D2-40, oncofetal M2A antigen

D2-40 is directed against the M2A antigen, a 40 kDa O-linked sialoglycoprotein found on the cell surface of testicular gonocytes, germ cell tumours lymphatic endothelium and mesothelial cells. D2-40 is immunoreactive with formalin-fixed paraffin embedded tissue.

Immunohistochemical expression

The lymphatic staining is in a thin linear pattern.

Mesothelial immunoreactivity is membranous and, to a lesser extent, cytoplasmic. Other antibodies are shown for comparison.

D2-40

Calretinin

WT1

CK5/6

rate of positivity

mean percentage of cells staining in positive cases

Mesothelioma

epithelioid

33/33¹, 32/45 (the lower sensitivity may have been bcause this was a tissue microarray based study.)⁵

92%¹

33/33¹

19/33¹

29/32¹

biphasic

15/16¹

epithelioid cells: 90%, sarcomatous cells: 26%¹

16/16¹

10/16¹

13/16¹

sarcomatous

3/4¹

52%¹

2/4¹

0/4¹

1/4¹

Reactive pleura

27/28¹

71%¹

Pulmonary adenocarcinoma

2/31 (weak cytoplasmic staining)¹

13%¹

7/31¹

2/31¹

12/31¹

Ovarian serous carcinoma

17/26 (membranous staining)¹

33%¹

8/26¹

23/26¹

9/26¹

Renal cell carcinoma

Conventional

0/16¹

2/16¹

0/16¹

5/16¹

Papillary

5/13 (weak cytoplasmic staining)¹

43%¹

1/13¹

1/13¹

7/13¹

Chromophobe

0/5¹

1/5¹

1/5¹

1/5¹

Breast carcinoma

3/16 (weak cytoplasmic staining)¹

13%¹

Prostatic adenocarcinoma

0/11¹

Urothelial carcinoma

0/7¹

Another study evaluated podoplanin alongside D2-40²:

D2-40

Podoplanin

Mesothelioma

epithelioid

25/29 (better differentiated tumours show thick continuous membrane staining. In poorly differentiated tumours, membrane staining is discontinuous or staining is dot-like. Intracytoplasmic globoid staining may occur.)

25/29 (staining was seen in the same 25 cases that showed positivity with D2-40)

biphasic

4/5 (positivity confined to the epithelioid component)

4/5 (positivity confined to the epithelioid component)

sarcomatoid

0/6

0/6

Adenocarcinoma

lung

0/24

0/24

ovary

0/17

0/17

endometrium

0/5

0/5

breast

0/10

0/10

pancreas

0/5

0/5

stomach

0/5

0/5

colon

0/10

0/10

kidney

0/10

0/10

prostate

0/5

0/5

thyroid

0/3

0/3

Adenomatoid tumour

2/2 (staining is of the apical cell membrane)

2/2

Squamous carcinoma of lung

0/10

0/10

Angiosarcoma

2/5

2/5

Synovial sarcoma

biphasic

4/6 (positivity confined to the epithelioid component)

4/6 (positivity confined to the epithelioid component)

monophasic

0/6

0/6

This study was based on cell blocks prepared from serous effusions³. In cases where tumours stained, the pattern was membranous.

up to 10% of tumour cells staining

more than 10% of tumour cells staining

reactive mesothelium

153/153 (8 purely reactive, 145 mesothelial cells in the presence of malignancy)

malignant mesothelioma

2/32 (24 pleura, 8 peritoneal)

30/32 (24 pleura, 8 peritoneal)

malignant mesothelioma biopsies

12/12

ovarian carcinoma

85/169 (138 serous [including 23 primary peritoneal and 8 tubal], 3 clear cell, 3 endometrioid, 7 undifferentiated, 6 combined, 2 undefined: 129 peritoneal, 40 pleural)

13/169 (138 serous [including 23 primary peritoneal and 8 tubal], 3 clear cell, 3 endometrioid, 7 undifferentiated, 6 combined, 2 undefined: 129 peritoneal, 40 pleural)

ovarian microarray of biopsies

5/72 72 cores of primary and metastatic ovarian carcinoma from 23 patients)

9/72 72 cores of primary and metastatic ovarian carcinoma from 23 patients)

breast carcinoma

11/44 (40 pleura, 3 peritoneal)

2/44 (40 pleura, 3 peritoneal)

adenocarcinoma of unknown origin

3/14 (7 pleural, 7 peritoneal)

3/14 (7 pleural, 7 peritoneal)

lung carcinoma

2/6 (all pleural)

0/6 (all pleural)

Other cancers (4 oesophagus, 4 cervix uteri, 2 endometrial, 3 gastric, 1 colon, 1 pancreas, 1 prostate, 1 melanoma)

7/17 (2/2 cervical, 1/2 endometrial, 1/3 gastric, 2/4 oesophageal, 1/1 pancreatic, 1/1 prostatic and 1/1 melanoma were positive)

2/17 (2/2 cervical, 1/2 endometrial, 1/3 gastric, 2/4 oesophageal, 1/1 pancreatic, 1/1 prostatic and 1/1 melanoma were positive)

Diagnostic utility

Diagnosis of lymphatic-derived tumours.
Diagnosis of lymphatic invasion by tumours.
Differentiation of mesothelioma from adenocarcinoma⁴. The initial evidence as to its usefulness in the differentiation of mesothelioma from serous ovarian tumours is inconsistent. Like other mesothelial markers, it appears least useful in sarcomatous mesotheliomas.

References

¹Chu AY, Litzky LA, Pasha TL, Acs G,Zhang PJ Utility of D2-40, a novel mesothelial marker, in the diagnosis of malignant mesothelioma. Mod Pathol 2005; 18:105-10

²Ordonez NG D2-40 and podoplanin are highly specific and sensitive immunohistochemical markers of epithelioid malignant mesothelioma. Hum Pathol 2005; 36:372-80

³Bassarova AV, Nesland JM,Davidson B. D2-40 is Not a Specific Marker for Cells of Mesothelial Origin in Serous Effusions. Am J Surg Pathol 2006; 30:878-82

⁴Ordonez NG. Immunohistochemical diagnosis of epithelioid mesothelioma: an update. Arch Pathol Lab Med 2005; 129:1407-14 FULL TEXT

⁵SienkoA, Zander DS, Killen D et al. D2-40 is a novel new marker of malignant mesothelioma (MM): tissue microarray study of 45 MM versus 409 lung carcinomas and primary non-mesothelial neoplasm of the pleura and chest wall. Mod Pathol 2005;18(suppl1):318A.

This page last revised 1.8.2006.

		D2-40		Calretinin	WT1	CK5/6
		rate of positivity	mean percentage of cells staining in positive cases	Calretinin	WT1	CK5/6
Mesothelioma	epithelioid	33/33¹, 32/45 (the lower sensitivity may have been bcause this was a tissue microarray based study.)⁵	92%¹	33/33¹	19/33¹	29/32¹
	biphasic	15/16¹	epithelioid cells: 90%, sarcomatous cells: 26%¹	16/16¹	10/16¹	13/16¹
	sarcomatous	3/4¹	52%¹	2/4¹	0/4¹	1/4¹
Reactive pleura		27/28¹	71%¹
Pulmonary adenocarcinoma		2/31 (weak cytoplasmic staining)¹	13%¹	7/31¹	2/31¹	12/31¹
Ovarian serous carcinoma		17/26 (membranous staining)¹	33%¹	8/26¹	23/26¹	9/26¹
Renal cell carcinoma	Conventional	0/16¹		2/16¹	0/16¹	5/16¹
	Papillary	5/13 (weak cytoplasmic staining)¹	43%¹	1/13¹	1/13¹	7/13¹
	Chromophobe	0/5¹		1/5¹	1/5¹	1/5¹
Breast carcinoma		3/16 (weak cytoplasmic staining)¹	13%¹
Prostatic adenocarcinoma		0/11¹
Urothelial carcinoma		0/7¹

		D2-40	Podoplanin
Mesothelioma	epithelioid	25/29 (better differentiated tumours show thick continuous membrane staining. In poorly differentiated tumours, membrane staining is discontinuous or staining is dot-like. Intracytoplasmic globoid staining may occur.)	25/29 (staining was seen in the same 25 cases that showed positivity with D2-40)
	biphasic	4/5 (positivity confined to the epithelioid component)	4/5 (positivity confined to the epithelioid component)
	sarcomatoid	0/6	0/6
Adenocarcinoma	lung	0/24	0/24
	ovary	0/17	0/17
	endometrium	0/5	0/5
	breast	0/10	0/10
	pancreas	0/5	0/5
	stomach	0/5	0/5
	colon	0/10	0/10
	kidney	0/10	0/10
	prostate	0/5	0/5
	thyroid	0/3	0/3
Adenomatoid tumour		2/2 (staining is of the apical cell membrane)	2/2
Squamous carcinoma of lung		0/10	0/10
Angiosarcoma		2/5	2/5
Synovial sarcoma	biphasic	4/6 (positivity confined to the epithelioid component)	4/6 (positivity confined to the epithelioid component)
Synovial sarcoma	monophasic	0/6	0/6

	up to 10% of tumour cells staining	more than 10% of tumour cells staining
reactive mesothelium		153/153 (8 purely reactive, 145 mesothelial cells in the presence of malignancy)
malignant mesothelioma	2/32 (24 pleura, 8 peritoneal)	30/32 (24 pleura, 8 peritoneal)
malignant mesothelioma biopsies		12/12
ovarian carcinoma	85/169 (138 serous [including 23 primary peritoneal and 8 tubal], 3 clear cell, 3 endometrioid, 7 undifferentiated, 6 combined, 2 undefined: 129 peritoneal, 40 pleural)	13/169 (138 serous [including 23 primary peritoneal and 8 tubal], 3 clear cell, 3 endometrioid, 7 undifferentiated, 6 combined, 2 undefined: 129 peritoneal, 40 pleural)
ovarian microarray of biopsies	5/72 72 cores of primary and metastatic ovarian carcinoma from 23 patients)	9/72 72 cores of primary and metastatic ovarian carcinoma from 23 patients)
breast carcinoma	11/44 (40 pleura, 3 peritoneal)	2/44 (40 pleura, 3 peritoneal)
adenocarcinoma of unknown origin	3/14 (7 pleural, 7 peritoneal)	3/14 (7 pleural, 7 peritoneal)
lung carcinoma	2/6 (all pleural)	0/6 (all pleural)
Other cancers (4 oesophagus, 4 cervix uteri, 2 endometrial, 3 gastric, 1 colon, 1 pancreas, 1 prostate, 1 melanoma)	7/17 (2/2 cervical, 1/2 endometrial, 1/3 gastric, 2/4 oesophageal, 1/1 pancreatic, 1/1 prostatic and 1/1 melanoma were positive)	2/17 (2/2 cervical, 1/2 endometrial, 1/3 gastric, 2/4 oesophageal, 1/1 pancreatic, 1/1 prostatic and 1/1 melanoma were positive)