This work was supported by National Institutes of Health grants HL105947-01 (YL), HL07829-16 (AM), HL090152 (AM).

The animal experiments were approved by the Animal Care Committee and Institutional Biosafety Committees of the University of Illinois at Chicago.
NOTE: All procedures involving pseudomonas should be performed with Biosafety Level 2 (BSL2) practices, which include but are not limited to: mask, eye protection, gown or jumpsuit, and double gloves. Work in certified Biosafety Cabinet. Treat instruments in contact with bacteria with bleach or chlorine dioxide based disinfectant. Use a sealed box f...

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The authors have nothing to disclose.

The Pseudomonas mouse lung injury model that we describe here mimics the whole process of inflammation, lung injury, repair, and resolution that occur following acute lung injury or pneumonia. It has unique advantages comparing with several other injury models in that it is clinically relevant and relatively safe and easy to handle.
The critical step in the procedure is that the injection of bacteria solution needs to be very slow. If injection is too fast, the mice are likely to die by choke....

Lungs are exposed to environmental pathogens and are susceptible to inflammation and injury1-3. During pathological conditions such as pneumonia or Adult Respiratory Distress Syndrome (ARDS), pathogens as well as inflammatory factors released by leukocytes induce injury and death of alveolar cells1-3. It is important to develop animal models of acute lung injury to facilitate the study of pathology of injury as well as mechanism of repair.
Currently, most people use hyperoxia and bleomycin induced mouse lung injury models4. However, the mechanisms of hyperoxia caused injury are not the same as most common lung injuries that occur during pneumonia or ARDS5. Bleomycin induced acute injury is rare in a clinical context4. Here we report a mouse lung injury model using intra-tracheal injection of P. aeruginosa 6,7. This model is clinically relevant, and mimics the processes that happen following pneumonia8.
As an opportunistic, nosocomial pathogen of immunocompromised individuals, P. aeruginosa typically infects the pulmonary tract, urinary tract, burns, wounds, and also causes other blood infections6. The bacteria release virulence factor exotoxin A, multiply and trigger immune responses6. Intra-trachael administration of P. aeruginosa reflects the situation in human exposure to the bacteria which cause pneumonia and the pathology is likely to be different from the recently reported influenza virus H1N1 induced lung injury model9. Since P. aeruginosa is an opportunistic pathogen, it is relatively safe to handle as compared to some of the more virulent pathogens. &#160;Here we used intra-tracheal injection to administer the bacteria because we observed that this method introduced more bacteria into the distal alveoli region of the lung compared with some other procedures such as using a catheter via mouth.
Compared with other acute lung injury models, the P. aeruginosa model described here is suitable for studying lung injury induced by bacteria and by excessive inflammation. Unlike other animal models that use P. aeruginosa to induce sepsis10,11, here we use intra-tracheal injection of these bacteria to induce localized acute lung injury.

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			<td height="26" style="height:27px;width:215px;">Name</td>
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			<td style="width:145px;">Comments</td>
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			<td style="width:145px;">Fisher</td>
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			<td height="26" style="height:27px;width:215px;">syringe</td>
			<td style="width:145px;">Fisher</td>
			<td style="width:145px;">14823434</td>
			<td style="width:145px;">1 ml</td>
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			<td height="26" style="height:27px;width:215px;">scissors</td>
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			<td style="width:145px;"></td>
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			<td height="26" style="height:27px;width:215px;">forceps</td>
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			<td style="width:145px;"></td>
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			<td height="26" style="height:27px;width:215px;">suture&#160;</td>
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			<td style="width:145px;">NC0147607</td>
			<td style="width:145px;"></td>
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			<td height="26" style="height:27px;">Eye gauge, glove, gown</td>
			<td></td>
			<td></td>
			<td></td>
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			<td height="26" style="height:27px;">Biosafety Cabinet</td>
			<td></td>
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			<td height="26" style="height:27px;">chlorine dioxide based sterilant&#160;</td>
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			<td>Clidox</td>
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			<td height="26" style="height:27px;">sheep blood agar plates&#160;</td>
			<td>Medex supply</td>
			<td>HL-1160</td>
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pseudomonas aeruginosa induced lung injury model

In order to study human acute lung injury and pneumonia, it is important to develop animal models to mimic various pathological features of this disease. Here we have developed a mouse lung injury model by intra-tracheal injection of bacteria Pseudomonas aeruginosa (P. aeruginosa or PA). Using this model, we were able to show lung inflammation at the early phase of injury. In addition, alveolar epithelial barrier leakiness was observed by analyzing bronchoalveolar lavage (BAL); and alveolar cell death was observed by Tunel assay using tissue prepared from injured lungs. At a later phase following injury, we observed cell proliferation required for the repair process. The injury was resolved 7 days from the initiation of P. aeruginosa injection. This model mimics the sequential course of lung inflammation, injury and repair during pneumonia. This clinically relevant animal model is suitable for studying pathology, mechanism of repair, following acute lung injury, and also can be used to test potential therapeutic agents for this disease.

Starting from 24 - 72 hr post P. aeruginosa injection, increased cellularity was observed in lung sections (Figure 1A-D). The lung started to recover from 96 hr post injury (Figure 1E). At 7 days post P. aeruginosa, normal alveoli morphology was largely restored (Figure 1F). Tunnel staining using lung sections prepared at 24 hr post P. aeruginosa showed cell death in alveoli cells (Figure 1G-I). In order to study the repair pro...

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Pseudomonas aeruginosa Induced Lung Injury Model

We have developed a mouse lung injury model by intra-tracheal injection of bacteria Pseudomonas aeruginosa. This model mimics lung injury during pneumonia and is clinically relevant.

Pseudomonas aeruginosa Induced Lung Injury Model

In order to study human acute lung injury and pneumonia, it is important to develop animal models to mimic various pathological features of this disease. ...

pseudomonas-aeruginosa-induced-lung-injury-model

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JoVE Journal

Immunology and Infection

18.3K Görüntüleme.  University of Illinois at Chicago. We have developed a mouse lung injury model by intra-tracheal injection of bacteria Pseudomonas aeruginosa. This model mimics lung injury during pneumonia and is clinically relevant. 