Methods for the in vitro determination of an individual disposition towards TH1‐ or TH2‐reactivity by the application of appropriate stimulatory antigens

• Published in: Clinical and experimental immunology Vol. 134; no. 1; pp. 78 - 85
• Main Authors: BARTH, H., BERG, P. A., KLEIN, R.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Science Ltd 01-10-2003; Blackwell; Oxford University Press; Blackwell Science Inc
• Subjects: Adult; Aged; Biological and medical sciences; Biomarkers - analysis; Clinical Studies; Cytokines - analysis; Cytokines - genetics; Enzyme-Linked Immunosorbent Assay - methods; Female; Flow Cytometry; Humans; Hypersensitivity, Delayed - immunology; Immunization; Immunological methods for diagnosis and exploration; Immunopathology; Interferon-gamma - analysis; Interferon-gamma - genetics; Interleukin-13 - analysis; Interleukin-4 - analysis; Interleukin-5 - analysis; Male; Medical sciences; Other methods; peripheral blood mononuclear cells; Predictive Value of Tests; purified protein derivative; Reverse Transcriptase Polymerase Chain Reaction; Rhinitis, Allergic, Seasonal - immunology; RNA, Messenger - analysis; Serologic Tests; T-Lymphocytes - immunology; tetanus toxoid; Tetanus Toxoid - administration & dosage; TH1; Th1 Cells - immunology; Th2 Cells - immunology; TH2 cytokines; Tumor Necrosis Factor-alpha - analysis; Tumor Necrosis Factor-alpha - genetics; type 1; type 2 reactivity; Human; Immunopathology; Allergy; Immunization; Immune response; type 1/type 2 reactivity peripheral blood mononuclear cells purified protein derivative tetanus toxoid TH1/ TH2 cytokines; Immunological investigation; Cytokine; Th1 lymphocyte; Th2 lymphocyte; ELISA assay; Profile
• ISSN: 0009-9104; 1365-2249
• DOI: 10.1046/j.1365-2249.2003.02265.x

SUMMARY
In this study we performed several methods for the determination of cytokines (RT‐PCR for the demonstration of cytokine mRNA and flow cytometry for the analysis of intracellular cytokines) and compared them with a recently established test system stimulating peripheral blood mononuclear cells (PBMC) with TH1‐ and TH2‐relevant recall antigens and analysing type 1 and type 2 cytokines by ELISA. Aim of the study was therefore to evaluate the reliability of TH1/TH2 cytokine profiles in two individuals with different types of an allergic/atopic disposition: one of them showed a strong TH1/type 1‐mediated tuberculin‐reaction (subject A), the other (subject B) revealed elevated IgE‐levels and eosinophil counts (TH2/type 2‐mediated). PBMC were incubated with the type 1‐antigen purified protein derivative (PPD) and the type 2‐antigen tetanus‐toxoid (TT) for seven days. From the comparison of ELISA with RT‐PCR and flow cytometry‐analysis it became evident that all three methods allowed the definition of subject A as a ‘type 1‐responder’. Subject B showed a pure type 2‐response in the ELISA method; PCR and flow cytometry analysis revealed the simultaneous production of type 1‐ and type 2‐cytokines resulting in a mixed type 1/type 2‐profile. Active immunization of subject A with TT at the end of the observation period of 12 months resulted in a transient shift from type 1‐ to a mixed type 1/type 2‐profile (simultaneous PPD‐induced IFN‐γ‐ and TT‐induced IL‐5 production). From this pilot study based on clear cut clinical criteria concerning either a humoral or cellular immunological reactivity towards allergens/antigens it is suggested that the determination of type 1/type 2‐cytokines by ELISA in supernatants of PBMC stimulated with type 1/type 2‐relevant antigens is a useful approach for a better classification of ‘type1‐’ or ‘type 2‐responder’.