RESUMO
CD23 is a low-affinity receptor for immunoglobulin E and expressed on various hemopoietic cells. Although human epidermal cultured Langerhans cells express CD23, the study to identify CD23 on murine Langerhans cells has so far failed. In this study, using highly enriched (> 95%) Langerhans cells from murine epidermis obtained by the panning method, we investigated whether murine Langerhans cells express CD23. As the result of a series of experiments using fluorescence activated cell sorter analysis and the polymerase chain reaction method, it was revealed that CD23 is expressed on cultured Langerhans cells, but not on freshly isolated Langerhans cells. Comparison of the DNA sequence of polymerase chain reaction products of CD23 from cultured Langerhans cells with that from spleen leukocytes demonstrated that there were the same sequences between the two polymerase chain reaction products. The expression of CD23 on cultured Langerhans cells was downregulated when Langerhans cells were cultured with keratinocyte-derived cytokines: interleukin-1alpha, interleukin-18, macrophage colony-stimulating factor, or granulocyte-macrophage colony-stimulating factor. Moreover, it was shown that murine IgE bound to cultured Langerhans cells and this binding was partially inhibited when Langerhans cells were cultured with monoclonal antibody against CD23 (B3B4). Thus this study revealed murine cultured Langerhans cells do express CD23 and the discrepancy from previous reports may be due to the influence of cytokines derived from keratinocytes. Furthermore, the finding that murine cultured Langerhans cells bind IgE through CD23 suggests that CD23 on murine Langerhans cells may be involved in IgE-mediated immune responses.
Assuntos
Células de Langerhans/química , Receptores de IgE/análise , Receptores de IgE/genética , Animais , Células Cultivadas , Células Epidérmicas , Feminino , Citometria de Fluxo , Expressão Gênica/imunologia , Imunoglobulina E/metabolismo , Queratinócitos/química , Células de Langerhans/citologia , Células de Langerhans/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase , RNA Mensageiro/análise , Receptores de IgE/metabolismoRESUMO
Transmissible spongiform encephalopathies are characterized by the accumulation of a proteinase-resistant isoform of the cellular prion-related protein (PrP(c)) within the central nervous system (CNS). The accumulation of scrapie-associated PrP (PrP(Sc)) within cells of the lymphoreticular system prior to its accumulation in the CNS is regarded as important for the development of neurological diseases after peripheral inoculation. Little, however, is known as to which cells are the targets for peripheral inoculation. Here, the presence of PrP(c) on murine Langerhans cells (LC), dendritic cells in the skin and mucosa, and keratinocytes (KC) is demonstrated by immunohistochemical staining, Western-blotting and FACS analysis. The expression of PrP(c) mRNA in freshly purified LC and KC was also detected by reverse transcriptase-polymerase chain reaction. The expression of PrP(c) on LC was slightly increased during culture. These data suggest that LC and KC may be the targets for peripheral infection with prions.