Nordic Immunohistochemical Quality Control
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1 Nordic Immunohistochemical Quality Control Immunohistochemistry in the classifiation of neoplasias of the alimentary tract & External Quality Assurance of Immunohistochemistry for GI cancer markers Mogens Vyberg, director, NordiQC
2 Carcinoma or GIST/sarcoma? Primary or secondary? GI tumours of uncertain origin
3 Nordic Immunohistochemical Quality Control Important IHC markers of the alimentary tract Epithelial markers CK-pan, CK20, CK7, CK5, EpCAM, Claudin4 Lower GI tract markers CEA, CDX2, SATB2, Cadherin 17 MMR-proteins pmlh1, pmsh2, pmsh6, ppms2 Neuroendocrine markers SYP, CGA Hepatocellular differentiation markers Glypican 3, can. CD66a/CD10, Arginase, GS6 Mesenchymal markers CD117, DOG1, SOX10, β-catenin
4
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6 PAN-CK Inappropriate retrieval AE1/AE3 + HIER TP Liver RCC FN AE1/AE3 + proteolysis TP 6 FN
7 IHC - NordiQC 2014 AE1/AE3 : Optimal results only obtained by HIER in NordiQC runs Dako: RTU HIER Leica: RTU Proteolysis Thermo: VMS: RTU Proteolysis Misleading data sheets + Wrong control material used Conc: Proteolysis or HIER Conc: HIER Conc: HIER Quanto Proteolysis UltraVision Proteolysis + HIER 7
8 EpCAM (Epithelial cell adhesion molecule, EP4, MOC31, ) +(-) Adenocarcinomas (of most types) Neuroendocrine neoplasms
9 EpCAM (Epithelial cell adhesion molecule, EP4, MOC31, ) -/+ Lobular breast carcinoma Hepatocellular carcinoma Squamous cell carcinoma Renal cell carcinoma Embryonal carcinoma Epithelioid mal. mesothelioma Adrenal cortical carcinoma Choroid plexus carcinoma
10 EpCAM protocol Colon adenocarcinoma Lab A optimal with EP4 Retrieval: TRS ph 6.1 DAKO Diva2 ph 6.2 Thermo CC1+Proteolysis VMS Clone BS14: TRIS-EDTA a.o. Lab B insufficient with EP4 Retrieval: Any other buffer without concomitant proteolysis
11 EpCAM antibodies renal cell carcinoma SPM491 Ber-EP4 Ab71916 with optimal protocols
12 Claudin 4 Integral membrane protein, which belongs to the claudin family. The protein is a component of tight junction. Never found in mesothelioma Claudin 4 EpCAM Tonsil
13 Claudin 4 vs. EpCAM Mal. mesothelioma Claudin 4 Mal. mesothelioma EpCAM
14 CDX-2 protein Drosophila caudal related homeobox gene 2 product: Nuclear transcription factor for intestinal differentiation Intestine all cell types incl. endocrine Intestinal metaplasia chronic gastritis Barrett s esophagus Pancreas/bil.tract colon pancreas
15 CDX-2 protein in adenocarcinoma Colorectum +( ) Mucinous ovar. +( ) Esoph./Stom. +/ Mucinous lung +/ Pancr./biliary /+ Prostate (+) Urothelial (+) Endometrioid (+) Yolk sac tumour +
16 ER CDX-2 Endometrioid carcinoma: ER & CDX-2
17 Quality assurance of immunohistochemical Cdx2 detection in carcinomas Results Mean H-score and proportion of positives N EPR* CONC EPR* RTU DAK- CDX2 AMT- 28 CDX2-88 High Ex % 100% 100% 98% 96% Low Ex rmab EPR2764Y (Ventana, CellMarque) 95% 48% 58% 19% 13%
18 CDX2 Normal colon 1 EPRCON EPRRTU DAK AMT 88
19 CDX2 Colon adenocarc. Optimized protocols EPRCON EPRRTU DAK AMT 88
20 Cadherin 17 Calcium dependent adhesion molecules CAD17 = Liver-Intestine (LI-) Cadherin Regulated by CDX2 Intestine (uniform) Pancreas/biliary tract (heterogenous)
21 Cadherin 17 + Adenocarcinoma, colon (incl. medullary MMR prot. defic.) Endocrine neoplasm of small intestine +/- or -/+ Adenocarcinomas of esophagus, stomach, pancreas / biliary tract -(+) Adenocarcinomas of lung, endometrium, ovary, breast Endocrine neoplasms of lung and pancreas Squamous cell carcinoma
22 CAD17 and CDX2 Colon adenocarcinoma Cadherin 17: rmab SP183, CM, 1:50, CC1M/16M/UV CDX2: rmab EPR2764Y, CM
23 CAD17 and CDX2 Colon adenocarcinoma Cadherin 17: rmab SP183, CM, 1:50, CC1M/16M/UV CDX2: rmab EPR2764Y, CM
24 SATB2 rmab SP281 CM Special Adenine Thymine-rich sequence-binding protein 2 Nuclear matrix-associated transcription factor of intestine, neurons and osteoblasts Colorectal adenocarcinomas positive in 90% Renal cell carcinoma positive in 30% Other carcinomas usually negative (10-20% focal) Neuroendocrine neoplasms usually negative
25 Colon adenocarcinoma rmab SP281, CM, 1:200 CC1_32M/16M/OP
26 Carcinoembryonic antigen (CD66e) Adhesion molecule espc. associated with intestine
27 Carcinoembryonic antigen (CD66e) in adenocarcinomas Colorectal + Medull. thyroid + Pancreas/biliary tract +/ Stomach +/ Lung +/ Ovary, mucinous +/ Ovary, non-muc. /+ Prostate Kidney Liver (!) Mesothelioma (!) metast. colon adenoc
28 Carcinoembryonic antigen which antibody?
29 Carcinoembryonic antigen which antibody? Normal liver Clone II-7 Clone TF3H8-1
30 Carcinoembryonic antigen which antibody? Mal. mesothelioma mab II-7 pab or TF3H8-1
31 MLH1 3-step polymer 2-step polymer Improper calibration of Ab titre Less succesful Ab clone G (8/9 insuff.) Less sensitive detection systems:
32 3-step polymer 2-step polymer MLH1 MLH1 loss False neg. internal control
33 MSH2 Improper calibration of Ab titre Poor Ab clone 25D12 (17/17 insuff.) Insufficient HIER 25D12 combined false negative and positive
34 Poor RTU formats: Chromogranin A Well diff endocrine carcinoma LK2H10 with optimized protocvol pab RTU Company 1 mab LK2H10 RTU Company 2 mab LK2H10 RTU Company 3
35 Poor RTU formats: Chromogranin A Small cell carcinoma LK2H10 REF pab RTU Company 1 mab LK2H10 RTU Company 2 mab LK2H10 RTU Company 3
36 Liver tumour of unknown origin hepatocellular carcinoma? Atrophic pancreas
37 Glypican 3 Hepatocellular carcinoma +/- Yolk sac tumour + Chorionic carcinoma + Merkel cell carcinoma +/- Colorectal adenocarcinoma -/+ Gastric adenocarcinoma -/+ Ovarian clear cell carcinoma -/+ Emb. carcinoma (+) Ovarian serous carcinoma (+) Liposarcoma +/- HCC
38 Glypican 3
39 Poor calibration of conc.: Glypican 3
40 CD66a (biliary glycoprotein-1) CEA-like cell adhesion molecule ( pcea ) Bile canaliculi Many epithelia Trophoblast
41 cancd66a (biliary glycoprotein-1) Hepatocelullar carcinoma ~70% canalicular staining HCC HCC 41
42 cancd10 ~70% pos. HCC HCC 42
43 Nordic Immunohistochemical Quality Control Immunohistochemistry in the classifiation of neoplasias of the alimentary tract & External Quality Assurance of Immunohistochemistry for GI cancer markers Mogens Vyberg, director, NordiQC
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