Asthma-like disease and chemokine dysregulation in the lung of T cell-deficient mice

Abstract

Rationale To elucidate the mechanism of Th2 cell-independent asthma-like disease, we investigated cytokine expression in the lungs of T cell-deficient adenosine deaminase knockout ( ada −/−) mice, as well as ada −/− rag1 −/− double knockout mice. Methods RNA and proteins were extracted from the lungs of ada −/− and ada −/− rag1 −/− mice and their control littermates. RNA was analyzed by gene chip assays (Affymetrix), RT-PCR, and real-time PCR. Proteins were analyzed by ELISA and by cytokine array membranes (RayBio). In situ hybridization was performed to localize the cytokine expressing cells. Results Microarray analyses of lung RNA suggested the involvement of certain cytokines and chemokines that have been shown to play a role in allergic asthma. They included TARC, eotaxin, MCP-1, -2, -3, -5, and KC/GRO. These results were validated by RT-PCR and real-time PCR. Cytokine array analyses also showed upregulation of these chemokines in ADA-deficient lungs. In situ hybridization demonstrated that TARC was expressed mainly in bronchial epithelial cells. Conclusions We demonstrate that certain cytokines, important in the pathogenesis of allergic asthma, are also upregulated in a Th2 cell-independent asthma model.