Hungarian Immunology

[Kinetic measurement on flow cytometer simultaneous monitoring of intracellular progresses]

MÉSZÁROS Gergő1, RÓNAI Katalin Zsuzsanna2, TOLDI Gergely1, KAPOSI Ambrus1, VÁSÁRHELYI Barna1, TRESZL András1

JANUARY 22, 2008

Hungarian Immunology - 2008;7(01-02)

[INTRODUCTION - Flow cytometry provides an opportunity for real-time monitoring of intracellular processes in several cell populations simultaneously. Cells stained with specific fluorescent dyes are sequentially measured during kinetic FACS measurements. Fluorescent light signals obtained in cells are recorded and analyzed to describe the alteration of the investigated parameter(s) over time. The use of kinetic FACS assays is not spread as there was no mathematic algorithm to characterize objectively the distribution of data and kinetic changes. MATERIALS, METHODS, RESULTS - We developed a new approach which fits functions to measured data sets, describes the statistical distribution and forms a basis for statistical comparison between individual kinetic measurements. We created two FACS assays on BD FACS Aria instrument. The first one monitors calcium flux, generation of reactive oxygen species and mitochondrial membrane potential, while the second one monitors mitochondrial calcium flux, nitric oxide generation and plasma membrane potential in CD4+ and CD8+ lymphocytes simultaneously before and after the administration of a lymphocyte activator. CONCLUSIONS - This technique may be used to investigate purposes (i.e. to test the impact of any agent (such as immunmodulatory drugs) on cellular processes in lymphocytes) and to diagnostic purposes (i.e. to test the alteration of lymphocyte activation characteristics in disease).]

AFFILIATIONS

  1. MTA-Semmelweis Egyetem, Gyermekgyógyászati és Nefrológiai Kutatócsoport
  2. Orvosi Vegytani, Molekuláris Biológiai és Patobiokémiai Intézet, Budapest

COMMENTS

0 comments

Further articles in this publication

Hungarian Immunology

[The role of endothelium, cell migration, chemokines and angiogenesis in inflammatory rheumatic diseases]

BESENYEI Tímea, PÁKOZDI Angéla, VÉGVÁRI Anikó, SZABÓ Zoltán, SZEKANECZ Zoltán

[Endothelial cells, leukocyte-endothelial interactions and angiogenesis are highly involved in the pathogenesis of inflammation and thus in that of inflammatory rheumatic diseases. As this research area is very progressive, one needs to review novel molecular mechanisms and new therapeutic approaches in this respect. Authors review the most important functions of endothelial cells, the process of leukocyte extravasation, tissue infiltration and their cellular and molecular basis. Endothelial cells themselves produce a number of inflammatory mediators including interleukin-1 (IL-1), IL-6, IL-8, chemokines and others. Among cell adhesion molecules, β1 and β3 integrins, as well as E-, L- and P-selectins and their respective ligands have been implicated in leukocyte-endothelial adhesion. In recent years, numerous inflammatory mediators, cytokines, chemokines and proteases have been implicated in angiogenesis and angiostasis. Hypoxia, the vascular endothelial growth factor (VEGF)-angiopoietin system and mechanisms driven by β3 integrins are of major importance during angiogenesis. Significant amount of data have become available of the regulation of cell adhesion, migration and neovascularisation. Adhesion, chemokine and angiogenesis research has important clinical, practical aspects for antirheumatic and anti-cancer therapy. VEGF antagonists, anti-integrin antibodies, chemokine and chemokine receptor inhibitors, as well as thalidomide are currently in the first line of development.]

Hungarian Immunology

[MCP-1 (monocyte chemoattractant protein-1) G/A and T-bet (T-helper promoter factor) C/G polymorphisms in primary Sjögren’s syndrome and systemic lupus erythematosus]

KOVÁCS Attila, KONCZ Ágnes, ENDREFFY Emőke, ARANKA László, PETRI Ildikó, ELLER József, SZALAI Csaba

[INTRODUCTION - Monocyte chemoattractant protein- 1 (MCP-1) is a β-chemokine involved in the attraction and accumulation of mononuclear granulocytes towards the site of inflammation. One of the transcriptional factors of T-cells is called T-bet. PATIENTS AND METHODS - The authors investigated the MCP-1-2518 G/A and T-bet 310 C/G (His33Gln) polymorphisms evaluating the distribution of the specific genotypes in 45 patients with primary Sjögren's syndrome (pSS), 51 patients with systemic lupus erythematosus (SLE), and in 320 healthy blood donors as the control group. MCP-1-2518 G/A and T-bet 310 C/G polymorphisms were detected with molecular genetic methods from the purified genomic DNA. RESULTS - The frequency of the MCP-1-2518 AG heterozygous genotype decreased tendentiously only in SLE patients, while the frequency of the MCP-1 AA homozygous genotype increased comparing to the control group (13.7% vs. 5.9%; Pearson’s χ2 test=6.125, ns.). Analyzing the genotype frequency for the MCP-1 wild (GG) and AA homozygous genotypes in pSS group, the MCP-1 AA homozygous genotype proved to be more frequent comparing to the control group (82.8%:17.2% vs. 90.7%:9.3%; Pearson’s χ2 test 1.755, ns). These relations showed only tendentious association in the SLE group (81.6%:18.7% vs. 90.7%:9.3%; Pearson’s χ2 2.811, p=0.094, ns.) There was not any significant correlation between the investigated MCP-1- 2518 G/A and the T-bet 310 C/G polymorphisms and the TNF-α -308 G/A and -238 allele polymorphisms. The frequency of T-bet was equal in relation with heterozygous (CG) to wild CC genotype in the investigated two autoimmune disorders. The GG homozygous genotype for T-bet could not be found in SLE and pSS groups, likely to be a protective factor. CONCLUSIONS - The above mentioned polymorphisms didn’t show any significant correlation with TNF-α -308 and -238 allele polymorphisms. The further research of the MCP-1 G/A and T-bet C/G polymorphisms is important, because of their possible prognostic importance for SLE and pSS.]

Hungarian Immunology

[Pseudolymphoma orbitae]

VÁNCSA Andrea, GERGELY Lajos, NEMES Zoltán, BÍRÓ Edit, ILLÉS Árpád, BAKÓ Gyula

[INTRODUCTION - Pseudolymphoma orbitae is a rare and difficult entity. The cooperation of the pathologist and clinician is needed to properly manage the patient. CASE REPORT - The authors report the case history of a 38 years old male patient. His disease started at the age of 30. He was previously treated with allergic rhinitis. No definitive diagnosis was made for eight years. Several surgical biopsies were made from nasal mucosa, but no specific histologyical diagnosis was applicable. At the age of 30 he developed an unilateral exophthalmus on the left side. Thyroid associated ophthalmopathy was ruled out several times with laboratory analysis. High dose methylprednisone therapy was repeatedly given with limited results. At the age of 34 orbital CT and MRI scan confirmed the pseudotumour orbitae already compressing the optical nerve. Laboratory analysis again ruled out thyroid associated ophthalmopathy. Churg-Strauss syndrome, Wegener’s granulomatosis or Sjögren’s syndrome could be ruled out. A bone marrow trephine biopsy excluded systemic hematological disease as well. A biopsy was performed from the retrobulbar mass again, which confirmed the lymphoid hyperplasia with B-cell dominance. High dose methylprednisone and local irradiation resulted only moderate decrease of the mass, so systemic chemotherapy was started using CVP (cyclophosphamide, vincristin, prednisone) then CHOP (CVP + anthrycycline) polychemotherapy for eight cycles and subcutaneous interferon-α for 20 months. CONCLUSIONS - This resulted a complete regression of the disease, and the patient is well for 48 months now.]

Hungarian Immunology

[EULAR]

SZEKANECZ Zoltán

Hungarian Immunology

[Mosaic of Autoimmunity]

SZEKANECZ Zoltán

All articles in the issue

Related contents

Ca&Bone

[The role of alfacalcidol in the prevention of osteopenia following renal transplantation]

BERCZI Csaba, ASZTALOS László, KINCSES Zsolt, BALOGH Ádám, LŐCSEY Lajos, BALÁZS György, LUKÁCS Géza

[AIM - The aim of this prospective study was the long-term evaluation of the effect of calcium and alfacalcidol treatment on calcium metabolism in patients with renal transplantation. METHODS - Patients were divided in two groups. Patients in Group 1 (n=159) received calcium substitution, while patients in Group 2 (n=81) were treated with alfacalcidol. Serum Ca, P, Mg, alkaline phosphatase (AP) and PTH levels were determined before and after transplantation regularly for three years. Femur neck and lumbar vertebral bone mineral densities (BMD) were measured at the same time after transplantation. RESULTS - After transplantation the mean serum calcium level significantly increased, while the mean serum phosphate level significantly decreased in both groups. After the operation the PTH levels decreased in both groups and it was found to be more pronounced in the alfacalcidol group.The majority of patients had osteopenia in the follow-up period. Between the third month and the third year after transplantation, BMD increased by 9.4% in Group1, and decreased by 4% in Group 2 at the lumbar spine. At 3 years the mean BMD value at the femoral neck was increased by 6.5% in Group 1, and by 6.7% in Group 2, compared to the 3-month values.The change in BMD was only significant at the lumbar spine, in Group 1 (p=0.019). During the follow-up period osteonecrosis was diagnosed in 6 patients in Group 1 and in 9 cases in Group 2. CONCLUSION - Alfacalcidol treatment decreased secondary hyperparathyroidism more rapidly and effectively, which was also indicated by the more pronounced decrease of serum PTH levels. During the 3 years follow-up period, BMD increased in both groups except for the lumbar spine in Group 2, however, the majority of the patients still had osteopenia.The study could not demonstrate a superiority of alfacalcidol over calcium supplementation in the prevention of posttransplantational osteopenia.]

LAM KID

[Calcium supplementation and the risk of cardiovascular disease - Real apprehension or picking the spin?]

SPEER Gábor

[Some data shows that calcium supplementation, a basic intervention for treating osteoporosis in postmenopausal women, may increase the risk of atherosclerotic vascular disease. Coronary artery calcified plaque is a marker for atheromatous plaque burden and predicts future risk of cardiovascular events. However, the deposition of calcium into the vascular wall is due to an active mechanism, involving such genes and proteins which play role in bone metabolism. In this work the data about the cardiovascular side effect of calcium supplementation are reviewed. Also, I demonstrate studies with the conclusions that calcium supplements with or without vitamin D do not increase the risk of cardiovascular events, especially that of myocardial infarction.]

LAM KID

[Osteoporotic patient’s use of prescription drugs - pilot study]

BATKA Gábor, SZENTANDRÁSSY Andrea Éva, SZEKERES László

[BACKGROUND - In Hungary, the number of the highest mortality hip fractures is between 12 000-15 000 per year. The cost of treating hip fractures is several times higher than that of preventive medical therapy. Thus, the compliance of patients with osteoporosis is of great importance. METHODS - Using the informatical database of St. András Rheumatology Hospital at Héviz, we collected one year’s prescription drugs for osteoporosis and compared them with the number of drugs obtained by the patients, determined from National Health Insurance data. RESULTS - In general, the patients obtained 75% of prescription drugs. From the 4354 boxes of prescribed antiporotics, 3637 contained bisphosphonate (not considering vitamin D and calcium). Within this group, 88% of combination preparations were obtained, which is a greater ratio than that of non-combination bisphosphonates (84%). CONCLUSIONS - On the basis of our results, we posit that prescription of a combination preparation somewhat improves the patients’ compliance. The low concordance of vitamin D and calcium preparations is worrying.]

Ca&Bone

[The role of calcium in the chemoprevention of colorectal cancer]

FUSZEK Péter, SPEER Gábor

[One of the most exciting research areas of the past decade has concerned the chemoprevention of colorectal cancer (CRC). Numerous clinical studies have been conducted on the preventive role of NSAIDs, high fibre intake, selenium, phytooestrogens, hormone replacement therapy, antioxidants, COX-inhibitors, folic acid and calcium, however, their results are controversial. Among the suggested chemopreventive agents, the preventive role of calcium is supported by the strongest evidence.This paper aims to review the available facts on the role of calcium. Recent studies suggest that appropriate calcium intake may partially counterbalance the effect of the genes that contribute to the development of CRC. Experimental data show that calcium directly influences the expression of several genes involved in tumorigenesis and that it is also involved in a number of signalling pathways that control cell proliferation, differentiation and apoptosis.These effects mostly arise through the activation of the calcium sensing receptor. The main goal of this review is to draw attention to the established chemopreventive role of calcium in CRC. Published data suggest that a lifelong daily calcium intake between 1200 to 1500 mg (even 2000 mg in high risk groups) would significantly decrease the incidence of CRC by inhibition of tumorigenesis.]

LAM KID

[The results of the Ca-Intake.hu study - Calcium supplementation as a part of basic therapy of osteoporosis is more than a routine step]

LAKATOS Péter, SPEER Gábor, DOMBAI Péter, ZAJZON Gergely

[Calcium intake is considered the base therapy of osteoporosis treatment. It is known that in case of inadequate calcium intake, specific anti-osteoporotic drugs are inefficient. In the present study, we aimed to investigate the alimentary and supplementary calcium intake among Hungarian osteoporotic patients, using a nationwide representative survey. Patients with osteoporosis were enrolled in the study. We determined the total alimentary calcium intake and the average supplementary dose. In some cases, total calcium intake was lower than recommended, in other cases it was significantly higher than that. In some cases, bone density showed a positive correlation with calcium intake. Vitamin D supplementation complied with current recommendations.]