Abstract:
BACKGROUND:A comprehensive understanding of the immune response induced by Mycobacterium bovis Bacille Calmette-Guérin in activation of protective T cells against tuberculosis is important to develop effective therapies to combat this disease. In this study, our experiments were designed to determine effects of transforming growth factor (TGF)-beta on M. bovis-induced T-cell activation and survival. METHODS:Fluorescence-activated cell sorter (FACS) analysis was used for detection of apo-ptotic cells by three different methods: 1). scattered light change during early phase of apoptosis; 2). detection of hypodiploid DNA, or 3). terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) technique. Quantification of positively stained population was based on samples stained with isotype control antibodies analyzed on a FACScan. RESULTS:TGF-beta added at initiation of culture did not alter percentage of viable cells. By contrast, TGF-beta added 72 h post-activation decreased percentage of viable cells. This effect was statistically significant (p <0.05). Furthermore, addition of anti-TGF-beta MoAb together with TGF-beta abolished the ability of this cytokine to decrease survival in post-activated human T cells. Role of TGF-beta on post-activated human T cells was further confirmed by staining apoptotic nuclei with propidium iodide, which detects late events of apoptosis, and by DNA fragmentation determined using TUNEL assay. Interestingly, TGF-beta did not promote Fas-mediated killing. Finally, TGF-beta increased apoptosis of CD4(+) T cells after mycobacterial stimulation. CONCLUSIONS:This study indicated an important role for TGF-beta in suppression of protective immune response against M. bovis by promoting elimination of post-activated T cells. Furthermore, results showed that TGF-beta had no direct effect on M. bovis-induced up-regulation of Fas (CD95).
journal_name
Arch Med Resjournal_title
Archives of medical researchauthors
Hernández-Garay M,Méndez-Samperio Pdoi
10.1016/s0188-4409(02)00463-0subject
Has Abstractpub_date
2003-01-01 00:00:00pages
20-5issue
1eissn
0188-4409issn
1873-5487pii
S0188440902004630journal_volume
34pub_type
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