Abstract
Adaptive immune responses begin with cognate antigen presentation-dependent specific interaction between T cells and antigen-presenting cells. However, there have been limited reports on the isolation and analysis of these cellular complexes of T cell-antigen-presenting cell (T/APC). In this study, we successfully isolated intact antigen-specific cellular complexes of CD8+ T/APC by utilizing a microfluidics cell sorter. Using ovalbumin (OVA) model antigen and OT-I-derived OVA-specific CD8+ T cells, we analyzed the formation of antigen-specific and antigen-non-specific T/APC cellular complexes and revealed that the antigen-specific T/APC cellular complex was highly stable than the non-specific one, and that the intact antigen-specific T/APC complex can be retrieved as well as enriched using a microfluidics sorter, but not a conventional cell sorter. The single T/APC cellular complex obtained can be further analyzed for the sequences of T cell receptor Vα and Vβ genes as well as cognate antigen information simultaneously. These results suggested that this approach can be applied for other antigen and CD8+ T cells of mice and possibly those of humans. We believe that this microfluidics sorting method of the T/APC complex will provide useful information for future T cell immunology research.
Highlights
T cells play important roles in various health conditions including infection, cancer, allergy, and autoimmunity [1,2,3,4,5,6,7]
We found that microfluidics-based cell sorting enabled the isolation of an antigen-specific T cell-antigen-presenting cell (T/antigen-presenting cells (APCs)) complex, as well as, detailed analysis of this immunological cellular complex
Our results suggested that the ability to sort an intact T/APC complex itself indicates that the isolated complex was very likely to be formed by the specific interaction between the cognate antigen peptide presented on the major histocompatibility complex (MHC) and the antigen-specific T cell receptor (TCR) recognition (Fig 4 and S2 Fig)
Summary
T cells play important roles in various health conditions including infection, cancer, allergy, and autoimmunity [1,2,3,4,5,6,7]. Each T cell expresses a unique T cell receptor (TCR), which recognizes antigenic peptides presented on the major histocompatibility complex (MHC) of antigen-presenting cells (APCs). When T cell-mediated immune responses begin, T cells usually interact with APCs, especially dendritic cells, which are one of the most efficient APCs for naïve T cells. When T cells recognize the cognate antigen peptide on the MHC of the dendritic. T/APC complex isolation and gene analysis decision to publish, or preparation of the manuscript
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