Editor-Communicated Paper

Free Access

The Role of Osteopontin in the Development of Granulomatous Lesions in Lung

Satoru Chiba

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Department of Cardiovascular Medicine, Hokkaido University Medical School, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Mohammod Mizanur Rashid,

Mohammod Mizanur Rashid

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Hiroshi Okamoto,

Hiroshi Okamoto

Department of Cardiovascular Medicine, Hokkaido University Medical School, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Hirotake Shiraiwa,

Hirotake Shiraiwa

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Shigeyuki Kon,

Shigeyuki Kon

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Masahiro Maeda,

Masahiro Maeda

Immuno-Biological Laboratories, Fujioka, Gunma, 375-0005 Japan

Search for more papers by this author

Masaaki Murakami,

Masaaki Murakami

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Manabu Inobe,

Manabu Inobe

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Akira Kitabatake,

Akira Kitabatake

Department of Cardiovascular Medicine, Hokkaido University Medical School, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Ann F. Chambers,

Ann F. Chambers

The London Regional Cancer Centre, London, Ontario, Canada

Search for more papers by this author

Toshimitsu Uede,

Corresponding Author

Toshimitsu Uede

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Address correspondence to Dr. Toshimitsu Uede, Institute of Immunological Science, Hokkaido University, Kita-15, Nishi-7, Kita-ku, Sapporo, Hokkaido 060-0815, Japan. Fax and Phone: 81-11-736-9836. E-mail: [email protected].Search for more papers by this author

Satoru Chiba,

Satoru Chiba

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Department of Cardiovascular Medicine, Hokkaido University Medical School, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Mohammod Mizanur Rashid,

Mohammod Mizanur Rashid

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Hiroshi Okamoto,

Hiroshi Okamoto

Department of Cardiovascular Medicine, Hokkaido University Medical School, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Hirotake Shiraiwa,

Hirotake Shiraiwa

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Shigeyuki Kon,

Shigeyuki Kon

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Masahiro Maeda,

Masahiro Maeda

Immuno-Biological Laboratories, Fujioka, Gunma, 375-0005 Japan

Search for more papers by this author

Masaaki Murakami,

Masaaki Murakami

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Manabu Inobe,

Manabu Inobe

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Akira Kitabatake,

Akira Kitabatake

Department of Cardiovascular Medicine, Hokkaido University Medical School, Sapporo, Hokkaido, 060-0815 Japan

Search for more papers by this author

Ann F. Chambers,

Ann F. Chambers

The London Regional Cancer Centre, London, Ontario, Canada

Search for more papers by this author

Toshimitsu Uede,

Corresponding Author

Toshimitsu Uede

Section of Immunopathogenesis, Institute of Immunological Science, Hokkaido University, Sapporo, Hokkaido, 060-0815 Japan

First published: 14 November 2013

https://doi.org/10.1111/j.1348-0421.2000.tb02501.x

Citations: 18

Share a link

Email
Wechat
Bluesky

Abstract

Osteopontin (OPN) has been shown to be expressed by cells in granulomas of various origins, but whether it plays a functional role in granuloma formation is not known. Here we used a cardiomyopathic hamster (T02) model, to test the hypothesis that OPN contributes functionally to granuloma development. We immunized cardiomyopathic and normal hamsters by subcutaneous injection of bovine serum albumin in complete Freund's adjuvant, and assessed various tissues for both OPN RNA expression and granuloma formation. Cardiomyopathic hamsters expressed OPN, and formed granulomatous lesions, in heart tissue in both immunized and untreated animals. In addition, immunization induced expression of OPN in lung and lymph nodes of cardiomyopathic (but not normal) hamsters, and also induced granuloma formation in these organs. To test whether OPN expression could play a functional role in inducing granulomas, we produced an adenoviral vector containing the murine OPN gene, and introduced this vector intratracheally into the lungs of normal hamsters. The OPN-containing vector, but not the control vector, induced pulmonary granuloma formation. These studies provided direct in vivo evidence that OPN can contribute functionally to the formation of granulomatous lesions, and suggest that OPN expression may be a common factor involved in formation of granulomas of various origin.

Abbreviations

BAL: bronchoalveolar lavage
BALF: BAL fluid
BSA-CFA: bovine serum albumin in complete Freund's adjuvant
DIG: digoxigenin
ELISA: enzyme linked immunoabsorbent assey
ETA-1: early T-lymphocyte acivation antigen-1
GM-CSF: granulocyte/macrophage colony stimulating factor
GST: glutathione S transferase
HE: hematoxylin and eosin
IFN-γ: interferon gamma
IL: interleukin
MCP-1: monocyte chemoattractant protein-1
MIP-1α: macrophage inflammatory protein la
NO: nitric oxide
NOS: nitric oxide synthase
OPN: osteopontin
PCR: polymerase chain reaction
PFU: plaque forming unit
PPD: protein purified derivatives
RT: reverse transcriptase
SDS: sodium dodecyl sulfate
TGF-β: tranforming growth factor beta.

References

1Ashkar, S., Weber, G.F., Panoutsakopoulou, V., Sanchirico, M.E., Jansson, M., Zawaideh, S., Rittling S.R., Denhardt, D.T., Glimcher, M.J., and Cantor, H. 2000. Eta-1 (ostopontin): an early component of type-1 (cell-mediated) immunity. Science 287: 860–864.
10.1126/science.287.5454.860
CAS PubMed Web of Science® Google Scholar
2Bajusz, E., Baker, J.R., Nixon, C.W., and Homburger, F. 1969. Spontaneous hereditary myocardial degeneration and congestive heart failure in a strain of Syrian hamsters. Ann. N.Y. Acad. Sci. 156: 105–129.
10.1111/j.1749-6632.1969.tb16721.x
CAS PubMed Web of Science® Google Scholar
3Brown, L.F., Berse, B., Van De Water, L., Papadopoulos Sergiou, A., Perruzzi, C.A., Manseau, E.J., Dvorak, H.F., and Senger, D.R. 1992. Expression and distribution of osteopontin in human tissues: widespread association with luminal epithelial surfaces. Mol. Biol. Cell. 3: 1169–1180.
10.1091/mbc.3.10.1169
CAS PubMed Web of Science® Google Scholar
4Braciak, T.A., Mittal, S.K., Graham, F.L., Richards, C.D., and Gauldie, J. 1993. Construction of recombinant human type 5 adenoviruses expressing rodent IL-6 genes: an approach to investigate in vivo cytokine function. J. Immunol. 151: 5145–5153.
CAS PubMed Web of Science® Google Scholar
5Burbach, J.A., Langworthy, T.A., Schlenker, E.H., and Frost Stotz, P.M. 1985. Lipidosis of pulmonary macrophages in the dystrophic hamster. Exp. Lung. Res. 9: 99–117.
10.3109/01902148509061531
CAS PubMed Web of Science® Google Scholar
6Carlson, I., Tognazzi, K., Manseau, E.J., Dvorak, H.F., and Brown, L.F. 1997. Osteopontin is strongly expressed by histiocytes in granulomas of diverse etiology. Lab. Invest. 77: 103–198.
CAS PubMed Web of Science® Google Scholar
7Chensue, S.W., Warmington, K.S., Hershey, S.D., Terebuh, P.D., Othmann, M., and Kunkel, S.L. 1993. Evolving T cell responses in murine schistosomiasis: Th2 cells mediate secondary granulomatous hypersensitivity and are regulated by CD8+ T cells in vivo. J. Immunol. 151: 1391–1400.
CAS PubMed Web of Science® Google Scholar
8Chensue, S.W., Warmington, K., Ruth, J., Lincoln, P., Kuo, M.C., and Kunkel, S.L. 1994. Cytokine responses during mycobacterial and schistosomal antigen-induced pulmonary granuloma formation: production of Th1 and Th2 cytokines and relative contribution of tumor necrosis factor. Am. J. Pathol. 145: 1105–1113.
CAS PubMed Web of Science® Google Scholar
9Craig, A.M., Smith, J.H., and Denhardt, D.T. 1989. Osteopontin, a transformation-associated cell adhesion phosphoprotein, is induced by 12-O-tetradecanoylphorbol-13 acetate in mouse epidermis. J. Biol. Chem. 264: 9682–9689.
CAS PubMed Web of Science® Google Scholar
10Denhardt, D.T., and Guo, X. 1993. Osteopontin: a protein with diverse functions. FASEB J. 7: 1475–1482.
10.1096/fasebj.7.15.8262332
CAS PubMed Web of Science® Google Scholar
11Gharaee Kermani, M., Denholm, E.M., and Phan, S.H. 1996. Costimulation of fibroblast collagen and transforming growth factor β1 gene expression by monocyte chemoatractant protein-1 via specific receptors. J. Biol. Chem. 271: 17779–17784.
10.1074/jbc.271.30.17779
CAS PubMed Web of Science® Google Scholar
12Giachelli, C.M., Lombardi, D., Johnson, R.J., Murry, C.E., and Almeida, M. 1998. Evidence for a role of osteopontin in macrophage infiltration in response to pathological stimuli in vivo. Am. J. Pathol. 152: 353–358.
CAS PubMed Web of Science® Google Scholar
13Grzych, J.M., Pearce, E., Cheever, A., Caulada, Z.A., Caspar, P., Heiny, S., Lewis, F., and Sher, A. 1991. Egg deposition in the major stimulus for the production of Th2 cytokines in murine Schistosmiasis mansoni. J. Immunol. 146: 1322–1327.
CAS PubMed Web of Science® Google Scholar
14Hogaboam, C.M., Chensue, S.W., Steinhauser, M.L., Huffnagle, G.B., Lukacs, N.W., Strieter, R.M., and Kunkel, S.L. 1997. Alteration of the cytokine phenotype in an experimental lung granuloma model by inhibiting nitric oxide. J. Immunol. 159: 5585–5593.
CAS PubMed Web of Science® Google Scholar
15Hogaboam, C.M., Gallinat, C.S., Bone Larson, C., Chensue, S.W., Lukacs, N.W., Strieter, R.M., and Kunkel, S.L. 1998. Collagen deposition in a non-fibrotic lung granuloma model after nitric oxide inhibition. Am. J. Pathol. 153: 1861–1872.
10.1016/S0002-9440(10)65700-8
CAS PubMed Web of Science® Google Scholar
16Hotta, H., Kon, S., Katagiri, Y.U., Tosa, N., Tsukamoto, T., Chambers, A.F., and Uede, T. 1999. Detection of various epitopes of murine osteopontin by monoclonal antibodies. Biochem. Biophys. Res. Commun. 257: 6–11.
10.1006/bbrc.1999.0412
CAS PubMed Web of Science® Google Scholar
17Hwang, S.M., Lopez, C.A., Heck, D.E., Gardner, C.R., Laskin, D.L., Laskin, J.D., and Denhardt, D.T. 1994. Osteopontin inhibits induction of nitric oxide synthetase gene expression by inflammatory mediators in mouse kidney epithelial cells. J. Biol. Chem. 269: 711–715.
CAS PubMed Web of Science® Google Scholar
18Inobe, M., Aoki, N., Linsley, P.S., Ledbetter, J.A., Abe, R., Murakami, M., and Uede, T. 1996. The role of the B7–1a molecule, an alternatively spliced form of murine B7–1 (CD80), on T cell activation. J. Immunol. 157: 582–588.
CAS PubMed Web of Science® Google Scholar
19Katagiri, Y.U., Murakami, M., Mori, K., Iizuka, J., Hara, T., Tanaka, K., Jia, W.Y., Chambers, A.F., and Uede, T. 1996. Non-RGD domains of osteopontin promote cell adhesion without involving αv integrins. J. Cell. Biochem. 62: 123–131.
10.1002/(SICI)1097-4644(199607)62:1<123::AID-JCB13>3.0.CO;2-O
CAS PubMed Google Scholar
20Katagiri, Y.U., Sleeman, J., Fujii, H., Herrlich, P., Hotta, H., Tanaka, K., Chikuma, S., Yagita, H., Okumura, K., Murakami, M., Saiki, I., Chambers, A.F., and Uede, T. 1999. CD44 variants but not CD44 cooperate with β1-containing integrins to permit cells to bind to osteopontin independently of arginie-glycine-aspartic acid, thereby stimulating cell motility and chemotaxis. Cancer Res. 59: 219–226.
CAS PubMed Web of Science® Google Scholar
21Kawashima, R., Mochida, S., Matsui, A., YouLuTuZ, Y., Ishikawa, K., Toshima, K., Yamanobe, F., Inao, M., Ikeda, H., Ohno, A., Nagoshi, S., Uede, T., and Fujiwara, K. 1999. Expression of osteopontin in kuppfer cells and hepatic macrophages and stellate cells in rat liver after carbon tetrachloride intoxication: a possible factor for macrophage migration into hepatic necrotic areas. Biochem. Biophys. Res. Commun. 256: 527–531.
10.1006/bbrc.1999.0372
CAS PubMed Web of Science® Google Scholar
22Kerr, J.M., Fisher, L.W., Termine, J.D., and Young, M.F 1991. The cDNA cloning and RNA distribution of bovine osteopontin. Gene 108: 237–243.
10.1016/0378-1119(91)90439-I
CAS PubMed Web of Science® Google Scholar
23Kunkel, S.L., Chensue, S.W., Strieter, R.M., Lynch, J.P., and Remick, D.G. 1989. Cellular and molecular aspects of granulomatous inflammation. Am. J. Respir. Cell. Mol. Biol. 1: 439–447.
10.1165/ajrcmb/1.6.439
CAS PubMed Web of Science® Google Scholar
24Mori, K., Kobayashi, S., Inobe, M., Jia, W.Y., Tamakoshi, M., Miyazaki, T., and Uede, T. 1994. In vivo cytokine expression in various T cell subsets of the autoimmune MRL-lpr/lpr mouse. Autoimmunity 17: 49–57.
10.3109/08916939409014658
PubMed Web of Science® Google Scholar
25Nakagawa, I., Murakami, M., Ijima, K., Chikuma, S., Saito, I., Kanegae, Y., Ishikura, H., Yoshiki, T., Okamoto, H., Kitabatake, A., and Uede, T. 1998. Persistent and secondary adnovirus-mediated hepatic gene expression using adenovirus vector containing CTLA4IgG. Human Gene Therapy 9: 1739–1745.
10.1089/hum.1998.9.12-1739
CAS PubMed Web of Science® Google Scholar
26Nau, G.J., Guilfoile, P., Chupp, G.L., Berman, J.S., Kim, S.J., Kornfeld, H., and Young, R.A. 1997. A chemoattractant cytokine associated with granulomas in tuberculosis and silicosis. Proc. Natl. Acad. Sci. U.S.A. 94: 6414–6419.
10.1073/pnas.94.12.6414
CAS PubMed Web of Science® Google Scholar
27Nigro, V., Okazaki, Y., Belstito, A., Piluso, G., Matsuda, Y., Politano, L., Nigro, G., Ventura, C., Abbondanza, C., Molinari, A.M., Acampora, D., Nishimura, M., Hayashizaki, Y., and Puca, G.A. 1997. Identification of the Syrian hamster cardiomyopathy gene. Human Mol. Genet. 6: 601–607.
10.1093/hmg/6.4.601
CAS PubMed Web of Science® Google Scholar
28Noda, M., Yoon, K., Prince, C.W., Butler, W.T., and Rodan, G.A. 1988. Transcriptional regulation of osteopontin production in rat osteosarcoma cells by type b transforming growth factor. J. Biol. Chem. 263: 13916–13921.
CAS PubMed Web of Science® Google Scholar
29Oldberg, A., Franzen, A., and Heinegard, D. 1986. Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence. Proc. Natl. Acad. Sci. U.S.A. 83: 8819–8823.
10.1073/pnas.83.23.8819
CAS PubMed Web of Science® Google Scholar
30O'Regan, A.W., Chupp, G.L., Lowry, J.A., Goetschkes, M., Mulligan, N., and Berman, J.S. 1999. Osteopontin is associated with T cells in sarcoid granulomas and has T cell adhesive and cytokine-like properties in vitro. J. Immunol. 162: 1024–1031.
10.4049/jimmunol.162.2.1024
CAS PubMed Web of Science® Google Scholar
31Patarca, R. Freeman, G.J. Singh, R.P. Wei, F.W. Durfee, T. Blattner, F. Regnier, D.C. Kozak, C.A. Mock, B.A. Morse, C. A. III, Jerrells, T.R. and Cantor, H. 1989. Structural and functional studies of the Eta-1 (early-T-lymphocyte-activation-1) gene: definition of a novel T cell-dependent response associated with genetic resistance to bacterial infection. J. Exp. Med. 170: 145–161.
10.1084/jem.170.1.145
CAS PubMed Web of Science® Google Scholar
32Patarca, R., Saavedra, R.A., and Cantor, H. 1993. Molecular and cellular basis of genetic resistance to bacterial infection: the role of the early T-lymphocyte activation- 1/osteopontin gene. Crit. Rev. Immunol. 13: 225–246.
CAS PubMed Web of Science® Google Scholar
33Scott, J.A., Weir, M.L., Wilson, S.M., Xuan, J.W., Chambers, A.F., and McCormack, D.G. 1998. Osteopontin inhibits inducible nitric oxide synthase activity in rat vascular tissue. Am. J. Physiol. 275: 2258–2265.
10.1152/ajpheart.1998.275.6.H2258
CAS PubMed Web of Science® Google Scholar
34Setoguchi, K., Takeya, M., Akaike, T., Suga, M., Hattori, R., Maeda, H., Ando, M., and Takahashi, K. 1996. Expression of inducible nitric oxide synthase and its involvement in pulmonary granulomatous inflammation in rats. Am. J. Pathol. 149: 2005–2022.
CAS PubMed Web of Science® Google Scholar
35Singh, K., Balligrand, J.L., Fischer, T., Smith, T.W., and Kelly, R.A. 1995. Glucocorticoids increase osteopontin expression in cardiac myocytes and microvascular endothelial cells. J. Biol. Chem. 270: 28471–28478.
10.1074/jbc.270.47.28471
CAS PubMed Web of Science® Google Scholar
36Straub, V., Duclos, F., Venzke, D.P., Lee, J.C., Cutshall, S., Leveille, C.J., and Campbell, K.P. 1998. Molecular pathogenesis of muscle degeneration in the d-sarcoglycan-deficient hamster. Am. J. Pathol. 153: 1623–1630.
10.1016/S0002-9440(10)65751-3
CAS PubMed Web of Science® Google Scholar
37Tsujii, M., Dimov, V.B., and Yoshida, T. 1995. In vivo expression of monokine and inducible nitric oxide synthase in experimentally induced pulmonary granulomatous inflammation. Am. J. Pathol. 147: 1001–1015.
PubMed Web of Science® Google Scholar
38Uede, T., Katagiri, Y., Iizuka, J., and Murakami, M. 1997. Osteopontin, a coordinator of host defense system: a cytokine or an extracellular adhesive protein?. Microbiol Immunol. 41: 641–648.
10.1111/j.1348-0421.1997.tb01906.x
CAS PubMed Web of Science® Google Scholar
39Ward, P.A., and Remick, D.G. 1988. Immune mechanisms in lung injury, p. 607–618. In A.P. Fishman (ed), Pulmonary diseases and disorders, McGraw-Hill Books, New York.
Google Scholar
40Watanabe, M., Kawaguchi, H., Onozuka, H., Mikami, T., Urasawa, K., Okamoto, H., Watanabe, S., Abe, K., and Kitabatake, A. 1998. Chronic effects of enalapril and amlodipine on cardiac remodeling in cardiomyopathic hamster hearts. J. Cardiovasc. Pharm. 32: 248–259.
10.1097/00005344-199808000-00012
CAS PubMed Web of Science® Google Scholar
41Williams, E.B., Halpert, I., Wickline, S., Davison, G., Parks, W.C., and Rottman, J.N. 1995. Osteopontin expression is increased in the heritable cardiomyopathy of Syrian hamsters. Circulation 92: 705–709.
10.1161/01.CIR.92.4.705
CAS PubMed Web of Science® Google Scholar
42Wrana, J.L., Zhang, Q., and Sodek, J. 1989. Full length cDNA sequence of porcine secreted phosphoprotein-I (SSP-I, osteopontin). Nucleic Acids Res. 17: 10119.
10.1093/nar/17.23.10119
CAS PubMed Web of Science® Google Scholar
43Xing, Z., Braciak, T., Jordana, M., Croitoru, K., Graham, F.L., and Gauldie, J. 1994. Adenovirus-mediated cytokine gene transfer at tissue sites: overexpression of IL-6 induces lymphocytic hyperplasia in the lung. J. Immunol. 153: 4059–4069.
CAS PubMed Web of Science® Google Scholar
44Xing, Z., Ohkawara, Y., Jordana, M., Graham, F.L., and Gauldie, J. 1996. Transfer of granulocyte-macrophage colony-stimulating factor gene to rat lung induces eosinophilia, monocytosis and fibrotic reactions. J. Clin. Invest. 97: 1102–1110.
10.1172/JCI118503
CAS PubMed Web of Science® Google Scholar
45Xing, Z., Tremblay, G.M., Sime, P.J., and Gauldie, J. 1997. Overexpression of granulocyte-macrophage colony-stimulating factor induces pulmonary granulation tissue formation and fibrosis by induction of tranforming growth factor-b1 and myofibroblast accumulation. Am. J. Pathol. 150: 59–66.
CAS PubMed Web of Science® Google Scholar
46Yang, Y., Ertle, J., and Wilson, J.M. 1994. MHC class I-restricted cytotoxic T lymphocytes to viral antigens destroy hepatocytes in mice infected with E1-deleted recombinant adenoviruses. Immunity 1: 433–442.
10.1016/1074-7613(94)90074-4
CAS PubMed Web of Science® Google Scholar
47Young, M.F., Kerr, J.M., Termine, J.D., Wewer, U.M., Wang, M.G., McBride, O.W., and Fisher, L.W. 1990. cDNA cloning, mRNA distribution and heterogeneity, chromosomal location, and RFLP analysis of human osteopontin (OPN). Genomics 7: 491–502.
10.1016/0888-7543(90)90191-V
CAS PubMed Web of Science® Google Scholar

Citing Literature

Volume44, Issue4

April 2000

Pages 319-332

The Role of Osteopontin in the Development of Granulomatous Lesions in Lung

Abstract

Abbreviations

References

Citing Literature

References

Information

About Wiley Online Library

Help & Support

Opportunities

Connect with Wiley

The Role of Osteopontin in the Development of Granulomatous Lesions in Lung

Abstract

Abbreviations

References

Citing Literature

References

Related

Information