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
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Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
| C57Bl/6J | The Jackson Laboratory | Stock 000664 | |
| Compact Scale | Ohaus Scale Corporation | 71142845 | |
| Small Animal Rectal Thermometer | Braintree Scientific | TH 5 | |
| Rectal Probe for Rodents | Braintree Scientific | RET 3 | |
| Ear Punch | Braintree Scientific | EP-S 901 | |
| Lipopolysaccharide from <em>E. coli</em> 0111:B4 | InvivoGen | LPS-EB | |
| 1x Phosphate Buffered Saline | Life Technologies | 10010-023 | |
| Isoflurane | Baxter | 40032609 | |
| Intratrachael Administration and Lung Inflation Stand | ICAP Manufacturing | n/a | |
| Rodent Intubation Stand | Braintree Scientific | RIS 100 | |
| Scissors (blunt/sharp) | Fisher Scientific | 13-806-2 | |
| forceps (straight) | Fisher Scientific | 22-327-379 | |
| forceps (45º, curved) | Fisher Scientific | 10-275 | |
| Scissors (blunt/blunt) | Fisher Scientific | 08-940 | |
| Pipette (200 µl Capacity) | Gilson | F123601 | |
| Ethanol | Sigma | 459844 | |
| 1 ml Syringe | BD Medical | 301025 | |
| 10 ml Syringe | BD Medical | 301604 | |
| 27 G x 0.5 in Needle | BD Medical | 305109 | |
| Refrigerated Microcentrifuge | Fisher Scientific | 13-100-676 | |
| 1.2 mm Tracheal Cannulae with Luer-adapter | Harvard Apparatus | 732836 | |
| Hank's Balanced Salt Solution | Life Technologies | 14025-076 | |
| 4-0 Silk Braided Surgical Suture | Ethicon | A183 | |
| Luer to Tube Connector Kits | Harvard Apparatus | 721406 | |
| Luer Stopcock Kit | Harvard Apparatus | 721664 | |
| Tygon formula E-3603 Laboratory Tubing | Sigma | R-3603 | |
| Formalin Solution, neutral buffered, 10% | Sigma | HT501128-4L | |
| Mouse IL-1β OptEIA ELISA Kit | BD Biosciences | 559603 | |
| Mouse IL-6 OptEIA ELISA Kit | BD Biosciences | 550950 | |
| Mouse TNF-α OptEIA ELISA Kit | BD Biosciences | 560478 | |
| Hemocytometer | Hausser Scientific | 3520 | |
| Hemocytometer Cover Glasses | Thermo Scientific | 22-021-801 | |
| Trypan Blue | Thermo Scientific | SV3008401 | |
| Cytology Funnel Clips | Fisher Scientific | 10-357 | |
| Cytology Funnels | Fisher Scientific | 10-354 | |
| Filter Cards | Fisher Scientific | 22-030-410 | |
| Microscope Slides | Fisher Scientific | 12-544-1 | |
| Cover Glasses | Fisher Scientific | 12-540A | |
| Cytospin Cytocentrifuge | Thermo Scientific | A78300003 | |
| Diff Quick Staining Kit | Fisher Scientific | 47733150 | |
| Permount Mounting Medium | Fisher Scientific | SP15-500 |
References
- 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).
- 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).
- 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).
- 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).
- Allen, I. C., et al. Analysis of NLRP3 in the development of allergic airway disease in mice. J. Immunol. 188, 2884-2893 (2012).
- Allen, I. C., et al. Characterization of NLRP12 during the in vivo host immune response to Klebsiella pneumoniae and Mycobacterium tuberculosis. PloS one. , (2013).
- 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).
- 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).
- 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).
- 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).
- Kline, J. N., et al. Variable airway responsiveness to inhaled lipopolysaccharide. Am. J. Respir. Crit. Med. 160, 297-303 (1999).
- 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).
Cite This Article
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).