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Microbial Immunity and Vaccines

Model of Differential Susceptibility to Mucosal Burkholderia pseudomallei Infection

Boping Liu, Ghee Chong Koo, Eu Hian Yap, Kim Lee Chua, Yunn-Hwen Gan
Boping Liu
1Department of Biochemistry
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Ghee Chong Koo
1Department of Biochemistry
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Eu Hian Yap
2Department of Microbiology, National University of Singapore, Singapore 119260
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Kim Lee Chua
1Department of Biochemistry
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Yunn-Hwen Gan
1Department of Biochemistry
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  • For correspondence: bchganyh@nus.edu.sg
DOI: 10.1128/IAI.70.2.504-511.2002
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  • FIG. 1.
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    FIG. 1.

    Bacterial loads in BALB/c and C57BL/6 mice. The bacterial loads in the lungs (A and D), livers (B and E), and spleens (C and F) of BALB/c (A, B, and C) and C57BL/6 (D, E, and F) mice at different time points after intranasal infection with virulent B. pseudomallei are shown. The logarithm of the number of CFU was plotted against time. Each symbol represents one mouse. The control mice are not represented, since no colonies were found in any of their organs. Filled symbols represent mice from one experiment, and open symbols represent animals from another experiment. Results from the two experiments performed under the same conditions are superimposed.

  • FIG. 2.
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    FIG. 2.

    Expression of IFN-γ mRNA in the lungs (A and D), livers (B and E), and spleens (C and F) of BALB/c (A, B, and C) and C57BL/6 (D, E, and F) mice as determined by RT-PCR. In each set of gels, the top panel depicts IFN-γ and the bottom panel depicts the corresponding GAPDH. Lanes M, 100-bp molecular size marker; lanes N, no-template control; lanes 1 to 3, three infected mice at 24 h postinfection; lanes 4, control mock-infected mouse at 24 h postinfection; lanes 5 to 7, three infected mice at 48 h postinfection; lanes 8, control mock-infected mouse at 48 h postinfection; lanes 9 to 11, three infected mice at 72 h postinfection; lanes 12, control mock-infected mouse at 72 h postinfection.

  • FIG. 3.
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    FIG. 3.

    Concentrations of IFN-γ (A and D), IL-1β (B and E), and TNF-α (C and F) in sera (A, B, and C) and nasal washes (D, E, and F) of BALB/c (white bars) and C57BL/6 (black bars) mice after intranasal infection with 50 CFU of virulent B. pseudomallei. Each bar represents pooled sera or nasal washes from three mice. The optical densities in the ELISA are expressed as picograms per milliliter corresponding to the linear portion of the standard curves.

  • FIG. 4.
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    FIG. 4.

    Serum IgG (A) and mucosal IgA (B) responses in C57BL/6 mice after intranasal inoculation with 50 CFU of virulent B. pseudomallei (•), 50,000 CFU of heat-killed B. pseudomallei (▪), or PBS (▴). Each symbol represents a 1:10 dilution of serum at the indicated days after the first inoculation at day 0, the second inoculation at day 14, and reinfection with 10,000 CFU of live bacteria at day 28 (A), or represents undiluted nasal wash at the indicated days after reinfection with 10,000 CFU live bacteria at day 28 (B), from three mice pooled together. Results represent means ± standard deviations of triplicates. OD, optical density.

Tables

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  • TABLE 1.

    Ten-day LD50s for C57BL/6 and BALB/c mice intranasally infected with B. pseudomallei strain KHW

    Mouse strainNo. dead/total at 10 days after intranasal infection with the following inoculation dose (CFU)a:LD50 (CFU)
    5/15015/45045/1,350135/4,050405/12,150
    BALB/c0/60/63/4b4/66/645
    C57BL/60/61/61/63/63/64,854
    • ↵ a Dose for BALB/c mice/dose for C57BL/6 mice.

    • ↵ b Two mice in this group died from the anesthetization procedure.

  • TABLE 2.

    Cytokine gene expression in infected mice

    CytokinemRNA expressiona at the indicated h postinfection in:
    C57BL/6 miceBALB/c mice
    LungLiverSpleenLungLiverSpleen
    244872244872244872244872244872244872
    IFN-γ+++++++++++−+++++++++++++++++−+++++−++++++++++−−++++++
    TNF-α−−−++−+−−++−+++++−+++++++++−−−−−−−−−+−−+−−+−−+−++−++−
    IL-1β++++++++++++++++++++++++++++++++++++++++++++++++++++++
    IL-4−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−
    IL-6+++++++++++++++++++++++++++++++++++−+++++−++++++++++++
    IL-10−−−−−−++++++++−+++−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−
    IL-12−−−−−−−−−−−−−−−−−−+−−++++++−−−−−−−−−−−−−−−−−−−−−−−−−−−
    • ↵ a +, presence of mRNA expression; −, absence of mRNA expression. Each symbol represents the result from one mouse.

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Model of Differential Susceptibility to Mucosal Burkholderia pseudomallei Infection
Boping Liu, Ghee Chong Koo, Eu Hian Yap, Kim Lee Chua, Yunn-Hwen Gan
Infection and Immunity Feb 2002, 70 (2) 504-511; DOI: 10.1128/IAI.70.2.504-511.2002

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Model of Differential Susceptibility to Mucosal Burkholderia pseudomallei Infection
Boping Liu, Ghee Chong Koo, Eu Hian Yap, Kim Lee Chua, Yunn-Hwen Gan
Infection and Immunity Feb 2002, 70 (2) 504-511; DOI: 10.1128/IAI.70.2.504-511.2002
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KEYWORDS

melioidosis
nasal mucosa

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