**3.1 Construction of genetically modified** *L. lactis* **secreting anti-inflammatory/ antioxidative stress protein HO-1**

To explore the anti-inflammatory therapeutic option other than corticosteroids in COPD, HO-1 was focused on because of its low serum level shown in patients with COPD [10]. The newly constructed HO-1 secreting *L. lactis* (**Figure 6**) was examined by oral administration in a dextran sulfate sodium-induced murine colitis model [17]. Since the favorable alleviation of disease symptoms was observed in this model, a further trial was planned for lung diseases by exploring another delivery method of intratracheal administration. **Figure 6** shows the vector constructed for the HO-1 secreting *L. lactis* NZ9000 system.

#### **3.2 HO-1 production in the lungs after nasally administering HO-1** *L. lactis*

HO-1 secreting *L. lactis* were nasally administered to the anesthetized mice. A total of 50 μL of saline containing 1.0 × 109 of *L. lactis* was migrated to the lungs through stable nasal breathing. Production of HO-1 derived from HO-1 *L. lactis* was confirmed by immunoblotting using anti-His antibody and anti-HO-1 antibody in lung homogenates (**Figure 7a**). Through the pulmonary trafficking of HO-1 *L. lactis*, serum HO-1 levels were significantly increased (**Figure 7b**).
