Immunostimulatory DNA inhibits IL-4-dependent IgE synthesis by human B cells.
BACKGROUND
Immunostimulatory sequence oligodeoxynucleotide (ISS-ODN) is a potent antiallergic immunomodulating agent in mice. However, few studies have addressed its antiallergic potential in human subjects.


OBJECTIVE
We sought to determine whether a phosphoro-thioate ISS-ODN could inhibit IL-4-dependent IgE synthesis by human B cells.


METHODS
Initially, nonatopic- and atopic-donor PBMCs were incubated with ISS-ODN or mutated oligodeoxynucleotide, and cytokine production and B-cell expression of IFN-gamma receptor and IL-4 receptor were measured by using ELISA and flow cytometry, respectively. In subsequent studies atopic-donor PBMCs were incubated with IL-4 alone or with ISS-ODN or mutated oligodeoxynucleotide. After 14 days, IgE production and IgM, IgG, and IgA production were determined by using ELISA. In select IgE studies cytokines were neutralized with mAbs.


RESULTS
ISS-ODN induced IL-12, IFN-alpha, IFN-gamma, IL-10, and IL-6 production from both nonatopic- and atopic-donor PBMCs. ISS-ODN also increased IFN-gamma receptor and inhibited IL-4 receptor expression on B cells from both donor populations. Furthermore, ISS-ODN inhibited IL-4-dependent IgE production by atopic-donor PBMCs. Neutralization of IL-12, IFN-alpha, IFN-gamma, and IL-10, but not IL-6, attenuated the inhibitory activity of ISS-ODN on IgE production. In contrast to its inhibition of IgE synthesis, ISS-ODN stimulated the production of IgM, IgG, and IgA.


CONCLUSIONS
These in vitro studies demonstrate that phos-phorothioate ISS-ODN elicits an innate immune response by PBMCs, which inhibits IL-4-dependent IgE synthesis. In addition, these results provide further support for consideration of ISS-ODN therapy for the treatment of allergic disease in clinical practice.
Horner
A A
AA
Department of Medicine, The Sam and Rose Stein Institute for Aging, University of California, San Diego, La Jolla 92093-0663, USA.
Widhopf
G F
GF
Burger
J A
JA
Takabayashi
K
K
Cinman
N
N
Ronaghy
A
A
Spiegelberg
H L
HL
Raz
E
E
D018926:Anti-Allergic Agents / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002478:Cells, Cultured; D004247:DNA / Q000276:immunology / Q000494:pharmacology*; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology; D006969:Hypersensitivity, Immediate / Q000276:immunology; D007070:Immunoglobulin A / Q000096:biosynthesis; D007073:Immunoglobulin E / Q000096:biosynthesis*; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D015847:Interleukin-4 / Q000276:immunology*; D009838:Oligodeoxyribonucleotides / Q000276:immunology / Q000494:pharmacology*; D013873:Thionucleotides / Q000276:immunology / Q000494:pharmacology*
D018926:Anti-Allergic Agents; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D009838:Oligodeoxyribonucleotides; D013873:Thionucleotides; D015847:Interleukin-4; D007073:Immunoglobulin E; D004247:DNA