Immunohistochemical co-localization of lymphatics and blood vessels in oral squamous cell carcinomas

BACKGROUND: Differentiating lymphatic vessels from blood vessels is difficult, partly due to the lack of a specific method for identifying lymphatics. A new lymphatic vessel-reactive antibody, D2-40 has recently become commercially available. We examined the selectivity of D2-40 for lymphatics in oral neoplastic lesions for discrimination from blood vessels. METHODS: Formalin-fixed, paraffin-embedded sections of oral lymphangiomas (n = 3), oral hemangiomas (n = 7), and oral squamous cell carcinomas (OSCC, n = 46) were double immunostained with D2-40 and anti-CD34 monoclonal antibodies (MoAb) using ENVISION-polymer technique with 5-bromo-4-chloro-3-indoxyl-phosphate (BCIP)/nitroblue tetrazolium chloride (NBT) and 3,3'-diaminobenzidine (DAB) as color reagents, respectively. Results: In the oral lymphangiomas and hemangiomas D2-40 was detected in all lymphatics, while all blood vessels were positive for CD34. In OSCC, number of vessels for lymphatics (P < 0.01) and for blood vessels in the perineoplastic areas were significantly greater than those in intratumoral areas. CONCLUSIONS: These results indicate that lymphatic proliferation might be much more extensive in the peritumoral area than intratumoral.