Ag85B DNA vaccine suppresses airway inflammation in a murine model of asthma.
BACKGROUND
In allergic asthma, Th2 lymphocytes are believed to play important roles in orchestrating airway eosinophilia and inflammation. Resetting the Th1/Th2 imbalance may have a therapeutic role in asthma. The mycobacterium tuberculosis 30-kilodalton major secretory protein (antigen 85B, Ag85B) can protect animals from M. tuberculosis infection by inducing a Th1-dominant response.


METHODS
In this study, the Ag85B gene was cloned into pMG plasmids to yield the pMG-Ag85B plasmid. The expression of Ag85B gene in murine bronchial epithelia cells was detected by Western blotting and immunohistochemical staining after intranasal immunization with reconstructed pMG-Ag85B plasmids. The protective effect of pMG-Ag85B plasmids immunization in airway inflammation was evaluated by histological examination and bronchoalveolar lavage (BAL). IL-4 and IFN-gamma levels in the BAL and supernatant from splenocyte culture were determined using ELISA kits.


RESULTS
The Ag85B gene was successfully expressed in murine bronchial epithelia cells by intranasal immunization with reconstructed pMG-Ag85B plasmids. Using a murine model of asthma induced by ovalbumin (OVA), pMG-Ag85B immunization significantly inhibited cellular infiltration across the airway epithelium with a 37% decrease in the total number of cells (9.6 +/- 2.6 x 10(5)/ml vs. 15.2 +/- 3.0 x 10(5)/ml, p < 0.05) and a 74% decrease in the number of eosinophils (1.4 +/- 0.2 x 10(5)/ml vs. 5.4 +/- 1.1 x 10(5)/ml, p < 0.01) compared with the OVA-sensitized control group. There was no difference in the number of neutrophils in BAL fluid between the pMG-Ag85B group, the OVA-sensitized control group and the empty pMG group. IL-4 production was significantly decreased in the BAL fluid (32.0 +/- 7.6 pg/ml vs. 130.8 +/- 32.6 pg/ml, p < 0.01) and in the splenocyte supernatant (5.1 +/- 1.6 pg/ml vs. 10.1 +/- 2.3 pg/ml, p < 0.05) in the pMG-Ag85B group compared with the OVA-sensitized control group, while IFN-gamma production was increased in the BAL fluid (137.9 +/- 25.6 pg/ml vs. 68.4 +/- 15.3 pg/ml, p < 0.05) and in the splenocyte supernatant (20.1 +/- 5.4 pg/ml vs. 11.3 +/- 3.2 pg/ml, p < 0.05).


CONCLUSIONS
In a murine model of asthma induced by OVA, intranasal immunization with pMG-Ag85B significantly reduced allergic airway inflammation with less eosinophil infiltration. This protective effect was associated with decreased IL-4 and increased IFN-gamma production in the BAL fluid and in the supernatant of cultured splenocytes.
Wu
Jian
J
Department of Respiratory Disease, Peking University First Hospital, Beijing, PR China. wjxst@hotmail.com
Xu
Jun
J
Cai
Chuang
C
Gao
Xinglin
X
Li
Li
L
Zhong
Nanshan
N
D000217:Acyltransferases / Q000008:administration & dosage / Q000494:pharmacology*; D000281:Administration, Intranasal; D000818:Animals; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000494:pharmacology*; D001249:Asthma / Q000473:pathology* / Q000517:prevention & control*; D001426:Bacterial Proteins / Q000008:administration & dosage / Q000494:pharmacology*; D015153:Blotting, Western; D016207:Cytokines / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D007150:Immunohistochemistry; D007249:Inflammation / Q000473:pathology* / Q000517:prevention & control*; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012137:Respiratory System / Q000473:pathology*; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D032581:Tuberculosis Vaccines / Q000008:administration & dosage / Q000494:pharmacology*; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000494:pharmacology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D016207:Cytokines; D012333:RNA, Messenger; D032581:Tuberculosis Vaccines; D019444:Vaccines, DNA; D000217:Acyltransferases; C121433:antigen 85B, Mycobacterium tuberculosis