Figure 9
CD11bhiCD11cintGr-1+F4/80+ suppressor cells (SCs) from wild-type (wt) mice suppress T helper type 2 (Th2), Th17, and Th1 immune responses in the lung. (a) Experimental design. Briefly, CD11bhiCD11cintGr-1+F4/80+ cells from wt bone marrow (BM) cultures were sorted on day 9 and added to wt BM-cultures before incubation with 1 μM DQ-OVA or with phosphate-buffered salinefor 24 h. The next day, 106 unpulsed or pulsed wt DCs±3 × 104 wt SCs were administered intratracheally (IT) into BALB/c wt recipient mice. After 10 days, mice were challenged IT with ovalbumin (OVA). Seventy-two hours after the injection, airway responsiveness was determined. Subsequently bronchoalveolar lavage (BAL) fluid, lung cells and tissues were collected for further analysis. BALB/c mice receiving unpulsed cells served as negative controls. Mice that received wt DCs without SCs served as positive controls. (b) Airway hyperresponsiveness (AHR) in response to IT administration of metacholine measured as airway resistance using Flexivent. (c) Total and differential cell counts in BAL fluid. (d) Histological examination of airway inflammation. Sections were stained with hematoxylin and eosin (original magnification × 200). (e) Histological examination of goblet cell hyperplasia. Sections were stained with periodic acid-Schiff for mucus production (original magnification × 200). Mucus producing airways are plotted relative to all analyzed airways (right panel). (f) Cytokine profiles of pulmonary cells harvested 72 h after OVA challenge. Supernatants were collected 72 h after in vitro cell culture. Values shown are the mean±s.e.m.; n=9–10 per group, *P<0.05, **P<0.01, ***P<0.001. GM-CSF, granulocyte macrophages colony-stimulating factor; IFN, interferon; IL, interleukin; TNF, tumor-necrosis factor.
