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Research ArticleChemotherapy, Antibiotics, and Gene Therapy

Premedication with Clarithromycin Is Effective against Secondary Bacterial Pneumonia during Influenza Virus Infection in a Pulmonary Emphysema Mouse Model

Tatsuhiko Harada, Yuji Ishimatsu, Atsuko Hara, Towako Morita, Shota Nakashima, Tomoyuki Kakugawa, Noriho Sakamoto, Kosuke Kosai, Koichi Izumikawa, Katsunori Yanagihara, Hiroshi Mukae and Shigeru Kohno
Journal of Pharmacology and Experimental Therapeutics September 2016, 358 (3) 457-463; DOI: https://doi.org/10.1124/jpet.116.233932
Tatsuhiko Harada
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Yuji Ishimatsu
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Atsuko Hara
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Towako Morita
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Shota Nakashima
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Tomoyuki Kakugawa
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Noriho Sakamoto
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Kosuke Kosai
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Koichi Izumikawa
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Katsunori Yanagihara
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Hiroshi Mukae
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Shigeru Kohno
Department of Infectious Diseases, Unit of Molecular Microbiology and Immunology (T.H., K.I.), Department of Cardiopulmonary Rehabilitation Sciences (Y.I.), Department of Respiratory Medicine (N.S., H.M.), and Department of Laboratory Medicine (K.K, K.Y.), Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan; Second Department of Internal Medicine, Nagasaki University School of Medicine, Nagasaki, Japan (T.H., A.H., T.M., S.N., T.K., N.S., H.M., S.K.)
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Article Figures & Data

Figures

  • Fig. 1.
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    Fig. 1.

    Experimental schedule of treatment. Mice were treated with clarithromycin (CAM; 200 mg/kg per day) or vehicle once daily. The postexposure medication group was treated from Day 1, and the premedication group was treated from Day –4.

  • Fig. 2.
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    Fig. 2.

    Survival curve of secondary bacterial pneumonia mice treated with or without CAM. (A) Survival curve of the postexposure medication group. Open triangle: mice treated with vehicle from Day 0 (control) (n = 6); solid triangle: mice treated with CAM from Day 0 (n = 10). (B) Survival curve of the premedication group. Open circle: mice treated with vehicle from Day –4 (control) (n = 9); solid circle: mice treated with CAM from Day –4 (n = 9); N.S., no significant difference versus control; *P < 0.05, significant difference versus control. S.P., Streptococcus pneumoniae.

  • Fig. 3.
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    Fig. 3.

    Representative hematoxylin and eosin–stained tissue sections of the vehicle control group (A, B) and CAM premedication group (C, D) showing pathologic inflammatory changes and bacterial proliferation on Day 6. Magnification: 40× (A, C) and 200× (B, D).

  • Fig. 4.
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    Fig. 4.

    Bacterial counts in lung homogenates (n = 7 mice/group). Data are presented as means ± S.E. *P < 0.05, significant difference versus control.

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    Fig. 5.

    Local antibacterial activity of CAM in the lung. Agar plates inoculated with (A) NU4471 (macrolide-resistant S. pneumoniae) and (B) ATCC49619 (macrolide-susceptible S. pneumoniae). Four paper discs were arranged as follows: upper left, lung homogenates of vehicle-treated mice; lower left, lung homogenates of CAM-treated mice; upper right, garenoxacin; and lower right, CAM.

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    Fig. 6.

    Influenza viral titers of lung homogenates at Day 4. Data are presented as means ± S.E., n = 4 mice. N.S., no significant difference between two groups; *P < 0.05, significant difference versus the vehicle control group.

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    Fig. 7.

    Anti-inflammatory effect of CAM on BALF on the basis of the number of (A) total cells and (B) neutrophils in each group. Data are presented as means ± S.E.; n = 5–9 mice. N.S., no significant difference between two groups; *P < 0.05, significant difference between two groups.

  • Fig. 8.
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    Fig. 8.

    Anti-inflammatory effect of CAM on cytokine levels in BALF: (A) IFN-α, (B) IFN-β, (C) IFN-γ, and (D) IL-10. Data are presented as means ± S.E., n = 5–9 mice. N.S, no significant difference between two groups; *P < 0.05, significant difference between two groups.

  • Fig. 9.
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    Fig. 9.

    Anti-inflammatory effect of CAM on IFN-γ levels in (A) lung homogenates and (B) serum samples. Data are presented as means ± S.E.; n = 5–9 mice. N.S., no significant difference; *P < 0.05, significant difference.

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Journal of Pharmacology and Experimental Therapeutics: 358 (3)
Journal of Pharmacology and Experimental Therapeutics
Vol. 358, Issue 3
1 Sep 2016
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Research ArticleChemotherapy, Antibiotics, and Gene Therapy

Effect of CAM on SBP during Influenza in Emphysema Mice

Tatsuhiko Harada, Yuji Ishimatsu, Atsuko Hara, Towako Morita, Shota Nakashima, Tomoyuki Kakugawa, Noriho Sakamoto, Kosuke Kosai, Koichi Izumikawa, Katsunori Yanagihara, Hiroshi Mukae and Shigeru Kohno
Journal of Pharmacology and Experimental Therapeutics September 1, 2016, 358 (3) 457-463; DOI: https://doi.org/10.1124/jpet.116.233932

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Research ArticleChemotherapy, Antibiotics, and Gene Therapy

Effect of CAM on SBP during Influenza in Emphysema Mice

Tatsuhiko Harada, Yuji Ishimatsu, Atsuko Hara, Towako Morita, Shota Nakashima, Tomoyuki Kakugawa, Noriho Sakamoto, Kosuke Kosai, Koichi Izumikawa, Katsunori Yanagihara, Hiroshi Mukae and Shigeru Kohno
Journal of Pharmacology and Experimental Therapeutics September 1, 2016, 358 (3) 457-463; DOI: https://doi.org/10.1124/jpet.116.233932
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