BioVector® 2B4 小鼠 T 淋巴细胞杂交瘤 / BioVector® 2B4 Mouse T Hybridoma Cell Line

通用定义 / General Definition:BioVector® 2B4 是一种经典的小鼠 T 淋巴细胞杂交瘤细胞系，在免疫学研究中具有里程碑意义。它是由通过鸽细胞色素 C (Pigeon Cytochrome c, PCC) 免疫的 B10.A 小鼠 T 细胞与缺失内源性 TCR 的融合受体细胞（如 BW5147）融合而成的。2B4 细胞系的主要特征是其 TCR 能够特异性识别由 I-Ek (MHC class II) 分子呈递的 PCC 肽段（特别是 81-104 残基）。它是研究 T 细胞抗原识别、TCR 与 MHC-肽复合物相互作用动力学以及 T 细胞早期信号转导（如 IL-2 分泌）的标准模型。

BioVector® 2B4 is a classic Mouse T-cell Hybridoma cell line with significant importance in fundamental immunology. It was generated by fusing T-lymphocytes from B10.A mice immunized with Pigeon Cytochrome c (PCC) with a TCR-negative fusion partner (such as BW5147). The defining characteristic of the 2B4 line is its T-cell receptor (TCR), which specifically recognizes the PCC peptide (residues 81-104) presented by the I-Ek (MHC class II) molecule. It serves as a gold-standard model for investigating T-cell antigen recognition, the biophysics of TCR-pMHC interactions, and early T-cell activation signaling, such as IL-2 production.

BioVector® 2B4 技术说明书 (Technical Datasheet)

中文版说明书 (Chinese Datasheet)

1. 产品基本信息

• 产品名称： BioVector® 2B4 小鼠 T 杂交瘤细胞

• 抗原特异性： 鸽细胞色素 C (PCC) 81-104 肽段

• MHC 限制性： I-Ek (Class II MHC)

• 反应指标： 受抗原刺激后分泌 IL-2

• 生长特性： 悬浮生长

• 细胞形态： 典型的淋巴母细胞样，圆形，成单个或小簇悬浮。

2. 培养条件

• 基础培养基： BioVector® RPMI 1640。

• 血清添加： 10% BioVector® 优质胎牛血清 (FBS)。

• 补充因子： 建议添加 1 mM 丙酮酸钠、2 mM L-谷氨酰胺和 50 uM beta-巯基乙醇。

• 培养环境： 37 摄氏度，5% CO2。

• 传代比例： 1:3 至 1:6；密度建议维持在 200,000 到 1,000,000 cells/mL。

3. 细胞应用

• 抗原呈递实验 (APC Assay)： 用于检测抗原呈递细胞（如 DC 或表达 I-Ek 的 B 细胞）的呈递效率。

• TCR 信号转导： 研究 TCR 结合配体后引发的 CD3 磷酸化、钙流及转录因子激活。

• 超抗原研究： 2B4 TCR 对某些细菌超抗原（如 SEB）具有反应性。

4. 注意事项

• 抗原要求： 实验通常使用 PCC 蛋白或合成的 PCC (81-104) 肽段。

• 信号敏感性： 长期连续传代可能导致 TCR 表达量下降或 IL-2 分泌能力减弱，建议定期使用流式细胞术检测 CD3/TCR 的表面表达。

English Datasheet

1. General Product Information

• Product Name: BioVector® 2B4 Mouse T Hybridoma Cell Line

• Antigen Specificity: Pigeon Cytochrome c (PCC) residues 81-104

• MHC Restriction: I-Ek (MHC class II)

• Response Marker: Secretion of IL-2 upon successful activation

• Growth Properties: Suspension

• Morphology: Lymphoblast-like; small round cells growing singly or in loose clusters.

2. Culture Conditions

• Basal Medium: BioVector® RPMI 1640.

• Serum Supplement: 10% BioVector® Fetal Bovine Serum (FBS).

• Supplements: 1 mM Sodium Pyruvate, 2 mM L-Glutamine, and 50 uM beta-mercaptoethanol are strongly recommended for maintaining T-cell vitality.

• Incubation: 37°C, 5% CO2.

• Subculturing: 1:3 to 1:6 ratio; keep cell density between 200,000 and 1,000,000 cells/mL.

3. Applications

• Antigen Presentation Assays: Measuring the efficiency of antigen processing and presentation by I-Ek-bearing APCs.

• TCR Signaling: Evaluating the biochemical cascades triggered by TCR engagement, including kinase activation and downstream cytokine gene transcription.

• Epitope Mapping: Testing variants of the PCC peptide to understand the structural basis of TCR binding and discrimination.

4. Key Usage Notes

• TCR Integrity: Hybridoma cells can sometimes lose the expression of their specific TCR chains over time. BioVector® recommends verifying surface TCR/CD3 expression by flow cytometry before initiating critical assays.

• Assay Ready Status: For activation assays, cells should be harvested in the log-phase of growth to ensure maximum IL-2 production potential.

注意 / Note: 2B4 细胞应在生物安全二级 (BSL-2) 条件下操作。IL-2 的检测通常采用 CTLL-2 生物活性测定或商业化的 ELISA 试剂盒。2B4 cells should be handled under Biosafety Level 2 (BSL-2) conditions. IL-2 production is typically quantified via CTLL-2 bioassay or standardized ELISA kits.

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