Abstract

An immunomodulatory protein (IPAF) was purified and cloned from Anoectochilus formosanus, an Orchidaceae herbal plant in Asia. The major targeting immune cells of IPAF and its modulating effects toward B lymphocytes were investigated. Rapid amplification of cDNA ends (RACE) was conducted to clone the IPAF gene, and the obtained sequence was BLAST compared on the NCBI database. MACS-purified mouse T and B lymphocytes were stimulated with IPAF and the cell proliferation, activation, and Igs production were examined. IPAF comprised a 25 amino acids signal peptide and a 138 amino acids protein which was homologous to the lectins from Orchidaceae plant. IPAF selectively induced the cell proliferation in mouse splenic B lymphocytes but not T lymphocytes. The IPAF-induced B cells exhibited increased CD69 and MHC class II expression, and a dose- and time-dependent enhancement in IgM production. These results suggested potential benefits of IPAF to strengthen the humoral immunity.

Highlights

  • Anoectochilus formosanus is an herbal plant widely used as dietary supplement and folk remedy in Asia

  • Purification and biochemical characteristics of immunomodulatory protein from A. formosanus (IPAF) To isolate the novel protein from A. formosanus, the crude protein solution was obtained by means of homogenization, sonication and, precipitation

  • Viewing on the fact that the IPAF sample examined in SDS-PAGE was reduced, it was possible that the observed 14 kDa band might be a subunit if the protein constituted two or more subunits

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Summary

Introduction

Anoectochilus formosanus is an herbal plant widely used as dietary supplement and folk remedy in Asia. It provided numerous health benefits such as hepatoprotection [1,2,3,4], anti-fatigue [5], antioxidative [6,7], anti-hyperglycemia [8], anti-hyperliposis [4], antiosteoporosis [9,10], anti-tumor [11,12], and immune modulation. Antigens recognized by B cells can be divided into two categories, T cell-dependent (TD) antigen and T cell-independent (TI) antigen. TI-2 antigen such as multivalent polysaccharide or antigen with repetitive structure could activate antigen-specific B cells through extensive cross-linking of BCRs [18]

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