Immune mechanisms in type-2 diabetic retinopathy

To enumerate CD4+CD25+ Treg cells and determine serum IL-6 and IL-17 in type 2 diabetes mellitus patients with retinopathy. 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. 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] [Q1-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]. 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 objects

The subjects having diabetic retinopathy were labeled as Group-I [30 healthy volunteers without diabetes], Group-II [30 T2DM patients without retinopathy] and Group-Ill [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-Ill [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

Negative association of serum IL-6 and IL-17 with type-II diabetes retinopathy

Diabetes mellitus [DM] is a health concern which leads to complications such as retinopathy. Pakistan has 6.9 million people living with DM and this toll will be doubled by 2025. To determine serum IL-6 and IL-17 of type 2 diabetes mellitus [T2DM] patients with retinopathy. In this cross-sectional case- control study, 212 subjects enrolled which were categorized into 3 groups. Group-I included 30 subjects without diabetes, group-II consisted of 30 subjects with T2DM without retinopathy and group-III consisted of 152 subjects with T2DM and retinopathy. Serum IL-6 and IL-17 levels were determined by ELISA. Data was analysed using SPSS 17.0 and one way ANOVA to observe group mean differences. Longer mean duration of disease was detected in group-III than group-II [p=0.007]. Highest IL-6 level was detected in group-II and highest IL-17 level was detected in group-I. For IL-6, significant differences were detected among groups in total, between Group-I and Group-III and between Group-II and Group-III [p<0.0001 each]. Regarding IL-17, significant differences were found among groups in total [p=0.002] and between Group-I and Group-III [p=0.001]. No significant difference in the percentages of HbA1c observed between groups. Age, gender and duration of diabetes contribute to T2DM retinopathy. Serum IL-6 and IL-17 were inversely associated with T2DM retinopathy

Effect of hepatitis C virus on C-reactive protein and interleukin-6 in hemodialysis patients

Patients with end-stage renal disease are at a high risk of hepatitis C virus [HCV] infections. These patients are on maintenance hemodialysis where they are exposed to dialysis fluid and dialysis membrane that generate an acute-phase response, which leads to inflammation, reflected in increased inflammatory markers like C-reactive protein [CRP] and interleukin-6 [IL-6]. The aim of this study was to investigate levels of IL-6 and CRP in patients on hemodialysis and to determine effects of HCV on these markers. A total of 43 patients [39.5% men and 60.5% women; age range, 21 to 65 years] on maintenance hemodialysis for a period of at least 3 months were included. Twenty-four of them were HCV positive. Serum IL-6 and CRP were assessed in all patients. Of HCV-positive patients, 11 [45.8%] had high levels of CRP, while 13 [54.2%] had low levels and increased levels of IL-6 [1064 +/- 544.2 pg/mL, P=.19]. Of 19 HCV-negative patients, 13 [68.4%] had high levels of CRP, while 6 [31.6%] had low levels, and all these patients had low levels of IL-6, as compared to HCVpositive patients. No significant correlation was observed between the levels of CRP or IL-6 and the duration of dialysis. In our study, we found high serum IL-6 and CRP levels in HCV-positive hemodialysis patients, compared with HCVnegative ones. However, we failed to show the significance of these differences. More studies with large sample sizes and evaluation of the other inflammatory markers are warranted

Quantification of serum IgA of coeliac disease patients

Celiac disease [CD] is a gluten - induced multi - organ disorder where small intestine is the primary target of inflammation. Onset of CD may occur at any age and its symptoms vary among individuals. Definitive diagnosis of CD is by intestinal biopsy but determination of anti - IgA tissue transglutaminase [tTG] and anti-gliadin antibodies has become key factors to decide for tissue biopsy. IgA - deficient CD patients may yield false - negative results, therefore total serum IgA level must be determined along with other serological markers to diagnose CD. The study included 42 CD patients who were positive for anti-tTg antibodies [Group A] and 40 subjects [Group B: disease control] presented with gastrointestinal complaints but were negative for anti-tTG antibodies. On the basis of age, Group A was further divided into: Sub-group- I comprised of patients between 1-6 years [n = 31] and Sub-group - II consisted of patients between 7-15 years [n=11]. Level of serum IgA was determined by nephlometry technique. Total serum IgA level was 38.77 +/- 31.21 mg/dl and 26.88 +/- 28.27 mg/ dl in CD patients and disease control group respectively and the difference in the level of serum IgA between these groups was not statistically significant [p=0.75]. Mean IgA level in sub-group -I and sub-group- II was 40.85 +/- 33.29 mg/dl and 32.92 +/- 24.85 mg/dl respectively and the difference in the level of serum IgA between these sub-groups was not statistically significant [p = 0.47]. In Group - A, mean level of IgA in males and females was 42.38 +/- 38.02 mg/dl and 34.41 +/- 20.36 mg/dl respectively and the difference in the level of IgA level was not statistically significant between these groups [p = 0.41]. Selective IgA deficiency [SIgAD] was found in CD and in patients of other gastrointestinal complaints. In order to detect CD in SIgAD, total serum IgA level should also be performed with IgG - antigliadin or IgG-anti-tTG antibodies

Detection of anti-Helicobacter pylori IgG antibodies in healthy subjects

Helicobacter pylori [H. pylori] are Gram - negative microaerophilic, spiral organisms. Factors such as family history of gastric disease, source of drinking water, number of siblings, sharing beds, and level of hygiene have been linked to acquisition of H. pylori infection. Most of the infected people do not have clinical symptoms. The study was planed to determine the level of anti-H. pylori IgG antibodies in the serum of healthy individuals. The study included 80 healthy subjects and was conducted in the Department of Immunology, University of Health Sciences Lahore. The studied population was divided on the basis of [a] eating food from outside home daily, twice a week or once a week, [b] using filtered or tap water for drinking, and [c] having family history of gastric ulcer or without family history of gastric ulcer. Level of anti H.pylori IgG antibody was determined by ELISA technique. Among 80 asymptomatic healthy individuals anti-H pylori IgG antibody was detected in 28 [35% subjects who did not have these antibodies [p-value < 0.001]. Mean level of anti-H. pylori IgG antibodies was 43 +/- 39.3 U/ml, 30.7 +/- 37.3 U/ml and 14.9 +/- 19.7 U/ml in subjects eating food from outside their homes once a week, twice a week and daily respectively. Statistically significant difference was observed in the level of H. pylori antibodies with different eating habits [p = 0.015]. However no statistically significant difference was observed in the level of anti-H. pylori antibodies between two genders, individuals using tap water and filtered water for drinking and with family history of gastric ulcer

Frequency of antineurtophil cytoplasmic antibody in glomerulonephritis

Aim of the study was to determine the frequency of antineutrophil cytoplasmic antibody [ANCA] [p-ANCA and c-ANCA] in clinically diagnosed glomerulonephritis. Autoimmune diseases including systemic vasculitis, affect a large number of people ill whom the leading cause of morbidity and mortality is glomerulonephritis that is often associated with chronic kidney disease. There are many risk factors for kidney diseases such as chronic inflammation, auto-immune diseases, immunosuppressive therapy, etc. Early phases of renal injury in autoimmune patients are clinically silent. For the detection of nephron damage, histopathological examination is gold standard but detection of antineutrophil cytoplasmic antibody [ANCA] can be used to find out early nephron damage. Design was analytical Cross-sectional. The study was conducted at the Department of Immunology, University of Health Sciences, Lahore in a period of November 2008 to October 2009. Study included 64 clinically diagnosed of glomerulonephritis. Levels of ANCA [MPO and PR3] were determined by ELISA technique. Out of which four [6.25%] patients showed positive reaction to myeloperoxidase [MPO] antigen while1 [1.56%] patient was positive for proteinase-3 [PR3] antigen. In 40-60 years of patients, sero-positivity for MPO and PR3 was 14% and 3.6% respectively, p-value for MPO and PR3 was <0.05 and >0.05 respectively. We concluded that glomerulonephritis is better related with MPO-ANCA than PR3-ANCA. The difference in the levels of MPO-ANCA in different age groups was significant but it was non-significant among different genders. Difference in the levels of PR3-ANCA was not significant for both age and gender

Evaluation of four ELISA based immunoassays for the detection of IgM antibodies against dengue virus

Dengue fever is a problem of serious concern which is mainly transmitted by mosquito, Aedes aegypti. These days' different ELISA kits are used for the diagnosis of dengue fever. It was a validation study. Study was conducted in the Department of Immunology at University of Health Sciences, Lahore during the period of August 2009 to October 2009. In this study four ELISA kits [Human, Nova Tech, Vircell and DRG] were used to determine IgM antibodies against dengue fever in forty four patients who were labelled positive for dengue fever by different commercially available ELISA kits. Human ELISA kit gave most accurate results with respect to the agreement, sensitivity, specificity and K value. Human ELISA kit was found most reliable for the diagnosis of dengue fever

Association of interleukin-4 and IgE levels with LDL oxidation in atherosclerosis

Atherosclerosis is an inflammatory and multifactorial disease, with a high prevalence rate in Pakistan. To find a relation between serum IL-4 and IgE levels with oxidized LDL in atherosclerosis. In this observational, cross sectional study 99 male patients, between forty and sixty years of age, with a history of ischemic heart disease [IHD] and established atherosclerotic plaques and angiography were recruited. The study was completed within three years [Jan 2007 to Jan 2009]. One hundred and on age and gender matched healthy subjects with no known history of IHD were also recruited. All the study participants were non-diabetics. Serum IL-4, IgE and oxidized LDL [ox-LDL] levels were measured by quantitative ELISA technique. Serum IL-4 levels were generally undetectable or very low, but were higher in the patient group compared to the control subjects. Similarly, oxidized LDL and serum IgE levels were also increased in the patient group compared to the control, but the differences were not statistically significant. Our study could not detect any relationship between IL-4 and IgE levels with LDL oxidation in atherosclerosis

Percentage of CD4[+] and CD8[+] T-lymphocytes in blood of tuberculosis patients

Tuberculosis is a fatal infectious disease, mainly caused by Mycobacterium tuberculosis. Spread of TB is controlled by cell-mediated immunity. Purpose of this study was to determine CD4[+] and CD8[+] T cell percentages in TB patients. 77 subjects consisted of 39 patients of active tuberculosis and 37 normal healthy individuals were recruited for the study. Among patients, 27 were at different stages of anti-tuberculous therapy while rests of the patients were not taking treatment. Sixteen patients were sputum positive for AFB while other patients were sputum negative for AFB. T cells percentages were determined by flow cytometer. In TB patients CD4[+] and CD8[+] T cells percentages were 34.4 +/- 9.8 and 32.0 +/- 9.8 while in controls these were 37.1 +/- 6.9 and 30.2 +/- 7.2 respectively but the difference was statistically insignificant. CD4[+] T cell percentage in newly diagnosed TB patients was 28.8 +/- 8.7 while it was 37.9 +/- 8.9 in TB patients who were on therapy and difference was statistically significant whereas difference in CD8[+] T-cell percentages was statistically insignificant. A negative correlation between CD8[+] Tcells percentage and the duration of ATT was found. CD4[+] and CD8[+] T-cells percentages may help to find out the immune status of TB patients before and after the completion of ATT

Level of interferon gamma in the blood of tuberculosis patients

Interferon gamma [IFN- gamma], 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- gamma also has diagnostic role in identification of active and latent tuberculosis. To determine the level of IFN- gamma in the blood of TB patients. 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- gamma concentration was determined in their blood by an ELISA technique. In TB patients, Mean +/- SD of IFN- gamma was 48.69 +/- 28.78 pg/ml while it was 12.99 +/- 5.70pg/ml in the control group [p <0.001]. Significant differences in the level of IFN- gamma 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- gamma in TB patients and the duration of anti-tuberculosis therapy. IFN- gamma level was high in both clinically diagnosed and confirmed TB patients as compared to a control group. Measurement of IFN- gamma production is helpful to diagnose active tuberculosis, but further research is required

Gated spect myocardial perfusion scintigraphy for identifying septal perfusion artifacts in left bundle branch block

To determine the role of gated Single Photon Emission Computed Tomography [SPECT] for accurate assessment of myocardial perfusion scintigraphy [MPS] of patients with left bundle branch block [LBBB]. Analytical study. Punjab Institute of Nuclear Medicine [PINUM], Faisalabad, Pakistan, from June 2002 to April 2003. MPS data of patients with LBBB was analyzed. Resting gated SPECT MPS was performed after an injection of 740 MBq 99mTc-MIBI in 10 normal and 25 subjects with LBBB [with low probability of coronary artery disease]. Visual and quantitative analyses were done on non-gated [NG], end diastolic [ED], end systolic [ES] images. Calculations included septal to lateral wall ratio [SLR], myocardial thickening [MT=% increase in counts during systole] at end systolic phase and myocardial thickening at peak level [% peak MT]. Septal hypoperfusion was noted in 19 [76%] patients on NG images and in only 1 [4%] patient on gated SPECT ED images. On NG images of LBBB group, SLR was lower than in controls [0.68 +/- 0.07 vs. 0.87 +/- 0.05, p<0.001]. SLR of LBBB patients approached to that of control group in gated SPECT ED data [0.86 +/- 0.06 vs 0.88 +/- 0.06, p=ns]. Myocardial thickening at ES for septum was markedly lower in LBBB group than in controls [21.83% ' 10.86 vs. 66.32% ' 20.15, p<0.001]. In patients with LBBB, reduced septal thickening results in artifactual septal perfusion defects. Gating the perfusion scintigraphy and reporting perfusion status on end diastolic frames in LBBB patients can eliminate these artifacts