Improving the in vitro antigen specific T cell proliferation assay: the use of interferon-alpha to elicit antigen specific stimulation and decrease bystander proliferation.
The measurement of the proliferative response of primed T cells to an antigenic stimulus (lymphocyte transformation assay: LTT) is commonly used for determining T cell immune responsiveness. However, the ratio between the spontaneous and the antigen-triggered response (stimulation index) is frequently quite low (<3-5) making the interpretation difficult. We modified the assay by the addition of interferon-alpha and the use of fresh autologous serum instead of human AB pool serum. These measures significantly enhanced the stimulation index following stimulation with tetanus toxoid, Candida albicans and tick-borne encephalitis (TBE) viral antigen in studies of sensitized patients. There was no concomitant increase in false positive results. Kinetic studies showed a reduced nonspecific background proliferation of non-stimulated cultures particularly between days 4 and 6 of culture. Furthermore, the positive effect of interferon-alpha were confirmed in studies of patients with contact allergy to nickel and gold. We conclude that this modified form of proliferation assay significantly increases the signal to noise ratio which can be attained. This may be of particular value when looking at T cell responses in immunocompromised patients or in diagnostic attempts to detect very low frequencies of antigen-specific T cells.
von Baehr
V
V
Institute of Medical Immunology, Charité Medical School, Campus Mitte, Humboldt University Berlin, D-10098, Berlin, Germany. baehr@ber.ladr.de
Mayer
W
W
Liebenthal
C
C
von Baehr
R
R
Bieger
W
W
Volk
H D
HD
D000328:Adult; D000941:Antigens / Q000008:administration & dosage; D000946:Antigens, Fungal / Q000008:administration & dosage; D000956:Antigens, Viral / Q000008:administration & dosage; D002176:Candida albicans / Q000276:immunology; D004669:Encephalitis Viruses, Tick-Borne / Q000276:immunology; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000008:administration & dosage; D007700:Kinetics; D008213:Lymphocyte Activation; D008297:Male; D008875:Middle Aged; D011994:Recombinant Proteins; D013601:T-Lymphocytes / Q000276:immunology*; D013745:Tetanus Toxoid / Q000008:administration & dosage
D000941:Antigens; D000946:Antigens, Fungal; D000956:Antigens, Viral; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D013745:Tetanus Toxoid