Molecular cloning, expression and bioactivity of B cell activating factor (BAFF) in African ostrich.

Abstract

B cell activating factor (BAFF), which belongs to the tumor necrosis factor (TNF) family, is testified to play a critical role in B cell survival, proliferation, maturation and immunoglobulin secretion. In the present study, the cDNA of open reading frame (ORF) in African ostrich (Struthio camelus) BAFF (designated OsBAFF) was cloned by reverse transcription-PCR (RT-PCR). The OsBAFF gene encodes a 288-amino acid protein containing a predicted transmembrane domain and a putative furin protease cleavage site like BAFFs from chicken (cBAFF), quail (qBAFF), duck (dBAFF), goose (gBAFF) and dove (doBAFF). RT-PCR analysis showed that the OsBAFF gene is strongly expressed in the bursa of Fabricius, thymus, spleen, and bone marrow. The soluble OsBAFF had been cloned into pET28a. SDS-PAGE and Western blotting analysis confirmed that the soluble fusion protein His-OsBAFF was efficiently expressed in Escherichia coli Rosset (DE3). In vitro, purified OsBAFF was not only able to promote the survival of African ostrich bursal lymphocytes, but also able to co-stimulate proliferation of mouse splenic B cells. The expression of OsBAFF in lymphocyte cells was higher than the control after LPS stimulation. These findings indicated that OsBAFF plays an important role in survival and proliferation of African ostrich bursal lymphocytes, which may provide valuable information for research into the immune system of African ostrich and OsBAFF may serve as a potential immunologic factor for enhancing immunological efficacy in African ostrich and any other birds.

• Authors: Keli Yang, Ke Xiao, Haibo Huang, Shun Lu, Juming Zhong, Abdur Rahman Ansari, Haseeb Khaliq, Hui Song, Huazhen Liu, Kemei Peng
• Journal: International immunopharmacology (Int Immunopharmacol) Vol. 28 Issue 1 Pg. 686-94 (Sep 2015) ISSN: 1878-1705 [Electronic] Netherlands
• PMID: 26256697 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 Elsevier B.V. All rights reserved.

Chemical References

• B-Cell Activating Factor
• DNA, Complementary
• Lipopolysaccharides
• lipopolysaccharide, E. coli O26-B6

Topics

• Amino Acid Sequence
• Animals
• B-Cell Activating Factor (genetics, immunology, pharmacology)
• B-Lymphocytes (immunology, metabolism)
• Base Sequence
• Blotting, Western
• Bursa of Fabricius (immunology)
• Cell Proliferation (drug effects)
• Cell Survival (immunology)
• Cells, Cultured
• Cloning, Molecular
• DNA, Complementary (genetics)
• Escherichia coli (genetics)
• Gene Expression Regulation
• Lipopolysaccharides (pharmacology)
• Mice
• Molecular Sequence Data
• Open Reading Frames (genetics)
• Organ Specificity
• Phylogeny
• Real-Time Polymerase Chain Reaction
• Struthioniformes (genetics, immunology)