Comparison of europium and chromium release assays: cytotoxicity in healthy individuals and patients with cervical carcinoma
Comparison of europium and chromium release assays: cytotoxicity in healthy individuals and patients with cervical carcinoma
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铕和铬释放测定的比较：健康个体和宫颈癌患者的细胞毒性
DOI：
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发表时间：
1997
期刊：
影响因子：
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通讯作者：
T. Whiteside
中科院分区：
文献类型：
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作者：
P. von Zons;P. Crowley;D. Friberg;M. Bell;U. Koldovsky;T. Whiteside
测量健康女性外周血中的自然杀伤 (NK) 和淋巴因子激活的杀伤 (LAK) 细胞活性，以将标准 51Cr 释放测定与基于时间分辨荧光的非放射性铕 (Eu3+) 释放测定进行比较。首先对 28 份外周血单核细胞样本进行配对测定，对这两种类型的细胞毒性测定进行比较，这些样本取自健康女性，这些女性的子宫颈抹片检查结果正常，或没有宫颈鳞状上皮内病变 (SIL) 的活检证据。靶细胞（NK 敏感 K562 和 NK 抗性 Raji 细胞系）仅用 Eu3+、仅 51Cr 或两种标记进行标记，并使用新鲜或白细胞介素 2 (IL-2) 激活的效应细胞在细胞毒性测定中进行比较。 Eu3+ 释放测定中的自发释放与 51Cr 释放测定中观察到的自发释放相当，但最大 Eu3+ 释放始终超过 51Cr。在 4 小时的测定中，Eu3+ 从靶细胞中的特异性释放比 51Cr 更快，始终导致活性水平高出 30% 至 40%。然而，根据 4 小时测定中 Eu3+ 和 51Cr 释放的测量结果，观察到细胞毒性水平之间存在显着的线性相关性 (P < 0.001)。然后使用 Eu3+ 释放测定来测量患有宫颈 SIL 或宫颈鳞状细胞癌 (SCC) 的女性外周血中 NK 和 LAK 的活性。研究发现，患有晚期 SIL（121 裂解单位 [LU]）或 SCC（93 LU）的女性的平均 NK 活性与对照组（101 LU）或正常宫颈活检患者（90 LU）相似，产生 IL-2 刺激的 NK 活性的能力也相似。然而，与正常对照相比，宫颈 SCC 患者在 IL-2 存在的情况下孵育 18 小时期间 LAK 活性降低（P < 0.05）。与患者样本平行进行的 51Cr 测定结果给出了可比较的结果。讨论了用于常规细胞毒性评估的 EU3+ 释放测定的优点。
Natural killer (NK) and lymphokine-activated killer (LAK) cell activities were measured in peripheral blood obtained from healthy women to compare a standard 51Cr release assay with a nonradioactive europium (Eu3+) release assay based on time-resolved fluorescence. The two types of cytotoxicity assays were first compared in paired determinations performed on 28 samples of peripheral blood mononuclear cells obtained from healthy women who had normal pap smears or no biopsy evidence of cervical squamous intraepithelial lesions (SIL). Target cells (NK-sensitive K562 and NK-resistant Raji cell lines) were labeled with Eu3+ only, 51Cr only, or both labels and compared in cytotoxicity assays using fresh or interleukin 2 (IL-2)-activated effector cells. Spontaneous release in the Eu3+ release assay was comparable to that observed in the 51Cr release assay, but maximum Eu3+ release always exceeded that of 51Cr. In 4-h assays, specific release of Eu3+ from target cells was more rapid than that of 51Cr, consistently resulting in 30 to 40% higher levels of activity. However, a significant linear correlation (P < 0.001) was observed between cytotoxicity levels based on measurements of Eu3+ and 51Cr release in 4-h assays. The Eu3+ release assay was then used to measure NK and LAK activities in the peripheral blood of women with cervical SIL or cervical squamous cell carcinoma (SCC). Mean NK activity of women with advanced SIL (121 lytic units [LU]) or SCC (93 LU) was found to be similar to that of controls (101 LU) or patients with normal cervical biopsies (90 LU), as was the ability to generate IL-2-stimulated NK activity. However, LAK activity during 18 h of incubation in the presence of IL-2 was reduced in patients with cervical SCC (P < 0.05) compared with that in normal controls. Results of 51Cr assays performed in parallel with patient samples gave comparable results. Advantages of EU3+ release assays for routine evaluation of cytotoxicity are discussed.