A Simple and Sensitive Method for Measuring Tumor-Specific T Cell Cytotoxicity

T cell therapies have demonstrated long-term efficacy and curative potential for the treatment of some cancers. However, their use is limited by damage to bystander tissues, as seen in graft-versus-host disease after donor lymphocyte infusion, or “on-target, off-tumor” toxicities incurred in some engineered T cell therapies. Non-specific immunosuppression and irreversible T cell elimination are currently the only means to control such deleterious responses, but at the cost of abrogating therapeutic benefits or causing secondary complications. On the basis of the physiological paradigm of immune inhibitory receptors, we designed antigen-specific inhibitory chimeric antigen receptors (iCARs) to preemptively constrain T cell responses. We demonstrate that CTLA-4– or PD-1–based iCARs can selectively limit cytokine secretion, cytotoxicity, and proliferation induced through the endogenous T cell receptor or an activating chimeric receptor. The initial effect of the iCAR is temporary, thus enabling T cells to function upon a subsequent encounter with the antigen recognized by their activating receptor. iCARs thus provide a dynamic, self-regulating safety switch to prevent, rather than treat, the consequences of inadequate T cell specificity.

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“…S5A) (36). All groups of T cells efficiently killed iPS-fib, demonstrating allogeneic cytotoxicity (Fig.…”

Section: Resultsmentioning

confidence: 99%

PD-1– and CTLA-4–Based Inhibitory Chimeric Antigen Receptors (iCARs) Divert Off-Target Immunotherapy Responses

2013

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“…The cytotoxicity was quantitated by a nonradioactive method recently developed in our own lab [13]. Briefly, immediately before the assay for luciferase activity, culturing medium was removed.…”

Section: Cytolytic Activity Of Chimeric T-effector Cells Against Her2mentioning

confidence: 99%

“…These modified T lymphocytes contain a chimeric antigen receptor comprising a single chain antibody specific to HER2 [11]. The cytolytic activity was measured with a recently developed nonradioactive method in our lab [13], which fully exploits the high sensitivity and the relative simplicity of luciferase quantitative assay. The results show that incubation of both clone 3 and 5 with the T-effector cells caused the progressive loss of the intracellular marker GFP-Luc.…”

Section: The Newly Established Cell Lines Can Be Efficientmentioning

confidence: 99%

Modification of a Popular Syngeneic Murine Mammary Tumor Model for Immunotherapy Studies

Rivera

Tao

et al. 2011

ISRN Immunology

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To facilitate the evaluation of immunotherapeutic intervention against malignant diseases, it is desirable to have a syngeneic tumor model that closely resembles the growth pattern of human tumors. Murine 4T1 breast cancer model is known for its metastatic properties that mimic its human counterpart. However, a drawback of this model is the lack of an identified tumor antigen to function as a therapeutic target for immunologic intervention. We used the piggyBac transposon system to stably transduce a tumor antigen, the human epidermal growth factor receptor 2 gene (HER2), into this tumor cell. In vitro characterization shows that the newly established cells have a similar growth pattern as the parental line. In vivo evaluation shows that host immune response was generated against the HER2 tumor antigen, despite the high homology between HER2 and its murine counterpart (neu gene). When implanted into immune-deficient mice, the HER2-expressing 4T1 cells readily formed sizable tumors, indicating that these cells are useful for evaluating the therapeutic effect of adoptively transferred cytotoxic T cells that are specifically raised or modified to target the HER2 tumor antigen.

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“…Luciferase quantitative assay, a simple and sensitive method, has been increasingly and widely used to measure the cytolytic effects of various effector cells (36)(37)(38). In the present study, we used this method to test the cytotoxicity of stimulated cells against K562-luc cells.…”

Section: Expanded Nk Cells Were Effectively Activated Through Immobilmentioning

confidence: 99%

Expansion of NK cells from PBMCs using immobilized 4-1BBL and interleukin-21

Liu

et al. 2015

International Journal of Oncology

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Abstract. Adoptive transfer of NK cells has been widely applied clinically for cancer immunotherapy. However, the difficulties to obtain a large number of activated NK cells impede the successful application of such therapy. In the present study, we implemented a novel method involving the use of immobilized human 4-1BBL and interleukin-21 to amplify NK cells from the peripheral blood mononuclear cells (PBMCs) of healthy donors. Following stimulation for 21 days, we achieved considerable expansion of NK cells with high purity and strong cytotoxicity. This is the first time solid phase cytokines were used to augment NK cells, and this method has the advantage of no need to introduce feeder cells, without prior purification of NK cells and it effectively stimulated and expanded NK cells. The strategy of cell proliferation and activation could lead to a safer and more effective application of NK cells clinically.

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A Simple and Sensitive Method for Measuring Tumor-Specific T Cell Cytotoxicity

Cited by 70 publications

References 15 publications

PD-1– and CTLA-4–Based Inhibitory Chimeric Antigen Receptors (iCARs) Divert Off-Target Immunotherapy Responses

PD-1– and CTLA-4–Based Inhibitory Chimeric Antigen Receptors (iCARs) Divert Off-Target Immunotherapy Responses

Modification of a Popular Syngeneic Murine Mammary Tumor Model for Immunotherapy Studies

Expansion of NK cells from PBMCs using immobilized 4-1BBL and interleukin-21

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