J Bacteriol Virol.  2014 Mar;44(1):52-58. 10.4167/jbv.2014.44.1.52.

Evaluation of Immune Response for Vi-CRM(197) Conjugated Vaccine against Salmonella enterica serovar Typhi in Mice

• Kim K
• Shim J
• Park S
• Park B
• Cho Y
• Hur B
• Kang SA
• Choi SK
• Baik Y
• Hahn TW

• KMID: 2135357
• DOI: http://doi.org/10.4167/jbv.2014.44.1.52

Abstract

Typhoid fever, a serious systemic infection caused by Salmonella enterica serovar Typhi, breaks out in developing countries. However, existing vaccines only induce relatively low protective effects with humoral responses and do not stimulate secondary immune response, especially to young people. The objective of this study is to evaluate the immunogenicity of the vaccine containing virulence capsular polysaccharide (Vi) conjugated with the optimal ratios of non-toxic variant of diphtheria toxin (CRM(197)) in mice. Six-week-old BALB/c female mice were injected intraperitoneally three times at intervals of 14 days and sera were collected on days 0, 14, 28, 42 and 56 post-injection. The efficacy of the vaccine was evaluated by comparing between negative control group injected with PBS and vaccine groups injected with Vi or Vi-CRM(197) conjugate of different ratio. Vi and CRM(197)-specific antibody responses were evaluated using enzyme-linked immunosorbent assay. The result showed that Vi-CRM(197)-1 group revealed the highest and significant Vi-specific IgG immune responses among the other groups and Vi group (p < 0.01). In conclusion, Vi-CRM(197)-1 conjugate vaccine induced the highest humoral immune response in mice and may be used as an effective vaccine to replace the existing typhoid vaccine for infants under 2 years old.

Keyword

Vi; CRM(197); Conjugated vaccine; Salmonella Typhi; Typhoid fever

MeSH Terms

Animals
Antibody Formation
Child, Preschool
Developing Countries
Diphtheria Toxin
Enzyme-Linked Immunosorbent Assay
Female
Humans
Immunity, Humoral
Immunoglobulin G
Infant
Mice*
Salmonella enterica*
Salmonella typhi*
Salmonella*
Typhoid Fever
Typhoid-Paratyphoid Vaccines
Vaccines
Virulence
Diphtheria Toxin
Immunoglobulin G
Typhoid-Paratyphoid Vaccines
Vaccines

Figure

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  Figure 1. SDS-PAGE and Western blot of recombinant CRM197 protein induced by 0.02% arabinose in E. coli. The total extracts (lane 1) of E. coli containing pBAD-CRM197 and purified CRM197 (lane 2) were separated by SDS-PAGE and stained with Coomassie blue, analyzed by Western blot of the duplicated gel of lane 1 using anti-His antibody and anti-mouse IgG AP conjugated antibody (lane 3).

• 

  Figure 2. Analysis of Vi-CRM197 conjugate levels by sandwich ELISA. For sandwich ELISA, plates were coated with anti-His antibody. Vi, CRM197 and Vi-CRM197 were added to each well. Anti-Vi polyclonal antibody was added and then anti-mouse IgG AP-conjugated antibody was added to each well. ***, Significant difference in Vi-CRM197 conjugate compared with the control group (p < 0.001).

• 

  Figure 3. Vi and CRM197-specific serum IgG responses in mice following dosing with Vi and Vi-CRM197 conjugates. BALB/c mice were immunized intraperitoneally three times at 14-day internals with 2.5 μg of Vi-CRM197. Values are reported as log2 geometric mean titer (GMT) and error bars indicate 95% confidence intervals. **p < 0.01 and *p < 0.05 versus control (PBS) immunized mice.

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