Journal: ESMO Open

Article Title: Positive tumour CD47 expression is an independent prognostic factor for recurrence in resected non-small cell lung cancer

doi: 10.1136/esmoopen-2020-000823

Figure Lengend Snippet: CD47 expression and tumour-associated macrophage infiltration. (A) Representative IHC images of different CD47 expression level scores in tumour and lung tissue (control respiratory disease sample). (B) CD47 pos expression in NSCLC is higher than in the control group (infection lung tissues) (33.0% vs 17.4%, p=0.038). (C) Tumour CD47 pos expression in female patients with NSCLC was higher than in male patients with NSCLC (43.5% vs 27.0%, p=0.020). (D) Tumour CD47 pos expression in never-smokers with NSCLC was higher than in smokers with NSCLC (46.1% vs 21.6%, p<0.001). (E) Tumour CD47 pos expression in LUAD was higher than in LUSC (52.0% vs 12.9%, p<0.001). (F) Representative immunofluorescence images of CD68 pos macrophages, CD163 pos macrophages, M2-TAMs (CD8 pos , CD163 pos ) and M1-TAMs (CD68 pos , CD163 neg ). (G) High M2/M1 ratios were more frequently observed in NSCLC compared with the control group (71.7% vs 45.7%, p=0.001). (H) M2 infiltration in the tumour area was higher than M1 infiltration in the tumour area (89 vs 57, p=0.001). (I) M2 infiltration in the tumour area in stage III NSCLC was higher than in stage I NSCLC (64.2% vs 29.2%, p<0.001). (J) High M2/M1 ratios were more frequently observed in stage III NSCLC than in stage I NSCLC (83.1% vs 57.3%, p<0.001). *p<0.05, **p<0.001. CD47 pos , cases with CD47 IHC scores of 2+ or 3+ were considered positive for tumour CD47 positive expression; CD47 neg , cases with CD47 IHC scores of 1+ or ve (0) were considered negative for tumour CD47 expression; if, immunofluorescence; IHC, immunohistochemistry; LUAD, lung adenocarcinoma; LUSC, lung squamous cell carcinoma; NSCLC, non-small cell lung cancer; NT, non-tumour area; TAMs, tumour-associated macrophages; T&NT, tumour area and non-tumour area.

Article Snippet: The following primary antibodies were used: rabbit anti-human PD-L1 (1:300, clone E1L3N, 13684; Cell Signalling Technology), mouse anti-human CD8 (1:2, clone C8/144B, IR623, DAKO, Glostrup, Denmark), rabbit anti-human CD68 (1:1600, clone D4B9C, 76437; Cell Signalling Technology) and mouse anti-human CD163 (1:100 clone 10D6, NB110-59935; Novus Biologicals, Littleton, Colorado, USA).

Techniques: Expressing, Control, Infection, Immunofluorescence, Immunohistochemistry

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: Antibodies and Applications

Article Snippet: Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: Immunohistochemistry-IF

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: Renal micromilieus induce CD209+ cells co-expressing CD14 and CD163. A: CD209+ cell subsets in RCC and NKC. Confocal image of an RCC tissue (original magnification, ×630; scale bar = 50 μm) showing single fluorescence channels for CD209 (green), CD14 (red), CD163 (blue), and nuclei (gray). Arrowhead: CD209+CD14−CD163− cells; triangular arrow: CD209+CD14−CD163+ cells; quadrangular arrow: CD209+CD14+CD163+ cells. Images are stack merges of three z-planes (z-step = 0.8 μm). B: CD209+ subsets, quantified from multiparameter-stained tissues. Tumor areas were subdivided into tumor center and periphery (see Supplemental Figure S1A at http://ajp.amjpathol.org). Frequency of CD209+ subsets are shown in the upper panel (n = 7; values are means with MD; ***P < 0.001; Mann-Whitney U-test corrected after multiple comparisons) and lower panel (n = 11: tumor center, n = 6: tumor periphery; n = 8: NKC; values are medians with interquartile ranges; Kruskal-Wallis test with Dunn's posttest; **P < 0.01). C: Histograms of flow cytometry of TILs depict relative expression levels of CD14 and CD209 among the three gated myeloid subsets. Shown is one example of at least 10 TIL analyses. Photomultiplier settings were optimized for lymphocytes within TILs. Thus, fluorescence profiles of the myeloid cells are shifted to the right due to their higher autofluorescence. D: Histograms depict the HLA-DR, CD40, CD80, and CD86 expression levels relative to isotype staining comparing the CD209+CD14+ ercDCs to CD209+CD14− cDCs.

Article Snippet: Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: Expressing, Fluorescence, Staining, MANN-WHITNEY, Flow Cytometry

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: In vitro generated ercDCs resemble cDCs in surface marker expression, endocytic activity, and antigen cross-presentation. A: CD209, CD14, and CD163 surface expression on in vitro generated myeloid cells. Cells are human blood monocytes after 7-day cultivation with IL-4/GM-CSF, cell-conditioned media (RCC-26, HK-2, Colo-357, LCL-26), or a cocktail of recombinant cytokines (CXCL8/IL-8, IL-6, VEGF) or from monocytes after infiltration into RCC-53 microtumors. Solid or dotted lines are specific and isotype stainings, respectively. One experiment of at least three is shown. B: Surface expression of costimulatory molecules and molecules associated with immunoinhibition on in vitro generated cDCs and ercDCs, assessed by flow cytometry. Solid lines represent the marker-specific staining; dotted lines, the corresponding isotype. C: Endocytic activity. cDCs and ercDCs were incubated with BSA-FITC at 37°C (solid lines) or 4°C (dotted lines) and analyzed by flow cytometry. D: Antigen cross-presentation. cDCs and ercDCs were incubated with indicated concentrations of pep70-MART peptide and then co-cultured with CTL-A42 for 24 hours. The extent of antigen cross-presentation was assessed by measuring the amount of IFN-γ secretion by the CTL-A42 using ELISA. Co-cultures of CTL-A42 with a melanoma line that endogenously expresses the MART-pMHC ligand served as control for CTL activity. Bars represent the mean of duplicate values + MD. All experiments are representatives of at least three independent experiments with similar results.

Article Snippet: Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: In Vitro, Generated, Marker, Expressing, Activity Assay, Recombinant, Flow Cytometry, Staining, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: ErcDCs are in contact with T cells without compromising CTL function. A and B: Confocal images of multiparameter-stained RCC tissue. A: Contacts of CD209+CD163+ cells (yellow, merge of CD209+ in green and CD163+ in red) with T cells (CD3+ blue) (arrows) (original magnification, ×630, scale bar = 50 μm). Image is a stack merge of 4 z-planes, z-step = 0.5 μm. B: CD3ε and CD3ζ on T cells. Arrowhead indicates T cell in direct contact; arrow indicates T cell without contact (blue, CD209; green, CD3ε; red, CD3ζ; gray, nuclei). Image (original magnification, ×630; scale bar = 25 μm) is a stack merge of 10 z-planes (z-step = 0.7 μm). C: CD3ε and CD3ζ expression on CTLs after co-culture with or without cDCs and ercDCs in vitro. Numbers are percentages of positive cells in respective quadrant. D: IFN-γ secretion and cytotoxicity of CTLs induced by target recognition in the presence of myeloid cells (generated from monocytes of the same HLA-A2− healthy donor). Left: IFN-γ secretion of CTL-JB4 (3 × 103) stimulated for 24 hours with RCC target cells (15 × 103, HLA-A2+) in the presence or absence of APCs (3 × 103; HLA-A2−). Right: Cytotoxicity of CTL-JB4 against cognate RCC cells determined by cell-mediated cytolysis, using a constant ratio of labeled RCC cells (2 × 103, HLA-A2+) to CTLs (10 × 103) in the presence of titrated numbers of HLA-A2− myeloid cells. Percentage of specific lysis values are the mean of duplicates ± MD. Shown is one of several experiments.

Article Snippet: Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: Staining, Expressing, Co-Culture Assay, In Vitro, Generated, Labeling, Lysis

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: Immune Cell Composition and Intercellular Contacts in Renal Tissue

Article Snippet: Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques:

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: Antibodies and Applications

Article Snippet: Clinicopathologic Features of Patients with Clear Cell RCC Antibodies Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: Immunohistochemistry-IF

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: Renal micromilieus induce CD209+ cells co-expressing CD14 and CD163. A: CD209+ cell subsets in RCC and NKC. Confocal image of an RCC tissue (original magnification, ×630; scale bar = 50 μm) showing single fluorescence channels for CD209 (green), CD14 (red), CD163 (blue), and nuclei (gray). Arrowhead: CD209+CD14−CD163− cells; triangular arrow: CD209+CD14−CD163+ cells; quadrangular arrow: CD209+CD14+CD163+ cells. Images are stack merges of three z-planes (z-step = 0.8 μm). B: CD209+ subsets, quantified from multiparameter-stained tissues. Tumor areas were subdivided into tumor center and periphery (see Supplemental Figure S1A at http://ajp.amjpathol.org). Frequency of CD209+ subsets are shown in the upper panel (n = 7; values are means with MD; ***P < 0.001; Mann-Whitney U-test corrected after multiple comparisons) and lower panel (n = 11: tumor center, n = 6: tumor periphery; n = 8: NKC; values are medians with interquartile ranges; Kruskal-Wallis test with Dunn's posttest; **P < 0.01). C: Histograms of flow cytometry of TILs depict relative expression levels of CD14 and CD209 among the three gated myeloid subsets. Shown is one example of at least 10 TIL analyses. Photomultiplier settings were optimized for lymphocytes within TILs. Thus, fluorescence profiles of the myeloid cells are shifted to the right due to their higher autofluorescence. D: Histograms depict the HLA-DR, CD40, CD80, and CD86 expression levels relative to isotype staining comparing the CD209+CD14+ ercDCs to CD209+CD14− cDCs.

Article Snippet: Clinicopathologic Features of Patients with Clear Cell RCC Antibodies Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: Expressing, Fluorescence, Staining, MANN-WHITNEY, Flow Cytometry

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: In vitro generated ercDCs resemble cDCs in surface marker expression, endocytic activity, and antigen cross-presentation. A: CD209, CD14, and CD163 surface expression on in vitro generated myeloid cells. Cells are human blood monocytes after 7-day cultivation with IL-4/GM-CSF, cell-conditioned media (RCC-26, HK-2, Colo-357, LCL-26), or a cocktail of recombinant cytokines (CXCL8/IL-8, IL-6, VEGF) or from monocytes after infiltration into RCC-53 microtumors. Solid or dotted lines are specific and isotype stainings, respectively. One experiment of at least three is shown. B: Surface expression of costimulatory molecules and molecules associated with immunoinhibition on in vitro generated cDCs and ercDCs, assessed by flow cytometry. Solid lines represent the marker-specific staining; dotted lines, the corresponding isotype. C: Endocytic activity. cDCs and ercDCs were incubated with BSA-FITC at 37°C (solid lines) or 4°C (dotted lines) and analyzed by flow cytometry. D: Antigen cross-presentation. cDCs and ercDCs were incubated with indicated concentrations of pep70-MART peptide and then co-cultured with CTL-A42 for 24 hours. The extent of antigen cross-presentation was assessed by measuring the amount of IFN-γ secretion by the CTL-A42 using ELISA. Co-cultures of CTL-A42 with a melanoma line that endogenously expresses the MART-pMHC ligand served as control for CTL activity. Bars represent the mean of duplicate values + MD. All experiments are representatives of at least three independent experiments with similar results.

Article Snippet: Clinicopathologic Features of Patients with Clear Cell RCC Antibodies Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: In Vitro, Generated, Marker, Expressing, Activity Assay, Recombinant, Flow Cytometry, Staining, Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: ErcDCs are in contact with T cells without compromising CTL function. A and B: Confocal images of multiparameter-stained RCC tissue. A: Contacts of CD209+CD163+ cells (yellow, merge of CD209+ in green and CD163+ in red) with T cells (CD3+ blue) (arrows) (original magnification, ×630, scale bar = 50 μm). Image is a stack merge of 4 z-planes, z-step = 0.5 μm. B: CD3ε and CD3ζ on T cells. Arrowhead indicates T cell in direct contact; arrow indicates T cell without contact (blue, CD209; green, CD3ε; red, CD3ζ; gray, nuclei). Image (original magnification, ×630; scale bar = 25 μm) is a stack merge of 10 z-planes (z-step = 0.7 μm). C: CD3ε and CD3ζ expression on CTLs after co-culture with or without cDCs and ercDCs in vitro. Numbers are percentages of positive cells in respective quadrant. D: IFN-γ secretion and cytotoxicity of CTLs induced by target recognition in the presence of myeloid cells (generated from monocytes of the same HLA-A2− healthy donor). Left: IFN-γ secretion of CTL-JB4 (3 × 103) stimulated for 24 hours with RCC target cells (15 × 103, HLA-A2+) in the presence or absence of APCs (3 × 103; HLA-A2−). Right: Cytotoxicity of CTL-JB4 against cognate RCC cells determined by cell-mediated cytolysis, using a constant ratio of labeled RCC cells (2 × 103, HLA-A2+) to CTLs (10 × 103) in the presence of titrated numbers of HLA-A2− myeloid cells. Percentage of specific lysis values are the mean of duplicates ± MD. Shown is one of several experiments.

Article Snippet: Clinicopathologic Features of Patients with Clear Cell RCC Antibodies Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: Staining, Expressing, Co-Culture Assay, In Vitro, Generated, Labeling, Lysis

Journal: The American Journal of Pathology

Article Title: Human Renal Cell Carcinoma Induces a Dendritic Cell Subset That Uses T-Cell Crosstalk for Tumor-Permissive Milieu Alterations

doi: 10.1016/j.ajpath.2011.03.011

Figure Lengend Snippet: Immune Cell Composition and Intercellular Contacts in Renal Tissue

Article Snippet: Clinicopathologic Features of Patients with Clear Cell RCC Antibodies Primary and secondary antibodies and their application are listed in . table ft1 table-wrap mode="anchored" t5 caption a7 Antibody Label Clone Species/isotype Company Application Primary antibody BDCA-1 L161 Mouse Immunotech IHC, IF BDCA-3 AD5-14H12 Mouse Miltenyi IHC, IF CD3ϵ polyclonal Rabbit Dako IHC CD3ϵ UCHT1 Mouse IgG1 Dako IHC, IF, FC CD3ϵ PB UCHT1 Mouse IgG1 Dako FC CD3ζ 8D3 Mouse IgG1 BD Biosciences IHC, IF, FC CD3ζ FITC G3 Mouse IgG2a Serotec FC CD8 C8/144B Mouse IgG1 Dako IHC CD8 PB RPA-T8 Mouse IgG1 BD Biosciences FC CD14 RMO52 Mouse IgG2a Immunotech IHC, IF, FC CD14 PB M5E2 Mouse IgG2a BD Biosciences FC CD15 FITC HI98 Mouse BD Biosciences FC CD16 FITC 3G8 Mouse IgG1 BD Biosciences FC CD40 FITC 5C3 Mouse IgG1 BD Biosciences FC CD45 PE-Cy7 HI30 Mouse BD Biosciences FC CD56 APC N901 Mouse IgG1 Beckman Coulter FC CD80 FITC BB1 Mouse IgM BD Biosciences FC CD83 HB15A Mouse IgG2b Immunotech IHC CD86 PE 2331/FUN-1 Mouse IgG1 BD Biosciences FC CD107a FITC H4A3 Mouse IgG1 BD Biosciences FC CD107b FITC H4B4 Mouse IgG1 BD Biosciences FC CD163 Ber-MAC3 Mouse IgG1 Dako IHC, IF, FC CD163 PE GHI/61 Mouse IgG1 BD Biosciences FC CD209/DC-SIGN DCN46 Mouse IgG2b BD Biosciences IHC, IF, FC CD209/DC-SIGN APC DCN46 Mouse IgG2b BD Biosciences FC CD274/B7-H1 FITC MIH1 Mouse IgG1 BD Biosciences FC CX 3 CR1 Polyclonal Rabbit eBioscience FC CD208/DC-LAMP 104.G4 Mouse IgG1 Immunotech IHC FOXP3 259D Mouse IgG1 BioLegend IHC, IF Granzyme B PE GB11 Mouse IgG1 Serotec FC HLA-ABC/MHC-I APC G46_2.6 Mouse IgG1 BD Biosciences FC HLA-DR/MHC-II FITC L243 Mouse IgG2a BD Biosciences FC HLA-DR/MHC-II PE G46-6 Mouse IgG2a BD Biosciences FC IFN-γ PE-Cy7 4S.B3 Mouse IgG1 BD Biosciences FC IL-2 APC MQ1-17H12 Rat IgG2a BD Biosciences FC Isotype FITC MOP-C21 Mouse IgG1 BD Biosciences FC Isotype PE MOP-C21 Mouse IgG1 BD Biosciences FC Perforin FITC δG9 Mouse IgG2b BD Biosciences FC TNF-α A700 Mab11 Mouse IgG1 BD Biosciences FC Secondary antibody Anti-AP AP Mouse IgG1 Dako IHC (APAAP) Anti-mouse Rabbit Dako IHC (APAAP) Anti-mouse HRP Goat Dianova IHC (doublestain) Anti-mouse IgG1 A488 Goat Molecular Probes FC Anti-mouse IgG1 A568 Goat Molecular Probes IF Anti-mouse IgG2a A568 Goat Molecular Probes IF Anti-mouse IgG2a A647 Goat Molecular Probes IF Anti-mouse IgG2b A488 Goat Molecular Probes IF Anti-rabbit A488 Goat Molecular Probes FC Anti-rabbit AP Goat Dianova IHC Anti-rabbit Cy5 Goat Dianova IF Open in a separate window A, Alexa Fluor; AP, alkaline phosphatase; APC, allophycocyanin; Cy, cyanin; FC, flow cytometry; HRP, horseradish peroxidase; IF, immunofluorescence; PB, Pacific blue; PE, phycoerythrin.

Techniques: