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Published on: December 7, 2019
Allergen-specific T cell responses to immunotherapy monitored by CD154 and intracellular cytokine expression
J D Campbell1, P Buchmann, S Kesting
1Dynavax Technologies, Berkeley, CA 94710, USA. DCampbell@dynavax.com
Insights
This study shows that measuring CD154 and intracellular cytokines in T cells can track immune responses to immunotherapy. This method effectively monitors changes in allergen-specific T cells and correlates with allergic symptom reduction.
Area of Science:
- Immunology
- Allergy Research
- Clinical Trial Methodology
Background:
- Measuring low numbers of intracellular cytokine-expressing antigen-specific T cells from peripheral blood mononuclear cells (PBMC) is crucial for monitoring immunotherapy.
- CD154 serves as a reliable marker for recently activated T cells, enabling sensitive detection of these responses.
Purpose of the Study:
- To assess the utility of CD154 and intracellular cytokine staining for measuring changes in allergen-specific T cell cytokine production within a clinical trial.
- To evaluate the effectiveness of this method in the context of allergic rhinitis immunotherapy.
Main Methods:
- Peripheral blood mononuclear cells (PBMC) were collected from subjects undergoing immunotherapy for ragweed allergy.
- Cryopreserved PBMC were stimulated with allergen (Amb a 1) and analyzed for CD154 and intracellular cytokine (IL-4, IL-5, IL-13, IFN-gamma) expression using flow cytometry.
- Subjects received either an Amb a 1-immunostimulatory oligodeoxynucleotide conjugate (AIC) or placebo treatment.
Main Results:
- Placebo group showed significant increases in T helper type 2 (Th2) cytokines (IL-4, IL-5, IL-13) after allergen exposure post-treatment.
- AIC-treated subjects did not exhibit increased Th2 cytokine expression post-treatment, indicating suppressed allergic T cell responses.
- Interferon-gamma (IFN-gamma) levels remained unchanged in both groups.
- Nasal symptom scores correlated with allergen-stimulated Th2 cytokine responses.
Conclusions:
- Combined CD154 and intracellular cytokine staining is a sensitive method for monitoring antigen-specific T cell subset frequencies in clinical studies.
- This technique can effectively track immune system changes during immunotherapy for allergic diseases.
- The measured antigen-specific cytokine expression showed a moderate correlation with reported allergic symptom severity.
Background:
A sensitive measurement of low numbers of intracellular cytokine-expressing antigen-specific T cells from peripheral blood mononuclear cells (PBMC) is possible using CD154 as a marker of recently activated T cells. This technique may have potential for monitoring peripheral blood T cell responses to immunotherapy.
Objective:
To evaluate the applicability of this method for measuring changes in cytokine production by allergen-specific T cells in a clinical trial setting.
Methods:
Ex vivo ragweed-specific CD154 and intracellular cytokine expression were evaluated using a subset of subjects in an environmental chamber study of allergic rhinitis immunotherapy. PBMC were collected and cryopreserved from Amb a 1-immunostimulatory oligodeoxynucleotide conjugate (AIC)-treated (n=17) and placebo-treated (n=15) ragweed-allergic subjects both after pre- and post-treatment ragweed exposures. In vitro allergen-stimulated CD3(+)CD4(+)CD154(+) T cell intracellular IL-4, IL-5, IL-13, and IFN-gamma expression were evaluated by flow cytometry.
Results:
Compared with the T helper type 2 (Th2) cytokine expression measured after pre-treatment ragweed exposures, placebo-treated subjects demonstrated a significantly elevated ragweed- and Amb a 1-specific T cell IL-4 and IL-13 co-expression (P=0.005 and P=0.022, respectively) and a significantly elevated ragweed-specific IL-5 expression (P<0.001) following post-treatment ragweed exposures. In contrast, AIC-treated subjects demonstrated no increases in allergen-specific Th2 cytokine expression following post-treatment ragweed exposures. IFN-gamma expression remained low and un-changed in both groups. Subject reported total nasal symptom scores demonstrated modest but significant correlations with Amb a 1- and ragweed-stimulated intracellular Th2 cytokine responses.
Conclusion:
Combined CD154 and intracellular cytokine staining in PBMC can be used to sensitively monitor changes in antigen-specific T cell subset frequencies in clinical studies. Antigen-specific cytokine expression moderately correlated with the reported levels of allergic symptoms.
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