Article Figures & Data
Figures
- FIG 1
Survival of Salmonella spp. in the presence of complement alone. Survival of S. Typhimurium D65, S. Enteritidis R11, S. Typhi Ty2, and S. Paratyphi A ATCC 9150 in the presence of guinea pig complement (GPC) (A) and baby rabbit complement (BRC) (B) in the absence of immune sera. The results are the mean ± standard deviation values from at least two independent experiments. The dashed line represents 100% of inoculum as a visual reference.
- FIG 2
Complement titrations. Shown is the survival of S. Typhimurium D65 (A), S. Enteritidis R11 (B), S. Typhi Ty2 (C), all grown to log phase (OD600, 0.4), and S. Paratyphi A ATCC 9150 (D) grown to stationary phase with BRC in the presence of immune (I) compared to nonimmune (NI) sera and decreasing complement concentrations. The results are shown as the mean ± standard deviation values from more than three independent experiments. *, P < 0.05, and **, P < 0.01, by Student's t test (two-tailed). The dashed lines represent 100% of inoculum as a visual reference.
- FIG 3
Survival of S. Typhimurium in the presence of complement from various animals and antibody. Shown is the survival of S. Typhimurium D65 in the presence of immune (I) antibody compared to nonimmune (NI) antibody with indicated complement sources. The results are the mean ± standard deviation values from more than three independent experiments. BRC is from 3- to 4-week-old rabbits. **, P < 0.01 by the Mann-Whitney test. The dashed line represents 100% of inoculum as a visual reference.
- FIG 4
LPS produced by Salmonella serovar. (A) LPS profiles of S. Typhimurium D65 (lane 1), S. Enteritidis R11 (lane 2), S. Typhi Ty2 (lane 3), and S. Paratyphi A ATCC 9150 (lane 4) after overnight culture. Ratio of long or medium O antigen (O-Ag) at OD600 of 0.4, 0.8, and 4.0 to overnight culture for S. Typhimurium D65 (B), S. Enteritidis R11 (C), S. Typhi Ty2 (D), and S. Paratyphi A ATCC 9150 (E). The results are the mean ± standard deviation values from more than three independent experiments. *, P < 0.05, **, P < 0.01, and ***, P < 0.001, by ANOVA, with Bonferroni's test for multiple comparison.
- FIG 5
SBA titer and presence of the gene rck. SBA titers in pooled sera from CVD 1941-immunized mice against clinical invasive rck-positive or rck-negative S. Enteritidis strains. The lines represent the mean titers from at least three independent experiments. P = 0.45, Student's t test, two-tailed. n.s., nonsignificant.
- FIG 6
Survival of S. Typhi strains that express various levels of Vi in the presence of BRC. Shown is the survival of S. Typhi strains Ty2 and CVD 909 grown to an OD600 of 0.4 and incubated with 12.5% BRC in the absence of immune serum. The results represent the mean ± standard deviation value from at least three independent experiments. c, constitutively expressed. *, P < 0.05 and **, P < 0.01, by Student's t test, two-tailed. The dashed line represents 100% of inoculum as a visual reference.
- FIG 7
Endpoint SBA titers for mice immunized with live attenuated typhoidal and nontyphoidal Salmonella vaccines. (A) Sera from mice immunized with S. Typhimurium CVD 1931 against the target strain S. Typhimurium D65. (B) Sera from mice immunized with S. Enteritidis CVD 1944 against the target strain S. Enteritidis S15. (C) Sera from mice immunized with S. Typhi CVD 910 against the target strain S. Typhi Ty2. (D) Sera from mice immunized with S. Paratyphi A CVD 1902 against the target strain S. Paratyphi A ATCC 9150. The solid lines indicate mean titers. The dashed lines indicate the detection limit of the assay. **, P < 0.01, and ****, P < 0.0001, by the Mann-Whitney test.
- FIG 8
Correlation of SBA titer with anti-LPS serum IgG titer. SBA and anti-LPS serum titers produced by mice immunized with S. Typhimurium CVD 1931 (A), S. Enteritidis CVD 1944 (B), S. Typhi CVD 910 (C), and S. Paratyphi A CVD 1902 (D). The data were analyzed using Spearman's correlation coefficient.
- FIG 9
SBA titers using attenuated or reference Salmonella strains. Shown in black are the SBA titers produced by mice immunized with S. Typhimurium CVD 1931 (n = 12, pooled) using S. Typhimurium strains D65 and D23580 as target strains; in gray are the SBA titers produced by mice immunized with S. Paratyphi A CVD 1902 (n = 10, pooled) using S. Paratyphi A strains ATCC 9150 and CVD 1901 as target strains; in white are the SBA titers produced by mice immunized with S. Typhi CVD 910 (n = 12, pooled) using S. Typhi strains Ty2 and CVD 915 as target strains. The results are the mean ± standard deviation values from at least two independent experiments.
Tables
- TABLE 1
Bacterial strains used in this study
Serovar or species Strain Isolation location or characteristics Reference(s) or source Salmonella Typhimurium D65 Clinical isolate from blood culture, Mali 43, 44 D23580 Clinical isolate from blood culture, Malawi 18, 45 CVD 1921 I77 ΔguaBA ΔclpP 15 CVD 1931 D65 ΔguaBA ΔclpX S. M. Tennant, P. Schmidlein, J. E. Galen, and M. M. Levine, unpublished data Salmonella Enteritidis R11 Clinical isolate from blood culture, Mali 43, 44 S01 Clinical isolate from blood culture, Mali 43, 44 S15 Clinical isolate from blood culture, Mali 43, 44 Q38 Clinical isolate from blood culture, Mali 43, 44 CVD 1941 R11 ΔguaBA ΔclpP 15 CVD 1944 R11 ΔguaBA ΔclpX 15 Salmonella Typhi Ty2 Wild-type 46 CVD 909 Ty2 ΔaroC ΔaroD ΔhtrA Ptac-tviA 11 CVD 910 Ty2 ΔguaBA ΔhtrA 26 CVD 915 Ty2 ΔguaBA 47 Ty21a Ty2 ΔgalE ΔilvD ΔviaB (Vi−) H2S− 48 Salmonella Paratyphi A ATCC 9150 Wild-type American Type Culture Collection, Manassas, VA CVD 1901 ATCC 9150 ΔguaBA 14 CVD 1902 ATCC 9150 ΔguaBA ΔclpX 14 Escherichia coli Bort O18:K1:H7, clinical isolate from blood culture, Walter Reed Army Medical Center 49 - TABLE 2
Optimized SBA assay conditions for typhoidal and nontyphoidal Salmonella
Salmonella serovar Growth phase No. of CFU % BRC Typhimurium Log (OD600 of 0.4) 100–350 25 Enteritidis Log (OD600 of 0.4) 100–350 25 Typhi Log (OD600 of 0.4) 100–350 12.5 Paratyphi A Stationary 100–350 12.5
Supplemental material
Files in this Data Supplement:
- Supplemental file 1 -
Fig. S1. Growth curve of Salmonella serovars S. Typhimurium D65, S. Enteritidis R11, S. Typhi Ty2, and S. Paratyphi A ATCC 9150. Fig. S2. Example of LPS gel used for densitometry analysis.
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- Supplemental file 1 -
