This study was to investigate the changes in nanostructure and biochemical properties of activated mature CD8+ T cells related to apoptosis at a molecular level. We confirmed the activation and apoptosis of CD8+ T cells by fluorescence-activated cell sorting and atomic force microscopy (AFM) and then performed Raman spectral measurements on activated mature CD8+ T cells and cellular deoxyribose nucleic acid (DNA). From AFM images, the activated mature CD8+ T cells result in an increased cell volume and cell perimeter (that is, a more irregular rim shape) over that observed in resting CD8+ T cells. In the activated mature CD8+ T cells, there were increases in protein spectra at 1002 and 1234 cm−1. In particular, to assess the apoptosis related DNA spectral signatures, we investigated the spectra of the cellular DNA isolated from resting and activated mature CD8+ T cells. Raman spectra at 765 to 786 cm−1 and 1053 to 1087 cm−1 were decreased in activated mature DNA. In addition, we analyzed Raman spectrum using the multivariate statistical method including principal component analysis. Raman spectra of activated mature DNA are especially well-discriminated from those of resting DNA. Our findings regarding the biochemical and structural changes associated with apoptosis in activated mature T cells and cellular DNA according to Raman spectroscopy provide important insights into allospecific immune responses generated after organ transplantation, and may be useful for therapeutic manipulation of the immune response.
Journal: TechConnect Briefs
Volume: 1, Advanced Materials: TechConnect Briefs 2016
Published: May 22, 2016
Pages: 33 - 36
Industry sector: Advanced Materials & Manufacturing
Topics: Materials Characterization & Imaging