Background Noninvasive sputum sampling has enabled the identification of biomarkers in asthmatic patients. Studies of discrete cell populations in sputum can enhance measurements compared with whole sputum in which changes in rare cells and cell-cell interactions can be masked. Objective We sought to enrich for sputum-derived human bronchial epithelial cells (sHBECs) and sputum-derived myeloid type 1 dendritic cells (sDCs) to describe transcriptional coexpression of targets associated with a type 2 immune response. Methods A case-control study was conducted with patients with mild asthma (asthmatic cases) and healthy control subjects. Induced sputum was obtained for simultaneous enrichment of sHBECs and sDCs by using flow cytometry. Quantitative PCR was used to measure mRNA for sHBEC thymic stromal lymphopoietin (TSLP) , IL33 , POSTN , and IL25 and downstream targets in sDCs (OX40 ligand OX40L , CCL17 , PPP1R14A , CD1E , CD1b , CD80 , and CD86 ). Results Final analyses for the study sample were based on 11 control subjects and 13 asthmatic cases. Expression of TSLP , IL33 , and POSTN mRNA was increased in sHBECs in asthmatic cases ( P  = .001, P  = .05, and P  = .04, respectively). Expression of sDC OX40L and CCL17 mRNA was increased in asthmatic cases ( P  = .003 and P  = .0001, respectively). sHBEC TSLP mRNA expression was strongly associated with sDC OX40L mRNA expression ( R  = 0.65, P  = .001) and less strongly with sDC CCL17 mRNA expression. sHBEC IL33 mRNA expression was associated with sDC OX40L mRNA expression ( R  = 0.42, P  = .04) but not sDC CCL17 mRNA expression. Conclusions Noninvasive sampling and enrichment of select cell populations from sputum can further our understanding of cell-cell interactions in asthmatic patients with the potential to enhance endotyping of asthmatic patients.