Summary

口咽部气管内PAMP管理和支气管肺泡灌洗的利用率来评估小鼠宿主的免疫反应

Published: April 02, 2014
doi:
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Summary

Abstract

Introduction

Protocol

Representative Results

Discussion

Disclosures

The authors have nothing to disclose.

Acknowledgements

Materials

C57Bl/6JThe Jackson LaboratoryStock 000664
Compact ScaleOhaus Scale Corporation71142845
Small Animal Rectal ThermometerBraintree ScientificTH 5
Rectal Probe for RodentsBraintree ScientificRET 3
Ear PunchBraintree ScientificEP-S 901
Lipopolysaccharide from <em>E. coli</em> 0111:B4InvivoGenLPS-EB
1x Phosphate Buffered SalineLife Technologies10010-023
IsofluraneBaxter40032609
Intratrachael Administration and Lung Inflation StandICAP Manufacturingn/a
Rodent Intubation StandBraintree ScientificRIS 100
Scissors (blunt/sharp)Fisher Scientific13-806-2
forceps (straight)Fisher Scientific22-327-379
forceps (45&ordm;, curved)Fisher Scientific10-275
Scissors (blunt/blunt)Fisher Scientific08-940
Pipette (200 &micro;l Capacity)GilsonF123601
EthanolSigma459844
1 ml SyringeBD Medical301025
10 ml SyringeBD Medical301604
27 G x 0.5 in NeedleBD Medical305109
Refrigerated MicrocentrifugeFisher Scientific13-100-676
1.2 mm Tracheal Cannulae with Luer-adapterHarvard Apparatus732836
Hank's Balanced Salt SolutionLife Technologies14025-076
4-0 Silk Braided Surgical SutureEthiconA183
Luer to Tube Connector KitsHarvard Apparatus721406
Luer Stopcock KitHarvard Apparatus721664
Tygon formula E-3603 Laboratory TubingSigmaR-3603
Formalin Solution, neutral buffered, 10%SigmaHT501128-4L
Mouse IL-1&beta; OptEIA ELISA KitBD Biosciences559603
Mouse IL-6 OptEIA ELISA KitBD Biosciences550950
Mouse TNF-&alpha; OptEIA ELISA KitBD Biosciences560478
HemocytometerHausser Scientific3520
Hemocytometer Cover GlassesThermo Scientific22-021-801
Trypan BlueThermo ScientificSV3008401
Cytology Funnel ClipsFisher Scientific10-357
Cytology FunnelsFisher Scientific10-354
Filter CardsFisher Scientific22-030-410
Microscope SlidesFisher Scientific12-544-1
Cover GlassesFisher Scientific12-540A
Cytospin CytocentrifugeThermo ScientificA78300003
Diff Quick Staining KitFisher Scientific47733150
Permount Mounting MediumFisher ScientificSP15-500

References

  1. Matute-Bello, G., et al. An official American Thoracic Society workshop report: features and measurements of experimental acute lung injury in animals. Am. J. respir. Cell Mol. Biol. 44, 725-738 (2011).
  2. Egger, C., et al. Administration of bleomycin via the oropharyngeal aspiration route leads to sustained lung fibrosis in mice and rats as quantified by UTE-MRI and histology. PloS one. 8, (2013).
  3. Rayamajhi, M., et al. Nonsurgical intratracheal instillation of mice with analysis of lungs and lung draining lymph nodes by flow cytometry. J. Vis. Exp. , (2011).
  4. Revelli, D. A., Boylan, J. A., Gherardini, F. C. A non-invasive intratracheal inoculation method for the study of pulmonary melioidosis. Front. Cell. Infect. Microbiol. 2, 164 (2012).
  5. Allen, I. C., et al. Analysis of NLRP3 in the development of allergic airway disease in mice. J. Immunol. 188, 2884-2893 (2012).
  6. Allen, I. C., et al. Characterization of NLRP12 during the in vivo host immune response to Klebsiella pneumoniae and Mycobacterium tuberculosis. PloS one. , (2013).
  7. Allen, I. C., et al. Expression and function of NPSR1/GPRA in the lung before and after induction of asthma-like disease. Am. J. Physiol. Lung Cell. Mol. Physiol. 291, 1005-1017 (2006).
  8. Kebaier, C., et al. Staphylococcus aureus alpha-hemolysin mediates virulence in a murine model of severe pneumonia through activation of the NLRP3 inflammasome. J. Infect. Dis. 205, 807-817 (2012).
  9. Roberts, R. A., et al. Analysis of the murine immune response to pulmonary delivery of precisely fabricated nano- and microscale particles. PloS one. 8, (2013).
  10. Willingham, S. B., et al. NLRP3 (NALP3, Cryopyrin) facilitates in vivo caspase-1 activation, necrosis, and HMGB1 release via inflammasome-dependent and -independent pathways. J. Immunol. 183, 2008-2015 (2009).
  11. Kline, J. N., et al. Variable airway responsiveness to inhaled lipopolysaccharide. Am. J. Respir. Crit. Med. 160, 297-303 (1999).
  12. Cressman, V. L., Hicks, E. M., Funkhouser, W. K., Backlund, D. C., Koller, B. H. The relationship of chronic mucin secretion to airway disease in normal and CFTR-deficient mice. Am. J. Respir. Cell Mol. Biol. 19, 853-866 (1998).
The Utilization of Oropharyngeal Intratracheal PAMP Administration and Bronchoalveolar Lavage to Evaluate the Host Immune Response in Mice

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Allen, I. C. The Utilization of Oropharyngeal Intratracheal PAMP Administration and Bronchoalveolar Lavage to Evaluate the Host Immune Response in Mice. J. Vis. Exp. (86), e51391, doi:10.3791/51391 (2014).

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