The major glycoprotein allergen from Arachis hypogaea, Ara h 1, is a ligand of dendritic cell-specific ICAM-grabbing nonintegrin and acts as a Th2 adjuvant in vitro.
Nonmammalian glycan structures from helminths act as Th2 adjuvants. Some of these structures are also common on plant glycoproteins. We hypothesized that glycan structures present on peanut glycoallergens act as Th2 adjuvants. Peanut Ag (PNAg), but not deglycosylated PNAg, activated monocyte-derived dendritic cells (MDDCs) as measured by MHC/costimulatory molecule up-regulation, and by their ability to drive T cell proliferation. Furthermore, PNAg-activated MDDCs induced 2- to 3-fold more IL-4- and IL-13-secreting Th2 cells than immature or TNF/IL-1-activated MDDCs when cultured with naive CD4+ T cells. Human MDDCs rapidly internalized Ag in a calcium- and glycan-dependent manner consistent with recognition by C-type lectin. Dendritic cell (DC)-specific ICAM-grabbing nonintegrin (DC-SIGN) (CD209) was shown to recognize PNAg by enhanced uptake in transfected cell lines. To identify the DC-SIGN ligand from unfractionated PNAg, we expressed the extracellular portion of DC-SIGN as an Fc-fusion protein and used it to immunoprecipitate PNAg. A single glycoprotein was pulled down in a calcium-dependent manner, and its identity as Ara h 1 was proven by immunolabeling and mass spectrometry. Purified Ara h 1 was found to be sufficient for the induction of MDDCs that prime Th2-skewed T cell responses. Both PNAg and purified Ara h 1 induced Erk 1/2 phosphorylation of MDDCs, consistent with previous reports on the effect of Th2 adjuvants on DCs.
Shreffler
Wayne G
WG
Jaffe Food Allergy Institute, Department of Pediatrics, Division of Allergy and Immunology, Mount Sinai School of Medicine, New York, NY 10029, USA. wayne.shreffler@mssm.edu
Castro
Russell R
RR
Kucuk
Z Yesim
ZY
Charlop-Powers
Zachary
Z
Grishina
Galina
G
Yoo
Steven
S
Burks
A Wesley
AW
Sampson
Hugh A
HA
D000276:Adjuvants, Immunologic / Q000378:metabolism / Q000502:physiology*; D000485:Allergens / Q000276:immunology / Q000378:metabolism*; D052179:Antigens, Plant; D010367:Arachis / Q000276:immunology*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002478:Cells, Cultured; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000378:metabolism*; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D037181:Lectins, C-Type / Q000378:metabolism*; D008024:Ligands; D008565:Membrane Proteins; D010940:Plant Proteins / Q000276:immunology / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018418:Th2 Cells / Q000276:immunology*; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D000276:Adjuvants, Immunologic; D000485:Allergens; D052179:Antigens, Plant; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D037181:Lectins, C-Type; D008024:Ligands; D008565:Membrane Proteins; D010940:Plant Proteins; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin; C500889:Ara h 1 protein, Arachis hypogaea