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

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.

Abbreviations

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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Correspondence to Michał Zimecki.

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Zimecki, M., Artym, J., Kocięba, M. et al. The effect of carbohydrate moiety structure on the immunoregulatory activity of lactoferrin in vitro. Cell Mol Biol Lett 19, 284–296 (2014). https://doi.org/10.2478/s11658-014-0196-2

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Keywords

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