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Protocol for In Vitro Activation of Jurkat E6-1 Cells Using Recombinant Human Galectin
Ruei-De Yan1, Ling Mao1, Yi-Jing Chen1
1Glycogenetics, Inc. 2F., No. 367, Sec. 3, Zhongzheng E. Rd., Dayuan Dist., Taoyuan, Taiwan.
Bio-Protocol
|August 11, 2026
Summary
Human galectins can modulate T-cell activation by crosslinking glycosylated surface receptors. This study provides a galectin toolset to investigate how glycosylation impacts adaptive immunity, confirming galectin-3 and galectin-8 induce immune responses.
Area of Science:
- Immunology
- Glycobiology
- Cellular Signaling
Background:
- T-cell activation is initiated by surface receptor engagement.
- Surface receptors are heavily glycosylated, suggesting lectin-mediated crosslinking can influence signaling.
- Galectins, as carbohydrate-binding proteins, can interact with these glycosylated receptors.
Purpose of the Study:
- To systematically evaluate recombinant human galectins for their ability to trigger immune responses.
- To establish a method for modulating Jurkat E6-1 cell activation using human galectins.
- To investigate the role of glycosylation in adaptive immunity through galectin-mediated crosslinking.
Main Methods:
- Screening of recombinant human galectins (galectin 1, 3, 7, 8 variants 1 and 2) for Jurkat E6-1 cell modulatory activity.
- Measuring cellular surface CD69 expression and Interleukin-2 (IL-2) mRNA levels.
- Utilizing a mechanism-based approach involving glycan crosslinking to induce receptor engagement.
Main Results:
- Galectin-3 and galectin-8 (variants 1 and 2) were confirmed to reproducibly induce CD69 and IL-2 expression on Jurkat E6-1 cells.
- Demonstrated the effectiveness of galectin-mediated crosslinking in modulating T-cell activation markers.
- Established a standardized workflow for evaluating extracellular galectin-driven immune activation.
Conclusions:
- Recombinant human galectins can be used as tools to modulate T-cell activation.
- This galectin-based approach provides a method to study the impact of glycosylation on adaptive immunity.
- The findings support the potential of galectins in understanding and manipulating immune responses.
