2017
DOI: 10.1128/iai.00593-17
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Antigen Localization Influences the Magnitude and Kinetics of Endogenous Adaptive Immune Response to Recombinant Salmonella Vaccines

Abstract: The use of recombinant attenuated Salmonella vaccine (RASV) strains is a promising strategy for presenting heterologous antigens to the mammalian immune system to induce both cellular and humoral immune responses. However, studies on RASV development differ on where heterologous antigens are expressed and localized within the bacterium, and it is unclear how antigen localization modulates the immune response. Previously, we exploited the plasmid-encoded toxin (Pet) autotransporter system for accumulation of he… Show more

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Cited by 6 publications
(5 citation statements)
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“…The purity of the neutrophils obtained by this method was >80%. To infect neutrophils, 5 × 10 5 enriched neutrophils were plated in a 96-well U-bottom plate, and STm that express GFP or mCherry ( 30 , 31 ) added at multiplicities of infection from 0.1 to 10. Plates were incubated at 37°C with light agitation for 2 h. After this, the cells were washed and incubated for 1 h in RPMI 1640 containing 50 μg/ml of gentamicin.…”
Section: Methodsmentioning
confidence: 99%
“…The purity of the neutrophils obtained by this method was >80%. To infect neutrophils, 5 × 10 5 enriched neutrophils were plated in a 96-well U-bottom plate, and STm that express GFP or mCherry ( 30 , 31 ) added at multiplicities of infection from 0.1 to 10. Plates were incubated at 37°C with light agitation for 2 h. After this, the cells were washed and incubated for 1 h in RPMI 1640 containing 50 μg/ml of gentamicin.…”
Section: Methodsmentioning
confidence: 99%
“…Second, secreted proteins are capable of inducing higher antibody responses than cell-bound proteins. 41 Cell-bound proteins are processed and presented by means of major histocompatibility complex class I, which selectively activates cytotoxic (CD8) T cells. Secreted proteins, however, get taken up by antigen-presenting cells and presented by means of major histocompatibility complex class II.…”
Section: Discussionmentioning
confidence: 99%
“…Whole cell lysates were resuspended in 2× Laemmli Buffer (Sigma) and boiled for 5 minutes. Proteins were visualized by Western immunoblotting after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) on precast polyacrylamide protein gels (NuPAGE 4%‐12% Bis‐Tris/MES, Invitrogen), as described previously . Proteins were transferred onto nitrocellulose membrane using the iBLOT (Life Technologies).…”
Section: Methodsmentioning
confidence: 99%
“…Proteins were visualized by Western immunoblotting after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) on precast polyacrylamide protein gels (NuPAGE 4%-12% Bis-Tris/MES, Invitrogen), as described previously. 16 Proteins were transferred onto nitrocellulose membrane using the iBLOT (Life Technologies). H pylori proteins were reacted with rabbit anti-CagA serum 17 (1:2000 dilution).…”
Section: Protein Analysesmentioning
confidence: 99%