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High expressions of activated surface markers and drug-resistance gene
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Abstract:
Objective:To analyze the proliferation and differential expressions of activated and inhibitory surface markers, drug-resistance gene ABCG2 and stem cell transcription factors in umbilical cord blood-derived cytokine induced killer (CIK) cells or peripheral blood-derived CIK (PB-CIK) cells in cancer patients, and to explore the superiority of umbilical cord blood-derived CIK (CB-CIK) cells in adoptive cellular immunotherapy. Methods: Cord blood mononuclear cells and peripheral blood mononuclear cells in cancer patients were isolated using Ficoll-plaque density gradients and incubated with cytokines. The proliferation, immunophenotypes, activated surface markers (CD28, CD27) and inhibitory surface marker (PD-1) were analyzed by flow cytometry, and the drug-resistance gene ABCG2 or stem cell transcription factors ( c-Myc, Nanog, Oct-4 and Sox2 ) were determined by RT-PCR. Results: CB-CIK cells and PB-CIK cells started to proliferate on the fourth day in vitro, and the proliferation index of CB-CIK cells was significantly higher than that of PB-CIK cells on day 10 (\[251.52±16.76\]% vs \[158.00±43.19\]%, P<005\]. Thirteen days after incubation, the proportion of CD3+CD56+ cells in CB-CIK cells was higher than that in PB-CIK cells (\[21.20±4.82\]% vs \[10.06±346\]%, P<0.05). When detecting the status of CIK cells, the proportions of activated CD4+CD28+, CD4+ CD27+ and CD8+CD27+ cells were significantly higher in CB-CIK cells than in PB-CIK cells (\[32.40±16.81\]% vs \[18.65±923\]%; \[27.48±13.53\]% vs \[0.98±0.55\]%; \[41.76±13.98\]% vs \[2.58±2.10\]%，P<0.05 or P<0.01), while the proportion of inhibitory CD8+PD-1+ cells was significantly lower in CB-CIK cells (\[3.25±2.13\]% vs \[8.05±9.23\]%, P<0.01). The stem cell transcription factors ( c-Myc, Nanog, Oct and Sox2 ) were expressed both in CB-CIK cells and PB-CIK cells. Moreover, no significant differences were found between the two kinds of CIK cells. Furthermore, only CB-CIK cells expressed a high level of drug-resistance gene ABCG2 . Conclusion: Compared with the PB-CIK cells, the CB-CIK cells showed increasing proliferation rates, higher expressions of activated surface markers, and lower expression of inhibitory surface marker. Besides, drug-resistant gene ABCG2 is highly expressed in CB-CIK cells, showing several advantages in applying to adoptive cellular tumor immunotherapy.
Keywords:
Project Supported:
Project support by the National Natural Science Foundation of China (No. 81171985, No. 81171986, No. 812111102)
Clc Number:
