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. 1981 Nov;46(2):435–442.

Characterization and demonstration of human liver-specific protein (LSP) and apo-LSP.

N Lebwohl, M A Gerber
PMCID: PMC1536408  PMID: 6802540

Abstract

Liver-specific protein (LSP) prepared by standard methods from five normal human livers showed significant variations in terms of quantitative yield, lipid/protein ratio and migration characteristics of different components on SDS-polyacrylamide gel electrophoresis. This heterogeneity is probably related to varying amounts of different molecular species in the LSP preparations. Delipidation and re-chromatography of the LSP preparation appeared to result in relative enrichment of apo-LSP which showed immunological identity with LSP. Rabbit antiserum to LSP gave a precipitin line of identity with standard antisera to human LSP (anti-LSP) from two other laboratories. After extensive absorption, anti-LSP showed selective reactivity with a surface membrane antigen on a human hepatocellular carcinoma cell line (PLC/PRF/5) that exhibits functional and morphological characteristics of differentiated hepatocytes. The antiserum did not react with cell lines derived from other organs as determined by the indirect fluorescent antibody technique. The surface staining of viable PLC/PRF/5 cells was eliminated by absorption with LSP and apo-LSP, but not with the equivalent kidney fractions. These findings support the concept of a liver-specific antigen and suggest that the PLC/PRF/5 cell line may serve as a source of homogeneous LSP.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alexander J. J., Bey E. M., Geddes E. W., Lecatsas G. Establishment of a continuously growing cell line from primary carcinoma of the liver. S Afr Med J. 1976 Dec 18;50(54):2124–2128. [PubMed] [Google Scholar]
  2. Behrens U. J., Paronetto F. Studies on "liver-specific" antigens. I. Evaluation of the liver specificity of "LSP" and "LP-2". Gastroenterology. 1979 Nov;77(5):1045–1052. [PubMed] [Google Scholar]
  3. Chisari F. V., Bieber M. S., Josepho C. A., Xavier C., Anderson D. S. Functional properties of lymphocyte subpopulations in hepatitis B virus infection. II. Cytotoxic effector cell killing of targets that naturally express hepatitis B surface antigen and liver-specific lipoprotein. J Immunol. 1981 Jan;126(1):45–49. [PubMed] [Google Scholar]
  4. Chisari F. V. Liver-specific protein in perspective. Gastroenterology. 1980 Jan;78(1):168–170. [PubMed] [Google Scholar]
  5. Frings C. S., Dunn R. T. A colorimetric method for determination of total serum lipids based on the sulfo-phospho-vanillin reaction. Am J Clin Pathol. 1970 Jan;53(1):89–91. doi: 10.1093/ajcp/53.1.89. [DOI] [PubMed] [Google Scholar]
  6. Gerber M. A., Garfinkel E., Hirschman S. Z., Thung S. N., Panagiotatos T. Immune and enzyme histochemical studies of a human hepatocellular carcinoma cell line producing hepatitis B surface antigen. J Immunol. 1981 Mar;126(3):1085–1089. [PubMed] [Google Scholar]
  7. Helenius A., Simons K. Removal of lipids from human plasma low-density lipoprotein by detergents. Biochemistry. 1971 Jun 22;10(13):2542–2547. doi: 10.1021/bi00789a019. [DOI] [PubMed] [Google Scholar]
  8. Hirschman S. Z., Gerber M., Garfinkel E. Purification of naked intranuclear particles from human liver infected by hepatitis B virus. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3345–3349. doi: 10.1073/pnas.71.9.3345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jensen D. M., McFarlane I. G., Portmann B. S., Eddleston A. L., Williams R. Detection of antibodies directed against a liver-specific membrane lipoprotein in patients with acute and chronic active hepatitis. N Engl J Med. 1978 Jul 6;299(1):1–7. doi: 10.1056/NEJM197807062990101. [DOI] [PubMed] [Google Scholar]
  10. Kakumu S., Arakawa Y., Goji H., Kashio T., Yata K. Occurrence and significance of antibody to liver-specific membrane lipoprotein by double-antibody immunoprecipitation method in sera of patients with acute and chronic liver diseases. Gastroenterology. 1979 Apr;76(4):665–672. [PubMed] [Google Scholar]
  11. Knowles B. B., Howe C. C., Aden D. P. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science. 1980 Jul 25;209(4455):497–499. doi: 10.1126/science.6248960. [DOI] [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Manns M., Meyer zum Büschenfelde K. H., Hütteroth T. H., Hess G. Detection and characterization of liver membrane autoantibodies in chronic active hepatitis by a solid-phase radioimmunoassay. Clin Exp Immunol. 1980 Nov;42(2):263–272. [PMC free article] [PubMed] [Google Scholar]
  14. McFarlane I. G., Wojcicka B. M., Williams R. Antigens of the human liver. Clin Exp Immunol. 1980 Apr;40(1):1–7. [PMC free article] [PubMed] [Google Scholar]
  15. McFarlane I. G., Wojcicka B. M., Zucker G. M., Eddleston A. L., Williams R. Purification and characterization of human liver-specific membrane lipoprotein (LSP). Clin Exp Immunol. 1977 Mar;27(3):381–390. [PMC free article] [PubMed] [Google Scholar]
  16. Mutchnick M. G., Kawanishi H., Hopf U. Absence of a liver-specific membrane protein in a strain of chang cells. Clin Immunol Immunopathol. 1978 Aug;10(4):398–402. doi: 10.1016/0090-1229(78)90151-4. [DOI] [PubMed] [Google Scholar]
  17. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  18. Ortona L., Laghi V., Cauda R., Nervo P. Immune response to rabbit liver-specific lipoprotein in acute viral hepatitis. Clin Exp Immunol. 1980 Dec;42(3):436–440. [PMC free article] [PubMed] [Google Scholar]
  19. Tsantoulas D., Perperas A., Portmann B., Eddleston A. L., Williams R. Antibodies to a human liver membrane lipoprotein (LSP) in primary biliary cirrhosis. Gut. 1980 Jul;21(7):557–560. doi: 10.1136/gut.21.7.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wilchek M., Bocchini V., Becker M., Givol D. A general method for the specific isolation of peptides containing modified residues, using insoluble antibody columns. Biochemistry. 1971 Jul 20;10(15):2828–2834. doi: 10.1021/bi00791a004. [DOI] [PubMed] [Google Scholar]

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