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【流式學習】γδT細胞:雖然我很少,但我很重要

作者:admin信息來源:達科為日期:2019年09月26日打印字體:  
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細胞表面抗原受體(T cell receptor, TCR )主要分為αβ TCR γδ TCR 兩大類,αβTCR細胞大家都很熟悉了,今天小達君主要跟大家分享一下γδT細胞。γδT細胞是一群固有免疫樣的T淋巴細胞,多數表型為 CD3+CD4CD8,主要分布在外周血和粘膜組織中,僅占人外周血 T 細胞的5%~10%。可不要小看γδT細胞,在受到刺激活化后一樣能發揮多種效應功能,包括殺傷感染的細胞和腫瘤細胞、分泌細胞因子和趨化因子、抗原遞呈功能及免疫調節功能。
 γδT細胞的亞群:

Human通常,人的γδT細胞根據其TCR基因重排過程中TCR δ鏈的不同主要分為兩大亞群:Vδ1T細胞亞群和Vδ2T細胞亞群,其中,與Vδ1T細胞對應的TCRγ鏈主要為VγI基因家族成員(Vγ2/3/4/5/8),主要分布于黏膜和皮下組織中,在不同的組織中數量不等;Vδ2 T 細胞是γδ T 細胞的主要細胞群,其TCRγ鏈主要為VγII基因家族成員(Vγ9),因此又叫 Vγ9Vδ2T 細胞,僅存在于靈長類動物和人體內,主要分布于外周血中,占外周血淋巴細胞的0.5% 10%

Mouse:小鼠的γδT細胞由于其在體內分布及功能特性的不同可分為不同的亞群。根據γδT細胞在發育過程中TCR基因重排時所選擇的鏈的不同,從結構上可分為Vγ1-Vγ7,七個亞群。
活化與極化:
以人的γδT細胞為例,用不同的細胞因子以及磷酸化的抗原刺激后,靜息的γδT細胞可以極化為FoxP3+γδTregγδT17γδTfhγδT1,或成為活化的Vδ1TVδ2T細胞亞群。

功能:
γδT細胞具有多種功能,抗感染、抗腫瘤,在CAR-T治療中也發揮重要作用。

流式檢測:
以健康人的冷凍保存的PBMC為樣本,通過多色流式染色分析γδT細胞的分化階段。
樣本類型:健康人的冷凍保存的PBMC
抗體:

設門策略:
(A)
  • 先通過FSC-A/SSC-A圈中淋巴細胞;

  • 通過FSC-A/W排除粘連體;

  • 通過SSC-A/W排除粘連體;

  • 通過EMA死活染料排除掉死細胞;

  • 圈中CD3T淋巴細胞;

  • CD3T淋巴細胞中接著分析CD4T細胞和CD8T細胞的比例;因為人外周血中γδT細胞的表型基本都是CD4CD8,所以圈出CD4CD8雙陰性T細胞,準備分析γδT表型;

  • 圈中泛γδT細胞;

  • 分析泛γδT細胞中的Vδ1T細胞和Vδ2T細胞 2個亞群的比例,以及Vδ1Vδ2T雙陰性細胞的比例;

(B)分析Vδ2T細胞中CD27CD28CD45RACD16等標記表達;
(C)分析Vδ1T細胞中CD27CD28CD45RACD16等標記表達。

γδT細胞的抗體清單:
BioLegend有很多檢測γδT細胞的流式抗體,不同種屬,多種熒光標記,滿足您的實驗需求。

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

參考文獻:
  • 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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