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目的研究4-1BBL分子在人急性单核细胞白血病细胞系U937和SHI-1的表达及促生长作用。方法用流式细胞术(FCM)和RT-PCR法检测4-1BBL分子在U937、SHI-1细胞系的表达;将激发型4-1BBL单抗(1F1)与U937、SHI-1细胞系分别进行培养,细胞计数分析2个细胞系的生长情况;收集U937细胞的培养上清,ELISA法检测细胞因子。结果FCM测得U937和SHI-1细胞有4-1BBL分子表达,RT-PCR法测得4-1BBLmRNA;细胞计数表明,1F1明显促U937、SHI-1细胞系生长(P<0·01);ELISA示1F1与U937细胞共培养48h,其上清液中IL-6含量(55·91±10·23)pg/ml,显著高于IgG1同型对照组的(12·14±3·8)pg/ml(P<0·01)。结论U937和SHI-1细胞系组成性表达4-1BBL分子;1F1与U937、SHI-1细胞的4-1BBL分子交联后,可促进U937、SHI-1细胞系生长,诱导U937细胞分泌细胞因子IL-6。
Objective To investigate the expression of 4-1BBL in human acute monocytic leukemia cell lines U937 and SHI-1 and its role in promoting growth. Methods The expression of 4-1BBL in U937 and SHI-1 cell lines was detected by flow cytometry (FCM) and RT-PCR. The induced 4-1BBL monoclonal antibody (1F1) and U937 and SHI-1 cell lines The growth of the two cell lines was analyzed by cell counting. The culture supernatant of U937 cells was collected and the cytokines were detected by ELISA. Results The expression of 4-1BBL in U937 and SHI-1 cells was detected by FCM. The 4-1BBL mRNA was detected by RT-PCR. The cell counts showed that 1F1 significantly promoted the growth of U937 and SHI-1 cell lines (P <0.01). ELISA showed that 1F1 and U937 cells co-cultured for 48h, the supernatant IL-6 content (55.91 ± 10.23) pg / ml, significantly higher than the IgG1 isotype control group (12.14 ± 3.8) pg / ml (P <0.01). CONCLUSIONS: The 4-1BBL molecule is constitutively expressed in U937 and SHI-1 cell lines. The cross-linking of 4-1BBL molecules in 1F1, U937 and SHI-1 cells can promote the growth of U937 and SHI-1 cell lines and induce the secretion of cytokines IL-6.