Abstract
The study was to investigate the changes in 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 and then performed Raman spectral measurements on activated mature CD8+ T cells and cellular deoxyribose nucleic acid (DNA). 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
Highlights
T cell activation is an essential event in immune responses
We investigated the identification and quantification of biochemical properties related to apoptosis of activated mature CD8þ T cell and cellular deoxyribose nucleic acid (DNA) using Raman spectroscopy and multivariate statistical methods
We observed the morphological changes in activated mature CD8þ T cells using atomic force microscopy (AFM)
Summary
T cell activation is an essential event in immune responses. The process of proper T cell activation is rigorously monitored and regulated by apoptosis. The well-known AICD receptor, has been shown to regulate immune homeostasis in mutational, transcriptional and clinical studies.[2,3,4,5] Following infection, Fas is rapidly expressed on the surfaces of activated T cells. These T cells are resistant to apoptosis for some days after activation. It has been shown that T cells only acquire sensitivity to Fas-mediated apoptosis at a few days after antigen-specific activation.[6,7,8] only activated mature T cells gain sensitivity to apoptosis and begin to undergo cell death. During this process, activated mature T cells rapidly exhibit classic characteristics of apoptosis such as membrane blebbing, chromatin condensation and formation of deoxyribose nucleic acid (DNA) fragments.[6,9] the process of T cell activation is being widely studied in the immunological and biochemical fields, the change in biochemical properties at a molecular level is still unknown due to the lack of a sensitive quantitative technique
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