The Abnormal Cytokeratin Expression In Hepatocytes In Hepatitis, Liver Cirrhosis And Hepatomas, Its Mechanisms And Significance

一, The Mechanism of the Morphogenesis of the Intrahepatic Bile Ducts in Rat Embryo, a cytokeratin-and Laminin-Immunohistochemical Study(—). The Histologic Recognition Spectnims of 4 MAbs and 1 self-made PAb against cytokeratins.In this research, we prepared a rabbit polyclonal anti-epidermal keratin (EK) antibody (RAK1) and worked out the different histologic recognition spectnims of RAK1 and 4 MAbs against cytokeratins (CKs) (privided by Dr. Cheng Ming).Human and bovine EK was extracted with the urea-denaturation procedures and further purified by 2-3 assembly-deassembly processes. The amino acid composition was analysed with amino acid analyser (Beckman 121MB) . The discontinuous SDS-PAGE showed that both of the EK preparations were adequately purified and consisted of 7 CK polypeptides with the molecular weight of 68-, 65-, 63-, 57-, 55-, 50- and 46- KD, seperately. Immunized Rabbits with EK and got the antisera. One of the antisera (RAK1) to bovine EK was shown to reach 1:16 of double diffusion dilution in Lee's modified Ouchterlony's agarose gel. The staining dilutions of RAK1 were determined to be 1 : 200 or more for indirect immunofluorescence (IIF) staining and 1 : 2000 or more for double peroxidase-anti-peroxidase ( PAP) staining in the frozen sections or in the paraffin sections of alcohol-fixed tissues.With IIF, double PAP and ABC staing method, we showed that the self-made PAb to EK, RAK1, and the EK MAb, K174, were almost identical in the histologic recognition spectrum. They could label many kinds of the simple and the complex epithelia, but not endothelium. In the large digestive glands, RAK1 and K174 could show definitely positive reaction in ducts and myoepithelial cells, but not in the acinar epithelia. The CK MAbs, HK2 and HK5, could label almost all kinds of simple epithelia except endothelium; But HK2 could react with stratified squamous epithelium and Hk5 could not. k27, the MAb to Ek. could only label the cornified or conifying epithelial cells in the suprabasal layers of squamous epithelium. Therefore, the 5 antibodies to CKs ( RAK1, K174, HK2, HK5 and K27) could be used not only to differentiate epithelial from non-epithelial tissues, but also to subclass epithelia.