Monoclonal antibodies to various morphologic components of human skin

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.     

Coexistence of varying combinations of neuropeptides with 5-hydroxytryptamine in neurons of the raphe pallidus et obscurus projecting to the spinal cord

The coexistence of varying combinations of substance P (SP), somatostatin (SOM), thyrotropin-releasing hormone (TRH) and met-enkephalin-Arg-Gly-Leu (ENK) with 5-hydroxytryptamine (5-HT) as semiquantitatively revealed by immunocytochemistry in neuronal perikarya of the raphe pallidus et obscurus in the guinea-pig was analyzed. SOM coexisted most frequently with 5-HT, followed by SP, ENK and TRH. Many 5-HT neurons were immunoreactive to 2 or more peptides such as SP/SOM, SOM/ENK, SP/ENK, SOM/TRH, SP/TRH or SOM/SP/ENK. Most of these neurons were shown to project to the spinal cord by retrograde HRP labeling combined with immunocytochemistry. After hemisection of the cervical spinal cord at the C5 level, ENK and 5-HT immunoreactive nerve terminals in the ipsilateral intermediolateral nucleus of the thoracic spinal cord were decreased in number. The results indicate that neurons in the raphe pallidus et obscurus projecting to the spinal cord can be classified into subpopulations according to which peptides coexist with 5-HT, and may have different functions.