title: A novel gold nanoparticle-doped polyaniline nanofibers-based cytosensor confers simple and efficient evaluation of T-cell activation.
abstract: 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 1×10(4) 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.
authors.affiliation: Department of Chemistry, Jinan University, Guangzhou, Guangdong, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Wang
authors.forename: Huai-Hong
authors.initials: HH
authors.lastname: Cai
authors.forename: Lu
authors.initials: L
authors.lastname: Zhang
authors.forename: Jiye
authors.initials: J
authors.lastname: Cai
authors.forename: Pei-Hui
authors.initials: PH
authors.lastname: Yang
authors.forename: Zheng W
authors.initials: ZW
authors.lastname: Chen
mesh_terms: D000814:Aniline Compounds / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D017097:Electric Impedance; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008213:Lymphocyte Activation; D057139:Nanofibers / Q000737:chemistry*; D015203:Reproducibility of Results; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000814:Aniline Compounds; C416807:polyaniline; D006046:Gold
keywords: Activation markers; Cytosensor; Dynamic analysis; Gold nanoparticles; Polyaniline nanofibers; T-cell activation