Ji Xiang-Bo, Luo Jun, Feng Xiu-Li, Xu Qiu-Liang, Man Teng, Zhao Dong, Li Xin-Feng, Zhang Gai-Ping, Chen Pu-Yan
Division of Key Lab of Animal Disease Diagnosis and Immunology of China's Department of Agriculture, Nanjing Agriculture University, Nanjing 210095, China.
Henan University of Animal Husbandry and Economy, Zhengzhou 450046, China.
Afr Health Sci. 2018 Dec;18(4):1292-1302. doi: 10.4314/ahs.v18i4.50.
In the recent past, many studies have been focused on extracts of BF and multiple biologically active factors and their effects on humoral immune system in chickens and birds. However, the mechanism of those immunomodulatory peptides on the B lineage cells proliferation and antibody production in chicken is fairly unknown. DT40 cell line, an avian leucosis virus-induced chicken pre-B cell line, expresses immunoglobulin M (IgM) isotype B cell reporter in the plasma membrane. There are many evidences suggesting that DT40 cells are best characterized as a bursal stem cell line. Because of the unique characteristics of DT40 cell line, it has been widely used to observe biological processes of pre-B lymphocyte cell within living cells.
The chicken B cell line DT40 was cultured in Roswell Park Memorial Institute (RPMI) 1640 medium and cytotoxicity was studied. Also, effect of BP5 on cell proliferation and cell cycle distribution of DT40 cells was studied. Also, the effect of BP5 on sIgM mRNA expression was studied by using real-time PCR.
To investigat the effects of Bursopentin (Cys-Lys-Arg-Val-Tyr, BP5) on a chicken promyelocyte cell line DT40, assays of cell proliferation, cell cycle distribution, detection of surface immunoglobulin G (sIgM) mRNA expression and gene microarray analysis were performed.
The results showed that BP5 displayed concentration-dependent effects on the proliferation, cell cycle, and sIgM mRNA expression in DT40 cells. And the analysis of expression profiles identified a signature set of 3022 genes (1254 up regulated genes, 1762 down regulated genes), which clearly discriminated the BP5-treated DT40 cells from control with high certainty (P≤0.02). The results of microarray analysis were confirmed by quantitative reverse transcription-polymerase chain reaction for 12 of the differentially expressed genes.
Theses findings showed the immuno-activity effect of BP5 on B lymphocyte and indicated that BP5 treatment regulated eight signaling pathways, in which Toll-like signaling pathway was the most significant enrichment pathway.
近年来，许多研究聚焦于法氏囊提取物、多种生物活性因子及其对鸡和鸟类体液免疫系统的影响。然而，这些免疫调节肽对鸡B淋巴细胞增殖和抗体产生的机制尚不清楚。DT40细胞系是一种禽白血病病毒诱导的鸡前B细胞系，在质膜上表达免疫球蛋白M（IgM）同种型B细胞报告基因。有许多证据表明DT40细胞最适合被表征为法氏囊干细胞系。由于DT40细胞系的独特特性，它已被广泛用于观察活细胞内前B淋巴细胞的生物学过程。
鸡B细胞系DT40在罗斯威尔公园纪念研究所（RPMI）1640培养基中培养，并研究其细胞毒性。此外，研究了BP5对DT40细胞增殖和细胞周期分布的影响。同时，采用实时PCR研究BP5对sIgM mRNA表达的影响。
为了研究法氏囊五肽（Cys-Lys-Arg-Val-Tyr，BP5）对鸡原髓细胞系DT40的影响，进行了细胞增殖、细胞周期分布测定、表面免疫球蛋白G（sIgM）mRNA表达检测和基因芯片分析。
结果表明，BP5对DT40细胞的增殖、细胞周期和sIgM mRNA表达具有浓度依赖性影响。表达谱分析确定了一组3022个基因的特征集（1254个上调基因，1762个下调基因），该特征集能够高度准确地区分BP5处理的DT40细胞与对照细胞（P≤0.02）。通过对12个差异表达基因进行定量逆转录-聚合酶链反应，证实了基因芯片分析的结果。
这些发现显示了BP5对B淋巴细胞的免疫活性作用，并表明BP5处理调节了八条信号通路，其中Toll样信号通路是最显著富集的通路。
