Abstract
The difficulty of identifying and differentiating lymphatic and blood microvessels in tissue sections can be overcome by a monoclonal antibody specific for lymphatic endothelium. Unfortunately, the only known antibody also reacts with the endothelium of some blood vessels. The technique of double immunization (passive, with an antiserum to blood endothelium, and active, with a suspension of lymphatic endothelial cells) was, therefore, used to increase the chances of recognizing specific lymphatic antigens by the mouse immune system. The monoclonal antibody obtained, LyMAb, a G1 immunoglobulin, reacted strongly with the endothelium of bovine thoracic duct, mesenteric collecting vessels and lymphatic vessels of gall-bladder and lymph nodes and moderately with those of the intestinal wall. Blood vessels (intercostal arteries, azygos vein and blood microvessels of all organs tested) were consistently negative. The antibody was species-specific and did not react with formalin-fixed, paraffin-embedded sections. Cross-reactivity was limited to some connective tissue fibres and scattered cells in the lymph node parenchyma, intestinal villi and hepatic lobules.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adamus G, Zam ZS, Emerson SS, Hargrave PA (1989) A practical method for rescuing desired hybridomas during monoclonal antibody production. In Vitro Cell Dev Biol 25: 1141-1146.
Alles JU, Bosslet K (1986) Immunohistochemical and immunochemical characterization of a newendothelial cell-specific antigen. J Histochem Cytochem 34: 209-214.
Auerbach R, Alby L, Grieves J, Joseph J, Lindgren C, Morrissey LW, Sidky YA, Tu M, Watt SL (1982) Monoclonal antibody against angiotensin-converting enzyme: its use as a marker for murine, bovine, and human endothelial cells. Proc Natl Acad Sci USA 79: 7891-7895.
Bardin N, George F, Mutin M, Brisson C, Horschowski N, Francés V, Lesaule G, Sampol J (1996) S-Endo 1, a pan-endothelial monoclonal antibody recognizing a novel human endothelial antigen. Tiss Antigens 48: 531-539.
Cui YC, Tai P-C, Gatter KC, Mason DY, Spry CJF (1983) A vascular endothelial cell antigen with restricted distribution in human foetal, adult and malignant tissues. Immunology 49: 183-189.
de Waal RMW, van Altena MC, Erhard H, Weidle UH, Nooijen PTGA, Ruiter DJ (1997) Lack of lymphangiogenesis in human primary cutaneous melanoma. Am J Pathol 150: 1951-1957.
Djoneidi M, Brodt P (1991) Isolation and characterization of rat lymphatic endothelial cells. Microcirc Endoth Lymphatics 7: 161-182.
Erhard H, Rietveld FJR, Brocker EB, de Waal RMW, Ruiter DJ (1996) Phenotype of normal cutaneous microvasculature. J Invest Dermatol 106: 135-140.
Ezaki T, Matsuno K, Fujii H, Hayashi N, Miyakawa K, Ohmori J, Kotani M (1990) Anewapproach for identification of rat lymphatic capillaries using a monoclonal antibody. Arch Histol Cytol 53: 77-86.
Jaffe EA, Hoyer LW, Nachman RL (1973) Synthesis of antiemophilic factor antigen by cultured human endothelial cells. J Clin Invest 52: 2757-2764.
Johnston MG, Walker MA (1984) Lymphatic endothelial and smoothmuscle cells in tissue culture. In Vitro 20: 566-572.
Jones BE, Yong LCJ (1987) Culture and characterization of bovine mesenteric lymphatic endothelium. In Vitro Cell Dev Biol 23: 698-706.
Kato S, Itonaga I, Ji RC, Miura M (1996) Enzyme triple staining for differentiation of lymphatics from venous and arterial capillaries. Lymphology 29: 15-19.
Kato S (2000) Organ specifity of the structural organization and fine distribution of lymphatic capillary networks: Histochemical study. Histol Histopathol 15: 185-197.
Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256: 495-497.
Leak LV, Burke JF (1966) Fine structure of the lymphatic capillary and the adjoining connective tissue area. Am J Anat 118: 785-809.
Pontecorvo G (1975) Production of mammalian somatic cell hybrids by means of polyethylene glycol treatment. Somatic Cell Genet 1: 397-400.
Russell Jones R, Spaull J, Spry C, Wilson Jones E (1986) Histogenesis of Kaposi's sarcoma in patients with and without acquired immune deficiency syndrome (AIDS). J Clin Pathol 39: 742-749.
Sacchi G, Fonzi L (1975) Osservazioni sui vasi linfatici della colecisti. Boll Soc It Biol Sper 51: 1574-1578.
Sacchi G, Weber E, Aglianò M, Raffaelli N, Comparini L (1997) The structure of superficial lymphatics in the human thigh: precollectors. Anat Rec 247: 53-62.
Sauter B, Foedinger D, Sterniczky B, Wolff K, Rappersberger K (1998) Immunoelectron microscopic characterization of human dermal lymphatic microvascular endothelial cells: differential expression of CD31, CD34, and type IV collagen with lymphatic endothelial cells vs blood capillary endothelial cells in normal human skin, lymphangioma, and hemangioma in situ. J Histochem Cytochem 46: 165-176.
Schlingemann RO, Dingjan GM, Emeis JJ, Blok J, Warnaar SO, Ruiter DJ (1985) Monoclonal antibody PAL-E specific for endothelium. Lab Invest 52: 71-76.
Solito R, Alessandrini A, Fruschelli M, Pucci AM, Gerli R (1997) An immunological correlation between the anchoring filaments of initial lymph vessels and the neighboring elastic fibers: a unified morphofunctional concept. Lymphology 30: 194-202.
Thalhamer J, Freund J (1985) Passive immunization: a method of enhancing the immune response against antigen mixtures. J Immunol Methods 80: 7-13.
Weber E, Lorenzoni P, Lozzi G, Sacchi G (1994a) Cytochemical differentiation between blood and lymphatic endothelium: bovine blood and lymphatic large vessels and endothelial cells in culture. J Histochem Cytochem 42: 1109-1115.
Weber E, Lorenzoni P, Lozzi G, Sacchi G (1994b) Culture of bovine thoracic duct endothelial cells. In Vitro Cell Dev Biol 30A: 287-288.
Zhu D, Pauli BU (1991) Generation of monoclonal antibodies directed against organ-specific endothelial cell surface determinants. J Histochem Cytochem 39: 1137-1142.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Weber, E., Lorenzoni, P., Cavina, N. et al. A Novel Monoclonal Antibody Specific for Lymphatic Endothelium. Histochem J 32, 653–658 (2000). https://doi.org/10.1023/A:1004159214913
Issue Date:
DOI: https://doi.org/10.1023/A:1004159214913