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. 2014 May 12;19(2):284–296. doi: 10.2478/s11658-014-0196-2

The effect of carbohydrate moiety structure on the immunoregulatory activity of lactoferrin in vitro

Michał Zimecki 1,✉, Jolanta Artym 1, Maja Kocięba 1, Maria Duk 2, Marian L Kruzel 3
PMCID: PMC6275861  PMID: 24820230

Abstract

The aim of this study was to evaluate the immunoregulatory effects of recombinant human lactoferrin (rhLF) in two in vitro models: (1) the secondary humoral immune response to sheep erythrocytes (SRBC); and (2) the mixed lymphocyte reaction (MLR). We compared the non-sialylated glycoform of rhLF as expressed by glycoengineered Pichia pastoris with one that was further chemically sialylated. In an earlier study, we showed that sialylated rhLF could reverse methotrexate-induced suppression of the secondary immune response of mouse splenocytes to SRBC, and that the phenomenon is dependent on the interaction of lactoferrin (LF) with sialoadhesin (CD169). We found that the immunorestorative activity of sialylated rhLF is also dependent on its interaction with the CD22 antigen, a member of the immunoglobulin superfamily that is expressed by B lymphocytes. We also demonstrated that only sialylated rhLF was able to inhibit the MLR reaction. MLR was inhibited by bovine lactoferrin (bLF), a glycoform that has a more complex glycan structure. Desialylated bLF and lactoferricin, a bLF-derived peptide devoid of carbohydrates, did not express such inhibitory activity. We showed that the interaction of LF with sialic acid receptors is essential for at least some of the immunoregulatory activity of this glycoprotein.

Keywords: Human recombinant lactoferrin, CD22, Sialic acid, Humoral immune response, Mixed lymphocyte reaction, CD169, Glycoproteins, Immune suppression

Full Text

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Abbreviations used

AFC

antibody-forming cells

bLF

bovine milk lactoferrin

ConA

concanavalin A

DegbLF

deglycosylated bovine lactoferrin

FCS

fetal calf serum

hLF

human lactoferrin

LF

lactoferrin

LPS

lipopolysaccharide

MLR

mixed lymphocyte reaction

MTX

methotrexate

PBMC

peripheral blood mononuclear cell

rhLF

recombinant human lactoferrin

rhLF A

non-sialylated recombinant human lactoferrin

rhLF B

sialylated recombinant human lactoferrin

SRBC

sheep red blood cells

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