Assessing T Cell Responses in Murine Glioma

Peripheral blood mononuclear cells (PBMCs) collected on day 1 post-tumor challenge or splenocytes collected at end-of-life from control mice and surviving mIGV-001 treated mice were used in FluoroSpot or ELISpot assays to measure IFN-γ production by T cells. PBMCs were stimulated using tumor lysate-pulsed DCs and splenocytes were incubated with major histocompatibility complex (MHC)-I H-2Db and H-2Kb GL261-specific peptides27 (link) (CHI Scientific). IFN-γ was measured using a murine IFN-γ capture FluoroSpot kit (CTL) or mouse IFN-γ ELISpot Kit (R&D Systems). IFN-γ counts were evaluated in an ImmunoSpot S6 UNIVERSAL Analyzer (CTL) or S6 Ultimate-V Analyzer (CTL).

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Cultrara C., Uhl C., Kirby K., Abed Elrazaq E., Zellander A., Andrews D.W., Scott C.B., Galluzzi L., Exley M.A, & Zilberberg J. (2023). A biologic-device combination product delivering tumor-derived antigens elicits immunogenic cell death-associated immune responses against glioblastoma. Journal for Immunotherapy of Cancer, 11(8), e006880.

Publication 2023

mouse IFN-γ ELISpot Kit

Assays Elispot Ifn γ Major histocompatibility complex Mouse ifn γ Murine Peripheral blood mononuclear cells T cells Tumor

Corresponding Organization :

Other organizations : Thomas Jefferson University, Cornell University, Lander Institute

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Variable analysis

independent variables

Stimulation of peripheral blood mononuclear cells (PBMCs) using tumor lysate-pulsed dendritic cells (DCs)
Incubation of splenocytes with major histocompatibility complex (MHC)-I H-2Db and H-2Kb GL261-specific peptides

dependent variables

IFN-γ production by T cells, measured using FluoroSpot or ELISpot assays

control variables

PBMCs collected on day 1 post-tumor challenge
Splenocytes collected at end-of-life from control mice and surviving mIGV-001 treated mice

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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