法氏囊九肽（BP9）在雏鸡免疫中对禽流感疫苗免疫反应及对禽类未成熟B细胞的作用。

The Roles of Bursal Nonapeptide (BP9) on AIV Vaccine Immune Response in Chick Immunization and on Avian Immature B Cell.

作者信息

Zheng Yang, Zong Man M, Chen Bo Y, Zhou Xiao H, Liu Zi N, Zhou Guang F, Chen Pu Y, Feng Xiu L

机构信息

Key Laboratory of Animal Microbiology of China's Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.

MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

Protein Pept Lett. 2019;26(12):940-948. doi: 10.2174/0929866526666190730101455.

DOI:10.2174/0929866526666190730101455

PMID:31362650

Abstract

BACKGROUND

Bursa of Fabricius plays the vital functions on B cell development and antibody production in poultry. The bursal-derived peptide plays the essential roles on avian immature B cell development.

OBJECTIVES

Here we explored the functions of the recently reported bursal nonapeptide (BP9) on the antibody production and the molecular basis of BP9 on avian immature B cell.

METHODS

Chicken were twice immunized with Avian Influenza Virus (AIV) inactivated vaccine plus with BP9 at three dosages, respectively. On two weeks after the second immunization, sera samples were collected from all experimental groups to measure AIV-specific Agglutination Inhibition (HI) antibody titers. Also, on 7th day after the second immunization, spleen lymphocytes were isolated from the immunized chicken to detect the lymphocyte viabilities. DT40 cells were treated with BP9 from 0.02 to 2 μg/mL for 4 and 20h to detect sIgM mRNA levels, and total RNAs from BP9-treated DT40 cells were collected to investigate the gene expression profiles of DT40 cells, and to analyze the enriched pathways and functional biological processes. Finally, nine gene expressions were validated with quantitative PCR (qPCR).

RESULTS

Our investigation proved the strong regulatory roles of BP9 on AIV-specific HI antibody titers and lymphocyte viabilities. BP9 promoted sIgM mRNA levels in DT40 cells, and upregulated 598 gene expressions and downregulated 395 gene expressions in DT40 cells with 0.2μg/mL BP9 treatment. Moreover, our findings verified the significantly enriched six pathways and various the biological functional processes of BP9 on avian immature B cell. Also, we found eight signaling pathways in the enriched biological processes of BP9-treated DT40 cells, and the expressions of nine selected genes with qPCR were identical to that of microarray data.

CONCLUSION

BP9 promoted the antibody production in the 21-old-day chicken immunization, and stimulated the sIgM expression in DT40 cells. Furthermore, we analyzed the gene expression profile and immune-related biological processes of DT40 cells treated with BP9, which provided some new insights into the mechanism on immature B cell development, and provided important references for adjuvant development on vaccine improvement and clinical application.

摘要

背景

法氏囊在禽类B细胞发育和抗体产生中发挥着至关重要的作用。法氏囊衍生肽在禽类未成熟B细胞发育中起着关键作用。

目的

在此，我们探讨了最近报道的法氏囊九肽（BP9）对抗体产生的作用以及BP9作用于禽类未成熟B细胞的分子基础。

方法

用禽流感病毒（AIV）灭活疫苗加三种剂量的BP9分别对鸡进行两次免疫。第二次免疫后两周，从所有实验组采集血清样本，检测AIV特异性血凝抑制（HI）抗体效价。此外，在第二次免疫后第7天，从免疫鸡中分离脾淋巴细胞，检测淋巴细胞活力。用0.02至2μg/mL的BP9处理DT40细胞4小时和20小时，以检测sIgM mRNA水平，并收集经BP9处理的DT40细胞的总RNA，以研究DT40细胞的基因表达谱，并分析富集的途径和功能生物学过程。最后，用定量PCR（qPCR）验证九个基因的表达。

结果

我们的研究证明了BP9对AIV特异性HI抗体效价和淋巴细胞活力具有强大的调节作用。BP9促进了DT40细胞中sIgM mRNA水平，并在0.2μg/mL BP9处理的DT40细胞中上调了598个基因表达，下调了395个基因表达。此外，我们的研究结果证实了BP9对禽类未成熟B细胞有显著富集的六个途径和各种生物学功能过程。我们还在经BP9处理的DT40细胞富集的生物学过程中发现了八个信号通路，并且用qPCR检测的九个选定基因的表达与微阵列数据一致。