The NTCC? CEM.NKR-CCR5-Luc cell line is a highly specialized tool in HIV/AIDS research, building upon the CEM.NKR-CCR5 cell line by adding a reporter gene.

Key Characteristics

• Parental Line: It is a derivative of the CEM.NKR-CCR5 cell line, which is in turn derived from the CEM.NKR line.

• Genetic Modification: This cell line has been genetically engineered to stably express two key components:

  1. CCR5: The human C-C chemokine receptor type 5, a critical coreceptor for R5-tropic HIV-1 entry.

  2. Luciferase Reporter Gene: This is the defining feature of the "Luc" variant. The luciferase gene is under the transcriptional control of the HIV-2 LTR (long terminal repeat).

• Mechanism of Action: When the CEM.NKR-CCR5-Luc cells are infected with HIV, the HIV Tat protein activates the HIV LTR promoter. This, in turn, drives the expression of the luciferase gene. By adding a luciferase substrate (like luciferin) to the culture, researchers can measure the amount of light produced, which directly correlates with the level of HIV replication. This provides a highly sensitive and quantifiable readout for infection.

• Susceptibility to HIV-1: Like its parent line, it expresses both CD4 and the coreceptors CCR5 and CXCR4. This makes it susceptible to a wide range of HIV-1 isolates, including both R5-tropic and X4-tropic strains.

Research Applications

The addition of the luciferase reporter gene makes the CEM.NKR-CCR5-Luc cell line extremely useful for high-throughput and quantitative assays, particularly for:

• HIV Neutralization Assays: This is its most common application. Researchers can use the cell line to test how well antibodies from patient serum or experimental drugs can neutralize HIV infection by measuring the reduction in luciferase activity.

• Antiretroviral Drug Screening: It is used to rapidly and quantitatively screen a large number of compounds for their ability to inhibit HIV replication. The luciferase readout provides a clear signal for a compound's antiviral effect.

• Studying HIV Entry Inhibitors: The cell line is an excellent model for studying drugs that specifically block HIV from entering cells by targeting the CCR5 or CXCR4 coreceptors.

• Vaccine Efficacy Testing: It is a valuable tool for assessing the effectiveness of HIV vaccine candidates by measuring their ability to elicit a neutralizing antibody response that prevents viral infection.

The use of this reporter system is a significant advancement over older methods, such as p24 antigen ELISA, because it is faster, more sensitive, and less labor-intensive, making it ideal for high-throughput screening.

NTCC? CEM.NKR-CCR5-Luc 細(xì)胞系是艾滋?。℉IV/AIDS）研究中一種高度專業(yè)化的工具，它在 CEM.NKR-CCR5 細(xì)胞系的基礎(chǔ)上，通過添加報(bào)告基因而進(jìn)一步優(yōu)化。

主要特點(diǎn)

• 親本細(xì)胞系: 它是 CEM.NKR-CCR5 細(xì)胞系的衍生品，而 CEM.NKR-CCR5 又源自 CEM.NKR 細(xì)胞系。

• 基因改造: 該細(xì)胞系經(jīng)過基因工程改造，可穩(wěn)定表達(dá)兩個(gè)關(guān)鍵組分：

  1. CCR5: 人類 C-C 趨化因子受體5型，是 R5 嗜性 HIV-1 進(jìn)入細(xì)胞的關(guān)鍵共受體。

  2. 熒光素酶報(bào)告基因 (Luc): 這是“Luc”變體的核心特征。該熒光素酶基因的表達(dá)受到 HIV-2 LTR（長末端重復(fù)序列）的轉(zhuǎn)錄控制。

• 作用機(jī)制: 當(dāng) CEM.NKR-CCR5-Luc 細(xì)胞被 HIV 感染后，HIV 的 Tat 蛋白會(huì)激活 HIV LTR 啟動(dòng)子。這反過來會(huì)驅(qū)動(dòng)熒光素酶基因的表達(dá)。通過向培養(yǎng)物中添加熒光素酶底物（如熒光素），研究人員可以測量產(chǎn)生的發(fā)光量，這與 HIV 復(fù)制水平直接相關(guān)。這種機(jī)制為感染提供了高度敏感和可量化的讀數(shù)。

• 對(duì) HIV-1 的敏感性: 像其親本細(xì)胞系一樣，它同時(shí)表達(dá) CD4 受體以及 CCR5 和 CXCR4 共受體。這使得它對(duì)廣泛的 HIV-1 分離株（包括 R5 嗜性和 X4 嗜性毒株）都具有敏感性。

研究應(yīng)用

熒光素酶報(bào)告基因的加入使得 CEM.NKR-CCR5-Luc 細(xì)胞系特別適合進(jìn)行高通量和定量分析，主要用于：

• HIV 中和試驗(yàn): 這是其最常見的應(yīng)用。研究人員可以通過測量熒光素酶活性的降低程度，來測試來自患者血清的抗體或?qū)嶒?yàn)性藥物中和 HIV 感染的效果。

• 抗逆轉(zhuǎn)錄病毒藥物篩選: 它被用來快速、定量地篩選大量化合物，以評(píng)估它們抑制 HIV 復(fù)制的能力。熒光素酶的讀數(shù)能清楚地顯示化合物的抗病毒效果。

• 研究 HIV 進(jìn)入抑制劑: 該細(xì)胞系是研究專門通過靶向 CCR5 或 CXCR4 共受體來阻斷 HIV 進(jìn)入細(xì)胞的藥物的優(yōu)秀模型。

• 疫苗效力測試: 它是評(píng)估 HIV 候選疫苗有效性的寶貴工具，通過測量它們引發(fā)中和抗體反應(yīng)、從而阻止病毒感染這些細(xì)胞的能力。

這種報(bào)告系統(tǒng)相比于傳統(tǒng)的 p24 抗原 ELISA 等舊方法是一個(gè)重大進(jìn)步，因?yàn)樗?、更靈敏、工作量更小，非常適合高通量篩選。

BioVector NTCC質(zhì)粒載體菌種細(xì)胞蛋白抗體基因保藏中心

BioVector NTCC Inc.

TEL: 400-800-2947, 189-0126-8599

E-mail: [email protected]

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