1.A serum-stable branched dimeric anti-VEGF peptide blocks tumor growth via anti-angiogenic activity.
Jung Wook KIM ; Tae Dong KIM ; Bok Sil HONG ; Oh Youn KIM ; Wan Hee YOON ; Chi Bom CHAE ; Yong Song GHO
Experimental & Molecular Medicine 2010;42(7):514-523
Angiogenesis is critical and indispensable for tumor progression. Since VEGF is known to play a central role in angiogenesis, the disruption of VEGF-VEGF receptor system is a promising target for anti-cancer therapy. Previously, we reported that a hexapeptide (RRKRRR, RK6) blocked the growth and metastasis of tumor by inhibiting VEGF binding to its receptors. In addition, dRK6, the D-form derivative of RK6, retained its biological activity with improved serum stability. In the present study, we developed a serum-stable branched dimeric peptide (MAP2-dRK6) with enhanced anti-VEGF and anti-tumor activity. MAP2-dRK6 is more effective than dRK6 in many respects: inhibition of VEGF binding to its receptors, VEGF- and tumor conditioned medium-induced proliferation and ERK signaling of endothelial cells, and VEGF-induced migration and tube formation of endothelial cells. Moreover, MAP2-dRK6 blocks in vivo growth of VEGF-secreting colorectal cancer cells by the suppression of angiogenesis and the subsequent induction of tumor cell apoptosis. Our observations suggest that MAP2-dRK6 can be a prospective therapeutic molecule or lead compound for the development of drugs for various VEGF-related angiogenic diseases.
Amino Acid Sequence
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Angiogenesis Inhibitors/*pharmacology
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Animals
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Cell Movement/drug effects
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Cell Proliferation/drug effects
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Colorectal Neoplasms/*pathology/secretion
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Endothelial Cells/cytology/drug effects/enzymology
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Enzyme Activation/drug effects
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Extracellular Signal-Regulated MAP Kinases/metabolism
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Humans
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Mice
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Mice, Nude
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Molecular Sequence Data
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Neovascularization, Pathologic/pathology/prevention & control
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Neovascularization, Physiologic/drug effects
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Peptides/chemistry/*pharmacology
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Protein Multimerization/*drug effects
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Protein Stability/drug effects
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Rats
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Serum
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Vascular Endothelial Growth Factor A/*antagonists & inhibitors/secretion
2.Vaccination with Klebsiella pneumoniae-derived extracellular vesicles protects against bacteria-induced lethality via both humoral and cellular immunity.
Won Hee LEE ; Hyun Il CHOI ; Sung Wook HONG ; Kwang Sun KIM ; Yong Song GHO ; Seong Gyu JEON
Experimental & Molecular Medicine 2015;47(9):e183-
The emergence of multidrug-resistant Klebsiella pneumoniae highlights the need to develop preventive measures to ameliorate Klebsiella infections. Bacteria-derived extracellular vesicles (EVs) are spherical nanometer-sized proteolipids enriched with outer membrane proteins. Gram-negative bacteria-derived EVs have gained interest for use as nonliving complex vaccines. In the present study, we evaluated whether K. pneumoniae-derived EVs confer protection against bacteria-induced lethality. K. pneumoniae-derived EVs isolated from in vitro bacterial culture supernatants induced innate immunity, including the upregulation of co-stimulatory molecule expression and proinflammatory mediator production. EV vaccination via the intraperitoneal route elicited EV-reactive antibodies and interferon-gamma-producing T-cell responses. Three vaccinations with the EVs prevented bacteria-induced lethality. As verified by sera and splenocytes adoptive transfer, the protective effect of EV vaccination was dependent on both humoral and cellular immunity. Taken together, these findings suggest that K. pneumoniae-derived EVs are a novel vaccine candidate against K. pneumoniae infections.
Animals
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Bacterial Vaccines/immunology/*microbiology/*therapeutic use
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Extracellular Vesicles/immunology/*microbiology
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Female
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Humans
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Immunity, Cellular
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Immunity, Innate
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Interferon-gamma/immunology
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Klebsiella Infections/immunology/*prevention &control
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Klebsiella pneumoniae/*immunology
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Mice, Inbred C57BL
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Vaccination
3.Acetyl salicylic acid inhibits Th17 airway inflammation via blockade of IL-6 and IL-17 positive feedback.
Hyung Geun MOON ; Chil Sung KANG ; Jun Pyo CHOI ; Dong Sic CHOI ; Hyun Il CHOI ; Yong Wook CHOI ; Seong Gyu JEON ; Joo Yeon YOO ; Myoung Ho JANG ; Yong Song GHO ; Yoon Keun KIM
Experimental & Molecular Medicine 2013;45(1):e5-
T-helper (Th)17 cell responses are important for the development of neutrophilic inflammatory disease. Recently, we found that acetyl salicylic acid (ASA) inhibited Th17 airway inflammation in an asthma mouse model induced by sensitization with lipopolysaccharide (LPS)-containing allergens. To investigate the mechanism(s) of the inhibitory effect of ASA on the development of Th17 airway inflammation, a neutrophilic asthma mouse model was generated by intranasal sensitization with LPS plus ovalbumin (OVA) and then challenged with OVA alone. Immunologic parameters and airway inflammation were evaluated 6 and 48 h after the last OVA challenge. ASA inhibited the production of interleukin (IL)-17 from lung T cells as well as in vitro Th17 polarization induced by IL-6. Additionally, ASA, but not salicylic acid, suppressed Th17 airway inflammation, which was associated with decreased expression of acetyl-STAT3 (downstream signaling of IL-6) in the lung. Moreover, the production of IL-6 from inflammatory cells, induced by IL-17, was abolished by treatment with ASA, whereas that induced by LPS was not. Altogether, ASA, likely via its acetyl moiety, inhibits Th17 airway inflammation by blockade of IL-6 and IL-17 positive feedback.
Animals
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Aspirin/pharmacology/*therapeutic use
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Cell Polarity/drug effects/immunology
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Feedback, Physiological/*drug effects
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Interferon-gamma/deficiency/metabolism
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Interleukin-17/*metabolism/pharmacology
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Interleukin-6/biosynthesis/*metabolism
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Lipopolysaccharides/pharmacology
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Lung/drug effects/metabolism/pathology
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Mice
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Mice, Inbred C57BL
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Pneumonia/*drug therapy/*immunology/pathology
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Th17 Cells/drug effects/*immunology/pathology
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Transforming Growth Factor beta1/pharmacology
4.Protective effects of basic fibroblast growth factor in the development of emphysema induced by interferon-gamma.
Byung Jae LEE ; Hyung Geun MOON ; Tae Seop SHIN ; Seong Gyu JEON ; Eun Young LEE ; Yong Song GHO ; Chun Geun LEE ; Zhou ZHU ; Jack A ELIAS ; Yoon Keun KIM
Experimental & Molecular Medicine 2011;43(4):169-178
Recent clinical evidence indicates that the non-eosinophilic subtype of severe asthma is characterized by fixed airway obstruction, which may be related to emphysema. Transgenic studies have demonstrated that high levels of IFN-gamma in the airways induce emphysema. Fibroblast growth factor 2 (FGF2), which is the downstream mediator of TGF-beta, is important in wound healing. We investigated the role of FGF2 in IFN-gamma-induced emphysema and the therapeutic effects of recombinant FGF2 in the prevention of emphysema in a severe non-eosinophilic asthma model. To evaluate the role of FGF2 in IFN-gamma-induced emphysema, lung targeted IFN-gamma transgenic mice were cross-bred with FGF2-deficient mice. A severe non-eosinophilic asthma model was generated by airway application of LPS-containing allergens twice a week for 4 weeks. To evaluate protective effects of FGF2, recombinant FGF2 (10 microg) was injected subcutaneously during allergen challenge in the severe asthma model. We found that non-eosinophilic inflammation and emphysema induced by transgenic overexpression of IFN-gamma in the airways were aggravated by the absence of FGF2. Airway challenge with LPS-containing allergens induced more inflammation in mice sensitized with LPS-containing allergens compared to challenge with allergens alone. In addition, LPS-induced lung inflammation and emphysema depended on IFN-gamma but not on IL-13. Interestingly, emphysema in the severe asthma model was significantly inhibited by treatment with recombinant FGF2 during allergen challenge, whereas lung inflammation was unaffected. Therefore, our present data suggest that FGF2 may help protect against IFN-gamma-induced emphysema, and that recombinant FGF2 may help lessen the severity of emphysema.
Animals
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Asthma/drug therapy/*prevention & control
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Bronchoalveolar Lavage Fluid
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Disease Models, Animal
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Emphysema/drug therapy/*prevention & control
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Enzyme-Linked Immunosorbent Assay
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Fibroblast Growth Factor 2/deficiency/*metabolism/*therapeutic use
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Flow Cytometry
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Inflammation/immunology
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Interferon-gamma/*biosynthesis/genetics
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Interleukin-13
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Lipopolysaccharides/administration & dosage/pharmacology
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Pulmonary Eosinophilia
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Recombinant Proteins/administration & dosage/therapeutic use
5.Aspirin attenuates the anti-inflammatory effects of theophylline via inhibition of cAMP production in mice with non-eosinophilic asthma.
Hyung Geun MOON ; You Sun KIM ; Jun Pyo CHOI ; Dong Sic CHOI ; Chang Min YOON ; Seong Gyu JEON ; Yong Song GHO ; Yoon Keun KIM
Experimental & Molecular Medicine 2010;42(1):47-60
Theophylline is commonly used to treat severe asthma and chronic obstructive pulmonary disease (COPD) characterized by non-eosinophilic inflammation. Acetyl salicylic acid (ASA) is one of the most widely used medications worldwide, but up to 20% of patients with asthma experience aggravated respiratory symptoms after taking ASA. Here we evaluated the adverse effect of ASA on the therapeutic effect of theophylline in mice with non-eosinophilic asthma. A non-eosinophilic asthma mouse model was induced by airway sensitization with lipopolysaccharide-containing allergen and then challenged with allergen alone. Therapeutic intervention was performed during allergen challenge. Theophylline inhibited lung inflammation partly induced by Th1 immune response. ASA attenuated the beneficial effects of theophylline. However, co-administration of the ASA metabolite salicylic acid (SA) showed no attenuating effect on theophylline treatment. The therapeutic effect of theophylline was associated with increase in cAMP levels, which was blocked by co-treatment of theophylline and ASA. ASA co-treatment also attenuated the anti-inflammatory effects of a specific phosphodiesterase 4 inhibitor. These results demonstrate that ASA reverses anti-inflammatory effects of theophylline, and that ASA exerts its adverse effects through the inhibition of cAMP production. Our data suggest that ASA reverses lung inflammation in patients taking theophylline, although clinical evidence will be needed.
Animals
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Anti-Inflammatory Agents/*therapeutic use
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Aspirin/*therapeutic use
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Asthma/*drug therapy/*metabolism
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Blotting, Western
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Bronchoalveolar Lavage Fluid
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Cyclic AMP/*metabolism
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Enzyme-Linked Immunosorbent Assay
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Immunoprecipitation
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Pulmonary Eosinophilia/*drug therapy/metabolism
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Theophylline/*therapeutic use
6.Role of inducible nitric oxide synthase on the development of virus-associated asthma exacerbation which is dependent on Th1 and Th17 cell responses.
Tae Seop SHIN ; Byung Jae LEE ; You Me TAE ; You Sun KIM ; Seong Gyu JEON ; Yong Song GHO ; Dong Chull CHOI ; Yoon Keun KIM
Experimental & Molecular Medicine 2010;42(10):721-730
Asthma is characterized by airway inflammation induced by immune dysfunction to inhaled antigens. Although respiratory viral infections are the most common cause of asthma exacerbation, immunologic mechanisms underlying virus-associated asthma exacerbation are controversial. Clinical evidence indicates that nitric oxide (NO) levels in exhaled air are increased in exacerbated asthma patients compared to stable patients. Here, we evaluated the immunologic mechanisms and the role of NO synthases (NOSs) in the development of virus-associated asthma exacerbation. A murine model of virus-associated asthma exacerbation was established using intranasal challenge with ovalbumin (OVA) plus dsRNA for 4 weeks in mice sensitized with OVA plus dsRNA. Lung infiltration of inflammatory cells, especially neutrophils, was increased by repeated challenge with OVA plus dsRNA, as compared to OVA alone. The neutrophilic inflammation enhanced by dsRNA was partly abolished in the absence of IFN-gamma or IL-17 gene expression, whereas unaffected in the absence of IL-13. In terms of the roles of NOSs, dsRNA-enhanced neutrophilic inflammation was significantly decreased in inducible NOS (iNOS)-deficient mice compared to wild type controls; in addition, this phenotype was inhibited by treatment with a non-specific NOS inhibitor (L-NAME) or an specific inhibitor (1400 W), but not with a specific endothelial NOS inhibitor (AP-CAV peptide). Taken together, these findings suggest that iNOS pathway is important in the development of virus-associated exacerbation of neutrophilic inflammation, which is dependent on both Th1 and Th17 cell responses.
Animals
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Asthma/*immunology/virology
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Imines/pharmacology
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Mice
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Mice, Inbred BALB C
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NG-Nitroarginine Methyl Ester/pharmacology
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Nitric Oxide Synthase Type II/antagonists & inhibitors/*metabolism
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RNA, Double-Stranded/metabolism
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Th1 Cells/*immunology
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Th17 Cells/*immunology
7.IL-12-STAT4-IFN-gamma axis is a key downstream pathway in the development of IL-13-mediated asthma phenotypes in a Th2 type asthma model.
You Sun KIM ; Seng Jin CHOI ; Jun Pyo CHOI ; Seong Gyu JEON ; Sun Young OH ; Byung Jae LEE ; Yong Song GHO ; Chun Geun LEE ; Zhou ZHU ; Jack A ELIAS ; Yoon Keun KIM
Experimental & Molecular Medicine 2010;42(8):533-546
IL-4 and IL-13 are closely related cytokines that are produced by Th2 cells. However, IL-4 and IL-13 have different effects on the development of asthma phenotypes. Here, we evaluated downstream molecular mechanisms involved in the development of Th2 type asthma phenotypes. A murine model of Th2 asthma was used that involved intraperitoneal sensitization with an allergen (ovalbumin) plus alum and then challenge with ovalbumin alone. Asthma phenotypes, including airway-hyperresponsiveness (AHR), lung inflammation, and immunologic parameters were evaluated after allergen challenge in mice deficient in candidate genes. The present study showed that methacholine AHR and lung inflammation developed in allergen-challenged IL-4-deficient mice but not in allergen-challenged IL-13-deficient mice. In addition, the production of OVA-specific IgG2a and IFN-gamma-inducible protein (IP)-10 was also impaired in the absence of IL-13, but not of IL-4. Lung-targeted IFN-gamma over-expression in the airways enhanced methacholine AHR and non-eosinophilic inflammation; in addition, these asthma phenotypes were impaired in allergen-challenged IFN-gamma-deficient mice. Moreover, AHR, non-eosinophilic inflammation, and IFN-gamma expression were impaired in allergen-challenged IL-12Rbeta2- and STAT4-deficient mice; however, AHR and non-eosinophilic inflammation were not impaired in allergen-challenged IL-4Ralpha-deficient mice, and these phenomena were accompanied by the enhanced expression of IL-12 and IFN-gamma. The present data suggest that IL-13-mediated asthma phenotypes, such as AHR and non-eosinophilic inflammation, in the Th2 type asthma are dependent on the IL-12-STAT4-IFN-gamma axis, and that these asthma phenotypes are independent of IL-4Ralpha-mediated signaling.
Allergens/immunology
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Animals
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Asthma/complications/*immunology/pathology/physiopathology
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Bronchial Hyperreactivity/complications/immunology/pathology
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Disease Models, Animal
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Interferon-gamma/*immunology
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Interleukin-12/*immunology
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Interleukin-12 Receptor beta 2 Subunit/metabolism
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Interleukin-13/deficiency/*immunology
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Interleukin-4/deficiency
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Methacholine Chloride
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Mice
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Mice, Transgenic
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Models, Immunological
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Organ Specificity
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Pneumonia/complications/immunology/pathology
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Receptors, Cell Surface/metabolism
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STAT4 Transcription Factor/*metabolism
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Signal Transduction/*immunology
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Th2 Cells/*immunology