Clinical relevance of detection of lymphovascular invasion in primary melanoma using endothelial markers D2-40 and CD34

Abstract

Immunohistochemistry (IHC) using endothelial markers may facilitate the detection of lymphovascular invasion (LVI) in primary melanoma; however, the clinical implications of enhanced detection are unknown. We evaluated whether the use of lymphatic endothelial marker D2-40 and panvascular marker CD34 increases LVI positivity relative to routine histology alone and then evaluated the prognostic relevance of LVI detected using these markers in terms of disease-free (DFS) and overall survival (OS). A total of 246 primary melanomas were assessed for LVI using D2-40, CD34, and routine histology. Associations between LVI positivity and clinicopathologic variables, DFS, and OS were compared using univariate and multivariate analyses. The use of endothelial markers increased the rate of LVI positivity (18% using D2-40 and/or CD34 vs. 3% by routine histology, P<0.0001). On univariate analysis, IHC-detected LVI was significantly associated with more adverse clinicopathologic variables (thickness, ulceration, mitoses, and nodular subtype) compared with LVI detected by routine histology (thickness and ulceration only). In a multivariate model controlling for stage, LVI detected using IHC markers remained a significant marker of both reduced DFS [hazard ratio (HR), 2.01; 95% confidence interval (CI): 1.27-3.18; P=0.003] and OS (HR, 2.08; 95% CI: 1.25-3.46; P=0.005). Results show that D2-40 and CD34 increase the detection of LVI in primary melanoma and that cases missed by routine histology have prognostic relevance.

FIGURE 1

Examples of LVI in primary melanoma detected using routine histology, panvascular marker CD34, and lymphatic endothelial marker D2-40. A, Evidence of LVI by routine histology in primary melanoma. B, Stain for CD34 highlights the presence of tumor embolus in the same case. C, CD34 stains both lymphatic channels (arrow) and blood vascular endothelium-lined spaces (arrowhead), whereas D2-40 (D) stains the lymphatic channels that contain melanoma cells (arrow) but not the blood vascular channels that contain only red blood cells (arrowhead). Magnification: × 200.

FIGURE 2

Rates of LVI in primary melanoma stratified by method of detection used: D2-40, CD34, or routine histology. *P < 0.0001; CD34 and/or D2-40 vs. routine histology; t test.

FIGURE 3

Representative case of LVI in primary melanoma detected using D2-40/S-100 dual IHC. Lymphatic endothelium (brown, arrow) surrounds an intratumoral melanoma cell that stains positive for S-100 (red, arrowheads), thus clearly differentiating it from surrounding lymphocytes (asterisk) that are negative for S-100. Magnification: × 400.

FIGURE 4

Kaplan-Meier survival curves showing DFS stratified by the presence of LVI in the primary melanoma as detected using endothelial markers D2-40 and/or CD34 (A) compared with routine histology (B). OS stratified by LVI positivity as detected using D2-40/and or CD34 (C).