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J Clin Invest. 1986 February; 77(2): 528–537.
PMCID: PMC423375

Evaluation of "at risk" alpha 1-antitrypsin genotype SZ with synthetic oligonucleotide gene probes.

Abstract

Alpha 1-antitrypsin (alpha 1AT), a 52,000-mol-wt serum glycoprotein produced by hepatocytes and mononuclear phagocytes, functions as the major inhibitor of neutrophil elastase. The alpha 1AT haplotype S is associated with childhood liver disease and/or adult emphysema when inherited with the Z haplotype to give the phenotype SZ. To accurately identify the SZ phenotype at the level of genomic DNA, four 32P-labeled 19-mer synthetic oligonucleotide probes were prepared; two to identify the M and S difference in exon III, and two to identify the M and Z difference in exon V. These probes were hybridized with various cloned DNAs and genomic DNAs cut with the restriction endonucleases BgII and EcoRI; the genomic DNAs represented all six possible phenotype combinations of the M, S, and Z haplotypes (MM, MS, MZ, SS, ZZ, and SZ). Using the four probes to evaluate 42 samples of genomic DNA, the "at risk" SZ and ZZ phenotypes were correctly identified in all cases, as were the "not at risk" phenotypes SS, MS, MM, and MZ, demonstrating that both exon III and exon V directed probes are necessary to properly identify all of the major "at risk" alpha 1AT genes. However, when used to evaluate a very rare family carrying a null allele, these four oligonucleotide probes misidentified the "at risk" null-null and S null phenotypes as "not at risk" MM and SM combinations. These observations indicate that oligonucleotide gene probes yielded reliable and accurate assessment of "at risk" alpha 1AT genotypes in almost all situations, but in the context of prenatal diagnosis and genetic counseling this approach must be used with caution and in combination with family studies so as not to misidentify rare genotypes that may be associated with a risk for disease.

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  • Travis J, Salvesen GS. Human plasma proteinase inhibitors. Annu Rev Biochem. 1983;52:655–709. [PubMed]
  • Gadek JE, Hunninghake GW, Fells GA, Zimmerman RL, Keogh BA, Crystal RG. Evaluation of the protease-antiprotease theory of human destructive lung disease. Bull Eur Physiopathol Respir. 1980;16 (Suppl):27–40. [PubMed]
  • Carrell RW, Jeppsson JO, Laurell CB, Brennan SO, Owen MC, Vaughan L, Boswell DR. Structure and variation of human alpha 1-antitrypsin. Nature. 1982 Jul 22;298(5872):329–334. [PubMed]
  • Fagerhol MK, Cox DW. The Pi polymorphism: genetic, biochemical, and clinical aspects of human alpha 1-antitrypsin. Adv Hum Genet. 1981;11:1–372. [PubMed]
  • Pierce JA, Eradio B, Dew TA. Antitrypsin phenotypes in St. Louis. JAMA. 1975 Feb 10;231(6):609–612. [PubMed]
  • Dykes DD, Miller SA, Polesky HF. Distribution of alpha 1-antitrypsin variants in a US white population. Hum Hered. 1984;34(5):308–310. [PubMed]
  • Evans HE, Bognacki NS, Perrott LM, Glass L. Prevalence of of alpha 1-antitrypsin Pi types among newborn infants of different ethnic backgrounds. J Pediatr. 1977 Apr;90(4):621–624. [PubMed]
  • Kueppers F, Christopherson MJ. Alpha1-antitrypsin: further genetic heterogeneity revealed by isoelectric focusing. Am J Hum Genet. 1978 Jul;30(4):359–365. [PubMed]
  • Eriksson S. Studies in alpha 1-antitrypsin deficiency. Acta Med Scand Suppl. 1965;432:1–85. [PubMed]
  • Morse JO. alpha1-antitrypsin deficiency (first of two parts). N Engl J Med. 1978 Nov 9;299(19):1045–1048. [PubMed]
  • Larsson C, Dirksen H, Sundström G, Eriksson S. Lung function studies in asymptomatic individuals with moderately (Pi SZ) and severely (Pi Z) reduced levels of alpha1-antitrypsin. Scand J Respir Dis. 1976;57(6):267–280. [PubMed]
  • Craig JR, Dunn AE, Peters RL. Cirrhosis associated with partial deficiency of alpha-1-antitrypsin: a clinical and autopsy study. Hum Pathol. 1975 Jan;6(1):113–120. [PubMed]
  • Sveger T. Liver disease in alpha1-antitrypsin deficiency detected by screening of 200,000 infants. N Engl J Med. 1976 Jun 10;294(24):1316–1321. [PubMed]
  • Yoshida A, Lieberman J, Gaidulis L, Ewing C. Molecular abnormality of human alpha1-antitrypsin variant (Pi-ZZ) associated with plasma activity deficiency. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1324–1328. [PubMed]
  • Yoshida A, Ewing C, Wessels M, Lieberman J, Gaidulis L. Molecular abnormality of PI S variant of human alpha1-antitrypsin. Am J Hum Genet. 1977 May;29(3):233–239. [PubMed]
  • Long GL, Chandra T, Woo SL, Davie EW, Kurachi K. Complete sequence of the cDNA for human alpha 1-antitrypsin and the gene for the S variant. Biochemistry. 1984 Oct 9;23(21):4828–4837. [PubMed]
  • Cox DW, Johnson AM, Fagerhol MK. Report of Nomenclature Meeting for alpha 1-antitrypsin, INSERM, Rouen/Bois-Guillaume-1978. Hum Genet. 1980;53(3):429–433. [PubMed]
  • Kidd VJ, Wallace RB, Itakura K, Woo SL. alpha 1-antitrypsin deficiency detection by direct analysis of the mutation in the gene. Nature. 1983 Jul 21;304(5923):230–234. [PubMed]
  • Kidd VJ, Golbus MS, Wallace RB, Itakura K, Woo SL. Prenatal diagnosis of alpha 1-antitrypsin deficiency by direct analysis of the mutation site in the gene. N Engl J Med. 1984 Mar 8;310(10):639–642. [PubMed]
  • Frants RR, Noordhoek GT, Eriksson AW. Separator isoelectric focusing for identification of alpha-1-antitrypsin (Pi M) subtypes. Scand J Clin Lab Invest. 1978 Sep;38(5):457–462. [PubMed]
  • Constans J, Viau M, Gouaillard C. Pi M4: an additional Pi M subtype. Hum Genet. 1980;55(1):119–121. [PubMed]
  • Jeppsson JO, Laurell CB, Fagerhol M. Properties of isolated human alpha1-antitrypsins of Pi types M, S and Z. Eur J Biochem. 1978 Feb 1;83(1):143–153. [PubMed]
  • Jeffreys AJ, Flavell RA. A physical map of the DNA regions flanking the rabbit beta-globin gene. Cell. 1977 Oct;12(2):429–439. [PubMed]
  • Lawn RM, Fritsch EF, Parker RC, Blake G, Maniatis T. The isolation and characterization of linked delta- and beta-globin genes from a cloned library of human DNA. Cell. 1978 Dec;15(4):1157–1174. [PubMed]
  • Courtney M, Buchwalder A, Tessier LH, Jaye M, Benavente A, Balland A, Kohli V, Lathe R, Tolstoshev P, Lecocq JP. High-level production of biologically active human alpha 1-antitrypsin in Escherichia coli. Proc Natl Acad Sci U S A. 1984 Feb;81(3):669–673. [PubMed]
  • Studencki AB, Wallace RB. Allele-specific hybridization using oligonucleotide probes of very high specific activity: discrimination of the human beta A- and beta S-globin genes. DNA. 1984;3(1):7–15. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Lai EC, Kao FT, Law ML, Woo SL. Assignment of the alpha 1-antitrypsin gene and a sequence-related gene to human chromosome 14 by molecular hybridization. Am J Hum Genet. 1983 May;35(3):385–392. [PubMed]
  • Geiger T, Northemann W, Schmelzer E, Gross V, Gauthier F, Heinrich PC. Synthesis of alpha 1-antitrypsin in rat-liver hepatocytes and in a cell-free system. Eur J Biochem. 1982 Aug;126(1):189–195. [PubMed]
  • Ciliberto G, Dente L, Cortese R. Cell-specific expression of a transfected human alpha 1-antitrypsin gene. Cell. 1985 Jun;41(2):531–540. [PubMed]
  • Perlmutter DH, Cole FS, Kilbridge P, Rossing TH, Colten HR. Expression of the alpha 1-proteinase inhibitor gene in human monocytes and macrophages. Proc Natl Acad Sci U S A. 1985 Feb;82(3):795–799. [PubMed]
  • Goedde HW, Hirth L, Benkmann HG, Pellicer A, Pellicer T, Stahn M, Singh S. Population genetic studies of serum protein polymorphisms in four Spanish populations. II. Hum Hered. 1973;23(2):135–146. [PubMed]
  • Martin JP, Sesboue R, Charlionet R, Ropartz C, Pereira MT. Genetic variants of serum alpha1-antitrypsin (Pi types) in Portuguese. Hum Hered. 1976;26(4):310–314. [PubMed]
  • Fagerhol MK. Serum Pi types in Norwegians. Acta Pathol Microbiol Scand. 1967;70(3):421–428. [PubMed]
  • Campra JL, Craig JR, Peters RL, Reynolds TB. Cirrhosis associated with partial deficiency of alpha-1 antitrypsin in an adult. Ann Intern Med. 1973 Feb;78(2):233–238. [PubMed]
  • Palmer PE, Gherardi GJ, Baldwin JM, Wolfe HJ. Adult liver disease in SZ phenotype alpha-1-antitrypsin deficiency. Ann Intern Med. 1978 Jan;88(1):59–60. [PubMed]
  • Chan CH, Steer CJ, Vergalla J, Jones EA. Alpha1-antitrypsin deficiency with cirrhosis associated with the protease inhibitor phenotype SZ. Am J Med. 1978 Dec;65(6):978–986. [PubMed]
  • Hutchison DC, Tobin MJ, Cook PJ. Alpha 1 antitrypsin deficiency: clinical and physiological features in heterozygotes of Pi type SZ. A survey by the British Thoracic Association. Br J Dis Chest. 1983 Jan;77(1):28–34. [PubMed]
  • Gishen P, Saunders AJ, Tobin MJ, Hutchison DC. Alpha 1-antitrypsin deficiency: the radiological features of pulmonary emphysema in subjects of Pi type Z and Pi type SZ: a survey by the British Thoracic Association. Clin Radiol. 1982 Jul;33(4):371–377. [PubMed]
  • Morse JO, Lebowitz MD, Knudson RJ, Burrows B. Relation of protease inhibitor phenotypes to obstructive lung diseases in a community. N Engl J Med. 1977 May 26;296(21):1190–1194. [PubMed]
  • Bruce RM, Cohen BH, Diamond EL, Fallat RJ, Knudson RJ, Lebowitz MD, Mittman C, Patterson CD, Tockman MS. Collaborative study to assess risk of lung disease in Pi MZ phenotype subjects. Am Rev Respir Dis. 1984 Sep;130(3):386–390. [PubMed]
  • Fisher RL, Taylor L, Sherlock S. alpha-1-antitrypsin deficiency in liver disease: the extent of the problem. Gastroenterology. 1976 Oct;71(4):646–651. [PubMed]
  • Hutchison DC. A survey of alpha 1-antitrypsin deficiency by the British Thoracic Association. Bull Eur Physiopathol Respir. 1980;16 (Suppl):315–319. [PubMed]
  • Gadek JE, Klein HG, Holland PV, Crystal RG. Replacement therapy of alpha 1-antitrypsin deficiency. Reversal of protease-antiprotease imbalance within the alveolar structures of PiZ subjects. J Clin Invest. 1981 Nov;68(5):1158–1165. [PMC free article] [PubMed]
  • Loebermann H, Tokuoka R, Deisenhofer J, Huber R. Human alpha 1-proteinase inhibitor. Crystal structure analysis of two crystal modifications, molecular model and preliminary analysis of the implications for function. J Mol Biol. 1984 Aug 15;177(3):531–557. [PubMed]
  • Wallace RB, Schold M, Johnson MJ, Dembek P, Itakura K. Oligonucleotide directed mutagenesis of the human beta-globin gene: a general method for producing specific point mutations in cloned DNA. Nucleic Acids Res. 1981 Aug 11;9(15):3647–3656. [PMC free article] [PubMed]
  • Itakura K, Rossi JJ, Wallace RB. Synthesis and use of synthetic oligonucleotides. Annu Rev Biochem. 1984;53:323–356. [PubMed]
  • Nei M, Li WH. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5269–5273. [PubMed]
  • Conner BJ, Reyes AA, Morin C, Itakura K, Teplitz RL, Wallace RB. Detection of sickle cell beta S-globin allele by hybridization with synthetic oligonucleotides. Proc Natl Acad Sci U S A. 1983 Jan;80(1):278–282. [PubMed]
  • Pirastu M, Kan YW, Cao A, Conner BJ, Teplitz RL, Wallace RB. Prenatal diagnosis of beta-thalassemia. Detection of a single nucleotide mutation in DNA. N Engl J Med. 1983 Aug 4;309(5):284–287. [PubMed]
  • Orkin SH, Markham AF, Kazazian HH., Jr Direct detection of the common Mediterranean beta-thalassemia gene with synthetic DNA probes. An alternative approach for prenatal diagnosis. J Clin Invest. 1983 Mar;71(3):775–779. [PMC free article] [PubMed]
  • Kazazian HH, Jr, Orkin SH, Markham AF, Chapman CR, Youssoufian H, Waber PG. Quantification of the close association between DNA haplotypes and specific beta-thalassaemia mutations in Mediterraneans. Nature. 1984 Jul 12;310(5973):152–154. [PubMed]
  • Rosatelli C, Falchi AM, Tuveri T, Scalas MT, Di Tucci A, Monni G, Cao A. Prenatal diagnosis of beta-thalassaemia with the synthetic-oligomer technique. Lancet. 1985 Feb 2;1(8423):241–243. [PubMed]
  • Studencki AB, Conner BJ, Impraim CC, Teplitz RL, Wallace RB. Discrimination among the human beta A, beta S, and beta C-globin genes using allele-specific oligonucleotide hybridization probes. Am J Hum Genet. 1985 Jan;37(1):42–51. [PubMed]
  • Bos JL, Toksoz D, Marshall CJ, Verlaan-de Vries M, Veeneman GH, van der Eb AJ, van Boom JH, Janssen JW, Steenvoorden AC. Amino-acid substitutions at codon 13 of the N-ras oncogene in human acute myeloid leukaemia. Nature. 315(6022):726–730. [PubMed]
  • Owen MC, Brennan SO, Lewis JH, Carrell RW. Mutation of antitrypsin to antithrombin. alpha 1-antitrypsin Pittsburgh (358 Met leads to Arg), a fatal bleeding disorder. N Engl J Med. 1983 Sep 22;309(12):694–698. [PubMed]
  • Fagerhol MK, Hauge HE. Serum Pi types in patients with pulmonary diseases. Acta Allergol. 1969 May;24(2):107–114. [PubMed]
  • Lieberman J, Gaidulis L, Klotz SD. A new deficient variant of alpha1-antitrypsin (MDUARTE). Inability to detect the heterozygous state by antitrypsin phenotyping. Am Rev Respir Dis. 1976 Jan;113(1):31–36. [PubMed]
  • Sproule BJ, Cox DW, Hsu K, Salkie ML, Herbert FA. Pulmonary function associated with the Mmalton deficient variant of alpha 1-antitrypsin. Am Rev Respir Dis. 1983 Feb;127(2):237–240. [PubMed]

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