Immunomodulatory effects of holothurian triterpene glycosides on mammalian splenocytes determined by mass spectrometric proteome analysis.
Spleen is a prime organ in which immuno-stimulation takes place in mammalians. Proteome analysis was used to investigate the elicited effects on mouse splenocytes upon exposure to holothurian triterpene glycosides. Cucumarioside A(2)-2, and Frondoside A, respectively, have been used to in-vitro stimulate primary splenocyte cultures. Differential protein expression was monitored by 2D gel analysis and proteins in spots of interest were identified by MALDI ToF MS and nano LC-ESI Q-ToF MS/MS, respectively. Differential image analysis of gels from control vs. gels from stimulated primary splenocyte cultures showed that approximately thirty protein spots were differentially expressed. Prime examples of differentially expressed proteins are NSFL1 cofactor p47 and hnRNP K (down-regulated), as well as Septin-2, NADH dehydrogenase [ubiquinone] iron-sulfur protein 3, and GRB2-related adaptor protein 2 (up-regulated). Immuno-analytical assays confirmed differential protein expression. Together with results from proliferation and cell adhesion assays, our results show that cellular proliferation is stimulated by holothurian triterpene glycosides. In conclusion, holothurian triterpene glycosides are thought to express their immuno-stimulatory effects by enhancing the natural cellular defense barrier that is necessary to fight pathogens and for which lymphocytes and splenocytes have to be recruited constantly due to limited lifetimes of B-cells and T-cells in the body.
Pacific Institute of Bioorganic Chemistry, Far East Division of the Russian Academy of Sciences, Vladivostock, 690022 Russia.
Dmitri L
DL
Aminin
Conelia
C
Koy
Pavel S
PS
Dmitrenok
Brigitte
B
Müller-Hilke
Dirk
D
Koczan
Brian
B
Arbogast
Aleksandra A
AA
Silchenko
Vladimir I
VI
Kalinin
Sergey A
SA
Avilov
Valentin A
VA
Stonik
Peter D
PD
Collin
Hans-Juergen
HJ
Thiesen
Max L
ML
Deinzer
Michael O
MO
Glocker
D000818:Animals; D002448:Cell Adhesion; D049109:Cell Proliferation; D002469:Cell Separation; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006027:Glycosides / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013154:Spleen / Q000166:cytology* / Q000378:metabolism; D013729:Terpenes / Q000737:chemistry*
D006027:Glycosides; D020543:Proteome; D013729:Terpenes