Fig. 1: Active immunisation with S. aureus vaccine candidates.
From: Epitope-specific immunity against Staphylococcus aureus coproporphyrinogen III oxidase
a SDS-PAGE analysis of coomassie stained E. coli lysate and purified rHis6-tagged CgoX protein (56 kDa) expressed in E. coli BL21(DE3). DE3 cells were left untreated (−) or induced with IPTG (+) for expression. b Left panel: antigen-specific IgG response of immunised BALB/c mice. Sera of mice (n = 12) immunised with S. aureus rCgoX formulated with Freund’s adjuvant (FA) were collected and pooled at day 0 (preimmune), 14 (initial) and 66 (2nd boost) after initial immunisation and analysed by ELISA for IgG binding to rCgoX-His (n = 2). Data are presented as mean ± s.d. Right panel: antigen-specificity of sera from immunised mice was analysed by WB. Sera collected and pooled at day 66 after initial immunisation with rCgoX-His were analysed for binding to rCgoX-His or His-tagged S. aureus rPephyd protein (39 kDa) used as control. Bound murine IgG were detected with anti-mIgG-HRP. c Survival of immunised mice after challenge with S. aureus (n = 11 for control, n = 12 for CgoX). BALB/c mice were immunised with rCgoX-His/FA and challenged i.v. with 3 × 107 S. aureus strain ATCC 29213. Control mice were immunised with BSA/FA. Significance was calculated by Log-rank (Mantel-Cox) test. d SDS-PAGE analysis of coomassie stained E. coli lysate and purified His6-tagged TPI protein (32 kDa) expressed in E. coli BL21(DE3). DE3 cells were left untreated (−) or induced (+) with IPTG for expression. e BALB/c mice (n = 12) were immunised with S. aureus antigen rTPI-His/FA and serum pools were analysed by ELISA for IgG binding to rTPI-His (n = 2). Data are presented as mean ± s.d. Right panel: Sera collected and pooled at day 66 after initial immunisation with rTPI-His were analysed by WB for binding to rTPI-His and rPephyd-His (39 kDa) used as control. Bound murine IgG were detected with anti-mIgG-HRP. f Survival of mice immunised with rTPI-His formulated with FA (n = 11) or BSA / FA (n = 12) and challenged i.v. with 3 × 107 S. aureus strain ATCC 29213. Significance was calculated by Log-rank (Mantel-Cox) test.
