Generation of Aβ-specific T cell lines and in vivo Transfer
β-淀粉样蛋白（Aβ）特异性T淋巴细胞系的建立和过继转移
发布: 2014年09月20日第4卷第18期 DOI: 10.21769/BioProtoc.1241 浏览次数: 9168
评审: Kanika GeraAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Mar 2013
Abstract
Amyloid-β (Aβ)-containing plaques accumulate in the brains of patients with Alzheimer’s disease (AD). Studies in transgenic mice which over-express amyloid precursor protein and presenilin 1 (APP/PS1 mice) have suggested that T cells that infiltrate the brain may influence the development of Aβ plaques and associated cognitive dysfuncation. Active immunization with Aβ peptides and adjuvants has been evaluated as a therapy for AD, based on the premise that it induces Aβ-specific antibodies that may help to clear the Aβ plaques. However, immunization with Aβ peptides and adjuvants also promotes the development of Aβ-specific T cells (McQuillan et al., 2010) and there is evidence that Aβ-specific T cell may influence the development of Aβ plaques and disease progression in AD patients. In the mouse model, Aβ-specific T cells that secrete IFN-γ (Th1 cells) have been shown to enhance the plaque burden (Browne et al., 2013). Adoptive transfer of Aβ-specific T cells that have been polarized in vitro to Th1, Th2, Th17 or Treg cells can be used to examine the function of these cells in vivo.
Materials and Reagents
- C57BL/6 mice (adult, >6 weeks old; typically 4 per experiment)
- Aβ1-42 peptide (Life Technologies, InvitrogenTM, catalog number: 03112 )
- CpG (CpG-oligodeoxynucleotide 1668; 5′-tccatgacgttccgatgct-3′; Sigma-Genosys)
- PBS (Sigma-Aldrich, catalog number: D8537 )
- IL-1 (Immunotools, catalog number: 12340013 )
- IL-2 (Immunotools, catalog number: 12340024 )
- IL-4(Immunotools, catalog number: 12340043 )
- IL-12 (Miltenyi, catalog number: 130-096-707 )
- IL-23 (Miltenyi, catalog number: 130-096-676 )
- Anti–IFNγ (BD, catalog number: 554430 )
- Dexamethasone (Sigma-Aldrich, catalog number: D4902 )
- High-performance liquid chromatography (HPLC)-grade water (sterile ddH2O)
- RPMI medium (Sigma-Aldrich, catalog number: R0883 )
- Penicillin-streptomycin (Sigma-Aldrich, catalog number: P4333 )
- L-glutamine (Sigma-Aldrich, catalog number: G7513 )
- FBS (Sigma-Aldrich, catalog number: F9665 )
- Complete RPMI medium (see Recipes)
Equipment
- Shaker capable of 200 rpm at 37 °C
- 24-well cell culture plates (Greiner Bio-one, catalog number: 662160 )
- 1 ml tuberculin syringes (BD, catalog number: 300013 )
- 70 μm nylon mesh filter (Corning, catalog number: 352350 )
- Tissue culture facilities including class II laminar flow hood
- Centrifuge
Procedure
文章信息
版权信息
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
McManus, R. M., Lynch, M. A. and Mills, K. H. (2014). Generation of Aβ-specific T cell lines and in vivo Transfer. Bio-protocol 4(18): e1241. DOI: 10.21769/BioProtoc.1241.
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分类
免疫学 > 免疫细胞功能 > 淋巴细胞
免疫学 > 免疫细胞分离 > 淋巴细胞
细胞生物学 > 细胞分离和培养 > 细胞分离
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