Expression of perforin in cord blood NK cells after IL-2/IL-15 stimulation and its relation with cytotoxicity.
- Author:
Yan-Feng WU
1
;
Bi-Hong ZHANG
;
Dan-Yang CEN
;
Jing WEI
;
Chun CHEN
Author Information
1. Center of Biologic Therapy, SUN Yat-Sen University, Guangzhou, Guangdong Province, China.
- Publication Type:Journal Article
- MeSH:
CD56 Antigen;
metabolism;
Cells, Cultured;
Cytotoxicity, Immunologic;
Fetal Blood;
cytology;
Flow Cytometry;
Humans;
Interleukin-15;
pharmacology;
Interleukin-2;
pharmacology;
K562 Cells;
Killer Cells, Natural;
cytology;
immunology;
metabolism;
Perforin;
metabolism
- From:
Journal of Experimental Hematology
2011;19(4):1015-1018
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to investigate the expression level of perforin in cord blood NK cells and the relation of perforin expression after IL-2, IL-15 stimulation to cytotoxicity of NK cells. NK cells were isolated from cord blood MNC by depleting CD3(+) cells and then enriching CD56(+) cells using immunomagnetic separation (CD3 and CD56 cell isolation kit, autoMACS, miltenyi). The purity was analysed by flow cytometry. According to the different combination of cytokines, there were two groups: group A (IL-2) and group B (IL-2 + IL-15). The cytotoxicity and perforin expression rate of fresh and different cultured CB-NK cells against K562/Jurkat cell lines were estimated by LDH release test (cytotoxic 96 non-radioactive cytotoxicity assay). The results showed that the purity of NK cells after separation was more than 90%. The cytotoxicity towards both tumor lines in group B was higher than that in group A (p < 0.05), and cytotoxicity in group A was higher than that of fresh NK cells (p < 0.05). Perforin expression rate of group A (84.55%) was higher than that of fresh NK cells (67.21%) (p < 0.05), and there was no significant difference between group A and B (84.55% versus 87.22%) Cytotoxic activity of CB-NK cells was positively correlated with perforin expression rate (r = 0.886, p < 0.05). It is concluded that IL-2 can enhance cytotoxicity of CB/BM-NK cells by increasing the perforin expression.
