Transcriptional Responses of Human Respiratory Epithelial Cells to Nontypeable Haemophilus influenzae Infection Analyzed by High Density cDNA Microarrays

  • Lee, Ji-Yeon (Department of Bioscience and Biotechnology, Sejong University) ;
  • Lee, Na-Gyong (Department of Bioscience and Biotechnology, Sejong University)
  • Published : 2004.08.01

Abstract

Nontypeable H. influenzae (NTHi), a Gram-negative obligate human pathogen, causes pneumonia, chronic bronchitis, and otitis media, and the respiratory epithelium is the first line of defense that copes with the pathogen. In an effort to identify transcriptional responses of human respiratory epithelial cells to infection with NTHi, we examined its differential gene expression using high density cDNA microarrays. BEAS-2B human bronchial epithelial cells were exposed to NTHi for 3 hand 24 h, and the alteration of mRNA expression was analyzed using microarrays consisting of 8,170 human cDNA clones. The results indicated that approximately 2.6% of the genes present on the microarrays increased in expression over 2-fold and 3.8% of the genes decreased during the 24-h infection period. Upregulated genes included cytokines (granulocyte-macrophage colony stimulating factor 2, granulocyte chemotactic protein 2, IL-6, IL-10, IL-8), transcription factors (Kruppel-like factor 7, CCAAT/enhancer binding protein $\beta$, E2F-1, NF-$\kappa$B, cell surface molecules (CD74, ICAM-1, ICAM-2, HLA class I), as well as those involved in signal transduction and cellular transport. Selected genes were further confirmed by reverse-transcription-PCR. These data expand our knowledge of host cellular responses during NTHi infection and should provide a molecular basis for the study of host-NTHi interaction.

Keywords

References

  1. Ann. N. Y. Acad. Sci. v.725 Interactions between respiratory epithelial cells and cytokines: Relationships to lung inflammation Adler, K. B.;B. M. Fischer;D. T. Wright;L. A. Cohn;S. Becker https://doi.org/10.1111/j.1749-6632.1994.tb00275.x
  2. Am. J. Respir. Cell. Mol. Biol. v.18 The effect of diesel exhaust particles on cell function and release of inflammatory mediators from human bronchial epithelial cells in vitro Bayram, H.;J. L. Devalia;R. J. Sapsford;T. Ohtoshi;Y. Miyabara;M. Sagai;R. J. Davies https://doi.org/10.1165/ajrcmb.18.3.2882
  3. Proc. Natl. Acad. Sci. USA v.97 The transcriptional responses of respiratory epithelial cells to Bordetella pertussis reveal host defensive and pathogen counter-defensive strategies Belcher, C. E.;J. Drenkow;B. Kehoe;T. R. Gingeras;N. McNamara;H. Lemjabbar;C. Basbaum;D. A. Relman https://doi.org/10.1073/pnas.230262797
  4. Infect. Immun. v.55 Antigenic diversity of lipooligosaccharides of nontypable Haemophilus influenzae Campagnari, A. A.;M. R. Gupta;K. C. Dudas;T. F. Murphy;M. A. Apicella
  5. Infect. Immun. v.68 Induction of proinflammatory cytokines from human respiratory epithelial cells after stimulation by nontypeable Haemophilus influenzae Clemans, D. L.;R. J. Bauer;J. A. Hanson;M. V. Hobbs;J. W. St. Geme Ⅲ;C. F. Marrs;J. R. Gilsdorf https://doi.org/10.1128/IAI.68.8.4430-4440.2000
  6. Am. J. Physiol. Lung Cell. Mol. Physiol. v.280 Identification of genes induced by inflammatory cytokines in airway epithelium Cooper, P.;S. Potter;B. Mueck;S. Yousefi;G. Jarai https://doi.org/10.1152/ajplung.2001.280.5.L841
  7. J. Biol. Chem. v.275 Analysis by high density cDNA arrays of altered gene expression in human intestinal epithelial cells in response to infection with the invasive enteric bacteia Salmonella Eckmann, L.;J. R. Smith;M. P. Housley;M. B. Dwinell;M. F. Kagnoff https://doi.org/10.1074/jbc.275.19.14084
  8. Infect. Immun. v.71 Microarray analysis of mRNA levels from RAW264.7 macrophages infected with Brucella abortus Eskra, L.;A. Mathison;G. Splitter https://doi.org/10.1128/IAI.71.3.1125-1133.2003
  9. J. Immunol. v.147 IL-10 inhibits cytokine production by activated macrophages Fiorentino, D.;A. Zlotnik;T. Mosmann;M. Howard;A. O'Garra
  10. Microbiol. Mol. Biol. Rev. v.62 Nontypeable Haemophilus influenzae: Pathogenesis and prevention Foxwell, A. R.;J. M. Kyd;A. W. Cripps
  11. J. Immunol. v.164 Haemophilus influenzae stimulates ICAM-1 expression on respiratory epithelia cells Frick, A. G.;T. D. Joseph;L. Pang;A. M. Rabe;J. W. St. Geme Ⅲ;D. C. Look https://doi.org/10.4049/jimmunol.164.8.4185
  12. J. Exp. Med. v.177 Interleukin-10 protects mice from lethal endotoxemia Howard, M.;T. Muchamuel;S. Andrade;S. Menon https://doi.org/10.1084/jem.177.4.1205
  13. Proc. Natl. Acad. Sci. USA v.97 Interaction of Pseudomonas aeruginosa with epithelia cells: Identification of differentially regulated genes by expression microarray analysis of human cDNAs Ichikawa, J. K.;A. Norris;M. G. Bangera;G. K. Geiss;A. B. van't Wout;R. E. Bumgarner;S. Lory https://doi.org/10.1073/pnas.160140297
  14. J. Microbiol. Biotechnol. v.12 cDNA microarray analysis of transcriptional response to hyperi in human gastric cancer cells Jeoung, D. L.;J. H. Kim;H. Lee;M. B. Baek;S. Lee;N.-I. Baek;H.-Y. Kim
  15. Pflugers Arch. v.443 Effects of hydrogen peroxide and hydroxyl radicals on the cytosolic side of a non-selective cation channel in the cultured human bronchial epithelial cell line 16HBE14o- Jeulin, C.;A. C. Dazy;F. Marano https://doi.org/10.1007/s00424-001-0733-5
  16. J. Immunol. v.152 Regulation of neutrophil-derived chemokine expression by IL-10 Kasama, T.;R. M. Strieter;N. W. Lukacs;M. D. Burdick;S. L. Kunkel
  17. Eur. Respir. J. v.9 Bacterial-induced release of inflammatory mediators by bronchial epithelial cells Khair, O. A.;R. J. Davies;J. L. Devalia https://doi.org/10.1183/09031936.96.09091913
  18. Infect. Immun. v.68 Expression of cytokine genes during pneumococcal and nontypeable Haemophilus influenzae acute otitis media in the rat Melhus, A.;A. F. Ryan https://doi.org/10.1128/IAI.68.7.4024-4031.2000
  19. Am. J. Respir. Crit. Care Med. v.155 Cigarette smoke induces interleukin-8 release from human bronchial epithelial cells Mio, T.;D. J. Romberger;A. B. Thompson;R. A. Robbins;A. Heires;S. I. Rennard https://doi.org/10.1164/ajrccm.155.5.9154890
  20. Rev. Infect. Dis. v.9 Nontypable Haemophilus influenzae: A review of clinical aspects, surface antigen, and the human immune response to infection Murphy, T. F.;M. A. Apicella https://doi.org/10.1093/clinids/9.1.1
  21. Am. Rev. Respir. Dis. v.146 Bacterial infection in chronic obstructive pulmonary disease Murphy, T. F.;S. Sethi https://doi.org/10.1164/ajrccm/146.4.1067
  22. Infect. Immun. v.68 Differential effects of virulent versus avirulent Legionella pneumophila on chemokine gene expression in murine alveolar macrophages determined by cDNA expression array technique Nakachi, N.;K. Matsunaga;T. W. Klein;H. Friedman;Y. Yamamoto https://doi.org/10.1128/IAI.68.10.6069-6072.2000
  23. Clin. Exp. Allergy v.31 Rhinovirus infection up-regulates eotaxin and eotaxin-2 expression in bronchial epithelial cells Papadopoulos, N. G.;A. Papi;J. Meyer;L. A. Stanciu;S. Salvi;S. T. Holgate;S. L. Johnston https://doi.org/10.1046/j.1365-2222.2001.01112.x
  24. J. Biol. Chem. v.274 Rhinovirus infection induces expression of its own receptor intercellular adhesion molecule 1 (ICAM-1) via increased NF-kappaB-mediated transcription Papi, A.;S. L. Johnston https://doi.org/10.1074/jbc.274.14.9707
  25. Nature Biotechnol. v.16 DNA chips: State-of-the art Ramsay, G. https://doi.org/10.1038/nbt0198-40
  26. FEMS Microbiol. Rev. v.23 Molecular determinants of the pathogenesis of disease due to non-typeable Haemophilus influenzae Rao, V. K.;G. P. Krasan;D. R. Hendrixson;S. Dawid;J. W. St. Geme Ⅲ https://doi.org/10.1111/j.1574-6976.1999.tb00393.x
  27. Proc. Natl. Acad. Sci. USA v.98 Activation of NF-${\kappa}$B by nontypeable Hemophilus influenzae is mediated by toll-like receptor 2-TAK1-dependent NIK-IKK ${\alpha}/{\beta}$-$I{\kappa}B{\alpha}$ and MKK3/6-p38 MAP kinase signaling pathways in epithelial cells Shuto, T.;H. Xu;B. Wang;J. Han;H. Kai;X. X. Gu;T. F. Murphy;D. J. Lim;J. D. Li https://doi.org/10.1073/pnas.151236098
  28. Infect. Immun. v.70 Host-pathogen interactions: Subversion and utilization of the NF-kappaB pathway during infection Tato, C. M.;C. A. Hunter https://doi.org/10.1128/IAI.70.7.3311-3317.2002
  29. Infect. Immun. v.69 Expression of cytokine and chemokine genes by human middle ear epithelial cells induced by formalin-killed Haemophilus influenzae of its lipooligosaccharide htrB and rfaD mutants Tong, H. H.;Y. Chen;M. James;J. Van Deusen;D. B. Welling;T. F. DeMaria https://doi.org/10.1128/IAI.69.6.3678-3684.2001
  30. Annu. Rev. Physiol. v.64 GM-CSF regulates pulmonary surfactant homeostasis and alveolar macrophage-mediated innate host defense Trapnell, B. C.;J. A. Whitsett https://doi.org/10.1146/annurev.physiol.64.090601.113847
  31. J. Microbiol. Biotechnol. v.10 Development of DNA chip microarray Yoon, S. H.;J. G. Choi;S. Y. Lee