Mycobacterium tuberculosis (Mtb) is an intracellular pathogen that infects alveolar macrophages following aerosol transmission. Lung macrophages provide a critical intracellular niche that is required for Mtb to establish infection in the human host. This parasitic relationship is made possible by the capacity of Mtb to block phagosome maturation following entry into the host macrophage, creating an environment that supports bacillary replication. Apoptosis is increasingly understood to play a role in host defense against intracellular pathogens including viruses, fungi, protozoa and bacteria. In the last 15 years an understanding of the role that macrophage apoptosis plays in TB has begun to emerge. Here we review the history and current state of the art of this topic and we offer a model of the macrophage-pathogen interaction that takes into the account the complexities of programmed cell death and the relationship between various death signaling pathways and host defense in TB.
Animals ; Apoptosis/*immunology ; Humans ; Macrophages/*cytology/*microbiology ; Mycobacterium tuberculosis/*immunology ; Tuberculosis, Pulmonary/*immunology

OBJECTIVETo detect the expression of Mycobacterium tuberculosis secreted protein Ag85B in paraffin-embedded tissues by immunohistochemistry (IHC), and to evaluate its application in the pathological diagnosis of tuberculosis.

METHODSOne hundred and five tuberculosis specimens (54 pulmonary tuberculosis, 51 lymph nodal tuberculosis) and 51 specimens of other diseases (8 lung cancer, 10 pulmonary abscess, 10 bronchiectasis, 7 lymphoma, 5 necrotizing lymphadenitis, 4 reactive hyperplasia lymphoid, and 7 sarcoidosis) were collected from January 2012 to July 2013 from Beijing Chest Hospital, Capital Medical University. One-step IHC was performed on paraffin-embedded tissues using antibody directed against Ag85B.

RESULTSIHC and Ziehl-Neelsen (ZN) acid-fast staining showed that distribution and intensity of Ag85B expression were concordant with the distribution and number of acid-fast bacilli. IHC showed significantly higher sensitivity than ZN staining (50.5%, 53/105 vs. 31.4%, 33/105; χ² = 7.877, P = 0.005). The combined sensitivity of IHC and ZN staining was 59.0%. Moreover, oil immersion was not necessary for IHC, allowing more rapid diagnosis.

CONCLUSIONIHC detection of Ag85B is a simple method with higher sensitivity than ZN staining, and demonstrated good value in the pathological diagnosis of tuberculosis.

Acyltransferases ; metabolism ; Antigens, Bacterial ; metabolism ; Biomarkers ; metabolism ; Bronchiectasis ; diagnosis ; immunology ; Humans ; Immunohistochemistry ; Lymphadenitis ; diagnosis ; immunology ; Mycobacterium tuberculosis ; immunology ; Sarcoidosis ; diagnosis ; Staining and Labeling ; Tuberculosis, Lymph Node ; diagnosis ; immunology ; Tuberculosis, Pulmonary ; diagnosis ; immunology

OBJECTIVETo verify efficacy of moxibustion apparatus on pulmonary tuberculosis (PT) and explore adjuvant treatment method for PT.

METHODSOne hundred cases of PT were randomly divided into a moxibustion group and a routine treatment group, 50 cases in each one. The regular antituberculous therapy (2HRZE/4HRE) was applied in both groups. In addition, the moxibustion apparatus was used at Bailao (EX-HN 15), Feishu (BL 13), Gaohuang (BL 43), Qihai (CV 6), Zhongfu (LU 1), Danzhong (CV 17), Guanyuan (CV 4), Zusanli (ST 36) and so on in the moxibustion group. The change of lesion area in chest radiography, degradation rate of bacte rium in the sputum, T-lymphocyte subsets and natural kill (NK) cells were observed before and after treatment in two groups.

RESULTSAfter the treatment for 3 months, there were 45 cases (90.0%) in the moxibustion group with more than 45% of focal absorption in chest radiography, which was obviously higher than 72.0% (36/50) in the routine treatment group (P < 0.01). The degradation rate of bacterium in the sputum in the moxibustion group was higher than that in the routine treatment group [82.0% (41/50) vs 60.0% (30/50), P < 0.01]. The CD3+, CD4+/CD8+ ratio of T-lymphocyte subsets and NK cells in the moxibustion group were significantly higher than those in the routine treatment group (P < 0.05, P < 0.01).

CONCLUSIONOn the basis of regular antituberculous therapy, moxibustion apparatus could significantly improve clinical effect, promote focal absorption and boost immunity, which is considered as an adjuvant treatment for PT.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; T-Lymphocyte Subsets ; immunology ; Tuberculosis, Pulmonary ; immunology ; therapy ; Young Adult

Aged ; Aged, 80 and over ; Antibodies, Bacterial ; blood ; Coal Mining ; Humans ; Male ; Middle Aged ; Mycobacterium tuberculosis ; immunology ; Pneumoconiosis ; complications ; Sensitivity and Specificity ; Tuberculosis, Pulmonary ; complications ; diagnosis

OBJECTIVETo establish a recombinant plasmid of CFP32 of Mycobacterium tuberculosis in E. coli, and to analyze its antigenicity.

METHODSRv0577 gene was amplified by polymerase chain reaction from genome of Mycobacterium tuberculosis, and then cloned into vector pMD18-T followed by the subclone into the expression vector pET21a. Recombinant CFP32 was expressed and purified. The antigenicity of the recombinant protein was analyzed by using Western-blot. The purified recombinant CFP32 protein was used as an antigen to screen the sera of 7 pulmonary TB patients (n = 97), as well as the other pulmonary disease patients (n = 25), and the clinically healthy controls (n = 38) by ELISA.

RESULTSRecombinant plasmid of CFP32 was established, and be expressed efficiently in E. coli BL21 (DE3). The relative molecular mass of the protein was about 300,000 by SDS-PAGE analysis. The protein purified by Ni-NTA was in a purity over 90%, which was confirmed by Western-blot analysis. ELISA analysis showed its sensitivity and specificity were 63.9% (62/97) and 96.8% (2/63) respectively.

CONCLUSIONThe recombinant expression plasmid pET21a CFP32 has been constructed and CFP32 proteins has been successfully expressed and be purified in E. coli and, ELISA analysis has identified the recombinant CFP32 as a candidate antigen for TB serodiagnosis.

Antigens, Bacterial ; blood ; Bacterial Proteins ; genetics ; immunology ; Cloning, Molecular ; Escherichia coli ; Gene Expression ; Humans ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Plasmids ; Recombinant Proteins ; Serologic Tests ; Tuberculosis, Pulmonary ; diagnosis ; microbiology

Recent increase in the incidence of lung cancer often makes it difficult to differentiate between lung cancer and tuberculosis (TB), due to their radiologic similarities. Fine needle aspiration biopsy (FNAB) has been widely employed for the diagnosis of lung cancer and TB, but the diagnostic accuracy of TB is not high enough. As a rapid screening test for tuberculosis, we evaluated serological tests using Mycobacterium tuberculosis PPD and lipoarabinomannan (LAM) antigens. A total of 95 patients with indication of FNAB cytology from initial CT findings were enrolled. 25 patients had TB, 76 thoracic malignancy, and six (7.9%) of the lung cancer patients also had TB, indicating much higher prevalence of TB in thoracic tumor patients. Antibodies to PPD were elevated in 18 (72.0%) of 25 TB patients and in 22 (31.4%) of 70 patients with thoracic malignancy. In contrast, only 3 (4.7%) of 64 healthy controls aged 40 or above were seropositive to PPD antigen. The prevalence of anti-PPD antibodies in thoracic tumor patients was therefore significantly greater than that amongst the healthy controls (p 0.001, chi-square test). However, no significant difference in the prevalence of anti-LAM antibodies was found between study subjects and controls. This study demonstrates that thoracic tumor patients have significantly elevated antibodies to PPD; therefore, high anti-PPD seroreactivity in thoracic tumor patients should be cautiously interpreted. A longitudinal investigation on seropositive thoracic tumor patients is required to determine the role of the serological test for TB in lung cancer patients.
Adult ; Aged ; Aged, 80 and over ; Antibodies, Bacterial/*analysis ; Biopsy, Needle ; Female ; Human ; Lipopolysaccharides/*immunology ; Lung Neoplasms/complications/diagnosis/*microbiology ; Male ; Middle Age ; Mycobacterium tuberculosis/*immunology ; Seroepidemiologic Studies ; Tuberculin/*immunology ; Tuberculosis, Pulmonary/complications/diagnosis

This study was performed to characterize the humoral immune responses with isotype profiles in Mycobacterium tuberculosis infection. PPD-Specific IgG and IgG subclasses were measured using ELISA in 212 patients with pulmonary tuberculosis. The values of PPD-specific IgG were significantly higher in pulmonary tuberculosis patients than those in the control group, and were correlated to the severity of illness (P < 0.01). The specificity and sensitivity of ELISA for IgG antibodies were 1.0 and 0.81, respectively as determined in 212 sera from tuberculosis patients and 44 from healthy controls. The positive predictive value was 1.0 (171/171), while negative predictive value was 0.52 (44/85). The values of PPD-specific IgG were significantly decreased after 2-4 months of treatment. Among the moderately and far advanced pulmonary tuberculosis patients, the values of PPD-specific IgG were significantly decreased in responders after 6 months of treatment. However, PPD-specific IgG in nonresponders was increased (P < 0.01). PPD-specific IgG subclass responses were evident to all four IgG subclasses. No changes of isotype response according to the severity of the disease were observed.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin G/*blood/classification ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Tuberculin/*immunology ; Tuberculosis, Pulmonary/*immunology

OBJECTIVETo explore the characteristics of cellular immunity in drug abusers with pulmonary tuberculosis.

METHODSSixty drug abusers with pulmonary tuberculosis and 60 non-drug abusers with pulmonary tuberculosis (control) were enrolled in this study. Three days after establishment of a definite diagnosis, peripheral blood was taken from the patients for lymphocyte subgroup (CD3(+), CD3(+)/CD4(+), CD3(+)/CD8(+) T lymphocyte subgroups and NK cells) examination by flow cytometry, and the CD4(+)/CD8+(+) ratio was calculated. The difference of cellular immunity between the drug abusers and control group was analyzed statistically.

RESULTSCD3(+) and CD3(+)/CD4(+) T lymphocytes subgroups and NK cells of the drug abusers were significantly lower than those of the control patients (P=0.037, 0.028 and 0.015), and the former patients had also significantly lower CD4(+)/CD8(+) ratio (P=0.021). The pulmonary tuberculosis types and CD3(+)/CD8(+) T lymphocyte subgroup were not significant different between the two groups (P=0.053 and 0.85).

CONCLUSIONDrug abuse might depress cellular immunity in patients with pulmonary tuberculosis, which further complicate the treatment of this disease.

Adolescent ; Adult ; CD3 Complex ; metabolism ; CD4 Antigens ; metabolism ; Female ; Humans ; Immunity, Cellular ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Substance-Related Disorders ; complications ; immunology ; T-Lymphocytes ; immunology ; metabolism ; Tuberculosis, Pulmonary ; complications ; immunology ; prevention & control ; therapy ; Young Adult

OBJECTIVEThe protection rate of inoculation with BCG vaccine is only 50 percent, and most of patients with tuberculosis had a history of BCG vaccine inoculation. Adenosine (ADO) has an immunomodulating effect; it promotes immune reaction by increasing number of macrophage and enhancing phagocytosis. The present study was designed to investigate if combined use of adenosine with BCG enhances the anti-Mycobacterium tuberculosis effect of macrophage in mice.

METHODSFifty BALB/C mice were divided randomly into 3 groups: BCG group (n = 21), BCG plus ADO group (n = 21) and control group (n = 8). The mice in BCG and BCG plus ADO groups were inoculated with 0.1 ml BCG intradermally and the mice in BCG plus ADO group were injected intraperitoneally with ADO 30 mg/(kg.d) for 5 days. The mice in BCG group and control group were injected with NS 0.1 ml/d for 5 days. Six weeks after the last injection, all mice were challenged with intravenous 1 x 10(6) CFU human Mycobacterium tuberculosis virulent strain. After challenging, lung and spleen specimens were taken at the 10th, 20th and 30th days from the mice of BCG and BCG plus ADO groups and at the 30th day from mice in control group. The pathological examinations of lung and spleen sections were performed after HE staining and acid-fast staining, and detection of cell apoptosis was also performed.

RESULTSConsolidation with neutrophil infiltration was found in most of the lung tissue taken at the day 30; there were a lot of tuberculous granulomas and Mycobacterium tuberculosis in the lungs of control group. The alveolar septum in BCG gradually became wide and in interstitium lymphocyte infiltration dominated, and there were less tuberculous granulomas but there were large number of Mycobacterium tuberculosis in the lungs from 10th to 30th days after challenging. The widening of alveolar septum and consolidation of lung tissue in BCG plus ADO group became milder with monocytes infiltration, and there were few tuberculosis granulomas and Mycobacterium tuberculosis in the lungs from 10th to 30th days after challenging.

CONCLUSIONADO could increase the number of monocyte-macrophages and promoted anti-bacterial effects of these cells.

Adenosine ; administration & dosage ; immunology ; Animals ; BCG Vaccine ; administration & dosage ; immunology ; Disease Models, Animal ; Drug Therapy, Combination ; Injections, Intradermal ; Injections, Intraperitoneal ; Macrophages ; drug effects ; immunology ; Mice ; Mice, Inbred BALB C ; Mycobacterium tuberculosis ; immunology ; Neutrophil Infiltration ; drug effects ; Phagocytosis ; drug effects ; Tuberculosis, Pulmonary ; immunology ; prevention & control

BACKGROUNDFactors of cell-mediated immunity and allergy together play their roles in the pathogenesis of pulmonary tuberculosis (PTB) and its prognosis. The purpose of this study was to investigate the computed tomographic demonstrations of HIV seropositive PTB and the relationship between its pathogenesis and CD4(+) T-lymphocyte count.

METHODSThe documented CT images of a total of 44 patients with HIV seropositive PTB, definitely diagnosed by etiological or pathological examinations, their clinical data and their CD4(+) T-lymphocyte count were retrospectively reviewed.

RESULTSThere were 15 cases of miliary tuberculosis, accounting for 34.1% of the total cases; 15 cases of nodular tuberculosis, 34.1%; 6 cases of ground-glass opacity, 13.6%; 5 cases of cord-liked fiber shadows, 11.4%; 16 cases of flaky and flocculating shadows, 36.4%; 5 cases of cavitation, 11.4%; 5 cases of tumor shadows, 11.4%; 2 cases of pleural thickening, 4.5% and 11 cases of pleural effusion, 25.0%; 1 case of calcification, 2.3%; 16 cases of lymphadenectasis, 36.4%. The foci were located around the pulmonary hilum, anterior segment of superior lobe, basal segment of inferior lobe, medial lobe and lingual lobe. CD4(+) T-lymphocyte count was closely related to the imaging demonstrations of HIV seropositive PTB.

CONCLUSIONSCT scanning can demonstrate various signs of PTB. CD4(+) T-lymphocyte level determines the variety of imaging demonstrations of HIV seropositive PTB and its prognosis.

Adolescent ; Adult ; Aged ; CD4 Lymphocyte Count ; CD4-Positive T-Lymphocytes ; immunology ; Child ; Female ; HIV Seropositivity ; diagnostic imaging ; immunology ; Humans ; Male ; Middle Aged ; Pleural Effusion ; Radiography ; Tuberculosis, Pulmonary ; diagnostic imaging ; immunology ; Young Adult