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Updated: May 30, 2026
Purification and Expansion of Mouse Invariant Natural Killer T Cells for in vitro and in vivo Studies
Published on: February 15, 2021
[Study on the stability of the anergic CD8(+);NKT cells in vitro]
Ye-lei Guo1, Yu Chen, Jiang Zhong
1Department of Immunology, General Hospital of PLA, Beijing, China.
Aim:
To study the CD8(+);NKT cell stability function.
Methods:
Splenic lymphocytes from C57BL/J mice were treated with Staphylococcal enterotoxin B (SEB) and then cultured in vitro. Day 10, 20, 30 and freezen effector cells were harvested and used. The cells were cultured in medium containing ConA and LPS for 72 hours and measured their response to mitogens. The inhibitory action of the effector cells were examined. The effector cells were co-cultured with normal lymphocytes and above mitogens for 72 hours. The cells proliferation was assessed with MTT method. The effector cells were cultured with heterogenic lymphocytes and were assessed with MTT method. The NKT cell subsets among these effector cells were analyzed by flow cytometry.
Results:
The response of these effector cells to ConA and LPS was significantly decreased compared with that of normal lymphocytes. The A values of cell proliferation were decreased from 0.67 and 0.61 to 0.30 and 0.31, 0.28 and 0.20, 0.26 and 0.24, 0.22 and 0.23 (P<0.05, n=3). The inhibitory ability of effector cells aginst the response of normal lymphocytes to ConA and LPS were clearly observed. They inhibited the response of normal lymphocytes to above mitogens. And the A values of cell proliferation were decreased from 0.67 and 0.61 to 0.33 and 0.39, 0.30 and 0.43, 0.36 and 0.43, 0.26 and 0.29(P<0.05, n=3). The response of effector cells to heterogenic lymphocytes was significantly decreased compared with normal lymphocytes. The A values of cell proliferation were decreased from 0.70 to 0.42, 0.42 and 0.54 on day 20, 30 and freezen effector cells respectively(P<0.05, n=3). The CD8(+);NKT cell subsets among these effector cells with tolerance function were significantly increased(P<0.05, n=3).
Conclusion:
The anergic CD8(+);NKT cells could culture in vitro. And the anergic function of these cells were still existing.
