气道变态反应性疾病，原发性流感感染，继发性细菌感染的三重小鼠模型

Sean   Roberts; Clare M Williams; Sreeja  Roy; Yoichi  Furuya

doi:10.21769/BioProtoc.3583

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
CN
- EN - English
- CN - 中文

Peer-reviewed

A Triple-challenge Mouse Model of Allergic Airway Disease, Primary Influenza Infection, and Secondary Bacterial Infection

气道变态反应性疾病，原发性流感感染，继发性细菌感染的三重小鼠模型

SR Sean Roberts

CW Clare M Williams

SR Sreeja Roy

YF Yoichi Furuya email

发布: 2020年04月20日第10卷第8期 DOI: 10.21769/BioProtoc.3583 浏览次数: 3571

评审: Kristin L. ShinglerAnonymous reviewer(s)

下载 PDF

Q&A

引用

Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of mBio, featuring study using the protocol.

Jul 2019

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

对EBV 和CMV特异性人CD8+T淋巴细胞的TCRβ克隆型分析

Nening M. Nanlohy [...] Debbie van Baarle

2015年10月05日 7967 阅读

利用多色受激发射损耗（STED）显微镜获取呼吸道合胞体病毒颗粒和感染细胞的高分辨率图像

Masfique Mehedi [...] Ursula J. Buchholz

2017年09月05日 9211 阅读

以小鼠霍乱弧菌感染为模型系统的TetR调控体内抑制技术鉴定基因工程菌的条件基因沉默

Franz G. Zingl [...] Stefan Schild

2020年10月05日 2665 阅读

Abstract

Asthma is a global problem that affects millions of individuals. An increased risk of respiratory viral and bacterial infections is one of the complications of asthma. We recently reported that mice with ovalbumin-induced allergic airway disease (AAD) are protected against influenza-Streptococcus pneumoniae co-infection. Here, we describe in detail a protocol on how to induce AAD and influenza-S. pneumoniae co-infection in mice and to evaluate the specific roles of asthma on immunity to viral and bacterial pathogens in the hope of translating findings to benefit asthmatic individuals.

Keywords: Asthma (哮喘)

Allergic airway disease (气道变态反应性疾病)

Influenza (流感)

Streptococcus pneumoniae (肺炎链球菌)

Co-infection (混合感染)

Background

The global prevalence of individuals with asthma is increasing, with 300 million presently suffering and an additional 100 million new incidences predicted by 2025 (Nunes et al., 2017). Because of the altered immune system, asthmatic individuals are believed to have an increased risk of susceptibility to influenza infection. Seasonal and pandemic influenza infection can result in concurrent bacterial infection which can lead to airway respiratory distress syndrome, a potentially life-threatening condition (Gilca et al., 2011). Whether or not asthmatics are more susceptible and have reduced protective anti-influenza immune responses, is controversial. During the recent 2009 H1N1 pandemic, asthmatics were more likely to be hospitalized due to influenza. However, clinical data also suggest that asthmatics were less likely to die or require ICU admission than those without asthma. The majority of deaths during influenza pandemics are not caused by the viral infection per se, but instead, are due to complications from secondary bacterial infections (Morens et al., 2008; MacIntyre et al., 2018). We previously reported that mice with AAD are resistant to a single infection with influenza virus or S. pneumoniae (Furuya et al., 2015; Sanfilippo et al., 2015). However, there was no murine model that addressed the influence of allergic airway disease (AAD) on immunity to influenza virus and bacterial co-infection. Thus, we have developed a triple challenge model in mice to test the hypothesis that asthmatics are protected from severe influenza because they are less likely to develop severe secondary bacterial pneumonia (Roberts et al., 2019).

Materials and Reagents

1.5 ml conical tubes (Fisher Scientific, catalog number: 05-408-129 ), autoclaved
50 ml tubes (Corning, catalog number: 352070 )
1 ml TB syringes with 25 G needle (BD, catalog number: 309626 )
1.2 ml cryovial (Corning, catalog number: 430487 )
Pipette tips
1,000 μl (VWR, catalog number: 16466-008 )
100 μl (VWR, catalog number: 53510-070 )
10 μl (VWR, catalog number: 89368-974 )
Glass coverslips (Corning, catalog number: 2975245 )
Adult 6- to 8-week-old BALB/c mice (Charles Rivers Bioscience or preferred vendor)
Bacterial stock: S. pneumoniae serotype 3 strain can be obtained from American Tissue Culture Collection (ATCC) (ATCC, catalog number: ATCC^® 6303 ) or S. pneumoniae serotype 2 strain D39 can be obtained from the National Center for Tissue Collection (National Center for Tissue Collection, catalog number: NCTC 7466 )
Influenza A virus (H1N1) can be obtained from ATCC: PR8 (ATCC, catalog number: ATCC VR-1469 ) or the 2009 pandemic virus (ATCC, catalog number: ATCC VR-1894 )
Albumin from chicken egg white (Sigma, catalog number: A2512 )
Blood agar plates (Fisher Scientific, catalog number: R01623 )
Aluminum hydroxide/Rehydragel (Fisher scientific, catalog number: NC1105454 )
1x Phosphate Buffered Saline (PBS) (Fisher Scientific, catalog number: 20-012-027 )
House dust mite (HDM) extract, lyophilized Dermatophagoides pteronyssinus (Stallergenes-Greer, item# B85)
Todd Hewitt broth (BD, catalog number: 249240 )
Glycerol (MilliporeSigma, catalog number: G7893 )
2 mg/ml ovalbumin (OVA) solution (see Recipes)
1 mg/ml HDM stock solution (see Recipes)
S. pneumoniae bacterial stock (see Recipes)

Equipment

250 ml beaker (Corning, catalog number: 1003250 )
1-L flask (Corning, catalog number: 4980-1L )
Stir bar (Fisher scientific, catalog number: 14-513-51 )
E-Z Anesthesia system (Euthanex Corp)
ABSL-2 biosafety cabinet
Pipette set (P20, P200, P1000) (Gilson, catalog number: F167300 )
Isoflurane liquid inhalation (Henry Schein, catalog number: 1182097 )
Centrifuge (Eppendorf, model: 5810R )
Microscope (Olympus, model: BX60 )
Microbiological Incubator (New Brunswick Scientific, Excella E24 incubator shaker series)
Dissection instruments
Forceps (Roboz surgical, catalog number: RS-8120 )
Scissors (Roboz surgical, catalog number: RS-6808 )
-80 °C freezer

Procedure

English

中文翻译

文章信息

版权信息

如何引用

Roberts, S., Williams, C. M., Roy, S. and Furuya, Y. (2020). A Triple-challenge Mouse Model of Allergic Airway Disease, Primary Influenza Infection, and Secondary Bacterial Infection. Bio-protocol 10(8): e3583. DOI: 10.21769/BioProtoc.3583.