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Cd163

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

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Lab products found in correlation

Pd l1, by Cell Signaling Technology (3 mentions) Lsm 700 microscope, by Zeiss (1 mentions) Cm1860, by Leica (1 mentions) Alexa fluor 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Cd79a, by Agilent Technologies (1 mentions) C reactive protein (crp), by Abcam (1 mentions) Automated staining system, by Roche (1 mentions) N cadherin, by Zymo Research (1 mentions) Bond max, by Leica (1 mentions) Cellsens standard software, by Olympus (1 mentions) Dp74 camera system, by Olympus (1 mentions) Cd68 pgm 1, by Agilent Technologies (1 mentions) Benchmark ultra platform, by Roche (1 mentions) Factor xiiia, by Cell Marque (1 mentions) Ventana benchmark ultra slide stainer, by Roche (1 mentions) Stat6, by Roche (1 mentions) Desmin, by Agilent Technologies (1 mentions) Desmin, by Roche (1 mentions) Cd163, by Roche (1 mentions)

10 protocols using cd163

1

Immunophenotyping of Tumor-Associated Macrophages

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Tumor chunks (n = 30) were fixed in 4% paraformaldehyde overnight, instilled in
30% sucrose, and embedded in optimal cutting temperature (OCT) medium, snap
frozen, and sectioned at 5 µm (Leica CM 1860). Tissue sections were
permeabilized and blocked prior to antigen retrieval with citrate buffer.
Sections were incubated with CD80 (1:500, #8679; ProSci) or CD163 (1:500, #163M;
Cell Marque) primary antibodies at 4°C overnight, Alexa Fluor 594–conjugated
secondary antibody (Life Technologies) for 2 hours at room temperature, and
4′6-diamidino-2-phenylindole (DAPI) nuclear stain for 10 minutes. Antifade
mounting medium was added before cover-slipping. Confocal images were obtained
for 3 different areas (Zeiss LSM 700 Microscope, 40× oil immersion lens).
Serum-treated sections were used as negative controls. After the minimum and
maximum threshold values were set to eliminate nonspecific staining and artifact
on ImageJ software (National Institutes of Health), expression levels for CD80
and CD163 were determined as percent area per region of interest on high-powered
field. Median values were compared between samples.
Nisenbaum E., Misztal C., Szczupak M., Thielhelm T., Peña S., Mei C., Goncalves S., Bracho O., Ma R., Ivan M.E., Morcos J., Telischi F., Liu X.Z., Fernandez-Valle C, & Dinh C.T. (2021). Tumor-Associated Macrophages in Vestibular Schwannoma and Relationship to Hearing. OTO Open, 5(4), 2473974X211059111.
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2

Tissue Immunohistochemistry of Ascending Aorta

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From specimens of ascending aorta that was ranging in length from 2 to 5 cm, the average three samples (from the bulge) were obtained, fixed in formalin for 24 hours, embedded in paraffin, and cut into 4.5 μm thick transversal step (step was 50 μm thick) serial sections. The sections were then stained with hematoxylin-eosin (HE) and Movat pentachrome [20 (link)]. The immunohistochemical staining was used to detect T killer – Tk cells (CD8; 1:50, Dako, Glostrup, Denmark), T helper – Th cells (CD4; 1:20, Cell Marque), B cells (CD79a; 1:20, Dako Glostrup, Denmark), plasma cells (CD138,1:30, Dako, Glostrup, Denmark), pro-inflammatory M1 macrophages (CD68;1:50, Dako, Glostrup, Denmark), and anti-inflammatory M2 macrophages (CD163; 1:40, Cell Marque) following the manufacturer’s instructions.
Milutinović A, & Zorc-Pleskovič R. (2021). Inflammatory cells in the ascending aortic aneurysm in patients with type 2 diabetes versus patients with hypertension. Bosnian Journal of Basic Medical Sciences, 22(2), 178-184.
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3

Comprehensive Immunohistochemical Analysis of Tumor Microenvironment

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Representative whole-section slides of the tumors were immunohistochemically stained for N-cadherin (1:100, mouse monoclonal; Zymed Laboratories Inc., San Francisco, CA, USA), neural cell adhesion molecule (1:100, mouse monoclonal; Leica Biosystems, Nussloch, Germany), S100 calcium-binding protein P (1:100, mouse monoclonal; R&D Systems, Minneapolis, MN, USA), CD3 (1:100, mouse polyclonal; Dako, Glostrup, Denmark), CD4 (prediluted, mouse monoclonal; Dako), CD8 (prediluted, mouse monoclonal; Leica Biosystems), CD68 (1:300, mouse monoclonal; Dako), CD163 (1:50, mouse monoclonal; Cell Marque, Rocklin, CA, USA), FOXP3 (1:100, mouse monoclonal; Abcam, Cambridge, MA, USA), C-reactive protein (rabbit monoclonal; Abcam), and programmed death-ligand 1 (mouse monoclonal; Dako) using an automated staining system (Ventana Medical Systems, Tucson, AZ, USA) according to the manufacturer’s instructions. The amount of immune cell infiltration was counted in 10 high-power field-equivalent area from microscopic images by QuPath software (University of Edinburgh, Edinburgh, UK).15 (link) Alcian blue staining was also performed to evaluate the presence and extent of intra/extracellular mucin production.
Chung T., Rhee H., Shim H.S., Yoo J.E., Choi G.H., Kim H, & Park Y.N. (2021). Genetic, Clinicopathological, and Radiological Features of Intrahepatic Cholangiocarcinoma with Ductal Plate Malformation Pattern. Gut and Liver, 16(4), 613-624.
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4

Immune Cell Profiling in Liver Lesions

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From a total of 53 patients with HCC with chronic HCV infection who underwent tumor resection at the University Medical Center of Cologne, different areas with cirrhosis, DNs, and hepatocellular carcinoma were studied by IHC. All immunostainings were conducted using the BOND MAX automated staining system (Leica Biosystems). Macrophages were stained with CD68 antibodies (Agilent Technologies, dilution 1:400) and tumor-associated macrophages with CD163 (Cell Marque, dilution 1:100). For IHC of B cells, CD20 antibodies (Agilent Technologies, dilution 1:1250) were used. For IHC of the T cell populations, a CD3 antibody (Thermo Fisher Scientific, dilution 1:50) and a CD8 antibody (Agilent Technologies, dilution 1:200) were used. Images were acquired using the Olympus DP74 camera system and the cellSens standard software (Olympus). Immunostained cells were counted in representative areas between 0.1–0.7 mm2 by means of the Image J (NIH) cell counter plugin tool (https://biii.eu/cell-counter-imagej) and expressed as cell numbers per mm2. A Kruskal-Wallis test was used to assess statistical significance.
Czauderna C., Poplawski A., O’Rourke C.J., Castven D., Pérez-Aguilar B., Becker D., Heilmann-Heimbach S., Odenthal M., Amer W., Schmiel M., Drebber U., Binder H., Ridder D.A., Schindeldecker M., Straub B.K., Galle P.R., Andersen J.B., Thorgeirsson S.S., Park Y.N, & Marquardt J.U. (2021). Epigenetic modifications precede molecular alterations and drive human hepatocarcinogenesis. JCI Insight, 6(17), e146196.
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5

Immunohistochemical Profiling of Tumor Markers

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Immunohistochemistry was performed as described previously (32 (link), 33 ) on 4-mm tissue microarrays sections using a standard avidin–biotin immunoperoxidase method with antibodies specific for CD163 (Cell Marque, prediluted), FOXP3 (Abcam, dilution 1:200), PD-L1 (Cell Signaling, dilution 1:600), and MCT4 (Santa Cruz, dilution 1:250). All stains were validated using recommended tissue controls, and evaluated for correct compartment specific staining by pathologists with experience in evaluating the indicated markers. Staining was performed on either a Dako or Ventana Benchmark automated immunohistochemical stainers.
Adams T.A., Vail P.J., Ruiz A., Mollaee M., McCue P.A., Knudsen E.S, & Witkiewicz A.K. (2017). Composite Analysis of Immunological and Metabolic Markers Defines Novel Subtypes of Triple Negative Breast Cancer. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 31(2), 288-298.
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6

Immunohistochemical Profiling of Tumor Markers

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Immunohistochemistry was performed as described previously (32 (link), 33 ) on 4-mm tissue microarrays sections using a standard avidin–biotin immunoperoxidase method with antibodies specific for CD163 (Cell Marque, prediluted), FOXP3 (Abcam, dilution 1:200), PD-L1 (Cell Signaling, dilution 1:600), and MCT4 (Santa Cruz, dilution 1:250). All stains were validated using recommended tissue controls, and evaluated for correct compartment specific staining by pathologists with experience in evaluating the indicated markers. Staining was performed on either a Dako or Ventana Benchmark automated immunohistochemical stainers.
Adams T.A., Vail P.J., Ruiz A., Mollaee M., McCue P.A., Knudsen E.S, & Witkiewicz A.K. (2017). Composite Analysis of Immunological and Metabolic Markers Defines Novel Subtypes of Triple Negative Breast Cancer. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, 31(2), 288-298.
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7

Immunohistochemical Profiling of the Tumor Microenvironment

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Hematoxylin staining and evaluation of the tissue architecture was used to define the stromal type. The TMAs were stained with the following antibodies: PD-L1 (Cell Signaling, dilution 1:600), CD163 (Cell Marque, prediluted), FOXP3 (Abcam, dilution 1:200), CD8 (). The tissues were also stained for CA9 indicative of hypoxia antibody (Cell Marque, dilution of 1:500). MCT4 was employed as a marker of glycolytic preference (Santa Cruz, dilution 1:250). Expression of IHC markers was categorized semi-quantitatively using published criteria. Stromal tumor infiltrating lymphocytes (TILs) were assessed based on criteria developed by Denkert and colleagues (30 ).
Knudsen E.S., Vail P., Balaji U., Ngo H., Botros I., Makarov V., Riaz N., Balachandran V., Leach S., Thompson D., Chan T.A, & Witkiewicz A.K. (2017). Stratification of Pancreatic Ductal Adenocarcinoma: Combinatorial Genetic, Stromal, and Immunological Markers. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(15), 4429-4440.
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8

Quantifying Tumor-Associated Macrophages

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To estimate the number of TAMs samples (M1 and M2 classes) from patients before surgery were used. We have chosen CD163 (1:200, pH 9.0, Cell Marque, Rocklin, CA, USA), CD68KP (RTU, pH 9.0, Dako Agilent), and CD68 PG-M1 (RTU, pH 9.0, Dako Agilent, Santa Clara, CA, USA) to evaluate the number of TAMs. We scored TAM by counting the number of positive-staining macrophages per mm2: 0—none, 1—low, and 2—high.
Szumera-Ciećkiewicz A., Bobak K., Spałek M.J., Sokół K., Wągrodzki M., Owczarek D., Kawecka M., Puton B., Koseła-Paterczyk H., Rutkowski P, & Czarnecka A.M. (2023). Predictive Biomarkers of Pathological Response to Neoadjuvant Chemoradiotherapy for Locally Advanced Soft Tissue Sarcomas. Cancers, 15(11), 2960.
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9

Comprehensive Immunohistochemistry Profiling

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Immunohistochemistry (IHC) was performed on the Ventana Benchmark Ultra platform. The antibodies were Caldesmon (Cell Marque E89, Rocklin, USA, ready-to-use (RTU)), CD99 (Agilent 12E7, Santa Clara, USA; RTU), CD163 (Cell Marque MRQ-26; RTU), collagen IV (Cell Marque CIV22; RTU), factor XIII (Cell Marque AC-1A1; RTU), tryptase (Cell Marque G3; RTU) and osteonectin (Leica 4A4, Wetzlar, Germany; 1:10 dilution). The stained slides were microscopically evaluated by a pathologist and quantified as stated in the results section under real-world conditions.
Dressler F.F., Diedrichs F., Sabtan D., Hinrichs S., Krisp C., Gemoll T., Hennig M., Mackedanz P., Schlotfeldt M., Voß H., Offermann A., Kirfel J., Roesch M.C., Struck J.P., Kramer M.W., Merseburger A.S., Gratzke C., Schoeb D.S., Miernik A., Schlüter H., Wetterauer U., Zubarev R., Perner S., Wolf P, & Végvári Á. (2024). Proteomic analysis of the urothelial cancer landscape. Nature Communications, 15, 4513.
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10

Immunohistochemical Analysis of Skin Tumor

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The excision specimen from the left foot comprising skin and subcutaneous tissue was received in buffered neutral formalin. The tumor in the specimen was extensively sampled and processed for routine hematoxylin and eosin–stained sections. Five-μm sections from formalin-fixed, paraffin-embedded tissues were stained with antibodies against CD31 (Neomarker), CD34 (DAKO), ERG (Cell Marque), STAT6 (Cell Marque), Factor XIIIa (Cell Marque), CD163 (Cell Marque), desmin (DAKO), and SMA (DAKO) using the Ventana Optiview detection kit on the Roche Ventana BenchMark ULTRA slide stainer (Roche Diagnostics) after antigen retrieval, if required. For CD34, STAT6, Factor XIIIa, CD163, and desmin stains, the ULTRA cell conditioning method on the Roche Ventana BenchMark ULTRA slide stainer was used for antigen retrieval. For ERG and CD31 stains, the pressure cooker with citrate buffer at pH6 method was employed. For SMA, no antigen retrieval treatment was required.
Leow M.K., Dogra S., Ge X., Chuah K.L., Liew H., Loke K.S, & McFarlane C. (2021). Paraneoplastic Secretion of Multiple Phosphatonins From a Deep Fibrous Histiocytoma Causing Oncogenic Osteomalacia. The Journal of Clinical Endocrinology and Metabolism, 106(5), e2299-e2308.
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