Jurkat JE6.1 NF-kB::eGFP hTLR5 (human Toll-like Receptor 5) Cell Line

Contamination with bacterial components is a potential issue with the biologics for research and therapeutic uses, as they can activate the so-called pattern-recognition receptors in mammalian cells and trigger immune responses, clouding the experimental results or even causing health hazards. Such responses are often mediated by the Toll-Like Receptors (TLRs), which recognize bacterial molecular patterns to activate, among other things, cytokine expression through the NF-kB pathway.

The current assessment method for bacterial contamination measures lipopolysaccharide (LPS), the active component of endotoxins. Endotoxin measurement relies on its reaction with the limulus amoebocyte lysate (LAL), derived from the horseshoe crab blood.3 While the method is well established, however, the use of primary lysate obtained from wild animals presents a major sustainability issue.4 Recombinant alternatives to the primary LAL method have recently been gaining acceptance, but all LAL-based methods measure only the endotoxin contamination, and other classes of bacterial components go undetected.

To address these unmet needs, the Jurkat JE6.1 cell line has been engineered to form a group of cell lines, each expressing a specific set of TLRs and harboring an eGFP expression cassette fused to the NF-kB Response Element. The result is that each of these strains expresses eGFP in response to a specific class of bacterial molecules.

SCC631, Jurkat JE6.1 NF-kB::eGFP hTLR5 cell line, expresses recombinant TLR5 on its surface and is designed to express eGFP in response to flagellin, a structural protein of flagella. It is not responsive to other bacterial components, such as LPS (ligand for SCC630, TLR4 expressing), and FSL-1 (ligand for SCC632, the TLR2/TLR6 cell line). It has also been tested negative for PAM3CSK4 (ligand for TLR2/TLR1).

Source
Jurkat JE6.1 was retrovirally (pBABE-MN) transduced with the NF-κB reporter construct, which is activated by the endogenous TLR5. SCC631 also expresses TLR6 endogenously, but it does not interact with TLR5.

Jurkat JE6.1 NF-kB::eGFP hTLR5 cell line expresses recombinant TLR5 on its surface and is designed to express eGFP in response to flagellin, a structural protein of flagella.

Jurkat JE6.1 T NF-kB::eGFP hTLR5 cells should be stored in liquid nitrogen. The cells can be cultured for at least 10 passages without significantly affecting cell marker expression and function.

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