Specialespecifikt kursus i Patologisk Anatomi Diagnostiske ...3 – Immunohistochemical classification of tumours based on keratin subtyping. Specialespecifikt kursus i Patologisk

3 – Immunohistochemicalclassification of tumours

based on keratin subtyping

Specialespecifikt kursus i Patologisk Anatomi

Diagnostiske metoder Immunhistokemi

2018

• Highly complex family of intermediate filaments• > 50 distinct types (excl. trichocytic keratins)• Types CK1-20 diagnostically most relevant:

Class I (type A - Acidic): CK9 (64 kDa) - CK20 (40 kDa)

Class II (type B - Basic/neutral): CK1 (68 kDa) - CK8 (52.5 kDa)

Cytokeratins – a multigene family

CK: renal cell carcinoma

1992

11?

(B) (A)

Moll et al. Cell 1982; 31:11

The Moll Cytokeratin catalogue 1982

• One CK class I and one CK class II ‘always’ paired• CK class I in a pair ~ 8 kDa smaller than class II

Modified from: Lane & Alexander Semin Cancer Biol 1990, 1:165

Pairing of cytokeratins

Neutral/Basic (B, class II)Acidic (A, class I)

110

413

514 17 19

720

818

Squamous epithelia:- suprabasal, keratinizing +++ - - + ++ (+) - - - -- suprabasal, non-keratinizing + +++ +++ + ++ (+) - - - -

- basal cells (1tonsil, 2mucosa) - - - +++ +++ (+)1 (++)12 (+)1 - (+)1

Transit. epith.: superficial cells - - - - - - +++ +++ ++ +++- intermed. / 3basal cells - (+)3 +++ (+++)3 - (++)3 +++ +++ (+) +++

Mesothelium - - - ++ ++ + +++ +++ - +++

Bronchus, breast, prost., cerv.:- basal/myoepithelial cells - - - +++ ++ +++ ++ - - -- luminal cells - - - + + + +++ +++ - +++

Biliary/pancr. ducts, lung alv.,endometr., renal collect. ducts - - - - - - +++ +++ - +++4Stomach (foveola), intestine - - - - - - +++ (+)4 +++ +++Hepatocytes, pancr. acini,prox. renal tubules - - - - - - - - - +++Endocrine cells (5Merkel, 6thyr.) - - - - - - (+++)5 (++)6 (+++)5 +++Smooth muscle (vasc., myom.),myofibrobl., 7sm.ves.endothelia - - - - - - + (++)7 - ++

Cytokeratin types and cell types

HMW Cytokeratin types - squamous epithelia


110

413

514 17 19

720

818




Mesothelium - - - ++ ++ + +++ +++ - +++




CK5: esophagus CK4: esophagus


CK17: tonsil CK17: skin

LMW Cytokeratin types - squamous epithelia


110

413

514 17 19

720

818



Transit. epith.: superficial cells - - - - - - +++ +++ ++ +++

- intermed. / 3basal cells - (+)3 +++ (+++)3 - (++)3 +++ +++ (+) +++Mesothelial cells - - - ++ ++ + +++ +++ - +++



LMW Cytokeratin types - squamous epithelia

CK19: tonsil CK19: esophagus

Cytokeratin types - urothelium


110

413

514 17 19

720

818




Mesothelium - - - ++ ++ + +++ +++ - +++



Cytokeratin types - urothelium

CK5 (+CK13) CK20

Cytokeratin types - mesotheliumNeutral/Basic (B, class II)Acidic (A, class I)

110

413

514 17 19

720

818

Squamous epithelia:- suprabasal, keratinizing +++ - - + ++ (+) - - - -- suprabasal, non-keratiniz.ep. + +++ +++ + ++ (+) - - - -



- intermed. / 3basal cells - (+)3 +++ (+++)3 - (++)3 +++ +++ (+) +++Mesothelium - - - ++ ++ + +++ +++ - +++Bronchus, breast, prost., cerv.:- basal/myoepithelial cells - - - +++ ++ +++ ++ - - -- luminal cells - - - + + + +++ +++ - +++


Cytokeratin types - mesothelium

CK5: mesothelium(adenocarcinoma neg.)CK19: mesothelium

Cytokeratin types - complex epithelia


110

413

514 17 19

720

818


- basal cells (1tonsil, 2mucosa) - - - - +++ (+)1 (++)12 (+)1 - (+)1


- intermed. / 3basal cells - (+)3 +++ (+++)3 - (++)3 +++ +++ (+) +++Mesothelium - - - ++ ++ + +++ +++ - +++



Cytokeratin types - complex epithelia

CK5 (14,17): breast CK18 (8,7): breast

Cytokeratin types - simple epithelia


110

413

514 17 19

720

818

Squamous epithelia:- suprabasal, keratinizing +++ - - + ++ -(+) - - - -- suprabasal, non-keratinizing + +++ +++ + ++ (+) - - - -

- basal cells (1tonsil, 2mucosa) - - - - +++ -(+)1 (++)12 -(+)1 - -(+)1





Cytokeratin types - simple epithelia (liver, kidney)

CK7(19): bile ducts CK7(19): renal col.



110

413

514 17 19

720

818








CK20: gastric foveolae CK20: colon



110

413

514 17 19

720

818








CK8: liver CK8: kidney



110

413

514 17 19

720

818








CK20: Merkel cells CK8: Pancreas

Islet of Langerhans

Cytokeratin types - mesenchymal cells


110

413

514 17 19

720

818







Cytokeratin types - mesenchymal cells

CK7: endotheliumCK8: lymph node fibroblastic reticulum cells


110

413

514 17 19

720

818

Squamous cell carcinoma (+) + + ++ ++ (+) (+) (+) - (+)Transitional cell tumour - + + + + + ++ ++ ++ ++Malignant mesothelioma - - - ++ ++ + ++ + - ++

Adenocarcinoma: complexepithelia (bronch., breast, prost.) - - - (+) (+) (+) ++ ++ - ++Adenocarc.: biliary tract,pancr., endom., ovary - - - (+) (+) (+) ++ ++ (+) ++Adenocarc.: stomach - - - - - - ++ + + ++Adenocarc.: intestine - - - - - - ++ + ++ ++Hepatocellular carcinomaRenal cell carcinoma - - - - - - - + - ++Endocrine tumours: carcinoids - - - - - - + + - ++- Merkel cell carcinoma - - - - - - + - ++ ++- Thyroid carcinoma - - - - - - + ++ - ++

Cytokeratins in epithelial neoplasias


110

413

514 17 19

720

818

Squamous cell carcinoma + + + ++ ++ + + + - +Transitional cell tumour - + + + + + ++ ++ ++ ++Malignant mesothelioma - - - ++ ++ + ++ + - ++


Cytokeratins in squamous cell carcinoma

CK1 CK10


CK4CK5

esophagus


CK13 CK4 (loss)



110

413

514 17 19

720

818

Squamous cell carcinoma + + + ++ ++ + + + - +Transitional cell tumour - + + ++ + ! + ! ++ ++ ++ ++Malignant mesothelioma - - - ++ ++ + ++ + - ++


Cytokeratins in urothelial carcinoma

CK20

CK20 CK5

CK5

Cytokeratins in urothelial carcinoma


110

413

514 17 19

720

818

Squamous cell carcinoma (+) + + ++ ++ (+) (+) (+) - (+)Transitional cell tumour - + + + + + ++ ++ ++ ++Malignant mesothelioma - - - ++ ++ + ++ + - ++Adenocarcinoma: complexepithelia (bronch., breast, prost.) - - - (+) (+) (+) ++ ++ - ++Adenocarc.: biliary tract,pancr., endom., ovary - - - (+) (+) (+) ++ ++ (+) ++Adenocarc.: stomach - - - - - - ++ + + ++Adenocarc.: intestine - - - - - - ++ + ++ ++Hepatocellular carcinomaRenal cell carcinoma - - - - - - - + - ++Endocrine tumours: carcinoids - - - - - - + + - ++- Merkel cell carcinoma - - - - - - + - ++ ++- Thyroid carcinoma - - - - - - + ++ - ++

Cytokeratins in malignant mesothelioma

CK5CK5


CK5

CK8 (loss)

CK7



110

413

514 17 19

720

818


Adenocarcinoma: complexepith. (bronch., breast, prost.) - - - (+) (+) (+) ++ ++ - ++Adenocarc.: biliary tract,pancr., endom., ovary - - - (+) (+) (+) ++ ++ (+) ++Adenocarc.: stomach - - - - - - ++ + + ++Adenocarc.: intestine - - - - - - ++ + ++ ++Hepatocellular carcinomaRenal cell carcinoma - - - - - - - + - ++Endocrine tumours: carcinoids - - - - - - + + - ++- Merkel cell carcinoma - - - - - - + - ++ ++- Thyroid carcinoma - - - - - - + ++ - ++

Cytokeratins in adenocarcinomas

CK5: Breast lob. carcinoma CK5: Breast duct. carc.


CK5: Breast lob. carcinoma CK5: prostate adenocarc.CK5: Prostate hyperplasia



110

413

514 17 19

720

818


Adenocarcinoma: complexepith. (bronch., breast, prost.) - - - (+) (+) (+) ++ ++ - ++Adenocarc.: biliary tract,pancr., endom., ovary - - - (+) ! (+) ! (+) ! ++ ++ (+) ! ++Adenocarc.: stomach - - - - - - ++ + + ++Adenocarc.: intestine - - - - - - ++ + ++ ++Hepatocellular carcinomaRenal cell carcinoma - - - - - - - + - ++Endocrine tumours: carcinoids - - - - - - + + - ++- Merkel cell carcinoma - - - - - - + - ++ ++- Thyroid carcinoma - - - - - - + ++ - ++


CK17: Pancreas adenocarcinoma

CK5: Biliary tract adenocarcinoma


CK17CK5: Serous ovarian carcinoma

CK20: Gall bladder adenocarcinoma



110

413

514 17 19

720

818


Adenocarcinoma: complexepith. (bronch., breast, prost.) - - - (+) (+) (+) ++ ++ - ++Adenocarc.: biliary tract,pancr., endom., ovary - - - (+) (+) (+) ++ ++ (+) ++Adenocarc.: stomach - - - - - - ++ + + ++Adenocarc.: intestine - - - - - - ++ + ! ++ ++Hepatocellular carcinomaRenal cell carcinoma - - - - - - - + - ++Endocrine tumours: carcinoids - - - - - - + + - ++- Merkel cell carcinoma - - - - - - + - ++ ++- Thyroid carcinoma - - - - - - + ++ - ++


CK20

CK7

CK20

CK7Colon: Typical Colon: Deviant



110

413

514 17 19

720

818


Adenocarcinoma: complexepith. (bronch., breast, prost.) - - - (+) (+) (+) ++ ++ - ++Adenocarc.: biliary tract,pancr., endom., ovary - - - (+) (+) (+) ++ ++ (+) ++Adenocarc.: stomach - - - - - - ++ + + ++Adenocarc.: intestine - - - - - - ++ + ++ ++Hepatocellular carcinomaRenal cell carcinoma - - - - - - - + - ++Endocrine tumours: carcinoids - - - - - - + + - ++- Merkel cell carcinoma - - - - - - + - ++ ++- Thyroid carcinoma - - - - - - + ++ - ++


CK7HCC: typical HCC: deviant

CK20

Mallory body

CK7

CK7


CK7: CCRCC CK7: CCRCC



110

413

514 17 19

720

818


Adenocarcinoma: complexepith. (bronch., breast, prost.) - - - (+) (+) (+) ++ ++ - ++Adenocarc.: biliary tract,pancr., endom., ovary - - - (+) (+) (+) ++ ++ (+) ++Adenocarc.: stomach - - - - - - ++ + + ++Adenocarc.: intestine - - - - - - ++ + ++ ++Hepatocellular carcinomaRenal cell carcinoma - - - - - - - + - ++Endocr. tumours: carcinoids - - - - - - + + - ++- Merkel cell carcinoma - - - - - - + - ++ ++- Thyroid carcinoma - - - - - - + ++ - ++

Cytokeratins in endocrine tumours

CK20: Merkel cell carc. CK19: Thyr. papill. carc.

Cytokeratins in endocrine tumours

CK20+ CK20 amb CK20-

CK7+Lung MBAC1

EsophagusBiliary tractPancreasOvary muc.Urothelium

Lung 1

Breast d./l.Thyroid p./f.

Endom. c.Cervix ad.Ovar. nonmuc

CK7 ambAppendixSmall intest.Rectum

StomachNeuroend.2

Germ cell t.5

Chr.phobe rc.6

CK7-ColonMerkel cell 2

Carcinoid 2

Liver cell c.Renal cell. c.6

Adrenal cort.c.Prostate

Coordinate cytokeratin 7/20 expression


CK7+Lung MBAC1


Lung 1




StomachNeuroend.2

Germ cell t.5

Chr.phobe rc.6


Carcinoid 2




CK7

CK20

CK7♀ 48, biopsy from vaginal ”polyp”


CK7

CK20

CK7


CK7

CK20

CK7CK7

CK20

cdx2


CK7 CK20



CK7+Lung MBAC1


Lung 1




StomachNeuroend.2

Germ cell t.5

Chr.phobe rc.6


Carcinoid 2





CK7+Lung MBAC1


Lung 1




StomachNeuroend.2

Germ cell t.5

Chr.phobe rc.6


Carcinoid 2




CK7CK7CK20

CDX2



CK7+Lung MBAC1


Lung 1




StomachNeuroend.2

Germ cell t.5

Chr.phobe rc.6


Carcinoid 2




CK20Urothelial carcinoma

CK7



CK7+Lung MBAC1


Lung 1




StomachNeuroend.2

Germ cell t.5

Chr.phobe rc.6


Carcinoid 2





CK7+Lung MBAC1


Lung 1




StomachNeuroend.2

Germ cell t.5

Chr.phobe rc.6


Carcinoid 2




Coordinate cytokeratin / vimentin expression in carcinomas

Thyroid, adrenal cortex ~ 100%Renal cell ~ 60%Serous, endometrioid ~ 50% Lung ~ 40%Prostate ~ 40%Pancreas, urothelium ~ 30%Breast ~ 10%Carcinoid ~ 10%

Coordinate cytokeratin-vimentin expression

Conc. Abs: CK types N Optimal Good Bord. Poor Suff.1 Suff. OPS2

mAb clone 34BE12

1/5/10/14 (19?) 89 1 6 75 7 8 % 40 %

mAb clone D5/16B4 5/6 17 6 7 3 1 76 % 82 %

mAb clone XM26 5 6 4 1 1 0 83 % 83 %

CK-HMW run 32, 2011

CK 5 or CK5/6 34BE12

Cytokeratins: Selection of antibodies

CK5/6: clone D5/16 B4CK5: XM-26

CK5/14/1/10/19?:34BE12

optimal too diluted HIER Prot.

5D3 35βH11C51

CAM5.2DC10

TS1

Cytokeratins – which clones

Cytokeratins: Selection of antibodies

optimalEsophagus

CK19: clone A53-B/A2.26 CK19: clone RCK108After proteolysis as recommended by the vendor

Papillary thyroid carcinoma

EP17/E30B22.1/B23.1

(CK8/18)DC10 (CK18)

CAM5.2 (CK8/7)

35BH11(CK8)

5D3 onBond

5D3 onBenchmark

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Specialespecifikt kursus i Patologisk Anatomi Diagnostiske ...3 – Immunohistochemical classification of tumours based on keratin subtyping. Specialespecifikt kursus i Patologisk