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Aim: To investigate effects of CpG oligodeoxynucleotide (CpG ODN) on the mRNA expression of transcription factors GATA binding protein 3 (GATA-3) and T-box expressed in T cells (T-bet) in asthmatic mice. Methods: An asthmatic mouse model was established and treated with CpG ODN. Total inflammatory cells and eosinophils in bronchoalveolar lavage fluid (BALF) were counted and inflammatory cell infiltration in lung tissue was evaluated. Interferon-γ and interleukin-4 concentrations in BALF and splenocyte culture supeatants were detected using an enzyme-linked immunosorbent assay. Transcription factor GATA-3 and Tbet mRNA expression in splenocytes and lung tissue were detected by reverse transcription-polymerase chain reaction. Results: Total inflammatory cells and eosinophils in BALF were reduced in the CpG ODN-treated group compared with the asthma group, and inflammatory cell infiltration in lung tissue was also significantly alleviated. CpG ODN treatment increased the interferon-γ concentration but decreased the interleukin-4 concentration in both BALF and splenocyte culture supeatants. GATA-3 mRNA expression was reduced in both lung tissue and splenocytes in the CpG ODN-treated group, while the mRNA ratio of T-bet to GATA-3 in splenocytes was increased. Conclusion: CpG ODN treatment inhibits airway inflammatory cell infiltration and regulates interferon-γ/interleukin-4 synthesis in asthmatic mice, possibly through a mechanism of downregulation of GATA-3 mRNA expression in both lung tissue and splenocytes.