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CD69 Expression on CD4⁺ T Lymphocytes after In Vitro Stimulation with Tuberculin Is an Indicator of Immune Sensitization against Mycobacterium tuberculosis Antigens

Bojana Avgustin,^1* Vladimir Kotnik,² Mojca koberne,² Tadej Malovrh,² Aleksandra Skralovnik-Stern,¹ and Marjeta Terelj¹

Institute of Microbiology and Immunology, Medical Faculty, University of Ljubljana,² Department of Pulmonary Diseases, University Medical Centre, Ljubljana, Slovenia¹

Received 17 February 2004/ Returned for modification 21 April 2004/ Accepted 16 September 2004

The expression of the CD69 antigen on CD4 T lymphocytes after in vitro stimulation with purified protein derivative (2 tuberculin units) was used to evaluate the tuberculin reactivities of 52 individuals from four experimental groups: Mycobacterium bovis BCG-vaccinated healthy individuals with a negative tuberculin skin test (TST) result (group A), BCG-vaccinated healthy individuals with a positive TST result (group B), patients with active tuberculosis (TB) before treatment (group C), and individuals with clinically inactive TB who had previously completed a prescribed course of chemotherapy (group D). The expression of CD69 on CD4 T lymphocytes was significantly higher in patients with active TB (16.2% ± 7.3%), individuals with clinically inactive TB (10.5% ± 7.4%), and healthy individuals with a positive TST result (15.5% ± 7.2%) than in healthy individuals with a negative TST result (3.8% ± 4.3%) (P < 0.005). We confirmed the correlation between CD69 antigen expression on T lymphocytes after stimulation with tuberculin and the TST induration diameter (Spearman rho = 0.783; P < 0.001), an assay for gamma interferon (the Quantiferon-TB assay; Spearman rho = 0.613; P < 0.001), and the lymphocyte BLAST transformation test (Spearman rho = 0.537; P < 0.001). Our results demonstrate the usefulness of the determination of CD69 on CD4 T lymphocytes after in vitro stimulation with tuberculin as a rapid indicator of immune sensitization against Mycobacterium tuberculosis.