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Published on: March 24, 2015
Interferon-gamma production in antigen specific T cell response: quantitation of specific mRNA and secreted protein
M Halminen1, P Klemetti, O Vaarala
1Turku Immunology Centre, University of Turku, Finland.
Insights
Interferon gamma (IFN-gamma) production assays effectively measure cellular sensitization to viruses like herpes simplex virus (HSV). These methods correlate well with T cell responsiveness, offering reliable immune assessment.
Area of Science:
- Immunology
- Cellular Biology
- Molecular Biology
Background:
- Cellular sensitization is crucial for adaptive immunity.
- Assessing T cell responsiveness is vital for understanding immune memory and response to pathogens.
- Existing methods for measuring immune response have limitations.
Purpose of the Study:
- To evaluate Interferon gamma (IFN-gamma) production assays as measures of cellular sensitization.
- To compare IFN-gamma detection via enzyme immunoassay (EIA) and reverse transcriptase polymerase chain reaction (RT-PCR) with the lymphocyte proliferation assay.
- To assess immune responses to herpes simplex virus (HSV) and tetanus toxoid (TT).
Main Methods:
- Detection of IFN-gamma in culture supernatant using enzyme immunoassay (EIA).
- Measurement of cellular IFN-gamma mRNA using reverse transcriptase polymerase chain reaction (RT-PCR).
- Comparison with the standard lymphocyte proliferation assay for T cell responsiveness.
Main Results:
- IFN-gamma assays and lymphocyte proliferation assays clearly distinguished between HSV seropositive and seronegative donors.
- Significant correlations were observed between IFN-gamma mRNA, secreted IFN-gamma, and proliferative response.
- While TT responses showed variability, individual IFN-gamma production levels remained constant, increasing after revaccination.
Conclusions:
- IFN-gamma production assays are reliable indicators of cellular sensitization and T cell responsiveness.
- These methods provide a robust way to assess immune status against viral and bacterial antigens.
- IFN-gamma assays can detect immune responses, even in individuals with initially low or absent proliferation responses after vaccination.
Abstract:
Interferon gamma (IFN-gamma) production as a measure of cellular sensitization was studied by detection of the cytokine in culture supernatant by enzyme immunoassay (EIA) and by measuring cellular mRNA using the reverse transcriptase polymerase chain reaction (RT-PCR) method. These assays were compared to the standard lymphocyte proliferation assay as a marker of T cell responsiveness to foreign antigens. When blood donors seropositive for herpes simplex virus (HSV) were compared to seronegative donors, all measurements of cellular sensitization separated the groups without overlap. There were significant correlations between the IFN-gamma mRNA titre and the secreted IFN-gamma (r = 0.57, P = 0.03), and the proliferative response and the secreted IFN-gamma (r = 0.78, P = 0.001), as well as between the IFN-gamma mRNA titre and the proliferative response (r = 0.78, P < 0.001). When tetanus toxoid (TT) responses were studied in immunized subjects, a wide range of responsiveness could be seen and correlation between various measurements was poor. However, constant individual levels of the cytokine production were demonstrated. Six people who had received their last TT booster vaccination more than 5 years ago were revaccinated and repeatedly studied. An increase in the levels of produced IFN-gamma could be seen in all subjects and two who lacked a lymphocyte proliferation response developed it after revaccination.
