Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1995 Feb;95(2):493–502. doi: 10.1172/JCI117690

Gene transfer into the airway epithelium of animals by targeting the polymeric immunoglobulin receptor.

T Ferkol 1, J C Perales 1, E Eckman 1, C S Kaetzel 1, R W Hanson 1, P B Davis 1
PMCID: PMC295497  PMID: 7860731

Abstract

Genes of interest can be targeted specifically to respiratory epithelial cells in intact animals with high efficiency by exploiting the receptor-mediated endocytosis of the polymeric immunoglobulin receptor. A DNA carrier, consisting of the Fab portion of polyclonal antibodies raised against rat secretory component covalently linked to poly-L-lysine, was used to introduce plasmids containing different reporter genes into airway epithelial cells in vivo. We observed significant levels of luciferase enzyme activity in protein extracts from the liver and lung, achieving maximum values of 13,795 +/- 4,431 and 346,954 +/- 199,120 integrated light units (ILU) per milligram of protein extract, respectively. No luciferase activity was detected in spleen or heart, which do not express the receptor. Transfections using complexes consisting of an irrelevant plasmid (pCMV lacZ) bound to the bona fide carrier or the expression plasmid (pGEMluc) bound to a carrier based on an irrelevant Fab fragment resulted in background levels of luciferase activity in all tissues examined. Thus, only tissues that contain cells bearing the polymeric immunoglobulin receptor are transfected, and transfection cannot be attributed to the nonspecific uptake of an irrelevant carrier-DNA complex. Specific mRNA from the luciferase gene was also detected in the lungs of transfected animals. To determine which cells in the lungs are transfected by this method, DNA complexes were prepared containing expression plasmids with genes encoding the bacterial beta-galactosidase or the human interleukin 2 receptor. Expression of these genes was localized to the surface epithelium of the airways and the submucosal glands, and not the bronchioles and alveoli. Receptor-mediated endocytosis can be used to introduce functional genes into the respiratory epithelium of rats, and may be a useful technique for gene therapy targeting the lung.

Full text

PDF
497

Images in this article

496Image
on p.496
497Image
on p.497
498Image
on p.498
499Image
on p.499
500Image
on p.500

Selected References

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

  1. Alton E. W., Middleton P. G., Caplen N. J., Smith S. N., Steel D. M., Munkonge F. M., Jeffery P. K., Geddes D. M., Hart S. L., Williamson R. Non-invasive liposome-mediated gene delivery can correct the ion transport defect in cystic fibrosis mutant mice. Nat Genet. 1993 Oct;5(2):135–142. doi: 10.1038/ng1093-135. [DOI] [PubMed] [Google Scholar]
  2. Bijsterbosch M. K., Van Berkel T. J. Uptake of lactosylated low-density lipoprotein by galactose-specific receptors in rat liver. Biochem J. 1990 Aug 15;270(1):233–239. doi: 10.1042/bj2700233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bijsterbosch M. K., Ziere G. J., Van Berkel T. J. Lactosylated low density lipoprotein: a potential carrier for the site-specific delivery of drugs to Kupffer cells. Mol Pharmacol. 1989 Sep;36(3):484–489. [PubMed] [Google Scholar]
  4. Brasier A. R., Tate J. E., Habener J. F. Optimized use of the firefly luciferase assay as a reporter gene in mammalian cell lines. Biotechniques. 1989 Nov-Dec;7(10):1116–1122. [PubMed] [Google Scholar]
  5. Brinster R. L., Chen H. Y., Messing A., van Dyke T., Levine A. J., Palmiter R. D. Transgenic mice harboring SV40 T-antigen genes develop characteristic brain tumors. Cell. 1984 Jun;37(2):367–379. doi: 10.1016/0092-8674(84)90367-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bukrinsky M. I., Haggerty S., Dempsey M. P., Sharova N., Adzhubel A., Spitz L., Lewis P., Goldfarb D., Emerman M., Stevenson M. A nuclear localization signal within HIV-1 matrix protein that governs infection of non-dividing cells. Nature. 1993 Oct 14;365(6447):666–669. doi: 10.1038/365666a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chintalacharuvu K. R., Tavill A. S., Louis L. N., Vaerman J. P., Lamm M. E., Kaetzel C. S. Disulfide bond formation between dimeric immunoglobulin A and the polymeric immunoglobulin receptor during hepatic transcytosis. Hepatology. 1994 Jan;19(1):162–173. [PubMed] [Google Scholar]
  8. Chowdhury N. R., Wu C. H., Wu G. Y., Yerneni P. C., Bommineni V. R., Chowdhury J. R. Fate of DNA targeted to the liver by asialoglycoprotein receptor-mediated endocytosis in vivo. Prolonged persistence in cytoplasmic vesicles after partial hepatectomy. J Biol Chem. 1993 May 25;268(15):11265–11271. [PubMed] [Google Scholar]
  9. Cotten M., Längle-Rouault F., Kirlappos H., Wagner E., Mechtler K., Zenke M., Beug H., Birnstiel M. L. Transferrin-polycation-mediated introduction of DNA into human leukemic cells: stimulation by agents that affect the survival of transfected DNA or modulate transferrin receptor levels. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4033–4037. doi: 10.1073/pnas.87.11.4033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cotten M., Wagner E., Zatloukal K., Phillips S., Curiel D. T., Birnstiel M. L. High-efficiency receptor-mediated delivery of small and large (48 kilobase gene constructs using the endosome-disruption activity of defective or chemically inactivated adenovirus particles. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6094–6098. doi: 10.1073/pnas.89.13.6094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Curiel D. T., Agarwal S., Wagner E., Cotten M. Adenovirus enhancement of transferrin-polylysine-mediated gene delivery. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8850–8854. doi: 10.1073/pnas.88.19.8850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Engelhardt J. F., Yang Y., Stratford-Perricaudet L. D., Allen E. D., Kozarsky K., Perricaudet M., Yankaskas J. R., Wilson J. M. Direct gene transfer of human CFTR into human bronchial epithelia of xenografts with E1-deleted adenoviruses. Nat Genet. 1993 May;4(1):27–34. doi: 10.1038/ng0593-27. [DOI] [PubMed] [Google Scholar]
  13. Engelhardt J. F., Yankaskas J. R., Ernst S. A., Yang Y., Marino C. R., Boucher R. C., Cohn J. A., Wilson J. M. Submucosal glands are the predominant site of CFTR expression in the human bronchus. Nat Genet. 1992 Nov;2(3):240–248. doi: 10.1038/ng1192-240. [DOI] [PubMed] [Google Scholar]
  14. Eppler K., Wyckoff E., Goates J., Parr R., Casjens S. Nucleotide sequence of the bacteriophage P22 genes required for DNA packaging. Virology. 1991 Aug;183(2):519–538. doi: 10.1016/0042-6822(91)90981-g. [DOI] [PubMed] [Google Scholar]
  15. Ferkol T., Kaetzel C. S., Davis P. B. Gene transfer into respiratory epithelial cells by targeting the polymeric immunoglobulin receptor. J Clin Invest. 1993 Nov;92(5):2394–2400. doi: 10.1172/JCI116845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ferkol T., Lindberg G. L., Chen J., Perales J. C., Crawford D. R., Ratnoff O. D., Hanson R. W. Regulation of the phosphoenolpyruvate carboxykinase/human factor IX gene introduced into the livers of adult rats by receptor-mediated gene transfer. FASEB J. 1993 Aug;7(11):1081–1091. doi: 10.1096/fasebj.7.11.8370479. [DOI] [PubMed] [Google Scholar]
  17. Fiedler M. A., Kaetzel C. S., Davis P. B. Sustained production of secretory component by human tracheal epithelial cells in primary culture. Am J Physiol. 1991 Oct;261(4 Pt 1):L255–L261. doi: 10.1152/ajplung.1991.261.4.L255. [DOI] [PubMed] [Google Scholar]
  18. Flotte T. R., Afione S. A., Solow R., Drumm M. L., Markakis D., Guggino W. B., Zeitlin P. L., Carter B. J. Expression of the cystic fibrosis transmembrane conductance regulator from a novel adeno-associated virus promoter. J Biol Chem. 1993 Feb 15;268(5):3781–3790. [PubMed] [Google Scholar]
  19. Flotte T. R., Solow R., Owens R. A., Afione S., Zeitlin P. L., Carter B. J. Gene expression from adeno-associated virus vectors in airway epithelial cells. Am J Respir Cell Mol Biol. 1992 Sep;7(3):349–356. doi: 10.1165/ajrcmb/7.3.349. [DOI] [PubMed] [Google Scholar]
  20. Gao L., Wagner E., Cotten M., Agarwal S., Harris C., Rømer M., Miller L., Hu P. C., Curiel D. Direct in vivo gene transfer to airway epithelium employing adenovirus-polylysine-DNA complexes. Hum Gene Ther. 1993 Feb;4(1):17–24. doi: 10.1089/hum.1993.4.1-17. [DOI] [PubMed] [Google Scholar]
  21. Hazinski T. A., Ladd P. A., DeMatteo C. A. Localization and induced expression of fusion genes in the rat lung. Am J Respir Cell Mol Biol. 1991 Mar;4(3):206–209. doi: 10.1165/ajrcmb/4.3.206. [DOI] [PubMed] [Google Scholar]
  22. Huling S., Fournier G. R., Feren A., Chuntharapai A., Jones A. L. Ontogeny of the secretory immune system: maturation of a functional polymeric immunoglobulin receptor regulated by gene expression. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4260–4264. doi: 10.1073/pnas.89.10.4260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hyde S. C., Gill D. R., Higgins C. F., Trezise A. E., MacVinish L. J., Cuthbert A. W., Ratcliff R., Evans M. J., Colledge W. H. Correction of the ion transport defect in cystic fibrosis transgenic mice by gene therapy. Nature. 1993 Mar 18;362(6417):250–255. doi: 10.1038/362250a0. [DOI] [PubMed] [Google Scholar]
  24. Johnson L. G., Olsen J. C., Sarkadi B., Moore K. L., Swanstrom R., Boucher R. C. Efficiency of gene transfer for restoration of normal airway epithelial function in cystic fibrosis. Nat Genet. 1992 Sep;2(1):21–25. doi: 10.1038/ng0992-21. [DOI] [PubMed] [Google Scholar]
  25. Jung G., Köhnlein W., Lüders G. Biological activity of the antitumor protein neocarzinostatin coupled to a monoclonal antibody by N-succinimidyl 3-(2-pyridyldithio)-propionate. Biochem Biophys Res Commun. 1981 Jul 30;101(2):599–606. doi: 10.1016/0006-291x(81)91301-2. [DOI] [PubMed] [Google Scholar]
  26. Kaetzel C. S., Robinson J. K., Chintalacharuvu K. R., Vaerman J. P., Lamm M. E. The polymeric immunoglobulin receptor (secretory component) mediates transport of immune complexes across epithelial cells: a local defense function for IgA. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8796–8800. doi: 10.1073/pnas.88.19.8796. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kalderon D., Richardson W. D., Markham A. F., Smith A. E. Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature. 1984 Sep 6;311(5981):33–38. doi: 10.1038/311033a0. [DOI] [PubMed] [Google Scholar]
  28. Lin W. C., Culp L. A. Selectable plasmid vectors with alternative and ultrasensitive histochemical marker genes. Biotechniques. 1991 Sep;11(3):344-8, 350-1. [PubMed] [Google Scholar]
  29. Overbeek P. A., Lai S. P., Van Quill K. R., Westphal H. Tissue-specific expression in transgenic mice of a fused gene containing RSV terminal sequences. Science. 1986 Mar 28;231(4745):1574–1577. doi: 10.1126/science.3006249. [DOI] [PubMed] [Google Scholar]
  30. Perales J. C., Ferkol T., Beegen H., Ratnoff O. D., Hanson R. W. Gene transfer in vivo: sustained expression and regulation of genes introduced into the liver by receptor-targeted uptake. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):4086–4090. doi: 10.1073/pnas.91.9.4086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rosenfeld M. A., Siegfried W., Yoshimura K., Yoneyama K., Fukayama M., Stier L. E., Päkkö P. K., Gilardi P., Stratford-Perricaudet L. D., Perricaudet M. Adenovirus-mediated transfer of a recombinant alpha 1-antitrypsin gene to the lung epithelium in vivo. Science. 1991 Apr 19;252(5004):431–434. doi: 10.1126/science.2017680. [DOI] [PubMed] [Google Scholar]
  32. Rosenfeld M. A., Yoshimura K., Trapnell B. C., Yoneyama K., Rosenthal E. R., Dalemans W., Fukayama M., Bargon J., Stier L. E., Stratford-Perricaudet L. In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium. Cell. 1992 Jan 10;68(1):143–155. doi: 10.1016/0092-8674(92)90213-v. [DOI] [PubMed] [Google Scholar]
  33. Stribling R., Brunette E., Liggitt D., Gaensler K., Debs R. Aerosol gene delivery in vivo. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11277–11281. doi: 10.1073/pnas.89.23.11277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Stutts M. J., Gabriel S. E., Olsen J. C., Gatzy J. T., O'Connell T. L., Price E. M., Boucher R. C. Functional consequences of heterologous expression of the cystic fibrosis transmembrane conductance regulator in fibroblasts. J Biol Chem. 1993 Sep 25;268(27):20653–20658. [PubMed] [Google Scholar]
  35. Sullivan D. A., Wira C. R. Variations in free secretory component levels in mucosal secretions of the rat. J Immunol. 1983 Mar;130(3):1330–1335. [PubMed] [Google Scholar]
  36. Timsit Y., Moras D. Groove-backbone interaction in B-DNA. Implication for DNA condensation and recombination. J Mol Biol. 1991 Oct 5;221(3):919–940. doi: 10.1016/0022-2836(91)80184-v. [DOI] [PubMed] [Google Scholar]
  37. Wagner E., Cotten M., Foisner R., Birnstiel M. L. Transferrin-polycation-DNA complexes: the effect of polycations on the structure of the complex and DNA delivery to cells. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4255–4259. doi: 10.1073/pnas.88.10.4255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wagner E., Plank C., Zatloukal K., Cotten M., Birnstiel M. L. Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin-polylysine-DNA complexes: toward a synthetic virus-like gene-transfer vehicle. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):7934–7938. doi: 10.1073/pnas.89.17.7934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wilson J. M., Grossman M., Wu C. H., Chowdhury N. R., Wu G. Y., Chowdhury J. R. Hepatocyte-directed gene transfer in vivo leads to transient improvement of hypercholesterolemia in low density lipoprotein receptor-deficient rabbits. J Biol Chem. 1992 Jan 15;267(2):963–967. [PubMed] [Google Scholar]
  40. Wu C. H., Wilson J. M., Wu G. Y. Targeting genes: delivery and persistent expression of a foreign gene driven by mammalian regulatory elements in vivo. J Biol Chem. 1989 Oct 15;264(29):16985–16987. [PubMed] [Google Scholar]
  41. Wu G. Y., Wilson J. M., Shalaby F., Grossman M., Shafritz D. A., Wu C. H. Receptor-mediated gene delivery in vivo. Partial correction of genetic analbuminemia in Nagase rats. J Biol Chem. 1991 Aug 5;266(22):14338–14342. [PubMed] [Google Scholar]
  42. Wu G. Y., Wu C. H. Receptor-mediated gene delivery and expression in vivo. J Biol Chem. 1988 Oct 15;263(29):14621–14624. [PubMed] [Google Scholar]
  43. Yoshimura K., Rosenfeld M. A., Nakamura H., Scherer E. M., Pavirani A., Lecocq J. P., Crystal R. G. Expression of the human cystic fibrosis transmembrane conductance regulator gene in the mouse lung after in vivo intratracheal plasmid-mediated gene transfer. Nucleic Acids Res. 1992 Jun 25;20(12):3233–3240. doi: 10.1093/nar/20.12.3233. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES