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Mycobacterium tuberculosis–Induced Activation Accelerates Apoptosis in Peripheral Blood Neutrophils from Patients with Active Tuberculosis

Departamento de Inmunología, Instituto de Investigaciones Hematológicas, Academia Nacional de Medicina; and División de Tisioneumonología, Hospital F. J. Muñiz, Buenos Aires, Argentina

Address correspondence to: Dr. María del Carmen Sasiain, IIHema, Inmunología, Academia Nacional de Medicina, Pacheco de Melo 3081 (1425) Buenos Aires, Argentina. E-mail: maleman{at}hematologia.anm.edu.ar

The activation of circulating polymorphonuclear neutrophils (PMN) from patients with active tuberculosis (TB-PMN) may be associated with induction of apoptosis. Spontaneous or Mycobacterium tuberculosis (MTB)-induced apoptosis of PMN were evaluated by microscopy, DNA content, and their binding to Annexin V at 0, 3, and 18 h. In addition, the expression of CD11b and of CD16 were evaluated as parameters of activation and apoptosis, respectively. Recently isolated TB-PMN showed a higher CD11b expression than normal PMN (N-PMN), but there were no features of apoptosis, even though an enhancement of Fas expression was observed. Spontaneous apoptosis was accelerated in TB-PMN at 3 h, but no differences were observed in TB- and N-PMN at 18 h of culture. When stimulated with MTB, both TB- and N-PMN steadily increased CD11b expression along the culture period. MTB induced apoptosis of N-PMN at 3 h with loss of CD16 expression. By contrast, MTB delayed the apoptotic rate of TB-PMN, preserving the CD16 receptor at 3 h, whereas it accelerated apoptosis at 18 h, increasing at the same time the expression of CD11b. Taken together, these data suggest that the acceleration of apoptosis observed in TB-PMN could be associated with the MTB-induced activation.

Abbreviations: Annexin V, AV • bronchoalveolar lavage fluid, BALF • fetal calf serum, FCS • granulocyte macrophage colony-stimulating factor, GM-CSF • interferon , IFN- • interleukin, IL • lipopolysaccharide, LPS • mean fluorescence intensity, MFI • Mycobacterium tuberculosis, MTB • polymorphonuclear neutrophils, PMN • propidium iodide, PI • radical oxygen intermediates, ROI • tumor necrosis factor-, TNF-

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