TY - JOUR
T1 - Associating structural characteristics to immunomodulating properties of carrot rhamnogalacturonan-I fractions
AU - Desai, Krishna
AU - Dobruchowska, Justyna M.
AU - Elbers, Kari
AU - Cybulska, Justyna
AU - Zdunek, Artur
AU - Porbahaie, Mojtaba
AU - Jansen, Erik
AU - Van Neerven, Joost
AU - Albers, Ruud
AU - Wennekes, Tom
AU - Mercenier, Annick
AU - Schols, Henk A.
PY - 2025/1/1
Y1 - 2025/1/1
N2 - Carrot rhamnogalacturonan-I (cRG-I) is a polydisperse polysaccharide with molecular weights of 7–250 kDa. Using size exclusion chromatography cRG-I was fractionated and pooled in fractions (PF1–6). All fractions contained the same RG-I monosaccharides and similar glycosidic linkages although in varying relative amounts. The main differences were in rhamnose substitution, arabinan- and galactan side chain length and in levels of acetylation and methyl esterification. Atomic force microscopy showed either spheric or elongated structures for cRG-I and its derived fractions. To gain insight in the structure-function relationship of cRG-I, the immunomodulatory effect of the six fractions and their saponified derivatives was assessed in vitro. All fractions, except PF2, dose-dependently stimulated TNFα, IL-6, IL-1β, IL-8 and IL-10 production in peripheral blood mononuclear cells (PBMCs) of three healthy donors. Cytokine levels were largely influenced by the Mw and degree of esterification of the individual fractions. Notably, the highest Mw fraction (100 kDa) displayed the most potent activity, which was strongly reduced after the removal of ester residues by saponification. In contrast, the 75 kDa Mw population (PF2) proved inactive while its saponified counterpart exhibited substantial immunomodulatory activity. This confirmed the role of ester residues on the immune profile of RG-I subpopulations.
AB - Carrot rhamnogalacturonan-I (cRG-I) is a polydisperse polysaccharide with molecular weights of 7–250 kDa. Using size exclusion chromatography cRG-I was fractionated and pooled in fractions (PF1–6). All fractions contained the same RG-I monosaccharides and similar glycosidic linkages although in varying relative amounts. The main differences were in rhamnose substitution, arabinan- and galactan side chain length and in levels of acetylation and methyl esterification. Atomic force microscopy showed either spheric or elongated structures for cRG-I and its derived fractions. To gain insight in the structure-function relationship of cRG-I, the immunomodulatory effect of the six fractions and their saponified derivatives was assessed in vitro. All fractions, except PF2, dose-dependently stimulated TNFα, IL-6, IL-1β, IL-8 and IL-10 production in peripheral blood mononuclear cells (PBMCs) of three healthy donors. Cytokine levels were largely influenced by the Mw and degree of esterification of the individual fractions. Notably, the highest Mw fraction (100 kDa) displayed the most potent activity, which was strongly reduced after the removal of ester residues by saponification. In contrast, the 75 kDa Mw population (PF2) proved inactive while its saponified counterpart exhibited substantial immunomodulatory activity. This confirmed the role of ester residues on the immune profile of RG-I subpopulations.
KW - Acetylated pectic polysaccharides
KW - Immunomodulation
KW - Rhamnogalacturonan
KW - Saponification
KW - Size fractionation
KW - Structural characterization
KW - Structure-function activity
U2 - 10.1016/j.carbpol.2024.122730
DO - 10.1016/j.carbpol.2024.122730
M3 - Article
AN - SCOPUS:85203638567
SN - 0144-8617
VL - 347
JO - Carbohydrate Polymers
JF - Carbohydrate Polymers
M1 - 122730
ER -