Name: a non-invasive and technically non-intensive method for induction and phenotyping of experimental bacterial pneumonia in mice
Uploaded: 2016-09-28T14:00:00
Description: Watch this Scientific Journal Video about A Non-invasive and Technically Non-intensive Method for Induction and Phenotyping of Experimental Bacterial Pneumonia in Mice at JoVE.com

This work was supported in part by the Intramural Research Program of the National Institutes of Health, National Institute of Environmental Health Sciences (Z01 ES102005).

Bütün deneyler Hayvan Bakımı ve NIEHS Kullanımı Komitesi tarafından incelendikten sonra Hayvan Refahı Yasası ve İnsani Laboratuvar Hayvanları Bakım ve Kullanımına İlişkin ABD Kamu Sağlığı Hizmetleri Politikası&#39;na uygun olarak yapıldı. K. 1. Hazırlık pneumoniae Kültür Dikkat: Bir biyogüvenlik düzeyi 2 (BSL2) başlık veya başka bir BSL2 belirlenmiş alandaki tüm adımları uygulayın ve enstitü BSL2 kurallarına ...

<ol>
	<li>Mizgerd, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+lower+respiratory+tract+infection.">Acute lower respiratory tract infection.</a> N Engl J Med. 358 (7), 716-727 (2008).</li><li>Waterer, G. W., Rello, J., Wunderink, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+community-acquired+pneumonia+in+adults.">Management of community-acquired pneumonia in adults.</a> Am J Respir Crit Care Med. 183 (2), 157-164 (2011).</li><li>Mizgerd, J. P., Skerrett, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+human+pneumonia.">Animal models of human pneumonia.</a> Am J Physiol Lung Cell Mol Physiol. 294 (3), L387-L398 (2008).</li><li>Revelli, D. A., Boylan, J. A., Gherardini, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+non-invasive+intratracheal+inoculation+method+for+the+study+of+pulmonary+melioidosis.">A non-invasive intratracheal inoculation method for the study of pulmonary melioidosis.</a> Front Cell Infect Microbiol. 2, 164 (2012).</li><li>Morales-Nebreda, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intratracheal+administration+of+influenza+virus+is+superior+to+intranasal+administration+as+a+model+of+acute+lung+injury.">Intratracheal administration of influenza virus is superior to intranasal administration as a model of acute lung injury.</a> J Virol Methods. 209, 116-120 (2014).</li><li>Aujla, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-22+mediates+mucosal+host+defense+against+Gram-negative+bacterial+pneumonia.">IL-22 mediates mucosal host defense against Gram-negative bacterial pneumonia.</a> Nat Med. 14 (3), 275-281 (2008).</li><li>Chen, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Th17+cells+mediate+clade-specific,+serotype-independent+mucosal+immunity.">Th17 cells mediate clade-specific, serotype-independent mucosal immunity.</a> Immunity. 35 (6), 997-1009 (2011).</li><li>Draper, D. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ATP-binding+cassette+transporter+G1+deficiency+dysregulates+host+defense+in+the+lung.">ATP-binding cassette transporter G1 deficiency dysregulates host defense in the lung.</a> Am J Respir Crit Care Med. 182 (3), 404-412 (2010).</li><li>Robinson, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influenza+A+exacerbates+Staphylococcus+aureus+pneumonia+by+attenuating+IL-1beta+production+in+mice.">Influenza A exacerbates Staphylococcus aureus pneumonia by attenuating IL-1beta production in mice.</a> J Immunol. 191 (10), 5153-5159 (2013).</li><li>Mizgerd, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+of+neutrophil+recruitment+elicited+by+bacteria+in+the+lungs.">Molecular mechanisms of neutrophil recruitment elicited by bacteria in the lungs.</a> Semin Immunol. 14 (2), 123-132 (2002).</li><li>Balamayooran, G., Batra, S., Fessler, M. B., Happel, K. I., Jeyaseelan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+neutrophil+accumulation+in+the+lungs+against+bacteria.">Mechanisms of neutrophil accumulation in the lungs against bacteria.</a> Am J Respir Cell Mol Biol. 43 (1), 5-16 (2010).</li><li>Quinton, L. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatocyte-specific+mutation+of+both+NF-kappaB+RelA+and+STAT3+abrogates+the+acute+phase+response+in+mice.">Hepatocyte-specific mutation of both NF-kappaB RelA and STAT3 abrogates the acute phase response in mice.</a> J Clin Invest. 122 (5), 1758-1763 (2012).</li><li>Gowdy, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Key+role+for+scavenger+receptor+B-I+in+the+integrative+physiology+of+host+defense+during+bacterial+pneumonia.">Key role for scavenger receptor B-I in the integrative physiology of host defense during bacterial pneumonia.</a> Mucosal Immunol. 8 (3), 559-571 (2015).</li><li>Madenspacher, J. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=p53+Integrates+host+defense+and+cell+fate+during+bacterial+pneumonia.">p53 Integrates host defense and cell fate during bacterial pneumonia.</a> J Exp Med. 210 (5), 891-904 (2013).</li><li>Brain, J. D., Knudson, D. E., Sorokin, S. P., Davis, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulmonary+distribution+of+particles+given+by+intratracheal+instillation+or+by+aerosol+inhalation.">Pulmonary distribution of particles given by intratracheal instillation or by aerosol inhalation.</a> Environ Res. 11 (1), 13-33 (1976).</li><li>Card, J. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender+differences+in+murine+airway+responsiveness+and+lipopolysaccharide-induced+inflammation.">Gender differences in murine airway responsiveness and lipopolysaccharide-induced inflammation.</a> J Immunol. 177 (1), 621-630 (2006).</li><li>Ivanov, I. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+intestinal+Th17+cells+by+segmented+filamentous+bacteria.">Induction of intestinal Th17 cells by segmented filamentous bacteria.</a> Cell. 139 (3), 485-498 (2009).</li><li>Hooper, L. V., Littman, D. R., Macpherson, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interactions+between+the+microbiota+and+the+immune+system.">Interactions between the microbiota and the immune system.</a> Science. 336 (6086), 1268-1273 (2012).</li></ol>

Bakteriyel pnömoni Murin modelleri, pulmoner konak savunma tepkisi içine kritik anlayışlar sağladı, gen hedefleme ve in vivo biyolojik ve farmakolojik girişimlerin ortak oldular. Büyük gelişmeler enfekte hava sahasında 10,11 nötrofillerin işe yöneten kemokinler ve adezyon moleküllerinin anlayışımızda özellikle yapılmıştır. Pnömoni in vivo modelleri, hücre bazlı veya alternatif yaklaşımlar aksine, aynı zamanda endokrin içine anahtar anlayışlar sağlad...

Toplum kökenli pnömoni aşılama iyileştirmeler ve antibiyotik stratejileri 1,2 rağmen son 40 yılda ölüm oranlarında küçük genel değişiklikle, ABD&#39;de enfeksiyon ölümlerin başta gelen nedenidir. halk sağlığı düzeyinde algılanabilir ilerleme olmamasına rağmen, son yıllarda dramatik gelişmeler öne akciğer enfeksiyonu fare modellerinin kullanımı ile mümkün bu adımların çoğu ile, pnömoni moleküler ve hücresel patogenezinin yapılmıştır. farenin genetik izlenebilir, murin ve insan bağışıklık sistemi ve ticari olarak temin edilebilir hale gelmiştir murin hedefli immünolojik reaktifler geniş bir dizi benzerliği bir araya alan hızlı bir ilerleme kolaylaştırmıştır. literatürde tarif bakteriyel pnömoni Fare modelleri genellikle patojen aşılama için dört teknik yolları üzerine dayanıyordu: i) aerozolizasyonuna; ii) geçişsizanasal dağıtım; iii) oral doğum; ve iv) cerrahi intratrakeal enjeksiyon (yani, trakeotomi) 3. Enfeksiyonun tüm yolları avantajları ve dezavantajları 3 var. oronazal florası, genel anestezi için gereksinimleri karışım özellikle, göreceli üst solunum yolunun maruz kalma potansiyeli distal akciğer teslim inokulum teslim patojenlerin lober dağılımı, teknik uzmanlık gereksinimleri ve prosedürel morbidite değişkenliği bu çapında büyük ölçüde değişiklik yaklaşımlar. Yaygın kullanılan oral enfeksiyon teknikleri veya doğrudan laringeal görselleştirme 3-5 altında bir &#39;kör&#39; (non-görüntülenmiştir) yaklaşımı ya vasıtasıyla endotrakeal (translaryngeal) kanül içerir. Her iki yöntem de, sağlam iken, önemli eğitim gerektiren ve aynı zamanda üst solunum travma riski taşımaktadır. Mevcut raporda, w, oral enfeksiyon teknik, yoğun olmayan non-invaziv, ucuz ve hızlı bir yöntem tarifburada bakteriler (verilen örnekte Klebsiella pneumoniae) aspirasyon (yani, inhalasyon) yoluyla akciğerlere iletilir bir anestezi fare orofarenks pipetlenir. Biz ve diğerleri başarıyla 6-9 aspirasyon pnömonisi tekniği kullandık. Bu çok yönlü ve kolayca öğrenilebilir akciğer dağıtım yöntemi, sitokinlere ve başka proteinlere, patojenle ilişkili moleküller (örneğin lipopolisakkarid), hücrelerin (örneğin, adoptif transfer) da dahil olmak üzere akciğer, pek çok ek olmayan kostik maddelerin iletimi için uzatılabilir ve toksinler (örneğin, bleomisin). Önemli bir teknik konuları ele ek olarak, aynı zamanda alt-bakteriyel açıklık ölçümünde (yani, koloni oluşturan birim [CFU] akciğer ve periferik organlarda miktar tayini), lökosit gibi pnömoni sonraki ana cevabın değerlendirilmesi için entegre, kantitatif bir yaklaşım tarif hava sahasında birikimi.

<table border="0" cellpadding="0" cellspacing="0" width="698">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">Klebsiella pneumoniae, serotype 2</td>
			<td style="width:169px;">ATCC</td>
			<td style="width:119px;">43816</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">Tryptic soy broth</td>
			<td style="width:169px;">Becton Dickenson</td>
			<td>211825</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Excel Safelet IV Catheters, 18G x 1 1/4&#34;</td>
			<td>Claflin Medical Equipment</td>
			<td style="width:119px;">MEDC-031122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">Hema 3 Solution 1</td>
			<td style="width:169px;">Fisher</td>
			<td style="width:119px;">23-122-937</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">Hema 3 Solution 2</td>
			<td style="width:169px;">Fisher</td>
			<td style="width:119px;">23-122-952</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">Hema 3 Fixative</td>
			<td style="width:169px;">Fisher</td>
			<td style="width:119px;">23-122-929</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">27&#189; gauge tuberculin syringes</td>
			<td style="width:169px;">Fisher</td>
			<td style="width:119px;">14-826-87</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">Lithium heparin plasma collectors</td>
			<td style="width:169px;">Fisher</td>
			<td style="width:119px;">2675187</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:243px;">L-shaped disposable spreaders</td>
			<td style="width:169px;">Lab Scientific</td>
			<td style="width:119px;">DSC</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:243px;">1x PBS, pH 7.4</td>
			<td style="width:169px;">prepared in-house</td>
			<td style="width:119px;">n/a</td>
			<td>Distilled water (5 L), NaCl (40 g), KCl (1 g), Na2HPO4 (5.75 g), KH2PO4 (1 g)&#160;&#160;&#160;</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:243px;">ACK lysis buffer</td>
			<td style="width:169px;">prepared in-house</td>
			<td style="width:119px;">n/a</td>
			<td>NH4Cl (4.145 g), KHCO3 (0.5 g), EDTA (18.6 mg), bring up to 500 ml with distilled water and pH to 7.4</td>
		</tr>
	</tbody>
</table>

a non-invasive and technically non-intensive method for induction and phenotyping of experimental bacterial pneumonia in mice

Toplum kökenli pnömoni, önemli bir halk sağlığı sorunu olmaya devam etmekle beraber, bakteriyel pnömoni fare modelleri son zamanlarda yatan hücresel ve moleküler patogenezi anlayışımızda önemli preklinik gelişmeler kolaylaştırmıştır. In vivo fare modelleri konak savunma tepkisi entegre fizyolojisini ve dayanıklılığını yakalamak bir şekilde alternatif basitleştirilmiş ex vivo yaklaşımlar ortaya değildir. Çeşitli yöntemler intrapulmoner aerosol içeren farelerde bakterilerin aşılanmasından, Burun içinden tatbikat, &quot;kör&quot; ve görsel koşullar altında oral endotrakeal kanülasyon ve deri endotrakeal kanülasyon için literatürde tarif edilmiştir. Tüm yöntemler göreceli yararları ve sınırlamaları vardır. Bu yazıda, detaylı fare ile aspirasyon (yani, inhalasyon) içeren bakterilerin intratrakeal teslimat için, non-invaziv teknik olarak yoğun olmayan, ucuz ve hızlı bir yöntem tarifise genel anestezi altında orofarenks pipetle bir bulaşıcı inokulum. Bu yöntem olmayan yakıcı biyolojik ve kimyasal maddeler, çok çeşitli akciğer verilmesi için kullanılan, ve hatta solunum prosedürlere az deneyimi olan Laboratuvar öğrenmek nispeten kolay olabilir. Aspirasyon pnömonisi yöntemi açıklayan ek olarak, aynı zamanda, özellikle, in vivo fare akciğer doğal immün yanıtının daha sonra analiz edilmesi için adım adım prosedürleri sağlayan bakteriyel açıklık ve enfekte solunum hücresel bağışıklık tepkisi ölçülmesi için yöntemler. pnömoni değerlendirmesine Bu entegre ve basit bir yaklaşım pulmoner doğuştan gelen bağışıklık üzerine genetik ve çevresel manipülasyon etkisi hızlı ve sağlam değerlendirme için izin verir.

C57BL / 6 fareleri K. 2000 CFU ile enfekte edilmiştir akciğerlere orofaringeal aspirasyon yoluyla pneumoniae 43816 (serotip 2). Bu dozda, fareler genelde klinik belirtiler uyuşukluk, karıştırdı kürk ve% 5-10 (Şekil 2A) kilo kaybı da dahil olmak üzere 12-24 saat sonrası enfeksiyon göstermeye başlar. 48-72 saat sonrası enfeksiyon içinde, farelerin birçok tipik azalmış aktivite ile hunched duruşlar% 20 kilo kaybı ve sonuçların ortala...

Watch this Scientific Journal Video about A Non-invasive and Technically Non-intensive Method for Induction and Phenotyping of Experimental Bacterial Pneumonia in Mice at JoVE.com

A Non-invasive and Technically Non-intensive Method for Induction and Phenotyping of Experimental Bacterial Pneumonia in Mice

Çeşitli yöntemler, farelerde bakteriyel pnömoni modelleme için literatürde tarif edilmiştir. Bu yazıda, orofarinksin pipetle bakteriyel inokulum aspirasyonu (yani, inhalasyon) yoluyla zatürree uyarılması için bir non-invaziv, ucuz, hızlı bir yöntem açıklanmaktadır. akciğer doğal bağışıklık yanıtının değerlendirilmesi için aşağı akış yöntemleri de ayrıntılı olarak açıklanmıştır.

Farelerde Deneysel Bakteriyel Pnömoni İndüksiyonunun ve fenotipleme bir Non-invaziv ve Teknik Olmayan yoğun Yöntem

Although community-acquired pneumonia remains a major public health problem, murine models of bacterial pneumonia have recently facilitated significant ...

The overall goal of this experimental protocol is to provide a non-invasive and technically non-intensive procedure for inducing bacterial pneumonia in mice and the subsequent quantification of the in vivo host defense response in the lungs. This method can help answer key questions in the fields of infectious disease and immunology, about the genes and molecular pathways that control the host response to infection. The main advantage of this lung infection method is its technical simplicity.Which allows even laboratories with little expertise in pulmonary procedures to study the pulmonary innate immune response. In a biosafety level two facility, thaw a glycerol stock of K.pneumoniae and transfer one milliliter of the bacteria into a 500 milliliter culture flask containing 50 milliliters of Tryptic Soy Broth. Incubate the suspension culture overnight at 37 degrees Celsius and 200 to 225 RPM.The next morning, dilute one to five milliliters of the bacterial culture in a new flask containing 50 milliliters of Tryptic Soy Broth, and incubate the culture, in the shaker, at 37 degrees Celsius for an additional 2.5 hours to achieve log phase. At the end of the second incubation, transfer one milliliter of bacteria from the second culture to a 1.5 milliliter tube for centrifugation. Remove the supernatant without disturbing the palette and gently resuspend the bacteria in one milliliter of sterile saline for three separate washes.After the third wash, resuspend the bacteria in another milliliter of sterile saline, and set up log-wise serial dilutions of the inoculum. Measure one milliliter of the one to 10 and one to 100 dilutions in disposable cuvettes, in duplicate, to determine the OD600 of the washed K.pneumoniae. An OD600 of 1.0 is roughly equivalent to four to seven times 10 to the eighth CFUs per milliliter.After calculating the concentration of the bacteria in each dilution, resuspend the desired inoculum CFU dose in 50 to 60 microliters of sterile saline per eight to 12 week old recipient animal. To deliver the bacteria to the animals, draw up the dosing inoculum into a P200 pipette tip, and place the anesthetized mouse in a semi-recumbent supine position, suspended by the maxillary incisors, from a rubber band stretched between the pegs of a slanted Plexiglas board. Using a pair of blunt, non-ridged forceps, gently pull the tongue to the side and deposit the dose into the animal's oral cavity taking care to avoid inducing trauma to the tongue or the oral pharynx.Keeping the tongue retracted, gently occlude the nose with a gloved finger, until the house inhales two to three times. When no liquid is visible in the oral cavity, transfer the mouse from the inoculation board to its cage, in the supine position, to prevent blockage of the nares while the mouse is recovering. Make a longitudinal cut just below the sternum.Then, lifting the sternum with forceps, nick the diaphragm, allowing the lung to fall back into the chest cavity. Continue the incision up through the chest cavity on either side of the vasculature and up through the neck. When the trachea is visible, carefully cut through the middle of the neck and push the tissue to the sides to avoid damaging the surrounding vasculiture.Now nick the trachea about one quarter of the way down from the head, and caudally insert a cannula attached to a one milliliter syringe pre-loaded with one milliliter of PBS into the trachea. Push the volume into the lungs, slowly, allowing all of the lobes to inflate. Then pull the volume back out with the syringe and pool the collected washes in a 15 milliliter tube, on ice.At the appropriate experimental end point, collect the blood from the left ventricle and transfer it into a heparinized tube to avoid clotting. Then harvest all of the lung lobes in a 15 milliliter tube containing five milliliters of PBS, and the spleen in a 15 milliliter tube containing two milliliters of PBS. Next, use individual, disposable homogenizers per sample, to macerate the tissues on ice, followed by serial dilution of the tissue slurries.Plate the dilutions, in duplicate, on separate sides of a single TSA plate, and allow the samples to, briefly, dry. Then invert the plates and culture the samples in a static 37 degrees Celsius incubator overnight, averaging the bacterial colonies from both sides of the plate and from each dilution the next morning. At 2000 CFU of K.pneumoniae, mice, typically, begin to demonstrate clinical symptoms 12 to 24 hours post infection.Within 48 to 72 hours, many of the animals exhibit symptoms of illness and morbidity that are typically preceded by an average of 20 percent weight loss. An LD50 dose, however, allows the detection of both the relatively increased and decreased survival within the experimental group, and may be preferred for longer term studies. Lower respiratory tract infection with K.pneumoniae is characterized by a robust influx of leukocytes into the airway within six hours of infection, that peaks by 24 hours and is mostly represented by neutrophils.Cytokines are detectable in the bronchoalveolar lavage fluid at both 24 and 48 hours post infection with K.pneumoniae, and provide insight into the lung microenvironment and its immune self-recruitment in response to bacterial invasion. The bacterial load in the lung, blood, and spleen, also dramatically increases between 24 and 48 hours of infection. Once mastered, the oropharyngeal aspiration method of lung infection can be done in just one to two minutes per mouse if it is performed properly.While attempting this procedure, it is important to remember not to use an excessive aspiration volume. A 50 to 60 microliter volume typically works well for an eight to 12 week old mouse. Following this procedure, a variety of methods can be performed on the lung and peripheral tissues to allow the quantification of the immune response in pathogen clearance.After watching this video you should have a good understanding of how to induce experimental bacterial pneumonia via oropharyngeal pathogen delivery and to determine the pathogen burden in mice. Don't forget that working with microbes can be hazardous and that basic BSL-2 precautions and practices should always be taken while preforming this procedure.

Research

JoVE Journal

Immunology and Infection

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Lökosit Homing Göç ve non-invaziv Görüntüleme In vivo</em

Two-photon (2P) microscopy is a high resolution imaging technique that has been broadly adapted by biologists.  The value of 2P microscopy is that it ...

İmmünoloji ve Enfeksiyon

Preparation of Mouse Pituitary Immunogen for the Induction of Experimental Autoimmune Hypophysitis

Deneysel Otoimmün hipofizitis İndüksiyon Fare Hipofiz İmmunogen hazırlanması

Autoimmune hypophysitis is a chronic inflammation of the pituitary gland caused or accompanied by autoimmunity1.  It has traditionally been considered a ...

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

SJL Farelerde Deneysel Otoimmün hipofizitis İndüksiyon

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse ...

Using Luciferase to Image Bacterial Infections in Mice

Farelerde Görüntü Bakteriyel Enfeksiyonlar Lusiferaz Kullanımı

Imaging is a valuable technique that can be used to monitor biological processes.  In particular, the presence of cancer cells, stem cells, specific ...

A Method for Labeling Vasculature in Embryonic Mice

Embriyonik Farelerde damarları Etiketleme Bir Yöntem

The establishment of a functional blood vessel network is an essential part of organogenesis, and is required for optimal organ function. For example, in ...

Non-invasive Imaging of Disseminated Candidiasis in Zebrafish Larvae

Zebra balığı Larva yılında Dissemine Kandidiyazis non-invaziv Görüntüleme

Disseminated candidiasis caused by the pathogen Candida albicans is a clinically important problem in hospitalized individuals and is associated with a 30 ...

Non-invasive Optical Imaging of the Lymphatic Vasculature of a Mouse

Bir Mouse Lenfatik damarları Non-invaziv Optik Görüntüleme

The lymphatic  vascular system is an important component of the circulatory system that  maintains fluid homeostasis, provides immune surveillance, and ...

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Non-Invasive Model of Neuropathogenic Escherichia coli Infection in the Neonatal Rat

Neuropathogenic olmayan İnvaziv Modeli<em&gt; Escherichia coli</em&gt; Yenidoğan Sıçan Enfeksiyon

Investigation of the interactions between animal host and bacterial pathogen is only meaningful if the infection model employed replicates the principal ...

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Efektör CD4 Evlatlık transferi ile Kemirgen Bağırsak Enflamasyon indüksiyonu<sup&gt; +</sup&gt; CD45RB<sup&gt; Yüksek</sup&gt; Bağışık yetmezliği olan farelere içine T Hücreleri

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages ...

Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of Streptococcus iniae Infection

Non-invasive Imaging of the Innate Immune Response in a Zebrafish Larval Model of Streptococcus iniae Infection

Bir Zebra balığı Larva Modeli Doğuştan Bağışıklık Tepki Non-invaziv Görüntüleme<em&gt; Streptococcus iniae</em&gt; Enfeksiyon

The aquatic pathogen, Streptococcus iniae, is responsible for over 100 million dollars in annual losses for the aquaculture industry and is capable of ...