調(diào)節(jié)性T細(xì)胞（regulatory T，Treg）是一類控制體內(nèi)免疫反應(yīng)性的T細(xì)胞亞群，根據(jù)來源的不同，Treg 細(xì)胞可分為自然性Treg (natural Treg, nTreg) 和誘導(dǎo)性 Treg（induced Treg, iTreg）。

       nTreg由胸腺分化而來，占外周血CD4+ T細(xì)胞的5%~10%，這類細(xì)胞本身缺乏增殖能力，但具有天然免疫抑制作用，不僅可抑制CD4+或CD8+ T細(xì)胞的活化和增殖，還可抑制初始T細(xì)胞和記憶性T細(xì)胞的功能。iTreg主要存在于外周血和腫瘤組織中，由抗原或抑制性因子如IL-2、TGF-β刺激初始CD4+ T細(xì)胞產(chǎn)生，對多種免疫細(xì)胞具有抑制作用。

       調(diào)節(jié)性T（regulatoryT，Treg）細(xì)胞對于調(diào)節(jié)免疫穩(wěn)態(tài)和抑制不同疾病環(huán)境中的免疫反應(yīng)具有至關(guān)重要的作用，它能通過抑制CD4+輔助性T細(xì)胞和CD8+細(xì)胞毒性T細(xì)胞的激活和分化，對自體和腫瘤表達(dá)的抗原產(chǎn)生反應(yīng)性。Treg細(xì)胞表達(dá)特異性轉(zhuǎn)錄因子Foxp3，因而，CD4+CD25+Foxp3+被認(rèn)為是Treg細(xì)胞的經(jīng)典標(biāo)記組合。

       盡管如此，選什么亞型，選什么來源，選什么熒光？依舊是小伙伴們?yōu)镃D4+CD25+Foxp3+抗體組合的選擇的一大困擾！

       作為專業(yè)的流式抗體與解決方案，艾美捷科技，為您推薦流式科研與診斷方案領(lǐng)導(dǎo)者EXBio的CD4+CD25+FOXP3+Treg調(diào)節(jié)性T細(xì)胞檢測試劑盒（TregFlowEx Kit）

      試劑盒組分：

      (CD4+CD25+FOXP3+) Treg調(diào)節(jié)性T細(xì)胞檢測試劑盒實(shí)驗(yàn)流程

      1.Surface staining：

      1.1 Add 10 ?l of CD4 FITC/CD25 PE reagent to tubes.

      1.2 Add 100 ?l of human blood.

      1.3 Mix well and incubate for 10 minutes in the dark in the refrigerator (2?8℃).

      1.4 (optional wash) Wash cells by adding 2 ml of cold PBS and centrifuge at 300 g for 5 minutes at 4℃.

      1.5 Aspirate supernatant and proceed immediately to the intracellular staining.

      2.Intracellular staining

      2.1 Add 1 ml of the diluted cold Fix and Lysing Solution.

      2.2 Mix and incubate for 10 minutes in the dark in the refrigerator (2?8℃).

      2.3 Add 0.5 ml of Permeabilizing Solution.

      2.4 Mix and incubate for 10 minutes in the dark in the refrigerator (2?8℃).

      2.5 Centrifuge the cells at 400 g for 5 minutes at 4℃. (Do not add any other buffer to the Fix/Perm mixture before centrifugation)

      2.6 Remove (decant) supernatant.

      2.7 Add 50 ?L of cold Blocking Buffer.

      2.8 Add 50 ?L of cold PBS.

      2.9 Mix and incubate for 5 minutes in the dark in the refrigerator (2?8℃).

      2.10 Add 5 μl of FoxP3 APC reagent.

      2.11 Mix and incubate for 30 minutes in the dark in the refrigerator (2?8℃).

      2.12. Wash cells twice with 2 ml of cold PBS, centrifugate at 400 g for 5 minutes at 4 ℃. Aspirate/decant supernatant.

      2.13. Add 300 ?l of 1% formaldehyde in PBS and store the stained samples at 2?8℃ until measured in flow cytometer. Analyze the processed samples within 4 hours.

      (CD4+CD25+FOXP3+) Treg調(diào)節(jié)性T細(xì)胞檢測試劑盒結(jié)果展示：

      使用TregFlowEx試劑盒中對人CD4陽性淋巴細(xì)胞進(jìn)行流式細(xì)胞術(shù)多色染色。 Treg細(xì)胞（CD4 + CD25 + FOXP3 +）位于右上象限。

      【發(fā)表文章】

      Cern?, V., et al. "Lower Functional and Proportional Characteristics of Cord Blood Treg of Male Newborns Compared with Female Newborns. Biomedicines 2021, 9, 170." (2021).

      Morizawa, Yosuke, et al. "Correlation of immune cells and cytokines in the tumor microenvironment with elevated neutrophil-to-lymphocyte ratio in blood: an analysis of muscle-invasive bladder cancer."Cancer investigation7 (2018): 395-405.

      Hrd?, Ji?í, et al. "Decreased allergy incidence in children supplemented with E. coli O83: K24: H31 and its possible modes of action."European journal of immunology12 (2018): 2015-2030.

      Ond?ejíková, Leona.Rozdílné vlastnosti T regula?ních buněk v pupe?níkové krvi novorozenc? zdrav?ch a alergick?ch matek. BS thesis. ?eské vysoké u?ení technické v Praze. Vypo?etní a informa?ní centrum., 2017.

      【干貨】CD家族流式抗體大全（高清大圖）