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【流式学习】γδT细胞:虽然我很少,但我很重要

作者:admin信息来源:达科为日期:2019年09月26日打印字体:  
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细胞表面抗原受体(T cell receptor, TCR )主要分为αβ TCR γδ TCR 两大类,αβTCR细胞大家都很熟悉了,今天小达君主要跟大家分享一下γδT细胞。γδT细胞是一群固有免疫样的T淋巴细胞,多数表型为 CD3+CD4CD8,主要分布在外周血和粘膜组织中,仅占人外周血 T 细胞的5%~10%。可不要小看γδT细胞,在受到刺激活化后一样能发挥多种效应功能,包括杀伤感染的细胞和肿瘤细胞、分泌细胞因子和趋化因子、抗原递呈功能及免疫调节功能。
 γδT细胞的亚群:

Human通常,人的γδT细胞根据其TCR基因重排过程中TCR δ链的不同主要分为两大亚群:Vδ1T细胞亚群和Vδ2T细胞亚群,其中,与Vδ1T细胞对应的TCRγ链主要为VγI基因家族成员(Vγ2/3/4/5/8),主要分布于黏膜和皮下组织中,在不同的组织中数量不等;Vδ2 T 细胞是γδ T 细胞的主要细胞群,其TCRγ链主要为VγII基因家族成员(Vγ9),因此又叫 Vγ9Vδ2T 细胞,仅存在于灵长类动物和人体内,主要分布于外周血中,占外周血淋巴细胞的0.5% 10%

Mouse:小鼠的γδT细胞由于其在体内分布及功能特性的不同可分为不同的亚群。根据γδT细胞在发育过程中TCR基因重排时所选择的链的不同,从结构上可分为Vγ1-Vγ7,七个亚群。
活化与极化:
以人的γδT细胞为例,用不同的细胞因子以及磷酸化的抗原刺激后,静息的γδT细胞可以极化为FoxP3+γδTregγδT17γδTfhγδT1,或成为活化的Vδ1TVδ2T细胞亚群。

功能:
γδT细胞具有多种功能,抗感染、抗肿瘤,在CAR-T治疗中也发挥重要作用。

流式检测:
以健康人的冷冻保存的PBMC为样本,通过多色流式染色分析γδT细胞的分化阶段。
样本类型:健康人的冷冻保存的PBMC
抗体:

设门策略:
(A)
  • 先通过FSC-A/SSC-A圈中淋巴细胞;

  • 通过FSC-A/W排除粘连体;

  • 通过SSC-A/W排除粘连体;

  • 通过EMA死活染料排除掉死细胞;

  • 圈中CD3T淋巴细胞;

  • CD3T淋巴细胞中接着分析CD4T细胞和CD8T细胞的比例;因为人外周血中γδT细胞的表型基本都是CD4CD8,所以圈出CD4CD8双阴性T细胞,准备分析γδT表型;

  • 圈中泛γδT细胞;

  • 分析泛γδT细胞中的Vδ1T细胞和Vδ2T细胞 2个亚群的比例,以及Vδ1Vδ2T双阴性细胞的比例;

(B)分析Vδ2T细胞中CD27CD28CD45RACD16等标记表达;
(C)分析Vδ1T细胞中CD27CD28CD45RACD16等标记表达。

γδT细胞的抗体清单:
BioLegend有很多检测γδT细胞的流式抗体,不同种属,多种荧光标记,满足您的实验需求。

Item

Cat

Size

Clone

APC anti-human  TCR γ/δ

331212

100 tests

B1

FITC anti-human  TCR γ/δ

331208

100 tests

B1

PE anti-human  TCR γ/δ

331210

100 tests

B1

PE/Cy7  anti-human TCR γ/δ

331222

100 tests

B1

PerCP/Cyanine5.5  anti-human TCR γ/δ

331224

100 tests

B1

Purified  anti-human TCR γ/δ

331202

100 ug

B1

LEAF™ Purified  anti-human TCR γ/δ

331204

500ug

B1

Biotin  anti-human TCR γ/δ

331206

100ug

B1

Alexa Fluor® 647  anti-human TCR γ/δ

331214

100 tests

B1

Brilliant Violet  421™ anti-human TCR γ/δ

331218

100 tests

B1

Brilliant Violet  510™ anti-human TCR γ/δ

331220

100 tests

B1

PE/Dazzle™ 594  anti-human TCR γ/δ

331226

100 tests

B1

APC/Fire™ 750  anti-human TCR γ/δ

331228

100 tests

B1

Purified anti-human  TCR Vγ9

331301

25ug

B3

FITC anti-human  TCR Vγ9

331306

100 tests

B3

PE anti-human  TCR Vγ9

331308

100 tests

B3

APC anti-human  TCR Vγ9

331310

100 tests

B3

PerCP anti-human  TCR Vδ2

331410

100 tests

B6

FITC anti-human  TCR Vδ2

331406

100 tests

B6

Pacific Blue™  anti-human TCR Vδ2

331414

100 tests

B6

Alexa Fluor® 700  anti-human TCR Vδ2

331416

100 tests

B6

APC anti-human  TCR Vδ2

331418

100 tests

B6

APC/Fire™ 750  anti-human TCR Vδ2

331420

100 tests

B6

PE/Cy7  anti-human TCR Vδ2

331422

100 tests

B6

PerCP/Cyanine5.5  anti-human TCR Vδ2

331424

100 tests

B6

Brilliant Violet  421™ anti-human TCR Vδ2

331428

100 tests

B6

Brilliant Violet  605™ anti-human TCR Vδ2

331430

100 tests

B6

Brilliant Violet  510™ anti-human TCR Vδ2

331432

100 tests

B6

PE/Dazzle™ 594  anti-human TCR Vδ2

331426

100 tests

B6

Brilliant Violet  711™ anti-human TCR Vδ2

331412

100 tests

B6

PE anti-human  TCR Vδ2

331408

100 tests

B6

Purified  anti-human TCR Vδ2

331402

100 ug

B6

Biotin  anti-human TCR Vδ2

331404

100 ug

B6

Purified  anti-mouse TCR γ/δ

107502

500ug

UC7-13D5

Brilliant Violet  421™ anti-mouse TCR γ/δ

118119

125ul

GL3

FITC anti-mouse  TCR γ/δ

118106

200 ug

GL3

Biotin  anti-mouse TCR γ/δ

118103

50 ug

GL3

LEAF™ Purified  anti-mouse TCR γ/δ

107506

500 ug

UC7-13D5

Alexa Fluor® 488  anti-mouse TCR γ/δ

107512

100 ug

UC7-13D5

APC anti-mouse  TCR γ/δ

118116

100 ug

GL3

PE anti-mouse  TCR γ/δ

118108

100 ug

GL3

FITC anti-mouse  TCR γ/δ

107504

500 ug

UC7-13D5

PE anti-mouse  TCR γ/δ

107508

100 ug

UC7-13D5

Purified  anti-mouse TCR γ/δ

118101

100 ug

GL3

LEAF™ Purified  anti-mouse TCR γ/δ

118114

500 ug

GL3

PerCP/Cyanine5.5  anti-mouse TCR γ/δ

118118

100 ug

GL3

Brilliant Violet  421™ anti-mouse TCR γ/δ

118120

50 ug

GL3

PE/Cy7  anti-mouse TCR γ/δ

118124

100 ug

GL3

Alexa Fluor® 488  anti-mouse TCR γ/δ

118128

100 ug

GL3

Brilliant Violet  605™ anti-mouse TCR γ/δ

118129

50 ug

GL3

Brilliant Violet  510™ anti-mouse TCR γ/δ

118131

50 ug

GL3

Alexa Fluor® 647  anti-mouse TCR γ/δ

118134

100 ug

GL3

APC/Fire™ 750  anti-mouse TCR γ/δ

118136

100 ug

GL3

APC anti-mouse  TCR Vγ1.1/Cr4 Antibody

141108

100 ug

2.11

FITC anti-mouse  TCR Vγ1.1/Cr4 Antibody

141104

100 ug

2.11

PE anti-mouse  TCR Vγ1.1/Cr4 Antibody

141106

100 ug

2.11

PerCP/Cyanine5.5  anti-mouse TCR Vγ1.1/Cr4 Antibody

141112

100 ug

2.11

Pacific Blue™  anti-mouse TCR Vγ1.1/Cr4 Antibody

141110

100 ug

2.11

Purified  anti-mouse TCR Vγ1.1/Cr4 Antibody

141102

100 ug

2.11

APC anti-mouse  TCR Vγ2 Antibody

137708

100 ug

UC3-10A6

FITC anti-mouse  TCR Vγ2 Antibody

137704

200 ug

UC3-10A6

PE anti-mouse  TCR Vγ2 Antibody

137706

200 ug

UC3-10A6

APC anti-mouse  TCR Vγ3 Antibody

137506

100 ug

536

PE anti-mouse  TCR Vγ3 Antibody

137504

100 ug

536

APC anti-mouse  TCR Vδ6.3 Antibody

154806

100 ug

C504.17C

FITC anti-mouse  TCR Vδ6.3 Antibody

154808

100 ug

C504.17C

PE anti-mouse  TCR Vδ6.3 Antibody

154804

100 ug

C504.17C

Purified  anti-mouse TCR Vδ6.3 Antibody

154802

500 µg

C504.17C

PE anti-mouse  TCR Vδ4 Antibody

134905

25 µg

GL2

PE anti-mouse  TCR Vγ1.1 + Vγ1.2 Antibody

142704

100 ug

4B2.9

Purified  anti-mouse TCR Vγ1.1 + Vγ1.2 Antibody

142702

100 ug

4B2.9

PE anti-rat TCR  γ/δ

202605

50 ug

V65

参考文献:
  • Gamma-delta(γδ) T cells: friend or foe in cancer development? J Transl Med.
  • Six-of-the-best:unique contributions of γδ T cells toimmunology. Nat Rev Immunol.
  • OMIP‐020:Phenotypic characterization of human γδ T‐cells by multicolor flow cytometry. Cytometry A.
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