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Cytotoxic T-cell (CTL) function assay

Aim:

A CTL assay indicates the presence and cytotoxic activity of T cells to a specific antigen and allows to examine the influence of a test item on this immune function. The CTL response against T-dependent antigens requires the cooperation of at least three different major categories of immune cells. These include professional antigen presenting cells (e.g., dendritic cells), CD4(+) T helper lymphocytes, and CD8(+) T effector lymphocytes. It is also among the few functional responses dependent on and, hence, capable of evaluating effective antigen presentation via both class I and class II molecules of the major histocompatibility complex (MHC).

Species:

Mice.

Design:

This assay allows the detection of cytolytic activity specific for a peptide epitope in-vivo in mice. One group of target cells is labeled with a higher concentration of CFSE dye and pulsed with the antigenic peptide epitope, while the other group of target cells is labeled with a lower concentration of CFSE and pulsed with an irrelevant peptide. They are mixed in a 1:1 ratio and injected i.v. into immunized and naïve mice. A deviation from the 1:1 ratio when the mice are sampled indicates specific lysis of the cells with the relevant epitope on the surface. Normal spleen cells are used as target cells.

Outcome:

Evaluating the influence of the test item on the presence and cytotoxic activity of T cells to a specific antigen.

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