A novel gold nanoparticle-doped polyaniline nanofibers-based cytosensor confers simple and efficient evaluation of T-cell activation

A rapid, easy assay for monitoring dynamics of T-cell activation should help to guide potential medical evaluation of immune responses or immunopathogenesis. Here, we report development of novel electrochemical cytosensors for dynamic analyses of T-cell activation markers on living cells. Gold nanoparticles-doped polyaniline nanofiber (Au/PANI-NFs) composite was greenly prepared by in situ one-step chemical inertness of PANI-NFs with gold nanoparticles to fabricate impedance-based electrochemical biosensors. Transmission electron micrographs indicated that the gold nanoparticles were uniformly anchored along with the structure of PANI-NF surface, displaying fibrillar morphology with a ∼60 nm diameter. Au/PANI-NFs-based cytosensors coated with anti-CD Ab molecules could provide biomimetic interface for multiple immunosensing of T-cell surface activation markers (CD69, CD25, and CD71). The dual signal amplification of Au nanoparticle and PANI-NFs-based electrochemical impedance spectroscopic (EIS) measurements enabled the cytosensors considerably sensitive, with a detection limit of 1×104 cells/ml of activated T-cells. The activation-targeted cytosensors detected early, middle and late stages for expression of activation markers CD69, CD25, and CD71 at 8 h, 24 h, and 36 h, respectively, after concanvalin A stimulation of T cells. The quantitative results consisted with those derived from flow cytometric analysis. Furthermore, activation-targeted cytosensor allowed for dynamic analysis of the immune inhibition of T-cell activation by immune regulatory drug icariin (ICA). Thus, Au/PANI-NFs-based cytosensors offer simple and fast approach for non-destructive, quantitative evaluation of T-cell activation markers, with considerable specificity, reproducibility, and low background noise.