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. 1992 Feb;87(2):251–255. doi: 10.1111/j.1365-2249.1992.tb02983.x

Study of CYP2D6 gene in children with autoimmune hepatitis and P450 IID6 autoantibodies.

A M Yamamoto 1, C Mura 1, M G Morales 1, O Bernard 1, R Krishnamoorthy 1, F Alvarez 1
PMCID: PMC1554279  PMID: 1346512

Abstract

Cytochrome P450 IID6 is an autoantigen recognized by the sera of children affected with a subtype of autoimmune hepatitis. It was hypothesized that a mutation in the CYP2D6 gene could explain the autoimmune response in these patients. To examine this question, genomic DNA from peripheral lymphocytes (n = 9) and liver (n = 1) of 10 patients with anti-LKM-1 antibody was analysed by Southern blot for genetic association studies between a particular CYP2D6 haplotype and autoimmune hepatitis. In addition, a region of CYP2D6, from the same genomic DNA, was amplified by polymerase chain reaction (PCR) and digested by BstNI, in a search for the most prevalent 29B mutation, described in subjects who do not express the P450 IID6. Total RNA and proteins, prepared from the liver of an anti-LKM-1+ patient, were analysed by Northern and Western (immunoblot) blots respectively. Our results do not reveal any major structural change in the DNA of this patient at the CYP2D6 locus that could explain their autoimmune response. Corroborating this observation, no changes were noted either in P450 IID6 mRNA size or in the corresponding protein. However, these data do not exclude the possibility of subtle changes in the protein due to point mutations in critical regions that might trigger an autoimmune response.

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Selected References

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

  1. Alvarez F., Bernard O., Homberg J. C., Kreibich G. Anti-liver-kidney microsome antibody recognizes a 50,000 molecular weight protein of the endoplasmic reticulum. J Exp Med. 1985 May 1;161(5):1231–1236. doi: 10.1084/jem.161.5.1231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Codońer-Franch P., Paradis K., Gueguen M., Bernard O., Costesec A. A., Alvarez F. A new antigen recognized by anti-liver-kidney-microsome antibody (LKMA). Clin Exp Immunol. 1989 Mar;75(3):354–358. [PMC free article] [PubMed] [Google Scholar]
  3. Daly A. K., Armstrong M., Idle J. R. Molecular genotyping to predict debrisoquine hydroxylation phenotype. Lancet. 1990 Oct 6;336(8719):889–890. doi: 10.1016/0140-6736(90)92413-c. [DOI] [PubMed] [Google Scholar]
  4. Gonzalez F. J., Skoda R. C., Kimura S., Umeno M., Zanger U. M., Nebert D. W., Gelboin H. V., Hardwick J. P., Meyer U. A. Characterization of the common genetic defect in humans deficient in debrisoquine metabolism. Nature. 1988 Feb 4;331(6155):442–446. doi: 10.1038/331442a0. [DOI] [PubMed] [Google Scholar]
  5. Gough A. C., Miles J. S., Spurr N. K., Moss J. E., Gaedigk A., Eichelbaum M., Wolf C. R. Identification of the primary gene defect at the cytochrome P450 CYP2D locus. Nature. 1990 Oct 25;347(6295):773–776. doi: 10.1038/347773a0. [DOI] [PubMed] [Google Scholar]
  6. Gueguen M., Meunier-Rotival M., Bernard O., Alvarez F. Anti-liver kidney microsome antibody recognizes a cytochrome P450 from the IID subfamily. J Exp Med. 1988 Aug 1;168(2):801–806. doi: 10.1084/jem.168.2.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gueguen M., Yamamoto A. M., Bernard O., Alvarez F. Anti-liver-kidney microsome antibody type 1 recognizes human cytochrome P450 db1. Biochem Biophys Res Commun. 1989 Mar 15;159(2):542–547. doi: 10.1016/0006-291x(89)90027-2. [DOI] [PubMed] [Google Scholar]
  8. Heim M., Meyer U. A. Genotyping of poor metabolisers of debrisoquine by allele-specific PCR amplification. Lancet. 1990 Sep 1;336(8714):529–532. doi: 10.1016/0140-6736(90)92086-w. [DOI] [PubMed] [Google Scholar]
  9. Jacqz-Aigrain E., Laurent J., Alvarez F. Dextromethorphan phenotypes in paediatric patients with autoimmune hepatitis. Br J Clin Pharmacol. 1990 Jul;30(1):153–154. doi: 10.1111/j.1365-2125.1990.tb03758.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kagimoto M., Heim M., Kagimoto K., Zeugin T., Meyer U. A. Multiple mutations of the human cytochrome P450IID6 gene (CYP2D6) in poor metabolizers of debrisoquine. Study of the functional significance of individual mutations by expression of chimeric genes. J Biol Chem. 1990 Oct 5;265(28):17209–17214. [PubMed] [Google Scholar]
  11. Kimura S., Umeno M., Skoda R. C., Meyer U. A., Gonzalez F. J. The human debrisoquine 4-hydroxylase (CYP2D) locus: sequence and identification of the polymorphic CYP2D6 gene, a related gene, and a pseudogene. Am J Hum Genet. 1989 Dec;45(6):889–904. [PMC free article] [PubMed] [Google Scholar]
  12. Manns M. P., Johnson E. F., Griffin K. J., Tan E. M., Sullivan K. F. Major antigen of liver kidney microsomal autoantibodies in idiopathic autoimmune hepatitis is cytochrome P450db1. J Clin Invest. 1989 Mar;83(3):1066–1072. doi: 10.1172/JCI113949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Manns M., Zanger U., Gerken G., Sullivan K. F., Meyer zum Büschenfelde K. H., Meyer U. A., Eichelbaum M. Patients with type II autoimmune hepatitis express functionally intact cytochrome P-450 db1 that is inhibited by LKM-1 autoantibodies in vitro but not in vivo. Hepatology. 1990 Jul;12(1):127–132. doi: 10.1002/hep.1840120120. [DOI] [PubMed] [Google Scholar]
  14. Martini E., Abuaf N., Cavalli F., Durand V., Johanet C., Homberg J. C. Antibody to liver cytosol (anti-LC1) in patients with autoimmune chronic active hepatitis type 2. Hepatology. 1988 Nov-Dec;8(6):1662–1666. doi: 10.1002/hep.1840080632. [DOI] [PubMed] [Google Scholar]
  15. McBride O. W., Merry D., Givol D. The gene for human p53 cellular tumor antigen is located on chromosome 17 short arm (17p13). Proc Natl Acad Sci U S A. 1986 Jan;83(1):130–134. doi: 10.1073/pnas.83.1.130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mogensen C. E. The glomerular permeability determined by dextran clearance using Sephadex gel filtration. Scand J Clin Lab Invest. 1968;21(1):77–82. doi: 10.3109/00365516809076979. [DOI] [PubMed] [Google Scholar]
  17. Pelloquin F., Lamelin J. P., Lenoir G. M. Human B lymphocytes immortalization by Epstein-Barr virus in the presence of cyclosporin A. In Vitro Cell Dev Biol. 1986 Dec;22(12):689–694. doi: 10.1007/BF02621085. [DOI] [PubMed] [Google Scholar]
  18. Puissant C., Houdebine L. M. An improvement of the single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Biotechniques. 1990 Feb;8(2):148–149. [PubMed] [Google Scholar]
  19. Skoda R. C., Gonzalez F. J., Demierre A., Meyer U. A. Two mutant alleles of the human cytochrome P-450db1 gene (P450C2D1) associated with genetically deficient metabolism of debrisoquine and other drugs. Proc Natl Acad Sci U S A. 1988 Jul;85(14):5240–5243. doi: 10.1073/pnas.85.14.5240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zanger U. M., Hauri H. P., Loeper J., Homberg J. C., Meyer U. A. Antibodies against human cytochrome P-450db1 in autoimmune hepatitis type II. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8256–8260. doi: 10.1073/pnas.85.21.8256. [DOI] [PMC free article] [PubMed] [Google Scholar]

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