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NK cells contribute to persistent airway inflammation and AHR during the later stage of RSV infection in mice

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Abstract

RSV can lead to persistent airway inflammation and AHR and is intimately associated with childhood recurrent wheezing and asthma, but the underlying mechanisms remain unclear. There are high numbers of NK cells in the lung, which not only play important roles in the acute stage of RSV infection, but also are pivotal in regulating the pathogenesis of asthma. Therefore, in this study, we assumed that NK cells might contribute to persistent airway disease during the later stage of RSV infection. Mice were killed at serial time points after RSV infection to collect samples. Leukocytes in bronchoalveolar lavage fluid (BALF) were counted, lung histopathology was examined, and airway hyperresponsiveness (AHR) was measured by whole-body plethysmography. Cytokines were detected by ELISA, and NK cells were determined by flow cytometry. Rabbit anti-mouse asialo-GM-1 antibodies and resveratrol were used to deplete or suppress NK cells. Inflammatory cells in BALF, lung tissue damage and AHR were persistent for 60 days post-RSV infection. Type 2 cytokines and NK cells were significantly increased during the later stage of infection. When NK cells were decreased by the antibodies or resveratrol, type 2 cytokines, the persistent airway inflammation and AHR were all markedly reduced. NK cells can contribute to the RSV-associated persistent airway inflammation and AHR at least partially by promoting type 2 cytokines. Therefore, therapeutic targeting of NK cells may provide a novel approach to alleviating the recurrent wheezing subsequent to RSV infection.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant Numbers: 81470208, 81170010).

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Authors and Affiliations

  1. Ministry of Education Key Laboratory of Child Development and Disorders; Key Laboratory of Pediatrics in Chongqing, CSTC2009CA5002; Chongqing International Science and Technology Cooperation Center for Child Development and Disorders, Children’s Hospital, Chongqing Medical University, Chongqing, 400014, People’s Republic of China

    Keting Zhao, Wei Li, Wei Tang, Sisi Chen & Luo Ren

  2. Department of Respiratory Medicine, Children’s Hospital, Chongqing Medical University, No. 136, Zhongshan 2nd Road, Yuzhong District, Chongqing, 400014, People’s Republic of China

    Xiaoru Long, Jun Xie, Na Zang, Yu Deng, Xiaohong Xie, Lijia Wang, Zhou Fu & Enmei Liu

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  1. Xiaoru Long
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Correspondence to Enmei Liu.

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Conflict of interest

No potential conflicts of interest exist with any companies/organizations whose products or services may be discussed in this article. The sponsor had no role in the design of the study, in the collection and analysis of the data, or in the preparation of the manuscript. All authors have no any relevant competing interests.

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Informed consent was obtained from all individual participants included in the study. All applicable international, national and/or institutional guidelines for the care and use of animals were followed.

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430_2016_459_MOESM1_ESM.docx

Supplementary data 1 Resveratrol decreased T cells in BALB/c mice. BALB/c mice were divided into three groups: control: mock-infected and PBS-treated mice; RSV: RSV-infected and PBS-treated mice; RSV+RES: RSV-infected and resveratrol-treated mice. Resveratrol was given from day 14 to day 20. Diseases parameters were assessed on day 21. Graphs are represented as the mean±SEM. Data are representative of two independent experiments performed on five animals per group. ***, P< 0.001 shown comparing the control group with the RSV group. ^^^ P< 0.001 shown comparing the RSV group with the RSV+RES group. (DOCX 73 kb)

430_2016_459_MOESM2_ESM.docx

Supplementary data 2 The persistent airway inflammation and AHR during the later stage of RSV infection were not decreased following basophils depletion in nude mice. Nude mice were divided into three groups: control: mock-infected and PBS-treated mice; RSV: RSV-infected and PBS-treated mice; RSV+anti-anti-FcεR1: RSV-infected and anti-FcεR1 (MAR1)-treated mice. PBS or antibodies were administrated on days 14, 17 and 20. Disease parameters were assessed on day 21. Levels of IL-4, IL-5, IL-13 (A), airway inflammatory cells in BALF (B) and AHR in response to methacholine (C) were not suppressed after basophils were depleted. Values are expressed as means ± SEM. Data are representative of two independent experiments performed on 3–5 animals per group. *, **, ***, P< 0.05, 0.01, 0.001 shown comparing the control group with the RSV group. ^,^^, P<0.05, 0.01 shown comparing the control group with the RSV+ anti-anti-FcεR1 group. (DOCX 102 kb)

430_2016_459_MOESM3_ESM.docx

Supplementary data 3 The persistent airway inflammation and AHR during the later stage of RSV infection were not decreased following basophils or CD8 + T cells depletion in BALB/c mice. BALB/c mice were divided into four groups: control: mock-infected and PBS-treated mice; RSV: RSV-infected and PBS-treated mice; RSV+anti-anti-FcεR1: RSV-infected and anti-FcεR1 (MAR1)-treated mice; RSV+anti-CD8: RSV-infected and anti-CD8 antibody-treated mice. PBS or antibodies were administrated on days 14, 17 and 20. Disease parameters were assessed on day 21. Levels of IL-4, IL-5, IL-13 (A), airway inflammatory cells in BALF (B) and AHR in response to methacholine (C) were not suppressed after basophils or CD8+ T cells were depleted. Values are expressed as means ± SEM. Data are representative of two independent experiments performed on 3–10 animals per group. *, **, ***, P< 0.05, 0.01, 0.001 shown comparing the control group with the RSV group. ^,^^, ^^^, P<0.05, 0.01, 0.001 shown comparing the control group with the RSV+ anti-anti-FcεR1 group. #, ##, ###, P<0.05, 0.01, 0.001 shown comparing the control group with the RSV+anti-CD8 group. (DOCX 113 kb)

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Long, X., Xie, J., Zhao, K. et al. NK cells contribute to persistent airway inflammation and AHR during the later stage of RSV infection in mice. Med Microbiol Immunol 205, 459–470 (2016). https://doi.org/10.1007/s00430-016-0459-9

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