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Local and Systemic Immune Responses to Influenza A Virus Infection in Pneumonia and Encephalitis Mous 时间:2017-08-30 14:54   来源:未知   作者:admin   点击: 次 Abstract:Objective. To compare local and systemic profiles between different disease pathologies (pneumonia and encephalitis) induced by influenza A virus (IAV). Methods. An IAV pneumonia model was created by intranasal inoculation of C57BL/6 mice with influenza A/WSN/33 (H1N1) virus. Lung lavage and blood collection were performed on day 3 after IAV inoculation. Similarly, an IAV encephalitis mouse model was created by direct intracranial IAV inoculation. Cerebrospinal fluid (CSF) and blood collection were conducted according to the same schedule. Cytokine/chemokine profiles were produced for each collected sample. Then the data were compared visually using radar charts. Results. Serum cytokine profiles were similar in pneumonia and encephalitis models, but local responses between the bronchoalveolar lavage fluid (BALF) in the pneumonia model and CSF in the encephalitis model differed. Moreover, to varying degrees, the profiles of local cytokines/chemokines differed from those of serum in both the pneumonia and encephalitis models. Conclusion. Investigating local samples such as BALF and CSF is important for evaluating local immune responses, providing insight into pathology at the primary loci of infection. Serum data alone might be insufficient to elucidate local immune responses and might not enable clinicians to devise the most appropriate treatment strategies.
1. Introduction
Cytokines and chemokines are key factors in the pathogenesis of influenza A virus (IAV) infection in human and animal models. Numerous studies of various IAV strains have revealed the involvement of various cytokines/chemokines in the pathology of this organism [1]. IAV infection causes various diseases in different organs, ranging from pneumonia [2–7] to encephalitis/encephalopathy [8–11]. To devise appropriate treatment strategies, the pathological events occurring at the primary loci of disease must be elucidated. Most reports describe studies that have used serum to investigate the cytokine/chemokine profiles of IAV-infected patients, especially pneumonia patients [2–7]. Nevertheless, it remains unclear whether these serum data correlate with the local immune response against IAV.
This background has prompted us to characterize the inflammatory mediator response in serum compared with local samples, that is, bronchoalveolar lavage fluid (BALF) and cerebrospinal fluid (CSF) in IAV pneumonia and IAV encephalitis mouse models, respectively. This report presents evidence that interpreting a serum cytokine/chemokine profile in terms of a local immune response against IAV infection can be misleading.
2. Materials and Methods
2.1. Ethics
   The Animal Use Committee of the Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences approved this study (number OKU-2012628), which was conducted in accordance with the National Institutes of Health Guidelines.
2.2. Experimental Animals
Eight-week-old male C57BL/6 mice were purchased from Charles River Laboratories Japan Inc. (Yokohama, Japan). The mice were housed in a specific pathogen-free animal facility at 25°C with a 12 hr light/dark cycle. They were fed a standard diet (Oriental MF; Oriental Yeast Co. Ltd., Tokyo, Japan).
2.3. Preparation of Viral Inocula
  Influenza A/WSN/33 (H1N1) virus was kindly provided by the Department of Microbiology, Kawasaki Medical University. This mouse-adapted H1N1 human IAV strain is not only pneumotropic after intranasal inoculation; it is also neurovirulent after inoculation into the CSF [12, 13]. We used this strain throughout this study. The virus was grown in 10-day-old embryonated chicken eggs. The virus titre was quantitated using a plaque assay with Madin-Darby canine kidney cells.