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The Treg-Protector
T regulatory cells (Tregs) are a heavily researched cell population due to their impressive immunosuppressive abilities. However, recovering high numbers of these cells from mice can be a difficult task due to mechanisms like NAD-induced cell death. To help in this endeavor, BioLegend is proud to release the unique Treg-Protector (anti-ARTC2 Nanobody). Injection of this nanobody into mice is shown to improve Treg recovery, as they display decreased Annexin V binding and maintain characteristic Treg marker expression like CD25. Learn more about Nanobodies in our blog below and request a free sample from sales@biolegend.com.
A C57BL/6 mouse was injected (i.v.) with 50 ug of Treg-Protector 15 minutes before harvesting the spleen to isolate lymphocytes (left). The same procedure was done with a control mouse that only received PBS (right). The cells obtained from each mouse were incubated for one hour at 37C and then were stained with CD3 BV605, CD4 FITC, CD25 PE/Dazzle 594, and Annexin V BV421. Data shown was gated on the CD3+ and CD4+ population. Note the higher frequency of live Tregs (CD4+, CD25+, and Annexin V- T cells) in the mouse that was treated with Treg-Protector (top).
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Adapted from: Koch-Nolte, F., University Medical Center Hamburg-Eppendorf
Damaged cells release NAD (Nicotinamide Adenine Dinucleotide). The free-floating NAD is bound by cells expressing the enzyme, ARTC2. ARTC2 then transfers ADP-ribose onto surface proteins like P2X7. This activates P2X7 for ion transport and causes downstream effects like the cleavage of L-selectin (CD62L) and CD27 and outer membrane expression of phosphatidylserine (PS). This last effect leads to apoptosis and thus, this process is termed 'NAD-induced cell death', or NICD. Injection of mice with the Treg-Protector blocks ARTC2 function and thus, prevents apoptosis and shedding of characteristic surface markers of this cell type, making it easier to recover more live Tregs.
Learn more about the Treg-Protector
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