SLC11A1 genetic variation and low expression may cause immune response impairment in TB patients.

Tuberculosis (TB) is caused by Mycobacterium tuberculosis. Host genetic factors are important for the detection of TB susceptibility. SLC11A1 is located in monocyte phagolysosomes that help to limit M. tuberculosis growth by transferring divalent cations across the membrane. Genetic variation in SLC11A1 may alter its expression and increase the susceptibility of individuals to TB. The current study aimed to provide insight into host genetic variations and gene expression in TB patients. A total of 164 TB patients and 85 healthy controls were enrolled in this study. SLC11A1 polymorphisms were detected by PCR-RFLP. Real-time qPCR was used for SLC11A1 gene expression, and ELISA was used for protein estimation. GTEx Portal was used for quantitative trait loci analysis, while the STRING (v.11) web platform was used for gene interactive network construction. Data were analyzed using SPSS, GraphPad Prism, Haploview, and SNPstats. SLC11A1 polymorphisms and combinatorial genotypes were strongly associated with TB susceptibility, which may explain the greater prevalence of tuberculosis in the local population. Polymorphisms in SLC11A1 have also been linked to gene expression variation. Furthermore, the expression of SLC11A1 was downregulated in TB patients, which may influence the function of other associated genes and may impair the immunological response to tuberculosis.

Immune mechanisms in type-2 diabetic retinopathy.

OBJECTIVE: To enumerate CDR+CD25+ Treg cells and determine serum IL-6 and IL-17 in type 2 diabetes mellitus patients with retinopathy. METHODS: The case-control study was conducted at the Department of Immunology, University of Health Sciences, Lahore, from November 2009 to January 2012 and comprised diabetic patients and healthy controls who were divided into three groups. Group 1 had controls, while Group 2 had diabetic patients without retinopathy and Group 3 had diabetic patients with retinopathy. Flowcytometre and enzyme-linked immunosorbent assay were used for CD4+CD25+ Tregs and serum IL-6 and IL-17 respectively. SPSS 20 was used for statistical analysis. RESULTS: Of the 212 subjects in the study, 30(14%) were Group 1, 30(14%) in Group 2 and 152(72%) in Group 3.There were 25 (83%) women in Group 2 and 101 (66%) in Group 3 compared to 9 (30%) in Group 1. Higher mean age was in Group 3 (50.88 ± 8.9 years) and Group 2 (49.46 ± 9.94 years) compared to Group 1 (34.66 ± 8.78 years) while longer mean disease duration was in Group 3 (10.51 ± 5.24 years) than Group 2 (7.76 ± 4.14 years). Highest median ratio of IL- 6 was in Group 1 (1468.62) (Q1-Q3: 1229.9-1543.35), followed by Group 2 (1455.32) (Q1-Q3:1214.22-158.9) and Group 3 (469.84) (Q1-Q3: 206.53-1231.33) whereas IL-17 was the highest in Group 1 (339.38) (QT-Q3: 159.89- 1174.93), followed by Group 3 (216.60) (Q1-Q3:141.87-410.25) and Group 2 (174.17) (Q1-Q3: 138.77-458.17). Higher percentage of Tregs was in Group 2 (3.07 ± 0.43) followed by Group 1 (2.91 ± 0.04) and Group 3 (2.88 ± 0.38). Significant difference was observed in gender, age, disease duration, level of IL-6 and IL-17 (p < 0.05 each), while no difference was found in glycated haemoglobin, CD4+CD25+ and Tregs (p > 0.05 each). CONCLUSION: Age, gender and duration of diabetes contributed to diabetic retinopathy, while CD4+CD25+ T cells and Treg cells did not. Serum IL-6 and IL-17 were inversely associated with diabetic retinopathy.

Enumeration of CD4(+)CD25(+)T regulatory cells in Type-II diabetes retinopathy.

Diabetes mellitus (DM) is a health concern because it leads to complications such as retinopathy. Pakistan has 6.9 million DM affected people that will be doubled by 2025. A study was designed to enumerate CD4(+)CD25(+)Treg cells in Pakistani type 2 diabetes mellitus (T2DM) patients. It was a cross-sectional case-control study that included 212 subjects. The subjects having diabetic retinopathy were labeled as Group-I (30 healthy volunteers without diabetes), Group-II (30 T2DM patients without retinopathy) and Group-III (152 T2DM patients with retinopathy). The percentage of CD4+CD25+ Treg cells was determined by Flowcytometry. Comparison of CD4(+)CD25(+)T cells among different groups was not significant and higher percentage of Treg cells was observed in Group-II (3.07%) compared to Group-III (2.88%). Age, gender and duration of diabetes may contribute while percentage of CD4(+)CD25(+)T cells and Treg cells were not associated with the development of DR in T2DM.

Inverse relationship of serum IL-17 with type-II diabetes retinopathy.

BACKGROUND: Diabetes mellitus (DM) is a health concern because it leads to complications such as retinopathy. Pakistan has 6.9 million DM affected people that will double by 2025. A study was designed to determine the level of IL-17 in the serum of Pakistani type 2 diabetes mellitus (T2DM) patients. METHODS: It was a cross-sectional case-control study that included 212 subjects. Subjects without diabetes were labeled as Group-I (30 healthy volunteers), Group-II (30 T2DM without retinopathy), and Group-III (152 T2DM with retinopathy). The serum level of IL-17 was determined by ELISA technique. Data was analysed using SPSS 17.0 and one way ANOVA to observe group mean differences. RESULTS: More females were in Group-II (83%) and Group-III (66%) compared to Group-I (30%). The age of subjects was higher in Group-III (50 years) and Group-II (49 years) compared to Group-I (34 years). Group-III had longer mean duration of disease (10.51 years) than Group-II (7.76 years). Group-I had increased levels of IL-17 followed by Group-II and Group-III. On comparison, statistically significant differences were observed among the three groups, and between Group-I and Group-III, but there was no significant difference between Group-I and Group-II, nor between Group-II and Group-III. Further, on comparison of age, gender, and duration of disease there were significant differences while there was no significant difference between the percentages of HbA1c. CONCLUSIONS: Age, gender, and duration of diabetes may contribute in the development of T2DM retinopathy while serum level of IL-17 was inversely associated with T2DM and retinopathy.

Decreased classical monocytes and CD163 expression in TB patients: an indicator of drug resistance.

Tuberculosis (TB) is a disease instigated by Mycobacterium tuberculosis. Peripheral blood monocytes represent highly efficient effector cells of innate immunity against TB. Little is known about monocyte subsets and their potential involvement in the development of M. tuberculosis drug resistance in patients with TB. This study was conducted to investigate alterations in monocyte subsets, CD163 expression on monocytes, and its serum level in patients without and with rifampicin resistance TB (RR-TB) and healthy controls. A total of 164 patients with TB (84 without RR-TB and 80 patients with RR-TB) and 85 healthy controls were enrolled in this study. The percentages of various monocyte subsets and surface expression of CD163 on monocytes were quantitatively determined using flow cytometry. The serum level of CD163 was determined by commercially available ELISA kits. Decreased frequency of classical monocytes was detected in patients with RR-TB. Non-classical monocytes were decreased in patients without RR-TB; however, intermediate monocytes were raised in patients with RR-TB. The serum level of CD163 was decreased in patients of RR-TB that showsed a positive correlation with the frequency of CD14++CD16-CD163+ and CD14++CD16+CD163+ monocytes. It is concluded that decreased classical monocytes and sCD163 in patients with RR-TB could be an indicator of drug resistance.

Frequency of HLA DQß1*0201 and DQß1*0301 Alleles and Total Serum IgE in Patients with Bronchial Asthma: A Pilot Study from Pakistan.

In Pakistan about 3.7% of the population is suffering from asthma, a chronic inflammatory disorder of airways. Asthma has wide spectrum of predisposing factors including environment and genetics. Many studies have been performed to determine association of asthma with serum IgE and major histocompatibility complex (MHC) alleles but conflicting results were reported. Therefore, present study was designed to determine frequency of HLA-DQß1*0201 and DQß1*0301 alleles in patients with bronchial asthma. This case control study included 85 asthmatic patients and 85 healthy controls. HLA-DQß1*0201 and DQß1*0301 alleles were detected by allele specific PCR and serum IgE was determined by ELISA. Median and inter-quartile range (IQR) of total IgE level were more increased in asthma patients (585.7 IU/mL and 247.2-848.1 IU/mL) compared to healthy controls (65.1 IU/mL and 28.1-181.3 IU/mL) (p<0.001). Frequency of HLA-DQß1*0201 and -DQß1*0301 alleles was more in healthy controls (32% and 38%, p=0.616) as compared to bronchial asthma patients (28% and 26%, p= 0.09). There was a significant association of IgE levels and HLA-DQß1*0201 allele. Patients positive for HLA-DQß1*0201 allele had low level of serum IgE 357.2 IU/mL (153.9-634.3 IU/mL) compared to the patients negative for this HLA allele i.e. 642.9 IU/mL (289.8-1299.5IU/mL) (p=0.005), whereas, HLA-DQß1*0301 allele was not associated with total serum IgE level (p=0.865). Our findings show that HLA-DQß1*0201 and -DQß1*0301 alleles were not associated with asthma; however, HLA-DQß1*0201 allele was associated with low levels of total serum IgE in the study population.

Level of interferon gamma in the blood of tuberculosis patients.

BACKGROUND: Interferon gamma (IFN-γ), a cytokine produced by a variety of cells is involved in the immune response against M. tuberculosis. It activates the production of other cytokines and molecules that kill mycobacterium. IFN-γ also has diagnostic role in identification of active and latent tuberculosis. OBJECTIVE: To determine the level of IFN-γ in the blood of TB patients. METHODS: Ninety-one subjects were selected, including 54 active TB patients and 37 healthy controls. Among 54 TB patients, 27 had confirmed TB and 27 were. Clinically diagnosed as having TB. IFN-γ concentration was determined in their blood by an ELISA technique. RESULTS: In TB patients, Mean + SD of IFN-γ was 48.69 + 28.78 pg/ml while it was 12.99 + 5.70 pg/ml in the control group (p <0.001). Significant differences in the level of IFN-γ were observed among confirmed TB patients, clinically diagnosed TB patients and the control group (Mean + SD 59.68 + 28.78, 36.85 + 24.76 and 12.99 + 5.70 pg/ml, respectively). Furthermore, a significant negative correlation was observed between the concentration of IFN-γ in TB patients and the duration of anti-tuberculosis therapy. CONCLUSION: IFN-γ level was high in both clinically diagnosed and confirmed TB patients as compared to a control group. Measurement of IFN-γ production is helpful to diagnose active tuberculosis, but further research is required.