CD69 Expression on CD4⁺T Lymphocytes after In Vitro Stimulation with Tuberculin Is an Indicator of Immune Sensitization againstMycobacterium tuberculosisAntigens

Abstract

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.The fast, early, and accurate diagnosis of Mycobacterium tuberculosis infection is a very important element of global health measures for the control of tuberculosis (TB). The identification of individuals with latent M. tuberculosis infection (LTBI) who will benefit from treatment is crucial to the goal of TB elimination (1, 2). Since the immune response to mycobacterial infection is predominantly cellular (11), assessment of whether a patient's T cells have been exposed to and sensitized by antigens specific to M. tuberculosis provides an approach to diagnosis (4). Delayed-type hypersensitivity skin testing by the tuberculin skin test (TST) with purified protein derivate (PPD) is the standard method of screening for TB and has been a convenient, cost-effective method for assessment of the cell-mediated immune response to a tuberculin. Although TST has been the "gold standard" for diagnostic screening and the detection of new or asymptomatic TB, it has a number of drawbacks, including the need for a return visit to allow reading of the results, problems with interpretation of the test results due to cross-reactivity with other mycobacterial species, the booster effect, false-negative results due to intercurrent immunosuppression, and the variability inherent in its application and reading (16). In addition, it is imprecise and can give only a partial assessment of the interrelationships among M. tuberculosis, host macrophages, and the surrounding cellular components of the immune system, particularly CD4 T lymphocytes (5, 22). Different types of...

Increased percentage of IFN-γ producing CD56+CD3+ cells in active tuberculosis patients upon CFP-10 stimulation of peripheral mononuclear cells

“…PPD or CFP-10 were increased in PTB and TSTþ HHC, compared to TSTÀ Non HHC, supporting that all of these responses differentiate these groups regardless of the stimuli used, and BCG vaccination. These findings are in agreement with immune response against PPD, CFP-10 or ESAT-6 determined by proliferation index [11,27], with IFN-g in cell culture supernatants [4,28], and with the percentages of IFN-g producing CD4 þ and CD8 þ lymphocytes where patients and latent infected individuals are differentiated from non-infected individuals [29,30], unlike the report by Cardoso et al in Brazil, who found that 8/10 TST-controls produced IFN-g in response to ESAT-6 [6].…”

“…Avgustin et al found an increased expression of CD69 on CD4 þ T lymphocytes stimulated with PPD in BCG-vaccinated TSTþ individuals, in patients with active and treated TB, compared to vaccinated TSTÀ individuals, enabling to differentiate infected individuals from non-infected ones [11]. Our results show that in Figure 3.…”

“…Its expression on CD4 þ T cells after PPD stimulus is higher in patients and TSTþ individuals than in TSTÀ individuals, indicating that CD69 expression may be a TB infection marker [11], however it did not differentiate TB disease from latent infection.…”

Involvement of CD244 in Regulating CD4+ T Cell Immunity in Patients with Active Tuberculosis

“…Antigen-responsive CD4 + T cells were identified by appearance of activation marker CD69 in the CD3 + CD4 + cell population (Fig. 1E) [29], [30]. In the cohort of TB patients, 30 were defined as new cases and 13 retreatment TB patients (Table 1).…”

Immunity towards tuberculosis infection: A review

“…There is a strong correlation between the replication of HIV and the production of proinflammatory cytokines in the lungs of patients with TB (Gold et al, 2004). It has been determined that the cytokine model mRNA could therefore indicate any superiority of Th-2 or the mediated inmmunosupression by the IL-10 ( Westenbrink et al, 2005); and it has been confirmed that immunity towards Mycobacterium is predominantly cellular (Avgustin et al, 2005). Th cells mature from a precursory state Th0 to Th1 phenotype or Th2 (Dovedi et al, 2005).…”