Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (24): 3870-3875.doi: 10.3969/j.issn.2095-4344.2015.24.019

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Effects of high expression of Csk-binding protein on morphology and biological function of Jurkat cells

Cong Bei-bei1, Gao Mei-hua1, Wang Bing1, Shao Zhi-wei2, Wang Li-na1, Zhang Wen1   

  1. 1 Department of Immunology, Medical College of Qingdao University, Qingdao 266000, Shandong Province, China
    2 Department of Neurosurgery, Medical College of Qingdao University, Qingdao 266000, Shandong Province, China
  • Online:2015-06-11 Published:2015-06-11
  • Contact: Gao Mei-hua, M.D., Professor, Department of Immunology, Medical College of Qingdao University, Qingdao 266000, Shandong Province, China
  • About author:Cong Bei-bei, Department of Immunology, Medical College of Qingdao University, Qingdao 266000, Shandong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81273206

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Abstract:

BACKGROUND: The linkage and synergistic effect of adaptor proteins can effectively regulate signal transduction of T cells, which can form a limit or amplification cascade to realize the complex immune function of T cells. C-terminal Src kinase (Csk)-binding protein (Cbp) is an adaptor protein, which mainly exert the negative feedback regulation of Src kinase activity. This negative feedback effect depends on Y317 of Cbp, which may be involved in the SH2 domain of Csk.
OBJECTIVE: To explore the effects of high expression of Cbp on ultrastructure and related biological function of Jurkat cells.
METHODS: The virus particles were constructed with expressing enhanced green fluorescent protein (EGFP) only and Cbp-EGFP fusion protein to transfect Jurkat cells. There were untransfected group (Jurkat group), negative control group (transfected with expression of EGFP virus only), and Cbp group (transfected with Cbp-EGFP virus).
RESULTS AND CONCLUSION: Confocal microscope showed that cell transfection efficiency was more than 95% and Cbp was located on the cell membrane. Optical microscope showed after transfection with Cbp-EGFP virus, more Jurkat cells shrunk, with poor size uniformity. Apoptosis detection showed that after transfection with Cbp-EGFP virus, the number of apoptotic and necrotic cells was greatly increased. Cbp mRNA expression was increased, Csk expression was decreased obviously and lymphocyte-specific protein tyrosine kinase expression was increased. So, in Jurkat cells, the high expression of Cbp can decrease the uniformity of cells and increase the necrosis cells, thus inhibiting the signal transduction.  

Key words: Apoptosis, T-Lymphocytes, Jurkat Cells, Adaptor Proteins, Signal Transducing

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