Abstract
Recombinant attenuated Salmonella vaccines (RASVs) have been used extensively to express and deliver heterologous antigens to host mucosal tissues. Immune responses can be enhanced greatly when the antigen is secreted to the periplasm or extracellular compartment. The most common method for accomplishing this is by fusion of the antigen to a secretion signal sequence. Finding an optimal signal sequence is typically done empirically. To facilitate this process, we constructed a series of plasmid expression vectors, each containing a different type II signal sequence. We evaluated the utilities of these vectors by fusing two different antigens, the α-helix domains of pneumococcal surface protein A (PspA) and pneumococcal surface protein C (PspC), to the signal sequences of β-lactamase (bla SS), ompA, and phoA and the signal sequence and C-terminal peptide of β-lactamase (bla SS+CT) on Asd+ plasmids under the control of the Ptrc promoter. Strains were characterized for level of expression, subcellular antigen location, and the capacity to elicit antigen-specific immune responses and protection against challenge with Streptococcus pneumoniae in mice. The immune responses to each protein differed depending on the signal sequence used. Strains carrying the bla SS-pspA and bla SS+CT-pspC fusions yielded the largest amounts of secreted PspA and PspC, respectively, and induced the highest serum IgG titers, although all fusion proteins tested induced some level of antigen-specific IgG response. Consistent with the serum antibody responses, RASVs expressing the bla SS-pspA and bla SS+CT-pspC fusions induced the greatest protection against S. pneumoniae challenge.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3241-3254 |
| Number of pages | 14 |
| Journal | Infection and Immunity |
| Volume | 76 |
| Issue number | 7 |
| DOIs | |
| State | Published - Jul 2008 |
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ASJC Scopus subject areas
- Immunology
Cite this
Analysis of type II secretion of recombinant pneumococcal PspA and PspC in a Salmonella enterica serovar typhimurium vaccine with regulated delayed antigen synthesis. / Xin, Wei; Wanda, Soo Young; Li, Yuhua; Wang, Shifeng; Mo, Hua; Curtiss, Roy.
In: Infection and Immunity, Vol. 76, No. 7, 07.2008, p. 3241-3254.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Analysis of type II secretion of recombinant pneumococcal PspA and PspC in a Salmonella enterica serovar typhimurium vaccine with regulated delayed antigen synthesis
AU - Xin, Wei
AU - Wanda, Soo Young
AU - Li, Yuhua
AU - Wang, Shifeng
AU - Mo, Hua
AU - Curtiss, Roy
PY - 2008/7
Y1 - 2008/7
N2 - Recombinant attenuated Salmonella vaccines (RASVs) have been used extensively to express and deliver heterologous antigens to host mucosal tissues. Immune responses can be enhanced greatly when the antigen is secreted to the periplasm or extracellular compartment. The most common method for accomplishing this is by fusion of the antigen to a secretion signal sequence. Finding an optimal signal sequence is typically done empirically. To facilitate this process, we constructed a series of plasmid expression vectors, each containing a different type II signal sequence. We evaluated the utilities of these vectors by fusing two different antigens, the α-helix domains of pneumococcal surface protein A (PspA) and pneumococcal surface protein C (PspC), to the signal sequences of β-lactamase (bla SS), ompA, and phoA and the signal sequence and C-terminal peptide of β-lactamase (bla SS+CT) on Asd+ plasmids under the control of the Ptrc promoter. Strains were characterized for level of expression, subcellular antigen location, and the capacity to elicit antigen-specific immune responses and protection against challenge with Streptococcus pneumoniae in mice. The immune responses to each protein differed depending on the signal sequence used. Strains carrying the bla SS-pspA and bla SS+CT-pspC fusions yielded the largest amounts of secreted PspA and PspC, respectively, and induced the highest serum IgG titers, although all fusion proteins tested induced some level of antigen-specific IgG response. Consistent with the serum antibody responses, RASVs expressing the bla SS-pspA and bla SS+CT-pspC fusions induced the greatest protection against S. pneumoniae challenge.
AB - Recombinant attenuated Salmonella vaccines (RASVs) have been used extensively to express and deliver heterologous antigens to host mucosal tissues. Immune responses can be enhanced greatly when the antigen is secreted to the periplasm or extracellular compartment. The most common method for accomplishing this is by fusion of the antigen to a secretion signal sequence. Finding an optimal signal sequence is typically done empirically. To facilitate this process, we constructed a series of plasmid expression vectors, each containing a different type II signal sequence. We evaluated the utilities of these vectors by fusing two different antigens, the α-helix domains of pneumococcal surface protein A (PspA) and pneumococcal surface protein C (PspC), to the signal sequences of β-lactamase (bla SS), ompA, and phoA and the signal sequence and C-terminal peptide of β-lactamase (bla SS+CT) on Asd+ plasmids under the control of the Ptrc promoter. Strains were characterized for level of expression, subcellular antigen location, and the capacity to elicit antigen-specific immune responses and protection against challenge with Streptococcus pneumoniae in mice. The immune responses to each protein differed depending on the signal sequence used. Strains carrying the bla SS-pspA and bla SS+CT-pspC fusions yielded the largest amounts of secreted PspA and PspC, respectively, and induced the highest serum IgG titers, although all fusion proteins tested induced some level of antigen-specific IgG response. Consistent with the serum antibody responses, RASVs expressing the bla SS-pspA and bla SS+CT-pspC fusions induced the greatest protection against S. pneumoniae challenge.
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U2 - 10.1128/IAI.01623-07
DO - 10.1128/IAI.01623-07
M3 - Article
VL - 76
SP - 3241
EP - 3254
JO - Infection and Immunity
JF - Infection and Immunity
SN - 0019-9567
IS - 7
ER -