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細胞池包括多個效應和記憶T細胞亞群的出現，通過抗原驅動的

膨脹和Naï幼稚T細胞的分化。的Naï已經CD4 + T細胞的抗原刺激的早期反應的特征

高水平的細胞因子的增殖和有限的劇目。進一步分化產生的細胞與不同的潛力

細胞因子的表達。根據局部細胞因子平衡，共刺激分子，抗原的水平，和遺傳

因素，Ⅰ型T輔助細胞（Th1，Th2和Th17細胞效應），和/或記憶細胞通過產生的免疫應答。

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.

功能性極化的CD4 + T細胞亞群是基于細胞分泌細胞因子的*的模式識別。作為一個

簽名細胞因子，Th1細胞選擇性地產生大量的干擾素γ（IFN-γ）。Th2細胞產生IL-4的選擇性，

Th17細胞表達高水平的IFN-γIL-17A。通過與其他效應分子的分泌，Th1細胞激活

巨噬細胞，自然殺傷（NK）細胞和CD8 + T細胞，并負責細胞免疫。Th1細胞提供

保護對細胞內的細菌，真菌，原生動物和病毒和參與一些自身免疫反應。IL-4

由Th2細胞產生特別強的驅動B細胞產生IgE的分泌細胞。IgE嗜堿性粒細胞和肥大作用

細胞介導的免疫反應。Th2細胞介導的防外寄生蟲也可能導致有害的過敏

反應性發展。通過對IL-17A和其他因子的分泌，招募和激活中性粒細胞和Th17細胞

調解對胞外細菌和真菌的免疫反應。Th17細胞也參與自身免疫反應。在

除了這些類型的T輔助細胞，

應注意，BD golgistop™蛋白轉運抑制劑的出現可能清楚的顏色范圍

（無色到淡黃色）。

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