Selective restimulation of antigen or allergen preactivated T cells using OKT3 F(ab)2 results in the secretion of TH-1 or TH-2-like cytokine patterns.
BACKGROUND
The synthesis of IgE is regulated by cytokines secreted from T-helper cells. The studies on cytokine secretion by peripheral blood mononuclear cells (PBMC) upon stimulation with antigen or allergen are difficult due to low levels of cytokines, especially of interleukin-4 (IL-4).


OBJECTIVE
In this study we tried to establish a culture system, which could enable the measurement of the cytokine profiles in specifically activated cultures.


METHODS
Three methods to potentiate cytokine secretion were evaluated: PBMC from bee venom or house dust mite (Dermatophagoides pteronyssinus) allergic patients as well as normal subjects were stimulated either with the major bee venom allergen phospholipase A2 (PLA) or with the major D. pteronyssinus allergen (Der p 1) or with the control antigens tetanus toxoid (TT) and purified protein derivate (PPD). After 7 days of culture the cells were restimulated either with plastic bound OKT3 F(ab)2 monoclonal antibodies (MoAbs), with the appropriate antigen + antigen presenting cells or with IL-2. The secretion of cytokines (IL-4, IFN gamma) was measured after restimulation of the cultures (day 8).


RESULTS
While OKT3 F(ab)2 was unable to activate resting T cells, it could restimulate preactivated cells. Restimulation with OKT3 F(ab)2 induced higher IL-4 and IFN gamma secretion than restimulation with IL-2 or antigen. TT and PLA stimulated a similar cytokine secretion profile in normal and PLA allergic subjects with substantial levels of both IL-4 and IFN gamma. In contrast, PPD induced virtually only IFN gamma secretion. Der p 1 stimulated mainly IL-4 secretion but also IFN gamma production in some mite allergic patients.


CONCLUSIONS
We have established a cell culture system, which combines antigen specificity with a strong cytokine inducing signal provided by anti-CD3 MoAbs. TH-1 and TH-2 characteristic cytokine patterns can be observed in short-term PBMC cultures already after 8 days of culture.
Jutel
M
M
Zieglerspital, Bern, Switzerland.
Wyss-Coray
T
T
Carballido
J M
JM
Blaser
K
K
Müller
U R
UR
Pichler
W J
WJ
D000485:Allergens / Q000494:pharmacology*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism*; D000939:Epitopes / Q000494:pharmacology; D006801:Humans; D007114:Immunization; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology*; D007376:Interleukin-2 / Q000276:immunology / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation; D016853:Muromonab-CD3 / Q000737:chemistry / Q000494:pharmacology*; D010741:Phospholipases A / Q000494:pharmacology; D054467:Phospholipases A2; D013745:Tetanus Toxoid / Q000494:pharmacology; D018417:Th1 Cells / Q000378:metabolism*; D018418:Th2 Cells / Q000378:metabolism*; D014373:Tuberculin / Q000494:pharmacology
D000485:Allergens; D016207:Cytokines; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007376:Interleukin-2; D016853:Muromonab-CD3; D013745:Tetanus Toxoid; D014373:Tuberculin; D010741:Phospholipases A; D054467:Phospholipases A2