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Infection and Immunity logoLink to Infection and Immunity
. 1995 Mar;63(3):799–803. doi: 10.1128/iai.63.3.799-803.1995

Secretion of human interleukin 2 by recombinant Mycobacterium bovis BCG.

D Kong 1, D Y Kunimoto 1
PMCID: PMC173073  PMID: 7868249

Abstract

The human interleukin 2 (huIL-2) gene was introduced into Mycobacterium bovis BCG by using the integrative vector pMV306. To express and secrete huIL-2 from BCG, two different plasmids, CI and CII, were made. In CI, the huIL-2-encoding region was under the control of the alpha-antigen promoter of BCG; in CII, the expression of huIL-2 was regulated by the heat shock protein 60 promoter. A signal peptide sequence isolated from the naturally secreted alpha-antigen of BCG was inserted between the promoter and huIL-2-encoding region to facilitate secretion. Both huIL-2 expression plasmids were integrated into the BCG genome when introduced into the BCG Pasteur strain by electroporation. Approximately 150 U of huIL-2 was secreted into the medium of a BCG-CII culture, while the BCG-CI cells secreted approximately one-sixth of that amount. When the IL-2-expressing BCG strain BCG-CII was injected intravenously into BALB/c mice, the number of BCG cells in the spleens of these mice was significantly less than the number in the control mice. The decreased number of IL-2-expressing BCG cells is likely due to the augmentation of the host immune response by the secreted huIL-2, although the exact mechanism is not known.

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Selected References

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  1. Aldovini A., Young R. A. Development of a BCG recombinant vehicle for candidate AIDS vaccines. Int Rev Immunol. 1990;7(1):79–83. doi: 10.3109/08830189009061766. [DOI] [PubMed] [Google Scholar]
  2. Aldovini A., Young R. A. Humoral and cell-mediated immune responses to live recombinant BCG-HIV vaccines. Nature. 1991 Jun 6;351(6326):479–482. doi: 10.1038/351479a0. [DOI] [PubMed] [Google Scholar]
  3. Belosevic M., Finbloom D. S., Meltzer M. S., Nacy C. A. IL-2. A cofactor for induction of activated macrophage resistance to infection. J Immunol. 1990 Aug 1;145(3):831–839. [PubMed] [Google Scholar]
  4. Bermudez L. E., Stevens P., Kolonoski P., Wu M., Young L. S. Treatment of experimental disseminated Mycobacterium avium complex infection in mice with recombinant IL-2 and tumor necrosis factor. J Immunol. 1989 Nov 1;143(9):2996–3000. [PubMed] [Google Scholar]
  5. Cockett A. T., Davis R. S., Cos L. R., Wheeless L. L., Jr Bacillus Calmette-Guerin and interleukin-2 for treatment of superficial bladder cancer. J Urol. 1991 Sep;146(3):766–770. doi: 10.1016/s0022-5347(17)37916-8. [DOI] [PubMed] [Google Scholar]
  6. Horton R. M., Hunt H. D., Ho S. N., Pullen J. K., Pease L. R. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene. 1989 Apr 15;77(1):61–68. doi: 10.1016/0378-1119(89)90359-4. [DOI] [PubMed] [Google Scholar]
  7. Jacobs W. R., Jr, Kalpana G. V., Cirillo J. D., Pascopella L., Snapper S. B., Udani R. A., Jones W., Barletta R. G., Bloom B. R. Genetic systems for mycobacteria. Methods Enzymol. 1991;204:537–555. doi: 10.1016/0076-6879(91)04027-l. [DOI] [PubMed] [Google Scholar]
  8. Jeevan A., Asherson G. L. Recombinant interleukin-2 limits the replication of Mycobacterium lepraemurium and Mycobacterium bovis BCG in mice. Infect Immun. 1988 Mar;56(3):660–664. doi: 10.1128/iai.56.3.660-664.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kawase I., Brooks C. G., Kuribayashi K., Olabuenaga S., Newman W., Gillis S., Henney C. S. Interleukin 2 induces gamma-interferon production: participation of macrophages and NK-like cells. J Immunol. 1983 Jul;131(1):288–292. [PubMed] [Google Scholar]
  10. Lee M. H., Pascopella L., Jacobs W. R., Jr, Hatfull G. F. Site-specific integration of mycobacteriophage L5: integration-proficient vectors for Mycobacterium smegmatis, Mycobacterium tuberculosis, and bacille Calmette-Guérin. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3111–3115. doi: 10.1073/pnas.88.8.3111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Malkovský M., Loveland B., North M., Asherson G. L., Gao L., Ward P., Fiers W. Recombinant interleukin-2 directly augments the cytotoxicity of human monocytes. Nature. 1987 Jan 15;325(6101):262–265. doi: 10.1038/325262a0. [DOI] [PubMed] [Google Scholar]
  12. Matsuo K., Yamaguchi R., Yamazaki A., Tasaka H., Terasaka K., Totsuka M., Kobayashi K., Yukitake H., Yamada T. Establishment of a foreign antigen secretion system in mycobacteria. Infect Immun. 1990 Dec;58(12):4049–4054. doi: 10.1128/iai.58.12.4049-4054.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Matsuo K., Yamaguchi R., Yamazaki A., Tasaka H., Yamada T. Cloning and expression of the Mycobacterium bovis BCG gene for extracellular alpha antigen. J Bacteriol. 1988 Sep;170(9):3847–3854. doi: 10.1128/jb.170.9.3847-3854.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Morgan D. A., Ruscetti F. W., Gallo R. Selective in vitro growth of T lymphocytes from normal human bone marrows. Science. 1976 Sep 10;193(4257):1007–1008. doi: 10.1126/science.181845. [DOI] [PubMed] [Google Scholar]
  15. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. doi: 10.1016/0022-1759(83)90303-4. [DOI] [PubMed] [Google Scholar]
  16. O'Donnell M. A., Aldovini A., Duda R. B., Yang H., Szilvasi A., Young R. A., DeWolf W. C. Recombinant Mycobacterium bovis BCG secreting functional interleukin-2 enhances gamma interferon production by splenocytes. Infect Immun. 1994 Jun;62(6):2508–2514. doi: 10.1128/iai.62.6.2508-2514.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rosenberg S. A. The development of new immunotherapies for the treatment of cancer using interleukin-2. A review. Ann Surg. 1988 Aug;208(2):121–135. doi: 10.1097/00000658-198808000-00001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Saunders C. W., Schmidt B. J., Mirot M. S., Thompson L. D., Guyer M. S. Use of chromosomal integration in the establishment and expression of blaZ, a Staphylococcus aureus beta-lactamase gene, in Bacillus subtilis. J Bacteriol. 1984 Mar;157(3):718–726. doi: 10.1128/jb.157.3.718-726.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sher N. A., Chaparas S. D., Pearson J., Chirigos M. Virulence of six strains of Mycobacterium bovis (BCG) in mice. Infect Immun. 1973 Nov;8(5):736–742. doi: 10.1128/iai.8.5.736-742.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smith K. A. Interleukin-2: inception, impact, and implications. Science. 1988 May 27;240(4856):1169–1176. doi: 10.1126/science.3131876. [DOI] [PubMed] [Google Scholar]
  22. Soloway M. S., Perito P. E. Superficial bladder cancer: diagnosis, surveillance and treatment. J Cell Biochem Suppl. 1992;16I:120–127. doi: 10.1002/jcb.240501323. [DOI] [PubMed] [Google Scholar]
  23. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  24. Stover C. K., Bansal G. P., Hanson M. S., Burlein J. E., Palaszynski S. R., Young J. F., Koenig S., Young D. B., Sadziene A., Barbour A. G. Protective immunity elicited by recombinant bacille Calmette-Guerin (BCG) expressing outer surface protein A (OspA) lipoprotein: a candidate Lyme disease vaccine. J Exp Med. 1993 Jul 1;178(1):197–209. doi: 10.1084/jem.178.1.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stover C. K., de la Cruz V. F., Fuerst T. R., Burlein J. E., Benson L. A., Bennett L. T., Bansal G. P., Young J. F., Lee M. H., Hatfull G. F. New use of BCG for recombinant vaccines. Nature. 1991 Jun 6;351(6326):456–460. doi: 10.1038/351456a0. [DOI] [PubMed] [Google Scholar]
  26. Strieter R. M., Remick D. G., Lynch J. P., 3rd, Spengler R. N., Kunkel S. L. Interleukin-2-induced tumor necrosis factor-alpha (TNF-alpha) gene expression in human alveolar macrophages and blood monocytes. Am Rev Respir Dis. 1989 Feb;139(2):335–342. doi: 10.1164/ajrccm/139.2.335. [DOI] [PubMed] [Google Scholar]
  27. Taniguchi T., Matsui H., Fujita T., Takaoka C., Kashima N., Yoshimoto R., Hamuro J. Structure and expression of a cloned cDNA for human interleukin-2. Nature. 1983 Mar 24;302(5906):305–310. doi: 10.1038/302305a0. [DOI] [PubMed] [Google Scholar]
  28. Wada S., Ikemoto S., Terada T., Tanaka H., Kamizuru M., Sakamoto W., Kishimoto T., Maekawa M. Combined therapy of interleukin 2 with cyclophosphamide or bacillus Calmette-Guérin against implanted bladder cancer cells in mice. Urol Int. 1991;47 (Suppl 1):104–107. doi: 10.1159/000282265. [DOI] [PubMed] [Google Scholar]
  29. Wiker H. G., Harboe M., Nagai S. A localization index for distinction between extracellular and intracellular antigens of Mycobacterium tuberculosis. J Gen Microbiol. 1991 Apr;137(4):875–884. doi: 10.1099/00221287-137-4-875. [DOI] [PubMed] [Google Scholar]
  30. Winter N., Lagranderie M., Rauzier J., Timm J., Leclerc C., Guy B., Kieny M. P., Gheorghiu M., Gicquel B. Expression of heterologous genes in Mycobacterium bovis BCG: induction of a cellular response against HIV-1 Nef protein. Gene. 1991 Dec 20;109(1):47–54. doi: 10.1016/0378-1119(91)90587-2. [DOI] [PubMed] [Google Scholar]
  31. Yokota T., Arai N., Lee F., Rennick D., Mosmann T., Arai K. Use of a cDNA expression vector for isolation of mouse interleukin 2 cDNA clones: expression of T-cell growth-factor activity after transfection of monkey cells. Proc Natl Acad Sci U S A. 1985 Jan;82(1):68–72. doi: 10.1073/pnas.82.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]

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