Jerrold J. Ellner

Apoptosis and T Cell Hyporesponsiveness in Pulmonary Tuberculosis

Mycobacterium tuberculosis (MTB)-induced T cell responses are depressed in peripheral blood mononuclear cells of persons with newly diagnosed pulmonary tuberculosis (TB), and levels of interferon (IFN)-gamma remain low even after completion of antituberculous therapy. Loss of MTB-reactive T cells through apoptotic mechanisms could account for this prolonged T cell hyporesponsiveness. T cell apoptosis was studied in TB patients and healthy control subjects. Both spontaneous and MTB-induced apoptosis (in CD4 and non-CD4 T cells) from TB patients was increased when compared with healthy control subjects, whereas coculture with control antigen (candida) had no effect on T cell apoptosis in either group of study subjects. An inverse correlation existed between increased MTB-induced T cell apoptosis and IFN-gamma and interleukin (IL)-2 immunoreactivities. Successful antituberculous chemotherapy resulted in a 50% reduction in both spontaneous and MTB-induced apoptosis, which coincided with 3- and 8-fold increases in levels of MTB-stimulated IL-2 and IFN-gamma, respectively. These data indicate that apoptotic pathways are operant during active MTB infection and may contribute to deletion of MTB-reactive T cells and the immunopathogenesis of this disease.

Augmentation of Apoptosis and Interferon-γ Production at Sites of Active Mycobacterium tuberculosis Infection in Human Tuberculosis

Pleural tuberculosis (TB) was employed as a model to study T cell apoptosis at sites of active Mycobacterium tuberculosis (MTB) infection in human immunodeficiency virus (HIV)-coinfected (HIV/TB) patients and patients infected with TB alone. Apoptosis in blood and in pleural fluid mononuclear cells and cytokine immunoreactivities in plasma and in pleural fluid were evaluated. T cells were expanded at the site of MTB infection, irrespective of HIV status. Apoptosis of CD4 and non-CD4 T cells in the pleural space occurred in both HIV/TB and TB. Interferon (IFN)-gamma levels were increased in pleural fluid, compared with plasma. Spontaneous apoptosis correlated with specific loss of MTB-reactive, IFN-gamma-producing pleural T cells. Immunoreactivities of molecules potentially involved in apoptosis, such as tumor necrosis factor-alpha, Fas-ligand, and Fas, were increased in pleural fluid, compared with plasma. These data suggest that continued exposure of immunoreactive cells to MTB at sites of infection may initiate a vicious cycle in which immune activation and loss of antigen-responsive T cells occur concomitantly, thus favoring persistence of MTB infection.

Sputum Cytokine Levels in Patients with Pulmonary Tuberculosis as Early Markers of Mycobacterial Clearance

ABSTRACT Sputum and serum from patients with active pulmonary tuberculosis (TB), healthy purified protein derivative-positive adults, and patients with bacterial pneumonia were collected to simultaneously assess local immunity in the lungs and peripheral blood. To determine whether cytokine profiles in sputum from TB patients and control subjects were a reflection of its cellular composition, cytospin slides were prepared in parallel and assessed for the presence of relative proportions of epithelial cells, neutrophils, macrophages, and T cells. Gamma interferon (IFN-γ) in sputum from TB patients was markedly elevated over levels for both control groups. With anti-TB therapy, IFN-γ levels in sputum from TB patients decreased rapidly and by week 4 of treatment were comparable to those in sputum from controls. Further, IFN-γ levels in sputum closely followed mycobacterial clearance. Although detected at fourfold-lower levels, IFN-γ immunoreactivities in serum followed kinetics in sputum. TNF-α, interleukin 8 (IL-8) and IL-6 also were readily detected in sputum from TB patients at baseline and responded to anti-TB therapy. In contrast to IFN-γ, however, TNF-α and IL-8 levels also were elevated in sputum from pneumonia controls. These data indicate that sputum cytokines correlate with disease activity during active TB of the lung and may serve as potential early markers for sputum conversion and response to anti-TB therapy.

Mycobacterium africanum Subtype II Is Associated with Two Distinct Genotypes and Is a Major Cause of Human Tuberculosis in Kampala, Uganda

ABSTRACT The population structure of 234 Mycobacterium tuberculosis complex strains obtained during 1995 and 1997 from tuberculosis patients living in Kampala, Uganda (East Africa), was analyzed by routine laboratory procedures, spoligotyping, and IS6110 restriction fragment length polymorphism (RFLP) typing. According to biochemical test results, 157 isolates (67%) were classified as M. africanum subtype II (resistant to thiophen-2-carboxylic acid hydrazide), 76 isolates (32%) were classified as M. tuberculosis, and 1 isolate was classified as classical M. bovis. Spoligotyping did not lead to clear differentiation of M. tuberculosis and M. africanum, but all M. africanum subtype II isolates lacked spacers 33 to 36, differentiating them from M. africanum subtype I. Moreover, spoligotyping was not sufficient for differentiation of isolates on the strain level, since 193 (82%) were grouped into clusters. In contrast, in the IS6110-based dendrogram, M. africanum strains were clustered into two closely related strain families (Uganda I and II) and clearly separated from the M. tuberculosis isolates. A further characteristic of both M. africanum subtype II families was the absence of spoligotype spacer 40. All strains of family I also lacked spacer 43. The clustering rate obtained by the combination of spoligotyping and RFLP IS6110 analysis was similar for M. africanum and M. tuberculosis, as 46% and 49% of the respective isolates were grouped into clusters. The results presented demonstrate that M. africanum subtype II isolates from Kampala, Uganda, belong to two closely related genotypes, which may represent unique phylogenetic branches within the M. tuberculosis complex. We conclude that M. africanum subtype II is the main cause of human tuberculosis in Kampala, Uganda.

Airborne infection with Bacillus anthracis--from mills to mail.

The lack of identified exposures in 2 of the 11 cases of bioterrorism-related inhalation anthrax in 2001 raised uncertainty about the infectious dose and transmission of Bacillus anthracis. We used the Wells-Riley mathematical model of airborne infection to estimate 1) the exposure concentrations in postal facilities where cases of inhalation anthrax occurred and 2) the risk for infection in various hypothetical scenarios of exposure to B. anthracis aerosolized from contaminated mail in residential settings. These models suggest that a small number of cases of inhalation anthrax can be expected when large numbers of persons are exposed to low concentrations of B. anthracis. The risk for inhalation anthrax is determined not only by bacillary virulence factors but also by infectious aerosol production and removal rates and by host factors.

Induction of the Antigen 85 Complex of Mycobacterium tuberculosis in Sputum: A Determinant of Outcome in Pulmonary Tuberculosis Treatment

Sputum quantitative culture, acid-fast smear, days-to-positive by BACTEC, and Mycobacterium tuberculosis antigen 85 complex were monitored during therapy in 42 patients with pulmonary tuberculosis (TB). By BACTEC, 4 patients were persistently positive on days 90-180, and treatment ultimately failed in 2 of these. Antigen 85 expression increased in subjects in whom disease persisted (persisters) from days 0 to 14 when the difference between persisters and nonpersisters was statistically significant (P = .002). Only antigen 85 complex values at day 14 suggested TB persistence at or after day 90. All subjects with day 14 antigen 85 complex values < 60 pg/mL responded rapidly to treatment and were cured. Of those with values > 60 pg/mL, in 33% TB persisted at or after day 90 and treatment failed in 17%. Biologic factors expressed early in therapy, not related to compliance or resistance, may exert a substantial influence on outcome. The antigen 85 complex is critical in cell wall biosynthesis and is induced by isoniazid in vitro. Its induction may represent an adaptive transition to a persistent state during therapy.

Absence of Mycobacterium avium Complex Disease in Patients with Aids in Uganda

The absence of disease due to Mycobacterium avium in Ugandan patients with AIDS, which we previously observed in a blood culture study, has been confirmed and our observations have been extended to 165 additional clinical isolates. Fourteen soil and water samples from the Ugandan environment have been cultured and revealed a high frequency of isolation of M. avium. The absence of M. avium complex disease in Uganda remains unexplained.

'Coinfection-helminthes and tuberculosis'

Purpose of reviewTuberculosis (TB) continues to be a significant problem and a major cause of morbidity and mortality in the developing world despite decades of intensive efforts to combat the disease. The poverty in these endemic areas is associated with an increased incidence of tropical helminthic infections. The purpose of this review is to bring to the fore, the urgent need to unravel the potential consequences of helminth coinfection to tuberculosis disease pathogenesis and transmission. Recent findingsThere is now strong experimental evidence that helminth-induced T helper (Th)2 and T regulatory (Treg) responses impinge on host resistance against Mycobacterium tuberculosis (Mtb) infection. Several studies show that Th1 response is reduced in helminth coinfected hosts. Emerging studies also indicate that helminth-induced alternatively activated macrophages contribute to enhanced susceptibility to TB. Despite studies showing an association between helminthes and diminished Th1 immunity, maternal antihelminthic treatment had no effect on an infants response to bacille Calmette-Guerin vaccination. SummaryWhether helminthes affect tuberculosis (TB) disease is still an open question and clinical trials are warranted to determine at the population level whether helminthes enhance TB incidence and transmission and diminish the protective immune response to vaccines. Consequently, mass deworming of infected individuals could contribute toward overall improvement of global public health.

Accessory function and properties of monocytes from healthy elderly humans for T lymphocyte responses to mitogen and antigen.

The waning of cell-mediated immunity during aging has been attributed primarily to defects in T lymphocyte properties and functions. We assessed the potential contribution of accessory dysfunction of monocytes from the elderly on responses of T cells to phytohemagglutinin (PHA) and to tetanus toxoid after in vivo boosting. Accessory function of monocytes from the elderly subjects for T lymphocyte responses to tetanus toxoid was comparable to the young. Expression of the cytokines interleukin-1, interleukin-6 and tumor necrosis factor, the cell adhesion molecules ICAM-1 and LFA-3 and the class II major histocompatibility molecule HLA-DR by monocytes from the elderly and young subjects was similar. T lymphocytes from the elderly responded poorly to PHA. Monocytes from the elderly had a decreased accessory function for PHA-stimulated T cells from young, third donors. Thus, although many accessory properties of monocytes from the elderly are normal, the monocyte and T lymphocyte defects in the elderly for mitogen may represent interactive factors in cell-mediated immunity during aging.

Latent tuberculosis infection--Revisiting and revising concepts.

Host- and pathogen-specific factors interplay with the environment in a complex fashion to determine the outcome of infection with Mycobacterium tuberculosis (Mtb), resulting in one of three possible outcomes: cure, latency or active disease. Although much remains unknown about its pathophysiology, latent tuberculosis infection (LTBI) defined by immunologic evidence of Mtb infection is a continuum between self-cure and asymptomatic, yet active tuberculosis (TB) disease. Strain virulence, intensity of exposure to the index case, size of the bacterial inoculum, and host factors such as age and co-morbidities, each contribute to where one settles on the continuum. Currently, the diagnosis of LTBI is based on reactive tuberculin skin testing (TST) and/or a positive interferon-gamma release assay (IGRA). Neither diagnostic test reflects the activity of the infectious focus or the risk of progression to active TB. This is a critical shortcoming, as accurate and efficient detection of those with LTBI at higher risk of progression to TB disease would allow for provision of targeted preventive therapy to those most likely to benefit. Host biomarkers may prove of value in stratifying risk of development of TB. New guidelines are required for interpretation of discordance between TST and IGRA, which may be due in part to a lack of stability (that is reproducibility) of IGRA or TST results or to a delay in conversion of IGRA to positivity compared to TST. In this review, the authors elaborate on the definition, diagnosis, pathophysiology and natural history of LTBI, as well as promising methods for better stratifying risk of progression to TB. The review is centered on the human host and the clinical and epidemiologic features of LTBI that are relevant to the development of new and improved diagnostic tools.