J Vet Sci.  2008 Sep;9(3):309-315. 10.4142/jvs.2008.9.3.309.

Cytokine response in Balb/c mice infected with Francisella tularensis LVS and the Pohang isolate

• Kim EJ
• Park SH
• Choi YS
• Shim SK
• Park MY
• Park MS
• Hwang KJ

• KMID: 1718585
• DOI: http://doi.org/10.4142/jvs.2008.9.3.309

Abstract

We investigated the immune response induced by the Francisella (F.) tularensis live vaccine strain (LVS) and the Pohang isolate. After the Balb/c mice were infected intradermally (i.d) with 2 x 10(4) cfu of F. tularensis LVS and Pohang, respectively, their blood and organs were collected at different times; 0, 3, 6, 24, 72, 96, 120 and 168 h after infection. Using these samples, RT-PCR and ELISA analysis were carried out for the comparative study of the cytokines, including TNF-alpha, INF-gamma, IL-2, IL-4, IL-10 and IL-12. In the Pohang-infected mice at 120 h, the liver showed a 53 times higher level of TNF-alpha and a 42 times higher level of IFN-gamma than the respective levels at the early time points after infection. The levels of TNF-alpha and IFN-gamma induced by LVS were 5 times lower than those induced by the Pohang isolate. Also, the organs from the Pohang-infected mice showed higher levels of TNF-alpha, IFN-gamma, IL-10 and IL-12 than the levels in the LVS-infected mice. The blood from the Pohang-infected mice at 120 h revealed about a 40 times increased level of IFN-gamma, and IL-10 was also increased by 4 times at 96 h compared to an early infection time point, while IL-4 was not induced during the whole infection period. These results suggest that F. tularensis may induce a Th1-mediated immune response to in vivo infection and the Pohang isolate has a higher capacity than the LVS to induce an acute immune response in Blab/c mice.

Keyword

Balb/c; F. tularensis; IFN-gamma; IL-12; TNF-alpha

MeSH Terms

Animals
*Bacterial Vaccines
Cytokines/*biosynthesis
Francisella tularensis/immunology/isolation & purification/*pathogenicity
Humans
Interferon-gamma/genetics/metabolism
Interleukins/genetics/metabolism
Korea
Liver/microbiology/pathology
Mice
Mice, Inbred BALB C
Polymerase Chain Reaction
Tularemia/*diagnosis/*immunology
Tumor Necrosis Factor-alpha/genetics/metabolism

Figure

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  Fig. 1 Francisella tularensis infection was induced decolorization of liver. Normal mouse liver (A), Francisella tularensis LVS (B), and Francisella tularensis Pohang (C) infected mouse liver after 120 h.

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  Fig. 2 PCR amplification of cytokines from liver after challenge with Francisella tularensis LVS (A) and Pohang (B) with 2 × 104 cfu. Samples were prepared at various times after inoculation. Lane 1: Negative control, 2: 3 h, 3: 6 h, 4: 24 h, 5: 48 h, 6: 72 h, 7: 96 h, 8: 120 h, 9: 168 h. Relative value means; cytokine expression level/negative control expression level. IL-2 and IL-4 were not expression within 168 h.

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  Fig. 3 Levels of TNF-α, IFN-γ, IL-10 and IL-12 of lung, kidney, lymph node from Francisella tularensis LVS and Pohang (2 × 104 cfu) infected mice. Relative value means; cytokine expression level/negative control expression level.

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  Fig. 4 Cytokine production of mice infected with Francisella tularensis Pohang and LVS. INF-γ (A), IL-10 (B) and IL-4 (C) production by mice after infection with Pohang and LVS.

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