Lymph vascular invasion in invasive mammary carcinomas identified by the endothelial lymphatic marker D2-40 is associated with other indicators of poor prognosis

Abstract

Background: Immunohistochemical studies of lymphatic vessels have been limited by a lack of specific markers. Recently, the novel D2-40 antibody, which selectively marks endothelium of lymphatic vessels, was released. The aim of our study is to compare lymphatic and blood vessel invasion detected by hematoxylin and eosin (H&E) versus that detected by immunohistochemistry, relating them with morphologic and molecular prognostic factors.

Methods: We selected 123 cases of invasive mammary carcinomas stratified into three subgroups according to axillary lymph node status: macrometastases, micrometastases, and lymph node negative. Lymphatic (LVI) and blood (BVI) vessel invasion were evaluated by H&E and immunohistochemistry using the D2-40 and CD31 antibodies, and related to histologic tumor type and grade, estrogen and progesterone receptors, E-cadherin, Ki67, p53, and Her2/neu expression.

Results: LVI was detected in H&E-stained sections in 17/123 cases (13.8%), and in D2-40 sections in 35/123 cases (28.5%) (Kappa = 0.433). BVI was detected in H&E-stained sections in 5/123 cases (4.1%), and in CD31 stained sections in 19/123 cases (15.4%) (Kappa = 0.198). LVI is positively related to higher histologic grade (p = 0.013), higher Ki67 expression (p = 0.00013), and to the presence of macrometastases (p = 0.002), and inversely related to estrogen (p = 0.0016) and progesterone (p = 0.00017) receptors expression.

Conclusion: D2-40 is a reliable marker of lymphatic vessels and is a useful tool for lymphatic emboli identification in immunostained sections of breast carcinomas with higher identification rates than H&E. Lymphatic vessel invasion was related to other features (high combined histologic grade, high Ki67 score, negative hormone receptors expression) associated with worse prognosis, probable reflecting a potential for lymphatic metastatic spread and aggressive behavior.

Figure 1

Lymphatic and blood vessels in sections of invasive breast cancer stained for D2-40 and CD31. Lymphatic vessel invasion stained for CD31(A) and D2-40 (B) in sections of breast tumors from the same case. In A, CD31 positive endothelium and in B, D2-40 positive endothelium (×400). Blood vessel invasion stained for CD31 (C) and D2-40 (D) in sections of breast tumors from the same case. In C, CD31 positive endothelium and in D, D2-40 negative endothelium (×400). In B and C, the endothelium of lymphatic vessels without invasion was also immunostained.

Figure 2

Lymphatic and blood vascular invasion in invasive breast cancer in H&E and immunostained slides. Lymphatic vascular invasion (arrow) seen in H&E (A) and D2-40 (B) stained sections of breast tumors from the same case; ×400; Blood vessel invasion (arrow) seen in H&E (C) and CD31 (D) stained sections; ×400.