BioVector? 2B4 小鼠 T 淋巴細(xì)胞雜交瘤 / BioVector? 2B4 Mouse T Hybridoma Cell Line

通用定義 / General Definition:BioVector? 2B4 是一種經(jīng)典的小鼠 T 淋巴細(xì)胞雜交瘤細(xì)胞系，在免疫學(xué)研究中具有里程碑意義。它是由通過(guò)鴿細(xì)胞色素 C (Pigeon Cytochrome c, PCC) 免疫的 B10.A 小鼠 T 細(xì)胞與缺失內(nèi)源性 TCR 的融合受體細(xì)胞（如 BW5147）融合而成的。2B4 細(xì)胞系的主要特征是其 TCR 能夠特異性識(shí)別由 I-Ek (MHC class II) 分子呈遞的 PCC 肽段（特別是 81-104 殘基）。它是研究 T 細(xì)胞抗原識(shí)別、TCR 與 MHC-肽復(fù)合物相互作用動(dòng)力學(xué)以及 T 細(xì)胞早期信號(hào)轉(zhuǎn)導(dǎo)（如 IL-2 分泌）的標(biāo)準(zhǔn)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 技術(shù)說(shuō)明書 (Technical Datasheet)

中文版說(shuō)明書 (Chinese Datasheet)

1. 產(chǎn)品基本信息

• 產(chǎn)品名稱： BioVector? 2B4 小鼠 T 雜交瘤細(xì)胞

• 抗原特異性： 鴿細(xì)胞色素 C (PCC) 81-104 肽段

• MHC 限制性： I-Ek (Class II MHC)

• 反應(yīng)指標(biāo)： 受抗原刺激后分泌 IL-2

• 生長(zhǎng)特性： 懸浮生長(zhǎng)

• 細(xì)胞形態(tài)： 典型的淋巴母細(xì)胞樣，圓形，成單個(gè)或小簇懸浮。

2. 培養(yǎng)條件

• 基礎(chǔ)培養(yǎng)基： BioVector? RPMI 1640。

• 血清添加： 10% BioVector? 優(yōu)質(zhì)胎牛血清 (FBS)。

• 補(bǔ)充因子： 建議添加 1 mM 丙酮酸鈉、2 mM L-谷氨酰胺和 50 uM beta-巰基乙醇。

• 培養(yǎng)環(huán)境： 37 攝氏度，5% CO2。

• 傳代比例： 1:3 至 1:6；密度建議維持在 200,000 到 1,000,000 cells/mL。

3. 細(xì)胞應(yīng)用

• 抗原呈遞實(shí)驗(yàn) (APC Assay)： 用于檢測(cè)抗原呈遞細(xì)胞（如 DC 或表達(dá) I-Ek 的 B 細(xì)胞）的呈遞效率。

• TCR 信號(hào)轉(zhuǎn)導(dǎo)： 研究 TCR 結(jié)合配體后引發(fā)的 CD3 磷酸化、鈣流及轉(zhuǎn)錄因子激活。

• 超抗原研究： 2B4 TCR 對(duì)某些細(xì)菌超抗原（如 SEB）具有反應(yīng)性。

4. 注意事項(xiàng)

• 抗原要求： 實(shí)驗(yàn)通常使用 PCC 蛋白或合成的 PCC (81-104) 肽段。

• 信號(hào)敏感性： 長(zhǎng)期連續(xù)傳代可能導(dǎo)致 TCR 表達(dá)量下降或 IL-2 分泌能力減弱，建議定期使用流式細(xì)胞術(shù)檢測(cè) CD3/TCR 的表面表達(dá)。

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 細(xì)胞應(yīng)在生物安全二級(jí) (BSL-2) 條件下操作。IL-2 的檢測(cè)通常采用 CTLL-2 生物活性測(cè)定或商業(yè)化的 ELISA 試劑盒。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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