Abstract
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.
Highlights
The bursa of fabricius (BF), or “cloacal thymus”, is the primary lymphoid organ in birds
The results showed that BP5 displayed concentration-dependent effects on the proliferation, cell cycle, and surface IgM (sIgM) mRNA expression in DT40 cells
In the growth medium (10% fetal bovine serum (FBS) plus 5% chicken serum (CHS)), the DT40 cells exhibited a rapid increase in number, and BP5 showed no significant effect on the growth curves of DT40 cells
Summary
The bursa of fabricius (BF), or “cloacal thymus”, is the primary lymphoid organ in birds It plays a central role in the proliferation and differentiation of the antibody-producing B lymphocyte lineage. Many studies have focused on BF extracts and multiple biologically active factors and their effects on the humoral immune system in chickens and other birds. DT40 cells are an avian leukosis virus-induced chicken pre-B cell line that express the immunoglobulin M (IgM) isotype B cell reporter in the plasma membrane. 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. Objectives: 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.
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