Inducing and stimulating the activation and proliferation of peripheral blood mononuclear lymphocyte subsets from rhesus macaque

doi:10. 3969 / j.issn.1671-7856. 2021. 05. 001

Home > Archive>Volume 31, Issue 5, 2021 >1-6. DOI:10. 3969 / j.issn.1671-7856. 2021. 05. 001

Inducing and stimulating the activation and proliferation of peripheral blood mononuclear lymphocyte subsets from rhesus macaque
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                        10. 3969 / j.issn.1671-7856. 2021. 05. 001
                    
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                        LI GuocuiLI Guocui
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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LU JiahanLU Jiahan
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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LU QiuhanLU Qiuhan
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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YANG ChenboYANG Chenbo
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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CONG ZheCONG Zhe
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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WEI QiangWEI Qiang
Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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Affiliation:Comparative Medicine Center, Peking Union College (PUMC)&Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences (CAMS); NHC Key Laboratory of Human Disease Comparative Medicine; Key Laboratory of Human Diseases Animal Models, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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Abstract:

Objective To investigate the effect of stimulants on the expression of T cell activation markers (CD69, CD38, HLA-DR) and a nuclear proliferation marker ( Ki67), and provide the basis for the study of T cell function after activation. Methods Peripheral blood was collected from healthy rhesus monkeys, then peripheral blood mononuclear cells ( PBMCs ) were separated and regular doses of PMA + Ionomycin, α-CD2 / α-CD3 / α-CD28, and phytohemagglutinin-P (PHA-P)+interleukin-2 ( IL-2) were added. Expression of CD4+ T cells and CD8⁺T cell surface markers, CD69, CD38, HLA-DR, and Ki67, were detected at different time points. Results Flow cytometry showed that when the stimulant, α-CD2 / α-CD3 / α-CD28, acted for 72 hours, expression of CD69 and Ki67 on CD4⁺T cells and CD8⁺T cells was higher compared with the other two stimulants, while the percentage of HLA-DR+ CD4⁺T cells and HLA-DR+ CD8⁺T cells and CD38⁺CD8⁺T cells was not different. Meanwhile, the percentage of CD38⁺CD4⁺T cells was not different between α-CD2 / α-CD3 / α-CD28 and PHA-P + IL-2 groups ( P> 0. 05). Expression of all four molecules was very low under PMA+Ionomycin stimulation. Conclusions CD2 / α-CD3 / α-CD28 had the best effect in activating T cells, followed by PHA-P+IL-2, and then PMA+Ionomycin. This provides the experimental basis for selecting appropriate stimulants on the basis of different experimental purposes.

Key words:PBMC; activation; PMA+Ionomycin; α-CD2 / α-CD3 / α-CD28; PHA-P+IL-2; CD69; CD38; HLA- DR; Ki67

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Received:December 29,2020
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Online: June 25,2021
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