1 2008 Vol: 34(11). DOI: 10.1590/S1806-37132008001100002

Influência do biofármaco DNA-hsp65 na lesão pulmonar induzida por bleomicina

OBJECTIVE: To evaluate the effects of immunization with a DNA-hsp65 vaccine in an experimental model of pulmonary fibrosis. METHODS: A total of 120 male C57BL/6 mice were distributed into four groups: SS, injected with saline (placebo) and then receiving intratracheal (IT) instillation of saline; SB, injected with saline (placebo) and then receiving IT instillation of bleomycin; PB, treated with plasmid only, without bacterial genome, and then receiving IT instillation of bleomycin; and BB, treated with the vaccine and then receiving IT instillation of bleomycin. Bleomycin was instilled 15 days after the last immunization, and the animals were killed six weeks thereafter. The left and right lungs were removed, the former for morphological analysis and the latter for hydroxyproline measurements. RESULTS: The proportion of deaths within the first 48 h after the IT instillation (deaths attributed to the surgical procedure) was higher in the SB group than in the SS group (57.7% vs. 11.1%). The mean area of pulmonary interstitial septa was greater in the SB and PB groups (53.1 ± 8.6% and 53.6±9.3%, respectively) than in the SS and BB groups (32.9 ± 2.7% and 34.3 ± 6.1%, respectively). The mean area of interstitial septa stained by picrosirius was greater in the SB, PB and BB groups than in the SS group (8.2 ± 4.9%, 7.2 ± 4.2% and 6.6 ± 4.1%, respectively, vs. 2.0±1.4%). The total hydroxyproline content in the lung was significantly lower in the SS group (104.9 ± 20.9 pg/lung) than in the other groups (SB: 160.4 ± 47.8 pg/lung; PB: 170.0 ± 72.0 pg/lung; and BB: 162.5 ± 39.7 pg/lung). CONCLUSIONS: Immunization with the DNA-hsp65 vaccine reduced the deposition of noncollagen matrix in a model of bleomycin-induced lung lesion.

Figure 1 Figure 2 Figure 3 Figure 4
  1. American Thoracic Society; European Respiratory Society. American Thoracic Society/European Respiratory Society International Multidisciplinary Consensus Classification of the Idiopathic Interstitial Pneumonias. This joint statement of the American Thoracic Society (ATS), and the European Respiratory Society (ERS) was adopted by the ATS board of directors, Juneand by the ERS Executive Committee, June 2001. Am J Respir Crit Care Med. 2002;165(2):. Review. No abstract available. Erratum in: Am J Respir Crit Care Med. 2002;166(3):426 , 277-304 (2001) .
  2. American Thoracic Society. Idiopathic pulmonary fibrosis: diagnosis and treatment. International consensus statement. American Thoracic Society (ATS), and the European Respiratory Society (ERS). Am J Respir Crit Care Med.161(2 Pt 1) , 646-64 (2000) .
  3. Nathan SD. Therapeutic management of idiopathic pulmonary fibrosis: an evidence-based approach. Clin Chest Med.27(1 Suppl 1):S, vi , 27-35 (2006) .
  4. Haslam PL. Evaluation of alveolitis by studies of lung biopsies. Lung.168 Suppl , 984-92 (1990) .
  5. Daniil Z, Kitsanta P, Kapotsis G, Mathioudaki M, Kollintza A, Karatza M, et al. CD8+ T lymphocytes in lung tissue from patients with idiopathic pulmonary fibrosis. Respir Res.6:81 , (2005) .
  6. Gross TJ, Hunninghake GW. Idiopathic pulmonary fibrosis. N Engl J Med.345(7) , 517-25 (2001) .
  7. Singh S, du Bois R. Autoantibodies in cryptogenic fibrosing alveolitis. Respir Res.2(2) , 61-3 (2001) .
  8. Wallace WA, Ramage EA, Lamb D, Howie SE. A type 2 (Th2-like) pattern of immune response predominates in the pulmonary interstitium of patients with cryptogenic fibrosing alveolitis (CFA). Clin Exp Immunol.101(3) , 436-41 (1995) .
  9. Martinez JA, King TE Jr, Brown K, Jennings CA, Borish L, Mortenson RL, et al. Increased expression of the interleukin-10 gene by alveolar macrophages in interstitial lung disease. Am J Physiol.273(3 Pt 1) , 676-83 (1997) .
  10. Hancock A, Armstrong L, Gama R, Millar A. Production of interleukin 13 by alveolar macrophages from normal and fibrotic lung. Am J Respir Cell Mol Biol.18(1) , 60-5 (1998) .
  11. Lowrie DB, Tascon RE, Colston MJ, Silva CL. Towards a DNA vaccine against tuberculosis. Vaccine.12(16) , 1537-40 (1994) .
  12. Tascon RE, Colston MJ, Ragno S, Stavropoulos E, Gregory D, Lowrie DB. Vaccination against tuberculosis by DNA injection. Nat Med.2(8) , 888-92 (1996) .
  13. Lowrie DB, Silva CL, Tascon RE. DNA vaccines against tuberculosis. Immunol Cell Biol.75(6) , 591-4 (1997) .
  14. Bonato VL, Lima VM, Tascon RE, Lowrie DB, Silva CL. Identification and characterization of protective T cells in hsp65 DNA-vaccinated and Mycobacterium tuberculosis-infected mice. Infect Immun.66(1) , 169-75 (1998) .
  15. Lowrie DB, Tascon RE, Bonato VL, Lima VM, Faccioli LH, Stavropoulos E, et al. Therapy of tuberculosis in mice by DNA vaccination. Nature.400(6741) , 269-71 (1999) .
  16. Junqueira LC, Bignolas G, Brentani RR. Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem J.11(4) , 447-55 (1979) .
  17. Frankel SK, Moats-Staats BM, Cool CD, Wynes MW, Stiles AD, Riches DW. Human insulin-like growth factor-IA expression in transgenic mice promotes adenomatous hyperplasia but not pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol.288(5) , 805-12 (2005) .
  18. Walter N, Collard HR, King TE Jr. Current perspectives on the treatment of idiopathic pulmonary fibrosis. Proc Am Thorac Soc.3(4) , 330-8 (2006) .
  19. Yildirim Z, Kotuk M, Iraz M, Kuku I, Ulu R, Armutcu F, et al. Attenuation of bleomycin-induced lung fibrosis by oral sulfhydryl containing antioxidants in rats: erdosteine and N-acetylcysteine. Pulm Pharmacol Ther.18(5) , 367-73 (2005) .
  20. Martinez JA, Ramos SG, Meirelles MS, Verceze AV, Arantes MR, Vannucchi H. Effects of quercetin on bleomycin-induced lung injury: a preliminary study. J Bras Pneumol.34(7) , 445-52 (2008) .
  21. Silva CL, Bonato VL, Coelho-Castelo AA, De Souza AO, Santos SA, Lima KM, et al. Immunotherapy with plasmid DNA encoding mycobacterial hsp65 in association with chemotherapy is a more rapid and efficient form of treatment for tuberculosis in mice. Gene Ther.12(3) , 281-7 (2005) .
  22. Lima KM, Bonato VL, Faccioli LH, Brandão IT, Santos SA, Coelho-Castelo AA et al. Comparison of different delivery systems of vaccination for the induction of protection against tuberculosis in mice. Vaccine.19:3518-25 , 25-26 (2001) .
  23. Smith RB. Avaliação da eficácia da injeção da vacina de DNA HSP65 do Mycobacterium leprae em pacientes com estádio avançado de carcinoma epidermóide de cabeça e pescoço, em ensaio clínico de fase I/II. [thesis]. São Paulo: Universidade de São Paulo , (2007) .
  24. Gurujeyalakshmi G, Giri SN. Molecular mechanisms of antifibrotic effect of interferon gamma in bleomycin-mouse model of lung fibrosis: downregulation of TGF-beta and procollagen I and III gene expression. Exp Lung Res.21(5) , 791-808 (1995) .
  25. Raghu G, Brown KK, Bradford WZ, Starko K, Noble PW, Schwartz DA, et al. A placebo-controlled trial of interferon gamma-1b in patients with idiopathic pulmonary fibrosis. N Engl J Med.350(2) , 125-33 (2004) .
  26. Roman J. Extracellular matrix in the pathogenesis of lung injury and repair. In: Schwarz M, King TE, editors. Interstitial lung disease. Hamilton: BC Decker; p , 277-99 (2003) .
  27. Selman M, King TE, Pardo A; American Thoracic Society; European Respiratory Society; American College of Chest Physicians. Idiopathic pulmonary fibrosis: prevailing and evolving hypotheses about its pathogenesis and implications for therapy. Ann Intern Med.134(2) , 136-51 (2001) .
  28. Shen AS, Haslett C, Feldsien DC, Henson PM, Cherniack RM. The intensity of chronic lung inflammation and fibrosis after bleomycin is directly related to the severity of acute injury. Am Rev Respir Dis.137(3) , 564-71 (1988) .