Hungarian Immunology

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

MÉSZÁROS Gergő, RÓNAI Katalin Zsuzsanna, TOLDI Gergely, KAPOSI Ambrus, VÁSÁRHELYI Barna, TRESZL András

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).]



Further articles in this publication

Hungarian Immunology

[Etanercept in early rheumatoid arthritis]


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



Hungarian Immunology

[Comparison of disease-modifying drugs in rheumatoid arthritis]

SZŰCS Gabriella, SZEKANECZ Zoltán

Hungarian Immunology

[Etoricoxib in the treatment of rheumatoid arthritis]


All articles in the issue

Related contents


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


[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.]


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


[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.]

Clinical Neuroscience

[Calcium ion is a common denominator in the pathophysiological processes of amyotrophic lateral sclerosis]

PATAI Roland, NÓGRÁDI Bernát, MESZLÉNYI Valéria, OBÁL Izabella, ENGELHARDT József István, SIKLÓS László

[Amyotrophic lateral sclerosis (ALS), the most frequent motor neuron disease is characterized by progressive muscle weakness caused by the degeneration of the motor neurons in the spinal cord and motor cortex. However, according to the recent observations, ALS is a rather complex syndrome which frequently involves symptoms of cognitive impairment. Therefore, ALS cases can be interpreted in a clinico-pathological spectrum spanning from the classical ALS involving only the motor system to the fronto-temporal dementia. The progression of the disease, however, manifested in the degeneration of the upper and lower motor neurons, is based on the same complex pathobiology. The main elements of the pathomechanism, such as oxidative stress, excitotoxicity, immune/inflammatory processes and mitochondrial dysfunction are well described already, which operate in orchestrated way and amplify the deleterious effect of each other. It is assumed that calcium ions act as a catalyst in this interaction, hence each of the individual mechanisms has strong, positive and reciprocal calcium dependence thus may combine the individual pathological processes into a unified escalating mechanism of neuronal destruction. This review provides an overview of the role of calcium in connecting and amplifying the major mechanisms which lead to degeneration of the motor neurons in ALS. ]


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


[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.]


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


[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.]