Kim Jae Cheol, Choi Joungbum, Lee Su Jin, Lee Yun A, Jeon Young Min, Kang Yong Won, Lee Jong Kyun
Immunology Laboratory, Seniors Inje Hospital, Holon Center, Inje, Korea.
J Korean Surg Soc. 2013 Nov;85(5):230-5. doi: 10.4174/jkss.2013.85.5.230. Epub 2013 Oct 25.
This study aimed to assess the cytolytic activity and the phenotype of circulating blood immune cells in cancer patients by using a simple preparation of peripheral blood mononuclear cells (PBMCs).
Peripheral blood was obtained from 94 diagnosed colorectal cancer (CRC) patients and 112 healthy donors. PBMCs were cocultured with K562 cells for 2 hours and lactate dehydrogenase released from the dead K562 cells was measured by using a spectrophotometer. Meanwhile, PBMCs were stained with fluorescence conjugated monoclonal antibodies (mAbs) and analyzed by flow cytometry.
The cytolytic activity of PBMCs were significantly different between CRC patient and healthy groups (8.82% ± 3.84% vs. 17.51% ± 8.57%; P < 0.001). However, no significant difference in the cytolytic activity was observed after surgery in the CRC patient group (before surgery, 8.82% ± 3.84% vs. after surgery, 9.95% ± 4.94%; P = 0.326). In addition, the percentage of peripheral blood natural killer cells was significantly higher in the preoperative patient group than in the healthy group (19.97% ± 11.51% vs. 15.60% ± 5.77%, P = 0.041). In contrast, the percentage of peripheral blood lymphocytes was lower in the preoperative patient group than in the healthy group (28.41% ± 8.31% vs. 36.4% ± 8.6%, P < 0.001).
These results demonstrate that circulating blood immune cells of CRC patients are functionally impaired and undergo an immunophenotypic perturbation, and show that a simple preparation of PBMCs can be useful to evaluate cellular immunity in cancer.
本研究旨在通过使用一种简单的外周血单个核细胞(PBMC)制备方法,评估癌症患者循环血液免疫细胞的细胞溶解活性和表型。
从94例确诊的结直肠癌(CRC)患者和112名健康供体中获取外周血。将PBMC与K562细胞共培养2小时,并使用分光光度计测量死亡的K562细胞释放的乳酸脱氢酶。同时,用荧光偶联单克隆抗体(mAb)对PBMC进行染色,并通过流式细胞术进行分析。
CRC患者组和健康组PBMC的细胞溶解活性存在显著差异(8.82%±3.84%对17.51%±8.57%;P<0.001)。然而,CRC患者组术后细胞溶解活性未观察到显著差异(术前8.82%±3.84%对术后9.95%±4.94%;P = 0.326)。此外,术前患者组外周血自然杀伤细胞的百分比显著高于健康组(19.97%±11.51%对15.60%±5.77%,P = 0.041)。相反,术前患者组外周血淋巴细胞的百分比低于健康组(28.41%±8.31%对36.4%±8.6%,P<0.001)。
这些结果表明,CRC患者的循环血液免疫细胞功能受损并经历免疫表型扰动,并表明简单的PBMC制备方法可用于评估癌症中的细胞免疫。