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Background@#Enzyme-linked immunosorbent assay (ELISA) kit is used abundantly in order to measure the quantity of agents and their antibodies. Introducing the production of the kit in our own country for research and diagnostic uses will be beneficial for economy and time.@*Goal@#Producing enzyme-linked immunosorbent assay for the determination of IL-2 cytokine in biological fluids.@*Materials and Methods@#Elisa kits made by R&D systems in USA were used in our study. Anti-IL-2 monoclonal antibody was diluted with concentrations of 0.5-3μg/ml in polystyrene well, and then incubated with conjugate solution of pH 7.2-7.4 for 12 hours with the purpose of measuring the optimal starting concentration for the conjugate antibody. Later, the reaction was blocked with PBS blocking buffer with 1% BSA (Bovine serum albumin) for an hour. After washing the wells, 100ul of standard solutions with concentrations of 7.81-2000pg/ul were added and incubated for 60 or 120 minutes. Subsequently, secondary antibody and streptavidin-HRP were added as substrate, and the reaction was terminated with stop solution. Absorbance was measured immediately at 450 nm optical densities, and then regression equation was calculated using standard curve.@*Result@#The light absorbance was insufficient when then dilution of starting concentration for the conjugate antibody was 0.5μg/ml and the light absorbance decreased as the concentration of conjugate antibody reduced. The light absorbance increased when the concentration was 2-4μg/ml, but the standard curve was constructed nonlinear. When standard curve was constructed with 1 ug/ml, the regression equation had R2=0.96, which identifies the optimal concentration. The results were analogous when the incubation times were 60 and 120 minutes with the concentration of 4μg/ml.@*Conclusion@#The optimal condition for determining IL-2 cytokine was when ELISA conjugate concentration was 1μg/ml (sample and conjugate) and incubation time was 120 minutes, and the regression equation from the standard curve was y=0.0013x+0.1901, R<sup>2</sup>=0.96.
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IntroductionAlthough systemic lupus erythematosus (SLE) is a relatively common autoimmune disorder, thebroad range of associated clinical symptoms means that its diagnosis can be diffi cult. Anti-ds-DNAautoantibodies are also considered to play a pathogenic role in inducing renal symptoms in SLE, anda strong correlation has been seen in lupus nephritis (LN) between disease activity and anti-ds-DNAautoantibody levels.GoalThe aim of the study was to compare the prevalence and levels of autoantibodies in the serum ofpatients with Systemic lupus erythematosus.Materials and MethodsThis patient-based descriptive study involved 39 patients with LN and 74 controls with primaryglomerulonephritis (GN). Face to face interview was used to obtain necessary information followed bythe physical examination and autoantibodies (anti-ds-DNA, anti-SS-A/Ro, anti-SS-B/La, c/p-ANCA, anti-Sm) measured by Enzyme-Linked Immunosorbent Assay (Germany, ORGENTEC Diagnostic GmbH).ResultsThe prevalence of anti-ds-DNA 48.7%, anti-SS-A/Ro 56.4%, anti-Smith 38.5%, anti-SS-B/La 12.8%were positive in lupus nephritis group (secondary GN), the prevalence of anti-ds-DNA 6.76%, anti-SS-A/Ro 6.76%, anti-Smith 2.7%, anti-SS-B/La 2.7% were positive in primary GN group.Conclusions: SLE associated with several autoantibodies (anti-ds-DNA, anti-SS-A/Ro, anti-Smith,and anti-SS-B/La) and each of which are very useful in distinguishing patients with SLE from otherautoimmune diseases and GN.
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Kidney transplantation is the best alternative treatment for end-stage renal disease and health-related quality of life and survival of the patients are improved compared with dialysis. Worldwide, more than 1.4 million patients with CKD receive renal replacement therapy with incidence growing by approximately 8% annually.1 Unfortunately, despite significant improvement in graft function, kidney transplants can still fail due to acute rejection and chronic allograft nephropathy.2 Kidney biopsy after transplantation, which has evaluated by Banff 09 classification is usefull method for diagnose of transplanted kidney disease.3,4Kidney graft rejection was diagnosed in 10 renal allograft biopsy specimens (bs) obtained from transplant patients followed up at our institute between 2015 and 2016. All specimens were evaluated as satisfactory which show more than 8 glomerulus under the light microscopy. Each renal cortical tissue was divided into two tips: one piece for routine H&E stain and special stains, including Masson’s trichrome, and PAS stain; another piece for immunofluorescence by frozen section, which were stained with IgA, IgM, IgG and complement component (C3, C4, C1q, C4d). All the renal biopsies were examined by the same pathologist.Out of 117 transplantations, 10 episodes of rejection selected. Among the 10 patients, 30% had an acute T cell rejection and 70% had a chronic allograft nephropathy. Interstitial inflammation (i1-7) was present in 7 bs (70%), tubulitis (t1-4,t2-2) in 6 bs (60%), transplant glomerulitis (g1-1, g2-2, g3-1) in 4 bs (40%), transplant interstitial fibrosis (ci1-2, ci2-2, ci3-2) in 6 bs (60%), tubular atrophy (ct1-6, ct2-2, ct3-1) in 9 bs (90%), mesangial matrix increase (mm1-5) in 5 bs (50%), vascular fibrosis intimal thickeness (cv1-3) in 3 bs (30%), arteriolar hyaline thickening (ah1-5) in 5 bs (50%), tubulitis (ti1-6, ti2-3, ti3-1) in 10 bs (100%) and peritubular capillaritis (ptc1-1, ptc2-2, ptc3-1) in 4 bs (40%). C4d deposition was present very mild in wall of the vessels and peritubular capillaries. Because of not good working Methenamin silver stain, we couldn’t demostrate glomerular basement membrane changes (cg) fully.We suggest that histopathological changes of transplant glomerulopathy might be accompanied by inflammation of the microvasculature, such as transplant glomerulitis and peritubular capillaritis. C4d deposition in the wall of the vessels and peritubular capillaritis is not always present in biopsy specimens of transplant glomerulopathy.
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Background Systemic Lupus Erythematous (SLE) is a multi-systemic autoimmune disease with numerous patterns of clinical and immunological manifestations. Renal disease in SLE occurs in 40–75% of patients, most often within five years of disease onset, and is one of the strongest predictors of a poor outcome. Anti-dsDNA antibodies are reported to be more prevalent in patients with SLE who have renal disease. Anti-Sm, anti SSA and anti SSB antibodies are also considered to play a pathogenic role in inducing renal symptoms in SLE, and a strong correlation has been seen in lupus nephritis (LN) between disease activity and anti-dsDNA antibody levels. Objective The aim of our study is to highlight the clinical and laboratory features in SLE patients. Methods This is a three year hospital based case-control study of patients with renal diseases, who were admitted to the nephrology and rheumatology units of the 1st central Hospital and 3rd central hospital, Mongolia. Standard methods were used for laboratory testing. Autoantibodies (C/P-ANCA, anti-dsDNA, anti-Sm, anti-SS-A/Ro, anti-SS-B/La, anti-Scl-70, anti-GBM) measured by Enzyme Immuno Assay (Germany, ORGENTEC Diagnostika GmbH). Renal function was evaluated by the eGFR (estimated glomerular filtration rate) using the Cockcroft-Gault formula. Result The study included 27 patients with lupus nephritis and 78 controls with other types of GN. There were 85.2% of female patients in the lupus nephritis group. Patients with LN were significantly younger than the controls (mean (SD) 31.9 (10.1) years vs. 37.1 (11.9) years; p=0.036). For the serology, a higher proportion of anti dsDNA (46.1%), anti Sm (29.6%), anti SSA (63%) and anti SSB (11.1%) were seen in the group with lupus nephritis (p=0.001; p=0.043; p<0.0001; p=0.096, respectively). The Pearson’s correlation analysis indicated that the level of anti-dsDNA (r=-0.249, p=0.021) and anti SSA (r=-0.195, p=0.048) were significantly correlated with the renal function (eGFR). All had dipstick proteinuria 1+/2+/3+, more than 10 red blood cells/hpf hematuria (n-12, 44.4%) in lupus nephritis group and renal function (mean eGFR (SD) 88.1 (51.3) ml/min vs. 112.3 (67) ml/min; p=0.05) was more decreased in lupus nephritis patients than controls. Conclusion Notably, rising titers of antibodies to dsDNA, SSA may indicate exacerbations of glomerulonephritis.
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INTRODUCTION: In the modern medical practice of Mongolia, autoimmune diseases have notbeen diagnosed in its early stage. The autoantibodies are useful in the patient’s early diagnosis,prognosing, and treatment of autoimmune diseases.GOAL: The aim of the study was to compare the prevalence and levels of autoantibodies in theserum of patients with autoimmune diseases.MATERIALS AND METHODS: This patient-based descriptive study involved 144 participants, withconfirmed diagnosis of autoimmune disease and glomerulonephritis (GN). Face to face interviewwas used to obtain necessary information followed by the physical examination and autoantibodies(anti-SS-A/Ro, anti-SS-B/La, anti-SCL-70, c/p-ANCA, anti-GBM, anti-Sm) measured by Enzyme-Linked Immunosorbent Assay (ELISA).RESULTS: The prevalence of anti-SS-A/Ro 38.6%, anti-Sm 25.7%, anti-SS-B/La 8.6%, c-ANCA7.14%, anti-SCL-70 1.4% were positive in autoimmune disease group (secondary GN), theprevalence of anti-SS-A/Ro 6.8%, anti-Sm 2.7%, anti-SS-B/La 2.7%, c-ANCA 1.4% were positivein primary GN group. A higher frequency of anti-SS-A/Ro 66.4%, anti-SS-B/La 22.8%, and anti-Sm38.4% was observed in the SLE group.CONCLUSIONS: Patients with autoimmune disease was significantly higherly younger and female.SLE associated with several auto antibodies (anti-SS-A/Ro, anti-SS-B/La, and anti-Sm) and eachof which are very useful in distinguishing patients with SLE from other autoimmune diseases.