Journal article
Recombinant Mycobacterium bovis BCG secreting functional interleukin-2 enhances gamma interferon production by splenocytes
Infection and immunity, Vol.62(6), pp.2508-2514
06/1994
DOI: 10.1128/iai.62.6.2508-2514.1994
PMCID: PMC186538
PMID: 8188376
Abstract
Mycobacterium bovis BCG was genetically engineered to express and secrete mouse interleukin-2 (IL-2) and rat IL-2. Genes encoding IL-2 were inserted into an Escherichia coli-BCG shuttle plasmid under the control of the BCG HSP60 promoter. To facilitate study of proteins produced in this system, the IL-2 gene product was expressed (i) alone, (ii) with the mycobacterial alpha-antigen secretion signal sequence at the amino terminus, (iii) with an influenza virus hemagglutinin epitope tag at the amino terminus, and (iv) with both the secretion signal sequence and the epitope tag. When expressed with the alpha-antigen signal sequence, biologically active IL-2 was secreted into the extracellular medium. Western blot (immunoblot) analysis of the intracellular IL-2 and extracellular IL-2 revealed that the secretion signal was appropriately cleaved from the recombinant lymphokine upon secretion. To assess the ability of recombinant BCG to stimulate cytokine production in a splenocyte population, mouse splenocytes were cultured together with wild-type or IL-2-producing BCG. IL-2-secreting BCG clones stimulated substantial increases in gamma interferon production, which could be reproduced by the addition of exogenous IL-2 to BCG. Levels of IL-6, IL-10, tumor necrosis factor alpha, and granulocyte-macrophage colony-stimulating factor were not significantly changed, while IL-4 and IL-5 remained undetectable (less than 50 pg/ml). The enhanced production of gamma interferon in response to IL-2-secreting BCG was strain independent. Recombinant BCG expressing mammalian cytokines provides a novel means to deliver cytokines and may augment the immunostimulatory properties of BCG in immunization and cancer therapy.
Details
- Title: Subtitle
- Recombinant Mycobacterium bovis BCG secreting functional interleukin-2 enhances gamma interferon production by splenocytes
- Creators
- Michael A O'Donnell - Division of Urology, Beth Israel Hospital, Boston, MA 02215Anna Aldovini - Beth Israel Deaconess HospitalRosemary B Duda - Beth Israel Deaconess HospitalHua Yang - Beth Israel Deaconess HospitalAkos Szilvasi - Beth Israel Deaconess HospitalRichard A Young - Beth Israel Deaconess HospitalWilliam C DeWolf - Beth Israel Deaconess Hospital
- Resource Type
- Journal article
- Publication Details
- Infection and immunity, Vol.62(6), pp.2508-2514
- DOI
- 10.1128/iai.62.6.2508-2514.1994
- PMID
- 8188376
- PMCID
- PMC186538
- NLM abbreviation
- Infect Immun
- ISSN
- 0019-9567
- eISSN
- 1098-5522
- Language
- English
- Date published
- 06/1994
- Academic Unit
- Urology
- Record Identifier
- 9984320851702771
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