Flow cytometry applications in the study of immunological lung disorders.

The use of flow cytometry in the clinical laboratory has grown substantially in the past decade. Flow cytometric analysis provides a rapid qualitative and quantitative description of multiple characteristics of individual cells. For example, it is possible to detect the cell size and granularity, aspects of DNA and RNA content and the presence of cell surface and nuclear markers which are used to characterize the phenotype of single cells. Flow cytometry has been used for the immunophenotyping of a variety of specimens including whole blood, bone marrow, serous cavity fluids, (cerebrospinal fluid) CSF, urine and all types of body fluids. The technique has also been applied to human bronchoalveolar lavage (BAL) fluid, peritoneal fluids and blood. In this review, we describe the current status of the application of flow cytometry as a diagnostic tool in various lung diseases. We focus on the analysis of BAL cell composition in chronic obstructive lung disease (COPD), asthma, lung cancer, sarcoidosis, tuberculosis and idiopathic eosinophilic pneumonia (IEP).

Elevated CXCL-8 expression in bronchoalveolar lavage correlates with disease severity in patients with acute respiratory distress syndrome resulting from tuberculosis.

BACKGROUND: Tuberculosis (TB) is a rare but known cause of acute respiratory distress syndrome (ARDS). The role of inflammatory cytokines in the progression of ARDS in TB patients is unknown. OBJECTIVES: In this study we investigated the possible link between the levels of inflammatory cytokines in bronchoalveolar lavage (BAL) in patients with TB or ARDS alone or in patients with TB-induced ARDS (ARDS + TB). METHODS: 90 patients were studied: 30 with TB alone, 30 with ARDS alone and 30 with ARDS + TB. BAL was collected by fiberoptic bronchoscopy and the concentrations of interleukin(IL)-6, CXCL8, TNF-α and IL-1ß and the amounts of total protein were measured by ELISA and bicinchoninic acid assay (BCA) methods respectively. The correlation between disease severity measured by Murray scores, SOFA and APACHE II analysis and BAL mediators and cells was also determined. RESULTS: CXCL8 levels in BAL were significantly higher in the ARDS + TB group compared to TB and ARDS alone groups. Disease severity in the ARDS + TB group as determined by Murray score correlated with BAL CXCL8 and neutrophils but not with IL-6, IL-1ß and TNF-α concentrations. In addition, CXCL8 levels and neutrophils were increased in non-miliary TB versus miliary TB. This difference in CXCL8 was lost in the presence of ARDS. CONCLUSIONS: BAL CXCL8 levels were significantly higher in patients with ARDS induced by TB and could suggest an important role of CXCL8 in the pathogenesis of this form of ARDS. This further suggests that CXCL8 inhibitors or blockers may be useful to control the onset and/or development of these combined diseases.

Allergic and nonallergic asthma in children: are they distinct phenotypes?

The aim of current study is to describe clinical similarities and differences between atopic and non-atopic asthma in children. In a cross-sectional study, 95 asthmatic children (75 allergics and 20 nonallergics) were included in the study. Demographic, clinical, and familial history were compared between two groups. There was no significant differences between variables like sex, age of onset (p=0.75), severity (p=0.70), and family history among the two groups (p=0.42). Patients with allergic asthma were significantly older than those with non- allergic asthma (11.28 ± 3.19 and 9.75 ± 2.35 years, respectively, p=0.02). The controversy lingers over the presence of a completely distinct phenotype of non-atopic asthma in children. Our study suggested that phenotypes of allergic and non-allergic asthma in children were not entirely distinct.

Comparison of spermatozoa quality in male smokers and nonsmokers of Iranian infertile couples.

BACKGROUND: Smoking has a negative effect on fertility and sperm quality. This study was designed to evaluate the effects of smoking on sperm quality and the related parameters such as sperm concentration, morphology and motility. MATERIALS AND METHODS: This cross-sectional study was conducted on 180 infertile men with at least one year history of idiopathic infertility, who admitted to the Avicenna Infertility Center, Tehran, Iran. A complete history including smoking habits and other diseases was obtained and semen analysis was performed for all participants. Statistical analysis was performed using SPSS software version 16 and t test and Mann-whitney tests with a significance level of α= 0.05. RESULTS: Comparison of sperm parameters in the two groups of smoker and nonsmoker subjects showed that active smoking (p=0.04) and cigarette consumption even in small amounts (p=0.03) decreased sperm concentration, However, no significant correlation was detected between smoking status and morphology or motility of sperms. CONCLUSION: This study failed to find a significant correlation between sperm analysis and smoking status except for sperm concentration, which was significantly decreased in the active smokers ,even in those consuming small amounts of tobacco. This finding propounds that tobacco consumption may negatively affect fertility.

Extensively drug-resistant tuberculosis: 2 years of surveillance in Iran.

BACKGROUND: Extensively drug-resistant (XDR) tuberculosis (TB) is a cause of concern, because it renders patients untreatable with available drugs. In this study, we documented the existence and transmission of XDR TB among patients with multidrug-resistant TB. These patients were referred to the National Research Institute of Tuberculosis and Lung Diseases (Tehran, Iran) for treatment and diagnosis from 2003 to 2005. METHODS: The sputum specimens from a total of 2030 patients with TB were digested, examined microscopically for acid-fast bacilli, and inoculated into Lowenstein-Jensen slants by standard procedures. Testing of susceptibility to first-line drugs was performed for 1284 Mycobacterium tuberculosis isolates. Subsequently, the strains that were identified as multidrug-resistant M. tuberculosis (113 isolates) were subjected to susceptibility testing for second-line drugs. Spoligotyping and restriction fragment-length polymorphism were performed for strains that were identified as XDR M. tuberculosis. RESULTS: A total of 12 (10.9%) of 113 multidrug-resistant M. tuberculosis strains were resistant to all 8 second-line drugs tested and, therefore, were denoted as XDR M. tuberculosis. Retrospective analysis of the cases of XDR TB showed that all of them belonged to 1 of 2 epidemiological clusters, either a single-family cluster (4 cases) or a cluster of close contacts (8 cases). The strains were identified as belonging to the M. tuberculosis superfamilies Haarlem 1 and East African Indian 3. CONCLUSIONS: The emergence of XDR TB cases in Iran highlights the need to reinforce the Iranian TB policy with regard to control and detection strategies.

Prevalence of Haarlem I and Beijing types of Mycobacterium tuberculosis strains in Iranian and Afghan MDR-TB patients.

OBJECTIVES: This survey identified the spoligopatterns of Mycobacterium tuberculosis strains with an international designation responsible for transmission and prevalence of Multi-Drug Resistance Tuberculosis (MDR-TB) among native and immigrant population of Tehran (2000-2005). METHODS: The spacer oligonucleotides typing was performed on 263 M. tuberculosis strains isolated from verified cases of MDR-TB. Clinical and demographical data of patients were collected using traditional methods. RESULTS: Classical epidemiological investigation revealed that out of 263 MDR-TB cases, 175, 66.5% were isolated from Afghan immigrants. In both communities, majority of MDR-TB cases had either previous history of TB (107, 40.6%) or had a close contact (84, 31.9%). By spoligotyping, 27 distinct patterns were observed, 253 clinical isolates were grouped in 17 clusters (62.9%) and 10 isolates displayed an orphan pattern (37%). Based on an international spoligotype database, Haarlem I (85, 33.5%), Beijing (52, 20.5%), Central Asia (32, 12.1%), and EAI (21, 8.3%) were the major identified super families. Although, 76.9% of the Beijing genotypes and 100% of ST(253) strains (that was prevalent through former Soviet Union) were isolated from Afghan patients only. The linkage patterns between 30 Iranian and Afghan patients were observed. CONCLUSION: The study highlighted the epidemic potential of Haarlem I and Beijing genotypes among MDR-TB cases in Tehran territory.

Evaluation of tuberculosis transmission in Tehran: using RFLP and spoligotyping methods.

OBJECTIVES: To evaluate the DNA polymorphism among Mycobacterium tuberculosis (MTB) strains isolated from new smear positive tuberculosis (TB) patients residing in Tehran capital city of Iran, during the year 2001. METHODS: IS6110-restriction fragment length polymorphism (RFLP) and spoligotyping analyses were performed on 129 M. tuberculosis strains. Additional patient's information was collected for further epidemiological analyses. Patients whose isolates had identical RFLP and spoligotyping patterns were considered a cluster. RESULTS: The results show that the IS6110 were polymorphic and the strains with 8 or 9 IS6110 copy number were more frequently defected (42%). Out of 129 available isolates, 56 (43%) belonged to clusters and 72 (57%) did not. The risk factors like age, sex, family history or close contact and intravenous drug abuse were associated with clustering. Whereas, unemployment (61%) and poor living conditions (83%) contributed to diseases development in both groups. Spoligotyping of M. tuberculosis strains resulted in 46 different patterns, out of which 38 patterns were unique and reported for the first time. We found one M. tuberculosis strains with a pattern characteristic of the Beijing family. CONCLUSION: In the studied time period both reactivation (57%) and recent transmission (43%) were contributing to annual new TB cases in Tehran.