Antigen Specific <em>In Vivo</em> Killing Assay using CFSE Labeled Target Cells

Abstract

Carboxyfluorescein diacetate succinimidyl ester (CFSE) can be used to easily and quickly label a cell population of interest for in vivo investigation. This labeling has classically been used to study proliferation and migration. In the method presented here, we have shortened the timeline after adoptive transfer to look at survival and killing of epitope specific CFSE labeled target cells [4][5][6] . The level of specific killing of a CD8 + T cell clone can indicate the quality of the response, as their quantity may be misleading. Specific CD8+ T cells can become functionally exhausted over time with a decline in cytokine production and killing 7,8 . Also, certain CD8 + T cell clones may not kill as well as others with differing TCR specificities 9 . For effective Cell Mediated Immunity (CMI), antigens must be identified that produce not only adequate numbers of responding T cells, but also functionally robust responding T cells. Here we assess the percent cell specific killing of two peptide specific T cell clones in BALB/c mice.

Consensus nomenclature for CD8+T cell phenotypes in cancer

“…For this, the selective elimination of adoptively transferred carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled target cells bearing a specific CD8 C T cell peptide has been examined in preclinical models. 38 While this technique has the major advantage of assessing CD8…”

Dengue vaccine–induced CD8+ T cell immunity confers protection in the context of enhancing, interfering maternal antibodies

“…The assay was performed according to a previously described protocol (83). Briefly, splenocytes were harvested from naive mice and pulsed with 1 μg/ml NS4B 99-107 or OVA SIIN-FEKL peptide.…”

Delivery of foreign cytotoxic T lymphocyte epitopes to tumor tissues for effective antitumor immunotherapy against pre-established solid tumors in mice

“…Booster injections were given on days 14 and 28 with ovalbumin and incomplete Freund’s adjuvant. To confirm the generation of SIINFEKL-specific CTLs, an in vivo CTL assay based on carboxyfluorescein succinimidyl ester (CFSE) labeling was carried out by injecting CFSE-labeled cells loaded with the CTLp to ovalbumin-immunized mice [24]. The viabilities of cells pulsed with the CTLp were significantly reduced compared to those not incubated with the epitope (Supplementary Figure S7), suggesting the successful production of SIINFEKL-specific CTLs in mice through immunization.…”