tumor-bearing mice were labeled with CFSE and mixed with APCs from WT or tumor-bearing Nf1 f/f ; DhhCre mice. Four days later, cells were harvested and assessed for CFSE dilution. Graph shows the percent of CD8 + T cells that had proliferated (CFSE low). ( B ) Quantification on proliferating CD8 T cells from WT mice cocultured with APC from WT or Nf1 f/f ; DhhCre mice. ( C ) CD8 T cell proliferation from Nf1 f/f ; DhhCre mice cocultured with APC from WT or Nf1 f/f ; DhhCre mice. (C) Representative images of gross dissections of mouse spinal cord with the associated neurofibromas of the dorsal root in Nf1 f/f ; DhhCre mice not observed in Rag1 −/− ; Nf1 f/f ; DhhCre mice that lack adaptive immune cells at 7 months of age. ( D ) Quantification of the number of tumors observed per mouse and tumor diameters between Nf1 f/f ; DhhCre and Rag1 −/− ; Nf1 f/f ; DhhCre mice. ( E ) Representative tissue histochemistry of DRGs/PNF in Nf1 f/f ; DhhCre or Rag1 −/− ; Nf1 f/f ; DhhCre stained with H&E, S100β, and Ki67. ( F ) Immunofluorescence of Iba-1 + macrophages in DRG or neurofibromas, and the number of Iba-1 + per field of view (FOV). ( G ) Toluidine blue stain for detection of mast cells, and the number of toluidine blue–positive mast cells per field of view. ( H ) Electron micrographs of the saphenous nerve of 7-month-old WT control, Nf1 f/f ; DhhCre neurofibroma, and Rag1 −/− ; Nf1 f/f ; DhhCre mice. ( I ) Quantification of intact Remak bundle. * P < 0.05; Student’s t-test: **adj. ** P < 0.001; *** P < 0.003, and **** P < 0.0001.

Techniques: Proliferation Assay, Purification, Selection, Labeling, Staining, Immunofluorescence, Control

Journal: Science Advances

Article Title: Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance

doi: 10.1126/sciadv.ado6342

Figure Lengend Snippet: ( A ) Schematic of CD8 T cell depletion. ( B ) Flow cytometric quantification in the proportion of CD4 or CD8 of the total blood CD3 T cells in Nf1 f/f ; DhhCre compared to that in Rag1 −/− ; Nf1 f/f ; DhhCre. ( C ) Percentage of CD3 T cells at 1 and 2 months (1Mo and 2Mo, respectively) after adoptive transfer. ( D ) Percentage of CD4 and CD8 T cells present in circulation after adoptive transfer. ( E ) Representative flow cytometric analysis of blood CD3 T cells for the markers CD44 and CD127 at 2 months old after total T cell AT. ( F ) Representative MRI image of Nf1 f/f ; DhhCre neurofibroma bearing mice and Rag1 −/− ; Nf1 f/f ; DhhCre that were adoptively transferred with T cells or PBS as control. ( G ) Representative images of gross dissection of spinal cord with the associated neurofibromas of the dorsal root in 7-month-old Rag1 −/− ; Nf1 f/f ; DhhCre after adoptive transfer of pan-T cells. ( H ) Representative flow cytometric analysis for CD44 and CD127 of blood from Rag1 −/− ; Nf1 f/f ; DhhCre mice at 2 months old after adoptive transfer (AT) of CD8 or CD4 T cells. ( I ) Representative MRI image of Rag1 −/− ; Nf1 f/f ; DhhCre that were adoptively transferred with CD4 T cells or CD8 T cells alone. ( J ) Quantification of T cell recipient Rag1 −/− ; Nf1 f/f ; DhhCre mice that developed neurofibromas after adoptively transferred with or without total T cells or CD4 or CD8 T cells. ( K ) Predicted gene expression of CCL5 and its receptor CCR5 in immune cells of PNF tissue. m.o., months old. ( L ) CCL5 protein expression determined by flow cytometry of PNF tissue. Student’s t-test: *adj. * P < 0.05, *** P < 0.003, and **** P < 0.0001..

Techniques: Adoptive Transfer Assay, Control, Dissection, Expressing, Flow Cytometry

Journal: Science Advances

Article Title: Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance

doi: 10.1126/sciadv.ado6342

Figure Lengend Snippet: ( A ) Experimental design of CD8 T cell depletion, in which, after adoptive transfer of CD8 T cells in Rag1 −/− ; Nf1 f/f DhhCre and confirming PNF development, CD8 antibody (Ab) or IgG control were administered once a week at 250 μg per mouse. ( B ) MRI images of Rag1 −/− ; Nf1 f/f ; DhhCre after CD8 T cell adoptive transfer confirming PNF development. ( C ) Flow cytometry of circulatory T cells after three treatment doses. ( D ) Gross dissection of mice after 2 months from treatment of CD8 antibody revealing reduced PNF formation. ( E ) Toluidine blue staining for mast cells of nerve or PNF sections. ( F ) Immunofluorescence of TCRβ for T cells and Iba-1 for macrophages in nerve or PNF. ( G ) Immunofluorescence of CCR5 (green) expressing F4/80 macrophages (red). ( H ) Flow cytometry of circulatory T cells after at the 6-month-old endpoint indicates reduced CD8 T cells mice treated with CD8 depletion antibody compared to that in IgG-treated control. Student’s t-test: *adj. * P < 0.05, ** P < 0.001, and *** P < 0.003.

Techniques: Adoptive Transfer Assay, Control, Flow Cytometry, Dissection, Staining, Immunofluorescence, Expressing

6q21 Amplification‐Associated NR3C1 Overexpression was Involved in Immune Escape of Tumor Cells in DCIS_Pure. A) Boxplot illustrating the immune scores across different stages of BRDC progression: Normal ( n = 19), DH ( n = 54), DCIS_Pure ( n = 30), DCIS_adjIDC ( n = 43), and IDC ( n = 78). The box represents the interquartile range (IQR) from the first (Q1) to the third quartile (Q3) of the distribution, and the line inside the box represents the median. The whiskers extend from Q1 to Q3 to the endpoints, which are defined as the most extreme data points within Q1 − 1.5 × IQR and Q3 + 1.5 × IQR, respectively. Student's t ‐test, **** p < 0.0001. B) Heatmap illustrating cell type compositions and activities of selected individual genes/proteins and pathways across the BRDC progression stages. The heatmap in the first section illustrated the immune/stromal signatures from xCell. The heatmap in the second section illustrates the protein abundances of sterol hormone receptors and immune‐related marker CD8A. ssGSEA scores based on global proteomic and phosphoproteomic data for biological pathways upregulated in different progression stages of BRDC are illustrated in the remaining sections. ANOVA test, * p < 0.05, *** p < 0.001, **** p < 0.0001. C) Comparison of the protein expression levels of sterol hormone receptors between the DH ( n = 54) and DCIS_Pure ( n = 30). The p value was calculated by Student's t ‐test. D) Volcano plot showing the correlation between the protein expression levels of NR3C1 TFs and their corresponding CNA. The red points show the cis ‐effect TFs (Spearman's rho > 0.2, p < 0.05). Histogram showing the fold changes of the protein expression levels of the cis ‐effect TFs in DCIS_Pure compared with DH. Student's t ‐test (upper left corner). E) Heatmaps showing copy number alteration (CNA) of PRDM1 , the protein abundances of PRDM1 and NR3C1, the immune score, and the pathway score of TNF‐α signaling via NF‐ĸB in DH and DCIS_Pure. Student's t ‐test, * p < 0.05, *** p < 0.001, **** p < 0.0001. F) Boxplot showing the transcriptional activities of NF‐ĸB1 and RelA in DH ( n = 54) and DCIS_Pure ( n = 30). Student's t ‐test, * p < 0.05, **** p < 0.0001. G) Volcano plot showing the correlations between the transcription factor activity of NF‐ĸB1 and RelA and the pathway scores by ssGSEA. The one highlighted in red represents the pathway of cytokine‐cytokine receptor interaction (Spearman's rho = 0.72, p = 1.61e‐14). H) Heatmaps showing the protein expression levels of CXCL12 and CX3CL1 (upper) and the mRNA levels of CXCL12 and CX3CL1 (bottom) in DH and DCIS_Pure. Student's t ‐test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. I) Spearman‐rank correlation of the protein expression levels of CX3CL1 and CD8A. J) Representative IHC images of NR3C1 and CD8 on DH, DCIS_Pure, and DCIS_adjIDC tissues. Scale bars = 50 µm. K) The paradigm of the immune alteration among the DH, DCIS_Pure, and DCIS_adjIDC.

Journal: Advanced Science

Article Title: Proteogenomic Landscape of Breast Ductal Carcinoma Reveals Tumor Progression Characteristics and Therapeutic Targets

doi: 10.1002/advs.202401041

Figure Lengend Snippet: 6q21 Amplification‐Associated NR3C1 Overexpression was Involved in Immune Escape of Tumor Cells in DCIS_Pure. A) Boxplot illustrating the immune scores across different stages of BRDC progression: Normal ( n = 19), DH ( n = 54), DCIS_Pure ( n = 30), DCIS_adjIDC ( n = 43), and IDC ( n = 78). The box represents the interquartile range (IQR) from the first (Q1) to the third quartile (Q3) of the distribution, and the line inside the box represents the median. The whiskers extend from Q1 to Q3 to the endpoints, which are defined as the most extreme data points within Q1 − 1.5 × IQR and Q3 + 1.5 × IQR, respectively. Student's t ‐test, **** p < 0.0001. B) Heatmap illustrating cell type compositions and activities of selected individual genes/proteins and pathways across the BRDC progression stages. The heatmap in the first section illustrated the immune/stromal signatures from xCell. The heatmap in the second section illustrates the protein abundances of sterol hormone receptors and immune‐related marker CD8A. ssGSEA scores based on global proteomic and phosphoproteomic data for biological pathways upregulated in different progression stages of BRDC are illustrated in the remaining sections. ANOVA test, * p < 0.05, *** p < 0.001, **** p < 0.0001. C) Comparison of the protein expression levels of sterol hormone receptors between the DH ( n = 54) and DCIS_Pure ( n = 30). The p value was calculated by Student's t ‐test. D) Volcano plot showing the correlation between the protein expression levels of NR3C1 TFs and their corresponding CNA. The red points show the cis ‐effect TFs (Spearman's rho > 0.2, p < 0.05). Histogram showing the fold changes of the protein expression levels of the cis ‐effect TFs in DCIS_Pure compared with DH. Student's t ‐test (upper left corner). E) Heatmaps showing copy number alteration (CNA) of PRDM1 , the protein abundances of PRDM1 and NR3C1, the immune score, and the pathway score of TNF‐α signaling via NF‐ĸB in DH and DCIS_Pure. Student's t ‐test, * p < 0.05, *** p < 0.001, **** p < 0.0001. F) Boxplot showing the transcriptional activities of NF‐ĸB1 and RelA in DH ( n = 54) and DCIS_Pure ( n = 30). Student's t ‐test, * p < 0.05, **** p < 0.0001. G) Volcano plot showing the correlations between the transcription factor activity of NF‐ĸB1 and RelA and the pathway scores by ssGSEA. The one highlighted in red represents the pathway of cytokine‐cytokine receptor interaction (Spearman's rho = 0.72, p = 1.61e‐14). H) Heatmaps showing the protein expression levels of CXCL12 and CX3CL1 (upper) and the mRNA levels of CXCL12 and CX3CL1 (bottom) in DH and DCIS_Pure. Student's t ‐test, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. I) Spearman‐rank correlation of the protein expression levels of CX3CL1 and CD8A. J) Representative IHC images of NR3C1 and CD8 on DH, DCIS_Pure, and DCIS_adjIDC tissues. Scale bars = 50 µm. K) The paradigm of the immune alteration among the DH, DCIS_Pure, and DCIS_adjIDC.

Article Snippet: Mutant TP53: mouse monoclonal anti‐P53 (1:800, Leica Biosystems, Cat# PA0067); ESR1: mouse monoclonal anti‐ESR1 (1:500, Abcam, Cat# ab241557); NR3C1: rabbit monoclonal anti‐NR3C1 (1:1000, Abcam, Cat# ab183127); CD8A: rabbit polyclonal anti‐CD8A (1:50, Invitrogen, Cat# PA5‐11453); TIAM1: rabbit polyclonal anti‐TIAM1 (1:100, Cloud‐clone, Cat# PAC778Hu01); AR: rabbit polyclonal anti‐AR (1:200, Gene Tech, Cat# GT245202); AKR1C1: rabbit polyclonal anti‐AKR1C1 (1:500, GeneTex, Cat# GTX105620).

Techniques: Amplification, Over Expression, Marker, Comparison, Expressing, Activity Assay

NR3C1 overexpression was involved in immune escape of tumor cells. A) Real‐time PCR analysis of PRDM1 in PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, *** p < 0.001. B) A representative western blot analysis of PRDM1 and NR3C1 in PRDM1 ‐knockdown MCF10DCIS.COM cells (left). Quantified western blot results (middle and right). Student's t ‐test, *** p < 0.001. C,D) Real‐time PCR analysis of CX3CL1 and CXCL12 in PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, ** p < 0.01, *** p < 0.001. E,F) ELISA analysis of the protein levels of CX3CL1 and CXCL12 in cell lysate supernatant of PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, **** p < 0.0001. G,H) ELISA analysis of the protein levels of CX3CL1 and CXCL12 in CM of PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, **** p < 0.0001. I) Real‐time PCR analysis of NR3C1 in PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, *** p < 0.001. J) A representative western blot analysis of NR3C1 in NR3C1 ‐knockdown MCF10DCIS.COM cells (left). Quantified western blot results (middle and right). Student's t ‐test, *** p < 0.001. K) A representative western blot analysis of p‐p65 in NR3C1 ‐knockdown MCF10DCIS.COM cells or treatment with 10 µ m BAY 11–7082 (left). Quantified western blot results (right). Student's t ‐test, * p < 0.05, ** p < 0.01. L,M) Real‐time PCR analysis of CX3CL1 and CXCL12 in NR3C1 ‐knockdown MCF10DCIS.COM cells or treatment with 10 µ m BAY 11–7082. Student's t ‐test, *** p < 0.001. N,O) ELISA analysis of the protein levels of CX3CL1 and CXCL12 in cell lysate supernatant of NR3C1 ‐knockdown MCF10DCIS.COM cells or treatment with 10 µ m BAY 11–7082. Student's t ‐test, *** p < 0.001, **** p < 0.0001. P,Q) ELISA analysis of the protein levels of CX3CL1 and CXCL12 in CM of NR3C1 ‐knockdown MCF10DCIS.COM cells or treatment with 10 µ m BAY 11–7082. Student's t ‐test, * p < 0.05, **** p < 0.0001. R) The purity of the isolated CD8+ T cells from PBMCs of BC patients were determined by flow cytometry. S) Two‐chamber migration of CD8+ T cells recruited by CM of NR3C1 ‐knockdown MCF10DCIS.COM cells for 12 h. Student's t ‐test, ** p < 0.01.

Journal: Advanced Science

Article Title: Proteogenomic Landscape of Breast Ductal Carcinoma Reveals Tumor Progression Characteristics and Therapeutic Targets

doi: 10.1002/advs.202401041

Figure Lengend Snippet: NR3C1 overexpression was involved in immune escape of tumor cells. A) Real‐time PCR analysis of PRDM1 in PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, *** p < 0.001. B) A representative western blot analysis of PRDM1 and NR3C1 in PRDM1 ‐knockdown MCF10DCIS.COM cells (left). Quantified western blot results (middle and right). Student's t ‐test, *** p < 0.001. C,D) Real‐time PCR analysis of CX3CL1 and CXCL12 in PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, ** p < 0.01, *** p < 0.001. E,F) ELISA analysis of the protein levels of CX3CL1 and CXCL12 in cell lysate supernatant of PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, **** p < 0.0001. G,H) ELISA analysis of the protein levels of CX3CL1 and CXCL12 in CM of PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, **** p < 0.0001. I) Real‐time PCR analysis of NR3C1 in PRDM1 ‐knockdown MCF10DCIS.COM cells. Student's t ‐test, *** p < 0.001. J) A representative western blot analysis of NR3C1 in NR3C1 ‐knockdown MCF10DCIS.COM cells (left). Quantified western blot results (middle and right). Student's t ‐test, *** p < 0.001. K) A representative western blot analysis of p‐p65 in NR3C1 ‐knockdown MCF10DCIS.COM cells or treatment with 10 µ m BAY 11–7082 (left). Quantified western blot results (right). Student's t ‐test, * p < 0.05, ** p < 0.01. L,M) Real‐time PCR analysis of CX3CL1 and CXCL12 in NR3C1 ‐knockdown MCF10DCIS.COM cells or treatment with 10 µ m BAY 11–7082. Student's t ‐test, *** p < 0.001. N,O) ELISA analysis of the protein levels of CX3CL1 and CXCL12 in cell lysate supernatant of NR3C1 ‐knockdown MCF10DCIS.COM cells or treatment with 10 µ m BAY 11–7082. Student's t ‐test, *** p < 0.001, **** p < 0.0001. P,Q) ELISA analysis of the protein levels of CX3CL1 and CXCL12 in CM of NR3C1 ‐knockdown MCF10DCIS.COM cells or treatment with 10 µ m BAY 11–7082. Student's t ‐test, * p < 0.05, **** p < 0.0001. R) The purity of the isolated CD8+ T cells from PBMCs of BC patients were determined by flow cytometry. S) Two‐chamber migration of CD8+ T cells recruited by CM of NR3C1 ‐knockdown MCF10DCIS.COM cells for 12 h. Student's t ‐test, ** p < 0.01.

Techniques: Over Expression, Real-time Polymerase Chain Reaction, Knockdown, Western Blot, Enzyme-linked Immunosorbent Assay, Isolation, Flow Cytometry, Migration

D-Nap-GFFY-T317 inhibits formation of urethane-induced lung tumors and atypical hyperplasia in WT but not IFNγ -/- mice. ( A ) WT or IFNγ -/- mice were randomly divided into 3 groups, and received following treatment: NC group, fed normal chow; TF group, fed normal chow containing T317 (5 mg/day/kg bodyweight); TH group, fed normal chow and s.c. injected D-Nap-GFFY once another day with dose of T317 at 10 mg/kg bodyweight or 5 mg/day/kg bodyweight. After one week of treatment, all the mice were i.p. injected urethane (1 g/kg bodyweight) once every 3 days for 8 times. After 126 days of the 1 st time urethane injection, all mice were sacrificed followed by collection of lung samples. ( B ) all lung samples were checked the tumor incidence on lung surface. ( C-D ) lung was photographed, the number of macroscopic external pulmonary nodules was counted. Arrows indicate the representative tumors. ( E-F ) after preparation, lung sections were conducted HE staining to determine tumor area with quantitative assay as % of whole section. ( G-L ) lung sections were conducted IHC staining to determine expression of Ki-67, TTF-1 and SPC with quantitative analysis of the average optical density (AOD) value (n ≥ 4). NEC: negative control, normal IgG was used to replace primary antibody. *P < 0.05, **P < 0.01, ***P < 0.001; NS: not significantly different. n = 14 for WT mice, n ≥ 8 for IFNγ -/- mice as indicated in Table .

Journal: Theranostics

Article Title: Encapsulation of LXR ligand by D-Nap-GFFY hydrogel enhances anti-tumorigenic actions of LXR and removes LXR-induced lipogenesis

doi: 10.7150/thno.53139

Figure Lengend Snippet: D-Nap-GFFY-T317 inhibits formation of urethane-induced lung tumors and atypical hyperplasia in WT but not IFNγ -/- mice. ( A ) WT or IFNγ -/- mice were randomly divided into 3 groups, and received following treatment: NC group, fed normal chow; TF group, fed normal chow containing T317 (5 mg/day/kg bodyweight); TH group, fed normal chow and s.c. injected D-Nap-GFFY once another day with dose of T317 at 10 mg/kg bodyweight or 5 mg/day/kg bodyweight. After one week of treatment, all the mice were i.p. injected urethane (1 g/kg bodyweight) once every 3 days for 8 times. After 126 days of the 1 st time urethane injection, all mice were sacrificed followed by collection of lung samples. ( B ) all lung samples were checked the tumor incidence on lung surface. ( C-D ) lung was photographed, the number of macroscopic external pulmonary nodules was counted. Arrows indicate the representative tumors. ( E-F ) after preparation, lung sections were conducted HE staining to determine tumor area with quantitative assay as % of whole section. ( G-L ) lung sections were conducted IHC staining to determine expression of Ki-67, TTF-1 and SPC with quantitative analysis of the average optical density (AOD) value (n ≥ 4). NEC: negative control, normal IgG was used to replace primary antibody. *P < 0.05, **P < 0.01, ***P < 0.001; NS: not significantly different. n = 14 for WT mice, n ≥ 8 for IFNγ -/- mice as indicated in Table .

Article Snippet: Rabbit anti-TTF-1, SPC, CD8a and VEGFR2 polyclonal antibodies were purchased from ABclonal Biotechnology Co., Ltd (Wuhan, Hubei, China).

Techniques: Injection, Staining, Immunohistochemistry, Expressing, Negative Control

(a) Schematic design of the pMHCI-IgG (top) and the TCB fusion proteins (bottom). Both formats consist of a full monoclonal antibody in an IgG format (light green: antibody light chain, dark green: antibody heavy chain variable domain and CH1, gray/black: constant domain of the antibody heavy chain in the knob and hole format or with charge mutations to drive heterodimerization). A single pMHCI complex consisting of the antigenic peptide (magenta), MHC class I heavy chain lacking the transmembrane domain (purple) and beta-2-microglobulin (blue) or a single anti-CD3 binding CrossMab Fab (crossed heavy and light chain in yellow and red) are fused to one of the antibody heavy chains. (b) Frequency of IFN-γ expressing CD8 T cells measured in flow cytometry after exposure to compound-treated target cells. Incubation of MC38-FAP tumor cells loaded with MCMV-derived M38 peptide (blue), T cell bispecific TCB-αFAP (green), pMHCI-IgG M38-αFAP (red) or pMHCI-IgG IE3-αFAP (pink) and unloaded tumor cells (cyan) with splenocytes from M38 vaccinated mice. Splenocytes were isolated nine days after start of vaccination. All graphs show mean of replicates (n = 2) with error bars indicating standard deviation. (c) Specific tumor cell lysis mediated by compounds after incubation with splenocytes from M38 vaccinated mice. Number of viable cells is measured with xCELLigence as electrical impedance mediated by adherent cells on the culture dish bottom. Color code: IE3-αFAP (purple), M38 peptide (blue), TCB-αFAP (green), M38-αFAP (red). Kinetics of cell lysis at a compound concentration of 25 nM is shown (left). Tumor cell elimination after 40 hours is shown for all concentrations tested (right). All graphs show mean of replicates (n = 3) with error bars indicating standard deviation. Two-sided t-test for significance evaluation was applied. P -values from 0.01 to 0.05 were considered as significant (*), p -values from 0.001 to < 0.01 were considered as very significant (**) and p -values < 0.001 were considered as extremely significant (***). (d) Relative expression of endogenous MHC class I compared to pMHCI-IgG delivered recombinant MHC class I. FAP expressing B16 and MC38 tumor cells were loaded with Ova257-264 peptide (light green) or pMHCI-IgG Ova257-264-αFAP (dark green) and SIINFEKL peptide-MHC class I complexes were detected with the monoclonal antibody anti-Ova257-264 – mouse H-2Kb antibody (25-D1.16) in flow cytometry

Journal: mAbs

Article Title: Vaccine-induced CD8 T cells are redirected with peptide-MHC class I-IgG antibody fusion proteins to eliminate tumor cells in vivo

doi: 10.1080/19420862.2020.1834818

Figure Lengend Snippet: (a) Schematic design of the pMHCI-IgG (top) and the TCB fusion proteins (bottom). Both formats consist of a full monoclonal antibody in an IgG format (light green: antibody light chain, dark green: antibody heavy chain variable domain and CH1, gray/black: constant domain of the antibody heavy chain in the knob and hole format or with charge mutations to drive heterodimerization). A single pMHCI complex consisting of the antigenic peptide (magenta), MHC class I heavy chain lacking the transmembrane domain (purple) and beta-2-microglobulin (blue) or a single anti-CD3 binding CrossMab Fab (crossed heavy and light chain in yellow and red) are fused to one of the antibody heavy chains. (b) Frequency of IFN-γ expressing CD8 T cells measured in flow cytometry after exposure to compound-treated target cells. Incubation of MC38-FAP tumor cells loaded with MCMV-derived M38 peptide (blue), T cell bispecific TCB-αFAP (green), pMHCI-IgG M38-αFAP (red) or pMHCI-IgG IE3-αFAP (pink) and unloaded tumor cells (cyan) with splenocytes from M38 vaccinated mice. Splenocytes were isolated nine days after start of vaccination. All graphs show mean of replicates (n = 2) with error bars indicating standard deviation. (c) Specific tumor cell lysis mediated by compounds after incubation with splenocytes from M38 vaccinated mice. Number of viable cells is measured with xCELLigence as electrical impedance mediated by adherent cells on the culture dish bottom. Color code: IE3-αFAP (purple), M38 peptide (blue), TCB-αFAP (green), M38-αFAP (red). Kinetics of cell lysis at a compound concentration of 25 nM is shown (left). Tumor cell elimination after 40 hours is shown for all concentrations tested (right). All graphs show mean of replicates (n = 3) with error bars indicating standard deviation. Two-sided t-test for significance evaluation was applied. P -values from 0.01 to 0.05 were considered as significant (*), p -values from 0.001 to < 0.01 were considered as very significant (**) and p -values < 0.001 were considered as extremely significant (***). (d) Relative expression of endogenous MHC class I compared to pMHCI-IgG delivered recombinant MHC class I. FAP expressing B16 and MC38 tumor cells were loaded with Ova257-264 peptide (light green) or pMHCI-IgG Ova257-264-αFAP (dark green) and SIINFEKL peptide-MHC class I complexes were detected with the monoclonal antibody anti-Ova257-264 – mouse H-2Kb antibody (25-D1.16) in flow cytometry

Article Snippet: CD8 T cells were stained with polyclonal rabbit anti-mouse CD8a (Synaptic Systems, 1:200) for 60 min.

Techniques: Binding Assay, Expressing, Flow Cytometry, Incubation, Derivative Assay, Isolation, Standard Deviation, Lysis, Concentration Assay, Recombinant

CD8 T cell infiltration in solid MC38-FAP tumors after a single treatment with compounds

Journal: mAbs

Article Title: Vaccine-induced CD8 T cells are redirected with peptide-MHC class I-IgG antibody fusion proteins to eliminate tumor cells in vivo

doi: 10.1080/19420862.2020.1834818

Figure Lengend Snippet: CD8 T cell infiltration in solid MC38-FAP tumors after a single treatment with compounds

Article Snippet: CD8 T cells were stained with polyclonal rabbit anti-mouse CD8a (Synaptic Systems, 1:200) for 60 min.

Techniques:

Journal: Frontiers in Oncology

Article Title: Changes in Tumor-Infiltrating Lymphocytes and Vascular Normalization in Breast Cancer Patients After Neoadjuvant Chemotherapy and Their Correlations With DFS

doi: 10.3389/fonc.2019.01545

Figure Lengend Snippet: Concentrations of primary antibodies.

Article Snippet: CD8 rabbit anti-human polyclonal antibody (17335-1-AP) , Proteintech, US , 1:1,000.

Techniques: Concentration Assay

(A) HE(x200). (B) Immunohistochemical (IHC, x400) CD8+, CD4+, FOXP3+Tregs, and PD-L1 of Tonsil (positive tissue) and breast tumor. (C) Immunofluorescence (IF, x20) Blue: DAPI; Red: CD105; Green: NG2 (The short arrow shows microvessels not covered by pericyte cells, the long arrow shows microvessels covered by pericyte cells).

Journal: Frontiers in Oncology

Article Title: Changes in Tumor-Infiltrating Lymphocytes and Vascular Normalization in Breast Cancer Patients After Neoadjuvant Chemotherapy and Their Correlations With DFS

doi: 10.3389/fonc.2019.01545

Figure Lengend Snippet: (A) HE(x200). (B) Immunohistochemical (IHC, x400) CD8+, CD4+, FOXP3+Tregs, and PD-L1 of Tonsil (positive tissue) and breast tumor. (C) Immunofluorescence (IF, x20) Blue: DAPI; Red: CD105; Green: NG2 (The short arrow shows microvessels not covered by pericyte cells, the long arrow shows microvessels covered by pericyte cells).

Article Snippet: CD8 rabbit anti-human polyclonal antibody (17335-1-AP) , Proteintech, US , 1:1,000.

Techniques: Immunohistochemical staining, Immunofluorescence

Changes of sTILs, PD-L1, MVD, and MPI in non-pCR group and pCR group after NAC.

Journal: Frontiers in Oncology

Article Title: Changes in Tumor-Infiltrating Lymphocytes and Vascular Normalization in Breast Cancer Patients After Neoadjuvant Chemotherapy and Their Correlations With DFS

doi: 10.3389/fonc.2019.01545

Figure Lengend Snippet: Changes of sTILs, PD-L1, MVD, and MPI in non-pCR group and pCR group after NAC.

Article Snippet: CD8 rabbit anti-human polyclonal antibody (17335-1-AP) , Proteintech, US , 1:1,000.

Techniques:

Univariate and multivariate analysis of pCR after NAC.

Journal: Frontiers in Oncology

Article Title: Changes in Tumor-Infiltrating Lymphocytes and Vascular Normalization in Breast Cancer Patients After Neoadjuvant Chemotherapy and Their Correlations With DFS

doi: 10.3389/fonc.2019.01545

Figure Lengend Snippet: Univariate and multivariate analysis of pCR after NAC.

Article Snippet: CD8 rabbit anti-human polyclonal antibody (17335-1-AP) , Proteintech, US , 1:1,000.

Techniques:

Relationship between the baseline ratios of populations and pCR.

Journal: Frontiers in Oncology

Article Title: Changes in Tumor-Infiltrating Lymphocytes and Vascular Normalization in Breast Cancer Patients After Neoadjuvant Chemotherapy and Their Correlations With DFS

doi: 10.3389/fonc.2019.01545

Figure Lengend Snippet: Relationship between the baseline ratios of populations and pCR.

Article Snippet: CD8 rabbit anti-human polyclonal antibody (17335-1-AP) , Proteintech, US , 1:1,000.

Techniques:

The relationship between the changes of sTILs, PD-L1, MVD, MPI, and DFS before and after NAC. (A) Survival analysis between pCR group and Non-pCR group. (B) Survival analysis of sTILs change. (C) Survival analysis of CD8+T change. (D) Survival analysis of CD4+T change. (E) Survival analysis of FOXP3+ Tregs change. (F) Survival analysis of PD-L1 change. (G) Survival analysis of MVD change. (H) Survival analysis of MPI change.

Journal: Frontiers in Oncology

Article Title: Changes in Tumor-Infiltrating Lymphocytes and Vascular Normalization in Breast Cancer Patients After Neoadjuvant Chemotherapy and Their Correlations With DFS

doi: 10.3389/fonc.2019.01545

Figure Lengend Snippet: The relationship between the changes of sTILs, PD-L1, MVD, MPI, and DFS before and after NAC. (A) Survival analysis between pCR group and Non-pCR group. (B) Survival analysis of sTILs change. (C) Survival analysis of CD8+T change. (D) Survival analysis of CD4+T change. (E) Survival analysis of FOXP3+ Tregs change. (F) Survival analysis of PD-L1 change. (G) Survival analysis of MVD change. (H) Survival analysis of MPI change.

Article Snippet: CD8 rabbit anti-human polyclonal antibody (17335-1-AP) , Proteintech, US , 1:1,000.

Techniques:

The relationship between DFS and sTILs, PD-L1, MVD, and MPI in Metastatic lymph node (LN). (A) Survival analysis of LN sTILs. (B) Survival analysis of LN CD8+ T cells. (C) Survival analysis of LN CD4+ T cells. (D) Survival analysis of LN FOXP3+ Tregs. (E) Survival analysis of LN PD-L1. (F) Survival analysis of LN MVD. (G) Survival analysis of LN MPI.

Journal: Frontiers in Oncology

Article Title: Changes in Tumor-Infiltrating Lymphocytes and Vascular Normalization in Breast Cancer Patients After Neoadjuvant Chemotherapy and Their Correlations With DFS

doi: 10.3389/fonc.2019.01545

Figure Lengend Snippet: The relationship between DFS and sTILs, PD-L1, MVD, and MPI in Metastatic lymph node (LN). (A) Survival analysis of LN sTILs. (B) Survival analysis of LN CD8+ T cells. (C) Survival analysis of LN CD4+ T cells. (D) Survival analysis of LN FOXP3+ Tregs. (E) Survival analysis of LN PD-L1. (F) Survival analysis of LN MVD. (G) Survival analysis of LN MPI.

Article Snippet: CD8 rabbit anti-human polyclonal antibody (17335-1-AP) , Proteintech, US , 1:1,000.

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1) Product Images from "Stimulator of interferon gene facilitates recruitment of effector CD8 T cells that drive neurofibromatosis type 1 nerve tumor initiation and maintenance"

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