HumanTh1/Th2/Th17 Phenotyping Kit

HumanTh1/Th2/Th17 Phenotyping Kit
Product Information
Material Number: 560751
Size: 50 tests
Description
Components:
51-9006615 Human Th1/Th2/Th17 Phenotyping Cocktail 1.0 ml
Containing the following:
Human CD4 PERCP-CY5.5 (clone: SK3)
Human IL-17A PE (clone: N49-653)
Human IFN-GMA FITC (clone: B27)
Human IL-4 APC (clone: MP4-25D2)
51-9006613 BD Cytofix™ Fixation Buffer 100 ml
51-2091KE BD Perm/Wash™ Buffer 25 ml
51-2092KZ BD GolgiStop™ Protein Transport Inhibitor (containing monensin) 0.7 ml
The peripheral CD4+ T cell pool includes multiple effector and memory T cell subsets that arise through antigen-driven
expansion and differentiation of naïve T cells. The early response of naïve CD4+ T cells to antigenic stimulation is characterized
by high level proliferation and a limited cytokine repertoire. Further differentiation yields cells with a more diverse potential for
cytokine expression. Depending upon the balance of local cytokines, costimulatory molecules, antigen levels, and genetic
factors, Type-1 T helper (Th1), Th2, and Th17 effector and/or memory cells are generated by immune responses.

外周血CD4+ T细胞池包括多个效应和记忆T细胞亚群的出现，通过抗原驱动的

膨胀和Naï幼稚T细胞的分化。的Naï已经CD4 + T细胞的抗原刺激的早期反应的特征

高水平的细胞因子的增殖和有限的剧目。进一步分化产生的细胞与不同的潜力

细胞因子的表达。根据局部细胞因子平衡，共刺激分子，抗原的水平，和遗传

因素，Ⅰ型T辅助细胞（Th1，Th2和Th17细胞效应），和/或记忆细胞通过产生的免疫应答。

Functionally-polarized CD4+ T cell subsets have been identified based on their distinctive patterns of cytokine secretion. As a
signature cytokine, Th1 cells selectively produce large amounts of interferon-gamma (IFN-γ). Th2 cells selectively produce IL-4,
and Th17 express high levels of IL-17A. Through secretion of IFN-γ and other effector molecules, Th1 cells activate
macrophages, natural killer (NK) cells, and CD8+ T cells and are responsible for cell-mediated immunity. Th1 cells provide
protection against intracellular bacteria, fungi, protozoa and viruses and are involved in some autoimmune responses. IL-4
produced by Th2 cells is particularly strong in driving B cells to generate IgE-secreting cells. IgE plays a role in basophil/mast
cell mediated immune reactions. Th2 cells mediate protection against extracellular parasites but may also cause harmful allergic
responsiveness to develop. Through the secretion of IL-17A and other factors, Th17 cells recruit and activate neutrophils and
mediate immune responses against extracellular bacteria and fungi. Th17 cells are also implicated in autoimmune responses. In
addition to these types of T helper Investigators should note that the appearance of BD GolgiStop™ Protein Transport Inhibitor may range in color from clear
(colorless) to light yellow.

功能性极化的CD4 + T细胞亚群是基于细胞分泌细胞因子的*的模式识别。作为一个

签名细胞因子，Th1细胞选择性地产生大量的干扰素γ（IFN-γ）。Th2细胞产生IL-4的选择性，

Th17细胞表达高水平的IFN-γIL-17A。通过与其他效应分子的分泌，Th1细胞激活

巨噬细胞，自然杀伤（NK）细胞和CD8 + T细胞，并负责细胞免疫。Th1细胞提供

保护对细胞内的细菌，真菌，原生动物和病毒和参与一些自身免疫反应。IL-4

由Th2细胞产生特别强的驱动B细胞产生IgE的分泌细胞。IgE嗜碱性粒细胞和肥大作用

细胞介导的免疫反应。Th2细胞介导的防外寄生虫也可能导致有害的过敏

反应性发展。通过对IL-17A和其他因子的分泌，招募和激活中性粒细胞和Th17细胞

调解对胞外细菌和真菌的免疫反应。Th17细胞也参与自身免疫反应。在

除了这些类型的T辅助细胞，

应注意，BD golgistop™蛋白转运抑制剂的出现可能清楚的颜色范围

（无色到淡黄色）。

Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Irritating to eyes and skin. Do not breathe vapor. In case of contact with eyes, rinse immediay with plenty of water and seek medical
advice. Wear suitable protective clothing.
The antibody was conjugated to APC under optimum conditions, and unconjugated antibody and free APC were removed.
The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were
removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
The antibody was conjugated with FITC under optimum conditions, and unreacted FITC was removed.

A. Stimulation of the Cells
Various in vitro methods have been reported for polarization or stimulation of T helper cells subsets of which PMA (Phorbol ester) plus
Ionomycin (Calcium Ionophore) has been particularly useful for quickly inducing and characterizing polyclonal cytokine-producing cells. For this
kit we recommend the stimulation of normal PBMCs at a concentration of 1-10 million cells per ml in media for 5 hours with PMA/Ionomycin (at
50 ng/ml and 1 μg/ml respectively) in the presence of BD GolgiStop™ Protein Transport Inhibitor (provided in the kit or Cat #554724). Add 4 μl
of BD GolgiStop™ for every 6 ml of cell culture and mix thoroughly. It is recommended that BD GolgiStop™ not be kept in cell culture for
longer than 12 hours.
Note: Kinetic studies need to be performed to determine the optimal incubation time for each experimental system. Depending on the donor, frequencies of cytokine
producing cells derived from activation of PBMC can vary widely for a specific cytokine. In particular, the number of IL-17 producing cells can be very low or even
negligible on PMA/Ionomycin stimulated PBMC. In these cases, Th17 polarization cultures should be considered. For specifics on polarization of Th17 cells please refer
to the references.
560751