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Degranulation Response in Cytotoxic Rat Lymphocytes Measured with a Novel CD107a Antibody
Amanda Sudworth1, Ke-Zheng Dai2, John T Vaage2
1Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway; Department of Immunology, University of Oslo, Oslo University Hospital, Rikshospitalet, Oslo, Norway.
Insights
Researchers developed a new antibody to measure degranulation in rat cytotoxic cells. This tool enables functional studies of natural killer (NK) cells and CD8 T cells, overcoming previous limitations in rat immunology research.
Area of Science:
- Immunology
- Cellular Biology
Background:
- CD107a expression is a key indicator of cytotoxic cell degranulation and function.
- Previous studies in rats were limited by the absence of a specific anti-rat CD107a antibody.
Purpose of the Study:
- To develop and characterize a novel antibody for detecting rat CD107a expression.
- To enable functional assessment of cytotoxic cells in rats.
Main Methods:
- Generation of a hybridoma (SIM1 clone) by immunizing Armenian hamsters with CD107a-expressing cells.
- Testing SIM1 antibody reactivity with rat natural killer (NK) cells and T cells.
- Measuring degranulation in response to target cell stimulation and receptor crosslinking.
Main Results:
- The SIM1 antibody specifically recognizes rat CD107a.
- SIM1 successfully measured degranulation in NK cells under various stimulation conditions, including low effector-to-target ratios.
- The antibody also stained activated CD8 T cells but not CD4 T cells.
Conclusions:
- A novel anti-rat CD107a antibody (SIM1) has been developed, facilitating degranulation studies in rats.
- This antibody is valuable for assessing the functional capacity of rat NK and CD8 T cells.
- The findings overcome a significant hurdle in rat immunology, enabling more detailed functional analysis of cytotoxic responses.
Abstract:
Measuring degranulation through CD107a expression has become an advantageous tool for testing the functional capacity of cytotoxic cells. Such functional studies have been hampered in the rat by the lack of a suitable anti-rat CD107a antibody. In this study, we report a novel hybridoma generated by immunizing Armenian inbred hamsters with transfected Chinese hamster ovary cells expressing CD107a. The SIM1 clone exhibited specific reactivity with CD107a and measured degranulation from natural killer (NK) cells stimulated with target cells or mAb crosslinking of their activating receptors. Degranulation in IL-2-activated NK cells could also be measured, when using low effector to target ratios. SIM1 also stained activated CD8, but not CD4 T cells. This report characterizes the degranulation response in cytotoxic rat cells with a new antibody against rat CD107a.
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