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Poly (Lactide-co-Glycolide) Microspheres in Respirable Sizes Enhance an In Vitro T Cell Response to Recombinant Mycobacterium tuberculosis Antigen 85B

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Abstract

Purpose

To investigate the use of poly (lactide-co-glycolide) (PLGA) microparticles in respirable sizes as carriers for Antigen 85B (Ag85B), a secreted protein of Mycobacterium tuberculosis, with the ultimate goal of employing them in pulmonary delivery of tuberculosis vaccine.

Materials and Methods

Recombinant Ag85B was expressed from two Escherichia coli strains and encapsulated by spray-drying in PLGA microspheres with/without adjuvants. These microspheres containing rAg85B were assessed for their ability to deliver antigen to macrophages for subsequent processing and presentation to the specific CD4 T-hybridoma cells DB-1. DB-1 cells recognize the Ag85B97–112 epitope presented in the context of MHC class II and secrete IL-2 as the cytokine marker.

Results

Microspheres suitable for aerosol delivery to the lungs (3.4–4.3 μm median diameter) and targeting alveolar macrophages were manufactured. THP-1 macrophage-like cells exposed with PLGA-rAg85B microspheres induced the DB-1 cells to produce IL-2 at a level that was two orders of magnitude larger than the response elicited by soluble rAg85B. This formulation demonstrated extended epitope presentation.

Conclusions

PLGA microspheres in respirable sizes were effective in delivering rAg85B in an immunologically relevant manner to macrophages. These results are a foundation for further investigation into the potential use of PLGA particles for delivery of vaccines to prevent M. tuberculosis infection.

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Acknowledgements

The authors greatly appreciate the donation of: E. coli JM109DE strain carrying Ag85B gene vector by Dr Douglas Kernodle in Vanderbilt University; and T-hybridoma cells DB1 by Dr W. Henry Boom in Case Western Reserve University. The work was supported by a grant NHLB1, HL67221. Dongmei Lu’s financial aid is from PHRMA foundation.

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Authors and Affiliations

  1. Molecular Pharmaceutics, School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, 27599-7360, USA

    Dongmei Lu, Lucila Garcia-Contreras & Anthony J. Hickey

  2. Medicinal Chemistry and Natural Product, School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, 27599-7360, USA

    Ding Xu & Jian Liu

  3. Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, 27599-7290, USA

    Sherry L. Kurtz & Miriam Braunstein

  4. Department of Microbial and Molecular Pathogenesis, The Texas A&M University System Health Science Center, College Station, 407 Reynolds Medical Building, College Station, Texas, 77843-1114, USA

    David N. McMurray

  5. School of Pharmacy, University of North Carolina at Chapel Hill, Kerr Hall 1310 CB# 7360, Chapel Hill, North Carolina, 27599, USA

    Anthony J. Hickey

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  1. Dongmei Lu
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  2. Lucila Garcia-Contreras
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Correspondence to Anthony J. Hickey.

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Lu, D., Garcia-Contreras, L., Xu, D. et al. Poly (Lactide-co-Glycolide) Microspheres in Respirable Sizes Enhance an In Vitro T Cell Response to Recombinant Mycobacterium tuberculosis Antigen 85B. Pharm Res 24, 1834–1843 (2007). https://doi.org/10.1007/s11095-007-9302-8

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