Enhanced priming of adaptive immunity by Mycobacterium smegmatis mutants with high-level protein secretion

Natalie Taylor, Faith Bahunde, Afton Thompson, Jae Sung Yu, William R. Jacobs, Norm L. Letvin, Barton F. Haynes, Sunhee Lee

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Mycobacteria have features that make them attractive as potential vaccine vectors. The nonpathogenic and rapidly growing Mycobacterium smegmatis can express both Mycobacterium tuberculosis antigens and heterologous antigens from other pathogens, and it has been used as a viable vector for the development of live vaccines. In order to further improve antigen-specific immunogenicity of M. smegmatis, we screened a random transposon mutant library for mutants displaying enhanced efficiency of protein secretion ("high secretors") and isolated 61 mutants showing enhanced endogenic and transgenic protein secretion. Sequence analysis identified a total of 54 genes involved in optimal secretion of insert proteins, as well as multiple independent transposon insertions localized within the same genomic loci and operons. The majority of transposon insertions occurred in genes that have no known protein secretion function. These transposon mutants were shown to prime antigen-specific CD8 + T cell responses better than the parental strain. Specifically, upon introducing the simian immunodeficiency virus (SIV) gag gene into these transposon mutant strains, we observed that they primed SIV Gag-specific CD8+ T cell responses significantly better than the control prime immunization in a heterologous prime/boost regimen. Our results reveal a dependence on bacterial secretion of mycobacterial and foreign antigens for the induction of antigen-specific CD8+ T cells in vivo. The data also suggest that these M. smegmatis transposon mutants could be used as novel live attenuated vaccine strains to express foreign antigens, such as those of human immunodeficiency virus type 1 (HIV-1), and induce strong antigen-specific T cell responses.

Original languageEnglish (US)
Pages (from-to)1416-1425
Number of pages10
JournalClinical and Vaccine Immunology
Volume19
Issue number9
DOIs
StatePublished - Sep 1 2012
Externally publishedYes

Fingerprint

Mycobacterium smegmatis
Adaptive Immunity
T-cells
CD8 Antigens
T-Lymphocytes
Antigens
Simian Immunodeficiency Virus
Viruses
Proteins
Vaccines
Genes
gag Genes
Heterophile Antigens
Attenuated Vaccines
Operon
Mycobacterium
Immunization
Sequence Analysis
HIV-1
Pathogens

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Microbiology (medical)

Cite this

Enhanced priming of adaptive immunity by Mycobacterium smegmatis mutants with high-level protein secretion. / Taylor, Natalie; Bahunde, Faith; Thompson, Afton; Yu, Jae Sung; Jacobs, William R.; Letvin, Norm L.; Haynes, Barton F.; Lee, Sunhee.

In: Clinical and Vaccine Immunology, Vol. 19, No. 9, 01.09.2012, p. 1416-1425.

Research output: Contribution to journalArticle

Taylor, N, Bahunde, F, Thompson, A, Yu, JS, Jacobs, WR, Letvin, NL, Haynes, BF & Lee, S 2012, 'Enhanced priming of adaptive immunity by Mycobacterium smegmatis mutants with high-level protein secretion', Clinical and Vaccine Immunology, vol. 19, no. 9, pp. 1416-1425. https://doi.org/10.1128/CVI.00131-12
Taylor, Natalie ; Bahunde, Faith ; Thompson, Afton ; Yu, Jae Sung ; Jacobs, William R. ; Letvin, Norm L. ; Haynes, Barton F. ; Lee, Sunhee. / Enhanced priming of adaptive immunity by Mycobacterium smegmatis mutants with high-level protein secretion. In: Clinical and Vaccine Immunology. 2012 ; Vol. 19, No. 9. pp. 1416-1425.
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