J Korean Surg Soc.  2013 Nov;85(5):230-235.

Evaluation of cytolytic activity and phenotypic changes of circulating blood immune cells in patients with colorectal cancer by a simple preparation of peripheral blood mononuclear cells

• Kim JC
• Choi J
• Lee SJ
• Lee YA
• Jeon YM
• Kang YW
• Lee JK

Abstract

PURPOSE
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).
METHODS
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.
RESULTS
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).
CONCLUSION
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.

Keyword

Blood cells; Natural killer cells; Immunologic cytotoxicity; Colorectal neoplasms

MeSH Terms

Antibodies, Monoclonal
Blood Cells
Colorectal Neoplasms*
Cytotoxicity, Immunologic
Fluorescence
Humans
Immunity, Cellular
K562 Cells
Killer Cells, Natural
L-Lactate Dehydrogenase
Lymphocytes
Phenotype
Tissue Donors
Antibodies, Monoclonal
L-Lactate Dehydrogenase

Figure

• 

  Fig. 1 Peripheral blood mononuclear cell (PBMC) cytolytic activity against K562 cells for two different methods in preoperative patients with colorectal cancer (n = 18) and healthy donors (n = 18). (A) 5 × 105 PBMCs were cocultured with 2 × 104 K562 cells in a ratio of 25:1 for 2 hours. (B) An unfixed number of PBMC isolated from 500 µL of blood was cocultured with 2 × 104 K562 cells for 2 hours. Values are expressed as mean ± standard deviation. *Designates that P-value against the healthy group is less than 0.001 (Student t-test).

• 

  Fig. 2 Peripheral blood mononuclear cell (PBMC) cytolytic activity against K562 cells in patients with colorectal cancer (n = 94) and in healthy donors (n = 112). An unfixed number of PBMCs derived from 500 µL of blood was cocultured with 2 × 104 K562 cells for 2 hours. Values are presented as mean ± standard deviation. *Signifies that P-value against the healthy group is less than 0.001 (Student t-test).

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