Cd163

Manufactured by Leica

Sourced in United States, Japan

CD163 is a cell surface glycoprotein that functions as a scavenger receptor. It is primarily expressed on the surface of monocytes and macrophages. CD163 plays a role in the clearance of hemoglobin-haptoglobin complexes from the bloodstream.

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Lab products found in correlation

Cleaved caspase 3, by Cell Signaling Technology (2 mentions) Clone 22c3, by Leica (1 mentions) Ab59028, by Abcam (1 mentions) Clone 15f9, by Abcam (1 mentions) Axio imager z2 upright epifluorescence microscope, by Zeiss (1 mentions) Tissue faxsfluo slide scanning system, by TissueGnostics (1 mentions) Envision kit, by Agilent Technologies (1 mentions) Envision detection system kit, by Agilent Technologies (1 mentions) Trilogy solution, by Cell Marque (1 mentions) Mouse anti human cd68, by Agilent Technologies (1 mentions) Ultraview detection kit, by Roche (1 mentions) Ventana automated immunostainer, by Roche (1 mentions) Cd163, by Roche (1 mentions) Anti ki67, by Abcam (1 mentions) Bond polymer kit, by Leica (1 mentions) Imagescope, by Leica (1 mentions) Optiview universal dab kit, by Roche (1 mentions) Bond max autostainer, by Leica (1 mentions) Ncl l cd163, by Leica (1 mentions) Autostainer system, by Agilent Technologies (1 mentions)

Spelling variants of this product

NCL-L-CD163 (15 citations) Anti-CD163 (13 citations) CD163 (clone 10D6, (9 citations) Anti-CD163 antibody (4 citations) CD163 (10D6; (4 citations) Anti-CD163 (clone 10D6, (4 citations) CD163 (clone Novacastra10D6; (3 citations) Mouse anti‐CD163 (2 citations) NCL-CD163 (2 citations) Anti-CD163 (10D6) antibody (2 citations)

18 protocols using cd163

Multiplex IF Analysis of Endemic BL

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Multiplex immunofluorescence (mIF) was carried out on 16 formalin fixed paraffin embedded (FFPE) endemic BL cases (validation cohort 2), belonging to set of samples previously studied and well characterized for EBV latency program [27 ].
Multiplex IF was applied to simultaneously detect the expression of: a) CD68 (Abcam, ab 955, 1:150) and CD163 (Leica Biosystem, 10D6, 1:200); b) PD-L1 (Dako, clone 22C3, 1:100) and CD163 (Leica Biosystem, 10D6, 1:200); c) PD-L1 and EBV-LMP2A (Abcam, clone 15F9, ab59028, 1:200). These double stainings use red and green or magenta and green chromogens. The colour assignment and staining location are: a) CD68 red/membranous; CD163 green/membranous; b) PD-L1, green/membranous and CD163, pink/membranous or PD-L1, green/membranous and CD163, red/membranous; c) PD-L1, red; LMP2A green/ membranous. The staining procedure was established according to previously published work [28 ]. Tissue sections from the same set of cases and without antibody/fluorophore were used as negative control. Multiplex IF staining reaction and image analysis (including quantification of antibodies expression) were performed using the Vectra 2.0 system (PerkinElmer, Waltham, MA) and Tissue FAXSFluo slide scanning system (TissueGnostics, Vienna Austria) based on a Zeiss Axio Imager Z2 upright epifluorescence microscope.

Granai M., Mundo L., Akarca A.U., Siciliano M.C., Rizvi H., Mancini V., Onyango N., Nyagol J., Abinya N.O., Maha I., Margielewska S., Wi W., Bibas M., Piccaluga P.P., Quintanilla-Martinez L., Fend F., Lazzi S., Leoncini L, & Marafioti T. (2020). Immune landscape in Burkitt lymphoma reveals M2-macrophage polarization and correlation between PD-L1 expression and non-canonical EBV latency program. Infectious Agents and Cancer, 15, 28.

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Quantitative Immunohistochemical Analysis of Macrophages in Atrial Tissue

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Assessment of CD68+ and CD163+ macrophage markers in atrial tissue samples was performed with the use of the EnVision Detection System Kit (Dako). Tissue sections (6 μm) underwent deparaffinization, rehydration, and antigen retrieval using Trilogy solution (Cell Marque) in a pressure cooker for 15 minutes. Slides were washed in distilled water and blocked in 1% hydrogen peroxide in the dark for 10 minutes. Nonspecific binding was blocked by incubation in PBS/5% bovine serum albumin/1% goat serum for 30 minutes. A mouse anti‐human CD68+ (Dako) or CD163+ (Novocastra) monoclonal antibodies (1:100 dilution in PBS) were applied for 1 hour. Following washes in PBS/0.05% Tween, the Dako EnVision Kit was used to complete the staining. This included an incubation in prediluted HRP‐conjugated secondary antibody for 25 minutes, followed by washes and incubation with diaminobenzidine colorimetric substrate for 5 minutes. Slides were counterstained with hematoxylin for nuclear visualization. Following dehydration and coverslip addition, slides were scanned with Aperio ScanScopeXT Slide Scanner (20‐fold magnification) and image analysis was performed with ImageScope software. A positive pixel count algorithm was used to quantify brown‐colored CD68+ or CD163+ within each scanned image.

Watson C.J., Glezeva N., Horgan S., Gallagher J., Phelan D., McDonald K., Tolan M., Baugh J., Collier P, & Ledwidge M. (2020). Atrial Tissue Pro‐Fibrotic M2 Macrophage Marker CD163+, Gene Expression of Procollagen and B‐Type Natriuretic Peptide. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 9(11), e013416.

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Immunohistochemical Analysis of Tumor Markers

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Immunohistochemical staining for IDO (Millipore, Billerica, MA, USA), CD68 (Dako, Carpinteria, California, USA), CD163 (Novocastra, Newcastle, UK), CD4 (Novocastra), CD8 (Dako), and FOXP3 (Abcam, Cambridge, UK) was performed on the TMA blocks following a standard protocol using a Ventana Automated Immunostainer (Ventana, Benchmark, Tuscan, AZ USA). After deparaffinization, heat-induced antigen retrieval was performed using citrate buffer, pH 6.0 (CC1 protocol, Ventana). Reactivity was detected using the Ultra-View detection kit (Ventana).

Choe J.Y., Yun J.Y., Jeon Y.K., Kim S.H., Park G., Huh J.R., Oh S, & Kim J.E. (2014). Indoleamine 2,3-dioxygenase (IDO) is frequently expressed in stromal cells of Hodgkin lymphoma and is associated with adverse clinical features: a retrospective cohort study. BMC Cancer, 14, 335.

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Immunohistochemical Profiling of Histiocytic Neoplasms

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Immunohistochemistry was performed on 3 μm thick formalin fixed paraffin embed (FFPE) sections using commercially available antibodies: CD163, CD68 PGM1, CD14, Factor XIIIa, Fascin, Ki-67, S100, CD1a, Langerin, and Braf-VE1 (Table 1).

Table 1

Immunohistochemistry of histiocytic neoplasms

Antibody (Source)	Clone	Dilution	Antigen Retrieval(Ventana proprietary reagents)	Detection(Ventana proprietary reagents)
CD163 (Novocastra)	10D6	1:200	uCC1 mild	iView DAB
CD68 (Dako)	PG-M1	1:100	uCC1 mild	iView DAB
CD14 (Cell Marque)	EPR3653	1:100	uCC1 standard	Optiview DAB
Factor XIIIα (GeneTex)	Polyclonal	1:250	Protease	iView DAB
Fascin (Dako)	55 K-2	1:500	uCC1 mild	iView DAB
Ki-67 (Dako)	MIB-1	1:25	uCC1 mild	iView DAB
S100 (Dako)	Polyclonal	1:3000	uCC1 mild	iView DAB
CD1α (Immunotech)	O1O	1:5	uCC1 mild	uV DAB
Langerin (Leica)	12D6	1:100	uCC1 standard	uV DAB
BRAF V600E (Ventana Medical Systems)	VE1	Pre-dilute	uCC1 standard	OptiView DAB

Picarsic J., Pysher T., Zhou H., Fluchel M., Pettit T., Whitehead M., Surrey L.F., Harding B., Goldstein G., Fellig Y., Weintraub M., Mobley B.C., Sharples P.M., Sulis M.L., Diamond E.L., Jaffe R., Shekdar K, & Santi M. (2019). BRAF V600E mutation in Juvenile Xanthogranuloma family neoplasms of the central nervous system (CNS-JXG): a revised diagnostic algorithm to include pediatric Erdheim-Chester disease. Acta Neuropathologica Communications, 7, 168.

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IHC Analysis of FFPE GC Tissues

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Formalin-fixed paraffin embedded (FFPE) GC tissues and paired adjacent normal tissues were sectioned, followed by staining with hematoxylin and eosin (H&E) for histopathological analysis. Immunohistochemistry (IHC) analysis was performed as described^{15 (link)}. In brief, sections were deparaffinized, rehydrated, followed by antigen retrieval. After blocking with 10% normal goat serum, slides were incubated with primary antibody at 4 °C overnight. Following primary antibodies were used in IHC analysis: anti-POU1F1 (1:100; Abcam, Cambridge, MA, USA); anti-Ki67 (1:500; Abcam, Cambridge, MA, USA); CD163 (1:200; Novocastra, Laboratories, Newcastle upon Tyne, UK); CD31 (1:100; Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). The immunoreactivity of target was visualized using labeled streptavidin biotin method. The staining intensity was quantitatively analyzed using ImageJ software.

Tang C., Lei X., Xiong L., Hu Z, & Tang B. (2021). HMGA1B/2 transcriptionally activated-POU1F1 facilitates gastric carcinoma metastasis via CXCL12/CXCR4 axis-mediated macrophage polarization. Cell Death & Disease, 12(5), 422.

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Quantifying Tumor-Infiltrating Immune Cells

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The presence of tumor-infiltrating immune cells was confirmed by immunohistochemistry using antibodies for CD3 (1:100, DAKO, Glostrup, Denmark), CD4 (RTU, Ventana, Tucson, AZ, USA), CD8 (1:100, Neomarkers, Fremont, CA, USA), FOXP3 (1:100, Abcam, Cambridge, UK), CD68 (1:100, DAKO), and CD163 (1:100, Novocastra, Newcastle, UK). Immunostaining for CD3, CD8, and FOXP3 was performed using a Bond polymer kit (Leica Microsystems) and Leica BOND-MAX autostainer (Leica Microsystems). CD4, CD68, and CD163 expression was detected immunohistochemically on a Ventana Bench mark XT autostainer (Ventana) with the OPTIVIEW universal DAB kit (Ventana).
All immunostained slides were scanned on an Aperio ScanScope^® CS instrument (Aperio Technologies, Inc., Vista, CA, USA) at 20 x magnifications. Each immunomarker-positive tumor-infiltrating immune cells quantified by computerized image analysis system, ImageScope™ (Aperio Technologies) (Figure1). CD3⁺, CD4⁺, CD8^+, and FOXP3⁺ lymphocytes were counted using the Nuclear v9 algorithm and CD68⁺ and CD163⁺ macrophages were counted using the Positive pixel count v9 algorithm. The density of immune infiltrates was obtained from the entire area of the tissue core.

Kwak Y., Koh J., Kim D.W., Kang S.B., Kim W.H, & Lee H.S. (2016). Immunoscore encompassing CD3+ and CD8+ T cell densities in distant metastasis is a robust prognostic marker for advanced colorectal cancer. Oncotarget, 7(49), 81778-81790.

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Quantitative Immunohistochemistry of Skin Cells

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Immunohistochemical analysis was performed on the sections obtained from formalin-fixed, paraffin-embedded tissue, using antibodies to CD1a (monoclonal mouse antihuman, dilution: 1:100; Dako, Glostrup, Denmark), langerin (CD207) (monoclonal mouse antihuman, dilution: 1:200, Dendritics, France), CD68 (monoclonal mouse antihuman, dilution: 1:1, Dako, Glostrup, Denmark), CD163 (monoclonal mouse anti-human, dilution: 1:50; Novocastra, Newcastle, UK), and dendritic cell lysosome-associated membrane glycoprotein (DC-LAMP) (CD208) (rabbit polyclonal, anti-human, dilution: 1:50; Novus, Centennial, USA). Cell populations were counted by 2 observers at 3 different high-power fields, which were chosen randomly in the follicular or non-follicular areas of the epidermis or dermis. Mean counts from 2 observers were used as data points for quantitative immunohistochemistry analysis. Statistical analysis was performed using the Mann–Whitney test with a 2-tailed p-value of < 0.05 considered statistically significant. These analyses were performed using Statcel version 4.0 software (OMS, Tokyo, Japan).

ITO A., SUGITA K., GOTO H, & YAMAMOTO O. (2020). Implication of Perifollicular Clusters and Folliculotropic Distribution of Dendritic Cells in the Pathogenesis of Seborrhoeic Dermatitis. Acta Dermato-Venereologica, 100(13), 5774.

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Tissue Microarray-Based Immunohistochemistry

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Tissue microarrays (TMAs) were obtained through the biobank of the National Center for Tumor Diseases Heidelberg. The tissue specimens provided by the National Center for Tumor Diseases tissue bank were used in accordance to the regulations of the tissue bank and under University of Heidelberg Ethics Committee approvals 206/2005 and 207/2005. Patient samples were collected between 1990 and 2005 with a mean follow-up time of 80.7 months (range 0.3 to 254.7 months). A cohort of 26 patients with varying TNM stages was retrieved from the archives of the Department of Pathology of the University of Heidelberg School of Medicine and reviewed by an expert pathologist (W.R.). The slides were processed as previously described [12] (link) and incubated with a PUMA antibody (Millipore ABC158, 1:100 dilution). Additional antibodies used for immunohistochemistry were directed against cleaved caspase-3 (Cell Signaling) and CD163 (Novocastra). Immunodetection was performed using the Histostain Plus IHC Detection Kit (Life Technologies). Slides were counterstained with hematoxylin.

Zhou X., Li J., Marx C., Tolstov Y., Rauch G., Herpel E., Macher-Goeppinger S., Roth W., Grüllich C., Pahernik S., Hohenfellner M, & Duensing S. (2015). Uncoupling of PUMA Expression and Apoptosis Contributes to Functional Heterogeneity in Renal Cell Carcinoma — Prognostic and Translational Implications. Translational Oncology, 8(6), 480-486.

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Immunohistochemical Staining of FFPE Samples

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FFPE blocks were cut into 3 μm thick sections and stained on a Dako autostainer system with the following antibodies: CD8 (1:100, Dako Cytomation, M7103), CD68 (1:5000, Dako Cytomation, M0814) and CD163 (1:1000, Novocastra, NCL-L-CD163). After standard deparaffinization with xylene and alcohol, the slides were treated with a sodium citrate solution (pH 6.0) at 95 °C for 20 min to retrieve the antigens, and incubated with the mentioned antibodies for 30 min at room temperature. The Dako detection system (FLEX + Mouse, K8002) was used accordingly. Fox P3 (1:25, BioLegend, 320,116) was stained with an automated Ventana BenchMark staining system and the UltraView Universal DAB detection kit (760−500). Counterstaining of all sections was done manually with hematoxylin.

Lang A., Jeron R.L., Lontzek B., Kiesel B., Mischkulnig M., Berghoff A.S., Ricken G., Wöhrer A., Rössler K., Lötsch-Gojo D., Roetzer-Pejrimovsky T., Berger W., Hainfellner J.A., Höftberger R., Widhalm G, & Erhart F. (2023). Mapping high-grade glioma immune infiltration to 5-ALA fluorescence levels: TCGA data computation, classical histology, and digital image analysis. Journal of Neuro-Oncology, 164(1), 211-220.

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Histological analysis of tumor samples

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Tumors were fixed in 10% formalin for 48 hours and paraffin embedded. 4 μm sections were stained with hematoxylin and eosin (H&E), Masson’s trichrome, or Picrosirius red at the University of Washington Histology Core. Primary antibodies used recognized CD163 (Novocastra, 10D6, 1:200), cleaved caspase 3 (Cell Signaling, D175, 1:200), and Ki67 (Thermo Fisher, clone SolA15, 1:200). Slides were scanned using the Nanozoomer Digital Pathology slide scanner (Hamamatsu; Bridgewater, New Jersey), and Visiopharm software (Hoersholm, Denmark) was used to identify regions of interest (ROI, i.e. tumor tissue, excluding normal tissue) sampled at 100%. The software was trained to detect immunoreactivity using a project-specific configuration based on a threshold of pixel values as we previously reported (6 (link)). The number of positively stained cells was measured in 3–5 non-overlapping 20X fields using NIS-Element imaging software (Nikon’s universal software platform, n=3–5 mice per group). Collagen was quantified from 2 tumor sections stained with Picrosirius red, and intensity of the red staining was assessed in a blinded manner across 3–5 20X fields as follows: 0, no staining detected; 1, light staining; 2, moderate; 3, moderate-high staining intensity; 4, high staining, as we previously described (6 (link)) (n= 3–6 animals per cohort).

Stromnes I.M., Burrack A.L., Hulbert A., Bonson P., Black C., Brockenbrough J.S., Raynor J.F., Spartz E.J., Pierce R.H., Greenberg P.D, & Hingorani S.R. (2019). Differential effects of depleting versus programming tumor-associated macrophages on engineered T cells in pancreatic ductal adenocarcinoma. Cancer immunology research, 7(6), 977-989.

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