Acidic polysaccharides of Arthrospira (Spirulina) platensis induce the synthesis of TNF-α in RAW macrophages
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Abstract
The study of the enhancement of the immune system by administration of algal cell components is a current research field of great interest for future development of algal biotechnology. Arthrospira (Spirulina) platensis is one of the key organisms, showing interesting results in the treatment of certain tumors, viral infection, and immunodeficiency. Polysaccharides from Arthrospira, together with phycocyanin, seem to be responsible for most of these positive effects. In this work, we isolated the acidic polysaccharide fraction from A. platensis and tested its capacity to induce the production of the proinflammatory cytokine tumor necrosis factor alpha in macrophages. For this purpose, we modified a previous isolation method developed by one of us, which includes several depigmentation steps, as well as differential partitioning with N-cetylpyridinium bromide (Cetavlon). Infrared spectroscopy of the acidic polysaccharide fraction indicates the presence of hydroxyl radicals, aliphatic residues, carbonyl groups, sulfate groups, and sulfate esters, as well as amine residues. Liquid chromatography confirmed the polysaccharidic nature of the fraction, revealing its high purity, essentially free of lipopolysaccharide (LPS) contamination (0.0017% w/w), and complying with international pharmacological standards. The results indicate that a very high production of tumor necrosis factor- α (TNF−α) occurred in macrophages in the presence of the polysaccharides in the range 5–100 μg mL−1, reaching values of 8 ng TNF-α mL−1 after 24 h and 30 ng TNF-α mL−1 after 48 h. These data demonstrate that acidic polysaccharides from Spirulina elicit TNF-α production levels comparable to LPS at ~100× higher concentration than LPS, but without significantly increasing the risk of septic shock or deleterious pyrogenesis.
Keywords
Spirulina Acidic polysaccharides Immune system Macrophages Tumor necrosis factor-αNotes
Acknowledgments
This work has been supported in part by a binational project (Acción Integrada HG2004-0025 to C.J. and to T.G.S.). We thank Dr. Ma Ángeles Vargas and Dr. Luis Alemany (Chemical Engineering Department, UMA) for their technical assistance in the FT-IR analysis, Prof. Francisco R. Sarabia and Miss Francisca Martín-Gálvez (Organic Chemistry Department, UMA) for HPLC analysis, Mr. Casimiro Cárdenas (Cell Culture Division, SCAI, UMA) for technical help during macrophage cultivation, and Mrs. Azucena Muñoz García (Pharmacology Section, CIMES, UMA) for LPS contamination assays.
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