IL-4 supplemented B-cell cultures of allergic children show reduced IgA and IgG production in response to additional stimulation with IL-10.
BACKGROUND
Cytokines play an important role in mediating immunoglobulin switch, the secretion of protective mucosal immunoglobulins, and the development of allergic diseases. This study investigates whether B cells from allergic and healthy children have different capacities to secrete immunoglobulins after stimulation with IL-4, IL-6, IL-10, IL-11, and IL13.


METHODS
We analyzed the peripheral venous blood of 44 healthy probands and of 109 allergic patients with a mean age of 13 years, allergic to grass pollen, birch pollen, and house dust mites. Lymphocytes were isolated by a density gradient and B cells were enriched by using a Magnetic Activated Cell Separator (MACS) and anti-CD19 microbeads. B Cells were co-cultured with human CDw32 (Fc gammaRII) expressing mouse Ltk fibroblasts and mouse anti-human CD40 monoclonal antibodies (CD40 system). The interleukins IL-4, IL-6, IL-10, IL-11, and IL-13 were supplemented in various combinations. After 14 days, concentrations of IgE, IgG, IgA, and IgM were measured in the supernatants with ELISA.


RESULTS
Suppression of IgA-, IgG, and IgM- synthesis was induced by stimulation of B cells with IL-4. After additional application of IL-10, IgA, IgG, and IgM synthesis was significantly increased. When cultures stimulated with IL-4 were additionally supplemented with IL-10, IgA, and IgG synthesis of B cells obtained from allergic individuals was significantly decreased compared to nonallergic individuals. IgE-secretion of B cells from allergic individuals was significantly increased compared to nonallergic individuals after stimulation with IL-4.


CONCLUSIONS
Our results implicate that IL-4 is essential for the regulation of immunoglobulin class switch to IgE and that IL-4 is an important cytokine for the development of allergic diseases. The capacity of B cells in allergic children to produce less IgA and IgG in response to additional stimulation with IL-10 of cultures supplemented with IL-4 could play an important role in mediating a mucosal immune system vulnerable to allergens. This phenomenon could contribute to the pathogenesis of allergic diseases.
Nies
J H
JH
Institute for Clinical Immunology, Friedrich-Schiller-University Jena, Germany.
Bär
C
C
Schlenvoigt
G
G
Fahlbusch
B
B
Zwacka
G
G
Markert
U R
UR
D000293:Adolescent; D001402:B-Lymphocytes / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D018929:Cell Culture Techniques; D002648:Child; D002669:Child Welfare; D002675:Child, Preschool; D004359:Drug Therapy, Combination; D005858:Germany; D006801:Humans; D007070:Immunoglobulin A / Q000096:biosynthesis* / Q000187:drug effects* / Q000276:immunology; D007073:Immunoglobulin E / Q000096:biosynthesis / Q000187:drug effects / Q000276:immunology; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000187:drug effects* / Q000276:immunology; D007075:Immunoglobulin M / Q000096:biosynthesis / Q000187:drug effects / Q000276:immunology; D016753:Interleukin-10 / Q000276:immunology / Q000627:therapeutic use*; D015847:Interleukin-4 / Q000276:immunology / Q000627:therapeutic use*; D011994:Recombinant Proteins / Q000276:immunology / Q000627:therapeutic use; D012130:Respiratory Hypersensitivity / Q000188:drug therapy* / Q000276:immunology; D013268:Stimulation, Chemical; D016896:Treatment Outcome
D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011994:Recombinant Proteins; D016753:Interleukin-10; D015847:Interleukin-4; D007073:Immunoglobulin E