Binding profiles and cytokine-inducing effects of fish rhamnose-binding lectins on Burkitt’s lymphoma Raji cells

• Published in: Fish physiology and biochemistry Vol. 40; no. 5; pp. 1559 - 1572
• Main Authors: Hosono, Masahiro, Sugawara, Shigeki, Matsuda, Atsushi, Tatsuta, Takeo, Koide, Yasuhiro, Hasan, Imtiaji, Ozeki, Yasuhiro, Nitta, Kazuo
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer-Verlag 01-10-2014; Springer Netherlands; Springer Nature B.V
• Subjects: Analysis of Variance; Animal Anatomy; Animal Biochemistry; Animal Physiology; Animals; Apoptosis; Biomedical and Life Sciences; Blotting, Western; Burkitt Lymphoma - immunology; Burkitt Lymphoma - metabolism; Carbohydrates; catfish; Catfishes - immunology; Catfishes - metabolism; cell death; cell growth; Cell Line, Tumor; cell viability; Cytokines; Cytokines - metabolism; DNA Primers - genetics; Eggs; Electrophoresis, Polyacrylamide Gel; Fish Proteins - immunology; Fish Proteins - metabolism; Flow Cytometry; fluorescence; Freshwater & Marine Ecology; Histology; Humans; immune response; interferon-gamma; interleukin-10; interleukin-1beta; lactose; Lectins; Lectins - immunology; Lectins - metabolism; Life Sciences; Lymphoma; Melibiose; Microscopy, Fluorescence; Morphology; Osmeriformes - immunology; Osmeriformes - metabolism; Osmerus; Phycoerythrin; Protein Binding; Real-Time Polymerase Chain Reaction; reverse transcriptase polymerase chain reaction; signal transduction; Signal Transduction - immunology; Silurus; Silurus asotus; Trihexosylceramides - immunology; Trihexosylceramides - metabolism; tumor necrosis factor-alpha; Zoology; Japan; Burkitt’s lymphoma; Lectin; Gb3; Domain structure; TNF-α; Fish eggs
• ISSN: 0920-1742; 1573-5168
• DOI: 10.1007/s10695-014-9948-1

Rhamnose-binding lectin (RBL) is one of the animal lectin categories which take part in the innate immune responses of fish. Osmerus lanceolatus lectin (OLL) from shishamo smelt eggs is an RBL composed of two tandem-repeated domains, both of which are considered to be a carbohydrate-recognition domain. SAL, catfish (Silurus asotus) egg RBL composed of three domains, binds to Burkitt’s lymphoma Raji cells through globotriaosylceramide (Gb3) carbohydrate chain and to reduce cell size and growth by altering membrane composition without causing cell death. In this experiment, we tried to compare the binding effects of these two RBLs on Raji cells. Flow cytometric and fluorescence microscopic analyses revealed that OLL also directly bound to and shrunk Raji cells with ten times less reactivity than SAL but reduced cell growth with decreasing cell viability. Anti-Gb3 antibody completely blocked the binding of SAL to Raji cells but not that of OLL. In addition, the direct bindings of OLL and SAL to Raji cells were comparably inhibited by melibiose, but lactose was more effective inhibitor for the binding of OLL than that of SAL. These results suggest that OLL has slightly different cell-binding property compared with SAL and binds not only to Gb3 but also to the other carbohydrate receptor-bearing β-galactoside chains. The quantitative RT-PCR analysis revealed that SAL induced the expression of TNF-α but not of IFN-γ, IL-1β, and IL-10. Thus, SAL-induced cytostatic effect on Raji cells might be partially caused by TNF-α-mediated signaling pathway.