Monoclonal Antibodies to Various Morphological Components of Human Skin

Abstract

Somatic cell hybrids were established from the mouse myeloma, P3x63Ag8.653 cells, and the spleen cells of a mouse hyperimmune to human epidermal cells. Indirect immunofluorescence test with hybridoma culture fluids displayed that 253 out of 263 hybridoma cultures secreted antibodies reactive with the frozen sections of human skin. The hybridomas secreting unique antibodies to skin components were cloned and designated as AHS-1 to -8. Monoclonal antibodies (MoAb) from AHS-1, AHS-2, and AHS-3 hybridomas did detect cytoplasmic antigens present in the epidermal layer, eccrine ducts and glands (except MoAb AHS-1), and hair follicles. Enzyme-linked immunosorbent assay showed that the antigen recognized by either MoAb AHS-1 or MoAb AHS-2, but not by MoAb AHS-3, shares the antigenic determinant with antigen(s) present in keratin polypeptides isolated from human callus. MoAb AHS-4 to -8 distinctly stained each morphologic component of the skin tissues: cell membranes of epidermal cells by MoAb AHS-4, cytoplasms in upper layer cells of epidermis by MoAb AHS-5, dermis by MoAb AHS-6, basement membrane zone by MoAb AHS-7, and eccrine ducts by MoAb AHS-8.