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Infection and Immunity, April 1999, p. 1789-1797, Vol. 67, No. 4
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Rapid Local Expression of Interleukin-12, Tumor Necrosis Factor Alpha, and Gamma Interferon after Cutaneous Francisella tularensis Infection in Tularemia-Immune Mice

Stephan Stenmark,1 Dan Sunnemark,2 Anders Bucht,3,4 and Anders Sjöstedt1,5,*

Department of Infectious Diseases, University of Umeå,1 and Department of Biomedicine4 and Department of Microbiology,5 Defence Research Establishment, Umeå, and Microbiology and Tumorbiology Centre2 and Department of Rheumatology,3 Karolinska Institute, Stockholm, Sweden

Received 18 August 1998/Returned for modification 8 October 1998/Accepted 5 January 1999

Francisella tularensis LVS is an effective live vaccine strain used for cutaneous vaccination against tularemia in man. In mice, injection of LVS causes invasive disease and subsequent development of immunity that is characterized by effective control of otherwise lethal doses of the organism. In the present investigation, it is shown that LVS-immune mice controlled an intradermal infection much more effectively than did naive mice; bacterial counts in skin samples were 1.5 to 2.0 log10 lower 24 h after injection and 6 log10 lower 72 h after injection in immune mice. Moreover, in contrast to naive mice, no bacteria were demonstrated in samples from livers and spleens of immune mice. By immunohistochemistry, skin samples from immune mice showed an intense staining for interleukin-12 (IL-12) and a moderate staining for tumor necrosis factor alpha (TNF-alpha ) at 24 h postinoculation, after which staining for both cytokines faded. In naive mice, the staining for IL-12 was weak at all time points and no staining for TNF-alpha was observed. No staining for gamma interferon (IFN-gamma ) was observed in any group before 72 h. At that time point, skin samples from immune mice showed moderate staining and skin samples from naive mice showed weak staining. Reverse transcriptase PCR showed an induction of mRNA of the three cytokines in the skin within the first day after injection. A quantitative analysis demonstrated higher IFN-gamma and TNF-alpha mRNA levels in immune mice at 24 h postinoculation. In conclusion, immunization with F. tularensis LVS conferred a capability to respond to cutaneous reinfection, with rapid local expression of IL-12, TNF-alpha , and IFN-gamma , and this expression was paralleled by containment and mitigation of the infection. The cytokine response may be part of a local barrier function of the skin, important to host protection against tularemia.

* Corresponding author. Mailing address: Department of Microbiology, Defence Research Establishment, S-901 82 Umeå, Sweden. Phone: 46-90-106665. Fax: 46-90-106806. E-mail: sjostedt{at}ume.foa.se.

Infection and Immunity, April 1999, p. 1789-1797, Vol. 67, No. 4
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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Copyright © 1999 by the American Society for Microbiology. All rights reserved.