A novel lamina lucida component of epithelial and endothelial basement membranes detected by LH39 monoclonal antibody.
The murine monoclonal antibody, LH39 was characterized in this study and appeared to bind to a novel basement membrane epitope. This antigen was expressed in the epithelial basement membrane of human tissue derived from all three germ cell layers and in basement membranes surrounding small blood vessels within the stroma of all organs examined. LH39 antigen could be first detected in fetal skin at the dermo-epidermal junction at 7 weeks estimated gestational age but was not present in the dermal vasculature until 16 weeks. When tested against tissue from a range of lower mammalian species, LH39 antigen appeared to be primate-specific. The epithelial basement membrane zone in organotypical cultures, where there is de novo synthesis of basement membrane components, contained abundant LH39 antigen in contrast to other basement membrane components, type IV collagen, laminin, and type VII collagen. Ultrastructural localization of LH39 epitope, using immunogold electron microscopy on unfixed freshly frozen tissue, was to the lamina lucida. No cross-reactivity could be detected between LH39 and laminin, fibronectin, and collagens I, III, IV, and V using the ELISA assay. In vitro studies with a range of proteolytic enzymes suggested that the antigen was non-collagenous in nature. LH39 precipitated a polypeptide with a molecular weight of 185 kD from extracts of metabolically labelled cultured keratinocytes, and polypeptides of 185 and 200 kD from the culture medium. The tissue distribution of LH39 antigen suggested that it may be an epitope within anchoring filaments. Potential applications of this antibody include the study of benign and malignant human vascular disorders, diseases and tumours associated with angiogenesis, epithelial neoplasms, and conditions of tissue regeneration and repair, such as wound healing.
Almeida
B M
BM
Department of Oral Medicine and Pathology, UMDS, Guy's Hospital, London, U.K.
Challacombe
S J
SJ
Eveson
J W
JW
Smith
C G
CG
Leigh
I M
IM
D000328:Adult; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis*; D001485:Basement Membrane / Q000276:immunology* / Q000648:ultrastructure; D004727:Endothelium / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D004848:Epithelium / Q000276:immunology; D005333:Fetus / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D016253:Microscopy, Immunoelectron; D011233:Precipitin Tests; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D000941:Antigens