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3.
Allergy ; 61(1): 56-63, 2006 Jan.
Article in English | MEDLINE | ID: mdl-16364157

ABSTRACT

BACKGROUND: Immunostimulatory oligodeoxynucleotides (CpG-ODN) usually contain phosphorothioate (PS) backbones for nucleotide protection, which may result in some nonspecific side-effects like prolongation of coagulation time. OBJECTIVE: The aim of the present study was to investigate the immunomodulatory potential of DNA molecules without PS backbones. Thus, we designed phosphorodiester (PO) molecules with a dumbbell-like covalently-closed structure (dSLIM-30L1). METHODS: We analyzed their effects on peripheral blood mononuclear cells (PBMC) from spontaneous high and low immunoglobulin (Ig)E producer (allergic and nonallergic donors) in comparison with linear CpG-ODN (lin-30L1) with PS backbones, using enzyme-linked immunosorbent assay and flow cytometry. RESULTS: We observed a decrease of spontaneous IgE levels in PBMC from high IgE producer of approximately 27% with both dSLIM-30L1 and lin-30L1. In addition, both molecules enhanced the production of IgA, IgM and IgG1/IgG2, but with a slightly different pattern. Both molecules stimulated the secretion of the T(H)1-like cytokines interleukin (IL)-2, interferon-gamma and IL-12p40 and the pro-inflammatory cytokine IL-6. The immunomodulatory potential of dSLIM-30L1 and lin-30L1 was also effective in PBMC from nonallergic donors, as was confirmed for IL-2, IL-12p40, IgG1/IgG2 and IgM. CONCLUSION: Our data show an inhibition of IgE production but also enhancement of the inflammatory cytokine response in PBMC from allergic and nonallergic donors by covalently-closed PO-based dSLIM-30L1 with a pattern similar to that of linear PS-based lin-30L1, while avoiding PS-modifications and thus PS-mediated side-effects. Whether such molecules are useful for the treatment of allergic diseases will need further clarification by appropriate in vivo studies.


Subject(s)
Cytokines/biosynthesis , Dermatitis, Atopic/immunology , Immunoglobulins/biosynthesis , Leukocytes, Mononuclear/immunology , Oligodeoxyribonucleotides/pharmacology , Polynucleotides/chemistry , Polynucleotides/pharmacology , Adult , Base Sequence , Case-Control Studies , Cells, Cultured , CpG Islands/immunology , Cytokines/immunology , Enzyme-Linked Immunosorbent Assay , Female , Flow Cytometry , Humans , Immunoglobulins/immunology , Immunologic Factors/chemistry , Immunologic Factors/pharmacology , Male , Middle Aged , Molecular Sequence Data , Oligodeoxyribonucleotides/chemistry , Probability , Reference Values , Sampling Studies , Sensitivity and Specificity
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