Włodzimierz Luczyński

Publications of Włodzimierz Luczyński

• [Polymorphism rs9939609 of FTO gene is related to the body mass index in children from podlaskie voievodship.]

  Authors: Magdalena Tercjak-Rećko, Włodzimierz Luczyński, Paweł Bernatowicz, Grzegorz Zalewski, Małgorzata Rembińska, Urszula Lachowska, Przemysław Rećko, Paweł Suchoń, Marcin Czaban, Justyna Sokal, Anna Kopalińska, Zuzanna Pajer, Karolina Bogdanowicz, Jacek Nikliński, Artur Bossowski

  Medycyna wieku rozwojowego. 01/2012; 16(1):53-60.

  Introduction: The presence of obesity and the features of metabolic syndrome plays a predictive role in cardiovascular diseases (CVD) in adults. It seems reasonable to seek new risk factors in theIntroduction: The presence of obesity and the features of metabolic syndrome plays a predictive role in cardiovascular diseases (CVD) in adults. It seems reasonable to seek new risk factors in the development of CVD. Defining the genetic background of obesity could help to select patients from a high risk group and help to introduce prevention and treatment, which, in consequence, lead to the lowering of morbidity and mortality. One of the genes probably related to the body weight is the Fat Mass and Obesity Associated Gene (FTO). The aim: of the study was an attempt to assess the relationship between the FTO polymorphism rs9939609 and body mass index in children from Podlaskie voievodship. Material and methods: 405 children aged 4-18 were selected for the study. The examination included body mass index, waist circumference, blood pressure and lipid profile analysis. FTO rs9939609 polymorphism was assessed using a discrimination allele method with the application of ABI 7900HT Fast Real-Time PCR System. Results: FTO rs9939609 polymorphism was related to the standarized body mass index and the AA genotype carriers had a higher risk of obesity. This polymorphism was also associated with waist circumference, systolic blood pressure and triglycerides concentration. It was not correlated with diastolic blood pressure and total HDL- and LDL-cholesterol concentrations. Conclusions: Our results demonstrate that rs9939609 FTO gene polymorphism is related to the body mass index in children. Our results should be confirmed in studies on a large cohort of healthy Polish children.

• [Endothelial progenitor cells as a new marker of endothelial function with respect to risk of cardiovascular disorders].

  Authors: Barbara Głowińska-Olszewska, Włodzimierz Luczyński, Artur Bossowski

  Postȩpy higieny i medycyny doświadczalnej (Online). 01/2011; 65:8-15.

  The discovery of endothelial progenitor cells (EPC), over a decade ago, has refuted the previous belief that vasculogenesis only occurs during embryogenesis. The results of several studies revealedThe discovery of endothelial progenitor cells (EPC), over a decade ago, has refuted the previous belief that vasculogenesis only occurs during embryogenesis. The results of several studies revealed altered number and impaired function of EPC in hyperlipidemia, hypertension, diabetes, obesity as well as in rheumatoid arthritis. The population of developmental age is characterized by higher counts of EPC compared to adults. However, among young patients with chronic disorders that affect the vascular system, the number of EPC decreases. The reduced circulating concentration of EPC has become a surrogate marker of endothelial function and has been implicated in the pathogenesis of many vascular diseases. This article aims to review the biology and pathophysiology of EPC in the conditions of cardiovascular risk factors. The potential possibilities of increasing EPC number and function as well as the use of EPC in the treatment of vascular pathology will also be discussed.

• Transformation of conventional T cells into regulatory T cells in children with metabolic syndrome.

  Authors: Włodzimierz Luczyński, Natalia Wawrusiewicz-Kurylonek, Artur Bossowski, Elżbieta Iłendo, Marek Hryszko, Adam Krętowski, Anna Stasiak-Barmuta

  Kardiologia polska. 01/2011; 69(12):1221-6.

  Much research has been done in the recent years to establish an association between obesity, metabolic syndrome and the immune system. Numerous data suggest that the decreased number and/or functionMuch research has been done in the recent years to establish an association between obesity, metabolic syndrome and the immune system. Numerous data suggest that the decreased number and/or function of regulatory T cells (Treg cells) can lead to chronic minimal inflammation present in patients with obesity and trigger formation of atherosclerotic plaque. To generate Treg cells from the peripheral blood in children meeting the diagnostic criteria of metabolic syndrome. A total of 25 children with metabolic syndrome and 25 controls were enrolled in the study. Peripheral blood was collected, CD4(+)/CD25(-) cells were separated and cultured for 4 weeks in the presence of a Treg expander (CD3/CD28) and interleukin-2. The expression of the transcription factor FoxP3 as a Treg marker was assessed before and after culture using reverse transcriptase polymerase chain reaction (RT-PCR) and flow cytometry. Before the culture we observed a slightly lower percentage of Treg cells in children with metabolic syndrome vs controls. After the culture we noted a significant increase in mRNA expression and in the percentage of FoxP3-positive cells. We observed no differences in the results between the children with metabolic syndrome and the controls. Our study shows that it is possible to generate Treg cells from peripheral blood of children with metabolic syndrome. In future, these findings could be used to develop a model of immunotherapeutic intervention for patients at risk of cardiovascular disease.

• [Generation of T regulatory cells in children with type 1 diabetes mellitus].

  Authors: Włodzimierz Luczyński, Natalia Wawrusiewicz-Kurylonek, Agnieszka Szypowska, Elżbieta Iłendo, Artur Bossowski, Adam Krętowski, Barbara Głowińska-Olszewska, Marek Hryszko, Maria Górska, Anna Stasiak-Barmuta

  Pediatric endocrinology, diabetes, and metabolism. 01/2011; 17(2):64-70.

  In spite of intensive research the pathogenesis of type 1 diabetes mellitus is not thoroughly understood. One of the ideas which currently has a great number of supporters is the theory of theIn spite of intensive research the pathogenesis of type 1 diabetes mellitus is not thoroughly understood. One of the ideas which currently has a great number of supporters is the theory of the participation of T regulatory cells in the mechanism of insufficient suppression of the immune response against pancreatic self -antigens. According to some authors, the infusion of T regulatory cells in autoimmune diseases could lead to long -term remission or even a complete cure. Aim of the study: The aim of our present study was to achieve T regulatory cells from conventional T lymphocytes isolated from a small amount of peripheral blood in children with type 1 diabetes mellitus and their comparison to the Tregs generated from the blood of control children. Additionally, we assessed the changes in the expression of selected genes essential for the function of these cells during Tregs generation. The examined group consisted of 20 children with type 1 diabetes mellitus, the control group consisted of 20 non -diabetic children. From the peripheral blood CD4+CD25 - cells were separated and cultured with T -reg expander and interleukin (IL) 2. Before and after the culture the cells were analysed according to the expression of transcription factor FoxP3 and other molecules/cytokines: OX40, 4 -1BB, GITR, ICOS -1, CTLA -4 and IFN -γ, IL -10 and TGF -β. We observed a significantly higher percentage of T regulatory cells after the culture (with no difference between diabetic and control children). Moreover, we observed a lower expression of mRNA for GITR molecule and a higher IL -10 expression in the cultures of diabetic children compared to the control ones. The cells cultured from the blood of control children were characterised by a higher increase in the expression of mRNA for ICOS -1 and a lower expression of mRNA for TGF -β in comparison to the cultures from diabetic children. The results of our investigations confirm the possibility of generating T regulatory cells from conventional T lymphocytes from peripheral blood of children with newly recognised type 1 diabetes mellitus.

• [Higher percentages of T regulatory cells in children at risk for developing type 1 diabetes mellitus].

  Authors: Włodzimierz Luczyński, Anna Stasiak-Barmuta, Małgorzata Myśliwiec, Agnieszka Nikołajuk, Agnieszka Brandt, Remigiusz Urban, Justyna Kos, Agnieszka Juchniewicz, Jolanta Jabłońska, Agnieszka Otocka, Barbara Głowińska-Olszewska, Bozena Florys, Mirosława Urban, Maria Górska, Anna Balcerska

  Pediatric endocrinology, diabetes, and metabolism. 01/2010; 16(1):7-10.

  The natural history of type 1 diabetes is concerned with the appearance of autoantibodies against antigens of pancreatic beta cells. The last decade revealed some evidence of the participation of TThe natural history of type 1 diabetes is concerned with the appearance of autoantibodies against antigens of pancreatic beta cells. The last decade revealed some evidence of the participation of T regulatory lymphocytes - cells which suppress immune response - in the pathogenesis of type 1 diabetes and prediabetes. was the assessment of T regulatory cells in the blood of children at risk for developing type 1 the diabetes mellitus. 85 subjects, siblings of children with type 1 diabetes, were enrolled into the study. The presence of anti-GAD65 antibodies was assessed. With the use of flow cytometry the following cell subpopulations were noted: CD4+, CD4+CD25high and CD4+CD25highCD127low with the coexpression of: CD28, CD45RO, CD54, CD62L and CD134 molecules. We did not observe any differences in white blood cell count, lymphocyte (including CD4+) count and the percentage between the examined and control groups. We noted higher percentages of T regulatory cells: CD4+CD25high, CD4+CD127low and CD4+CD25highCD127low in children with the presence of anti-GAD65 antibodies as compared to the control children. Higher percentages of T regulatory cells in the blood of children with the presence of anti-GAD65 antibodies may suggest an intensive regulatory response present in patients at risk for developing type 1 diabetes.

• [The role of FTO gene polymorphism in the pathogenesis of obesity].

  Authors: Magdalena Tercjak, Włodzimierz Luczyński, Natalia Wawrusiewicz-Kurylonek, Artur Bossowski

  Pediatric endocrinology, diabetes, and metabolism. 01/2010; 16(2):109-13.

  Both environmental and genetic factors play a role in the pathogenesis of obesity. At present, researchers are examining the genetic background of overweight. Over 100 genes are suspected toBoth environmental and genetic factors play a role in the pathogenesis of obesity. At present, researchers are examining the genetic background of overweight. Over 100 genes are suspected to influence the obesity. One of those genes is FTO (fat mass and obesity-associated gene). In the manuscript, the relationship between FTO gene polymorphism (AA allele) and overweight, obesity and their consequences are discussed. It was proved, in studies on large number of people, that FTO gene polymorphism is related to higher body mass index, weight and abdominal circumference. Some authors showed that FTO gene polymorphism influences the food intake, energy expanditure and insulin resistance. The expression of FTO gene product was noticed in both, the peripheral as well as central nervous system (hypothalamus). It is possible that FTO gene product is involved in the DNA and RNA reparation and other metabolic processes, however, its function is not yet clear. The assessment of FTO gene polymorphism can allow us to separate patients with the tendency for higher weight. The elucidation of the role of FTO in the pathogenesis of obesity can result in the development of new therapeutic options for this epidemic of XXI century.

• [Asymmetric dimethylarginine (ADMA) in children with diabetes type 1].

  Authors: Barbara Głowińska-Olszewska, Włodzimierz Luczyński, Jolanta Jabłońska, Agnieszka Otocka, Bożena Florys, Artur Bossowski

  Pediatric endocrinology, diabetes, and metabolism. 01/2010; 16(3):137-41.

  Asymmetric dimethylarginine (ADMA), a naturally occurring product of asymmetric methylation of proteins, is an endogenous inhibitor of endothelial nitric oxide synthase. ADMA is now recognized as anAsymmetric dimethylarginine (ADMA), a naturally occurring product of asymmetric methylation of proteins, is an endogenous inhibitor of endothelial nitric oxide synthase. ADMA is now recognized as an independent marker of endothelial dysfunction and atherosclerosis. Data concerning ADMA level in type 1 diabetes (DM1) are controversial. The aim of the study was to evaluate ADMA level in children with DM1, without clinical evidence of vascular complications, with particular attention to additional cardiovascular risk factors (hypertension, obesity, hyperlipidemia). The study group included 72 children with DM1, aged mean 15±3 yrs (8-20 yrs), 33 boys and 39 girls, with diabetes duration time mean 6.6±3.5 yrs (1-14 yrs), HBA1c mean level 8.2±2.3% (5.6-15%). The control group consisted of 41 (19 boys and 22 girls) healthy children, aged mean--14.8±2.6 yrs, from 8 to 18 yrs, gender matched, with no family history of cardiovascular disease. ADMA level was determined in plasma using ELISA kit (DLD Diagnostica, Hamburg, Germany) ADMA level was similar in children with diabetes and in the control group: 0.69±0.33 vs. 0.7±0.27 μmol/L, ns. We did not find differences in ADMA level in diabetic children with the presence of additional diseases being cardiovascular risk factors. In the group of 13 children with hypertension ADMA level was the highest: 0.79±0.25 μmol/L, but the difference was statistically insignificant in comparison to children with diabetes without hypertension and in comparison to healthy controls. Children with DM1, without clinically evident vascular complications, have ADMA levels similar to healthy children. A possible relationship between ADMA and hypertension in these patients requires further investigation.

• [The role of T-regulatory cells in the pathogenesis of immunological disturbances accompanying obesity and atherosclerosis].

  Authors: Włodzimierz Luczyński, Artur Bossowski, Barbara Głowińska-Olszewska, Justyna Kos, Anna Stasiak-Barmuta

  Postȩpy higieny i medycyny doświadczalnej (Online). 01/2010; 64:156-60.

  Obesity and atherosclerosis, and their consequences, including cardiovascular disease, are plagues of the 21st century. Chronic inflammation, whose mechanism is not well understood, underlies theObesity and atherosclerosis, and their consequences, including cardiovascular disease, are plagues of the 21st century. Chronic inflammation, whose mechanism is not well understood, underlies the pathophysiological bases of both processes. T lymphocytes, macrophages, and the proinflammatory cytokines produced by these cells play key roles in the immunological disturbances accompanying obesity and atherosclerosis. It was recently shown that T-regulatory cells can play a role in these processes. T-regulatory cells are a small subpopulation of T cells which are responsible for inhibition of the immune response. In this review, experiments conducted in mice and human models on the role of diminished number and/or function of T-regulatory lymphocytes in the pathogenesis of immune disturbances accompanying obesity and atherosclerosis are discussed. The results of studies using T-regulatory cells to stabilize and decrease atherosclerotic lesions in blood vessel walls are also summarized. The results of experiments performed so far are encouraging and give some hope for the future use of T-regulatory cells in the therapy of obesity and atherosclerosis.

• [Overweight, obesity and metabolic syndrome in children with type 1 diabetes melllitus].

  Authors: Włodzimierz Luczyński, Agnieszka Szypowska, Artur Bossowski, Anna Ramotowska, Przemysław Rećko, Małgorzata Rembińska, Magdalena Tercjak, Bernadetta Blecharczyk, Urszula Lachowska, Paweł Suchoń, Karolina Wiśniewska, Paweł Bernatowicz, Barbara Głowińska-Olszewska

  Pediatric endocrinology, diabetes, and metabolism. 01/2010; 16(2):83-8.

  Obesity can be an additional risk factor for developing cardiovascular diseases in patients with diabetes. Aim of the study was the assessment of overweight, obesity and other elements of theObesity can be an additional risk factor for developing cardiovascular diseases in patients with diabetes. Aim of the study was the assessment of overweight, obesity and other elements of the metabolic syndrome in children with type 1 diabetes mellitus. 300 children treated with insulin at least one year were enrolled in the study. In the examined group anthropometric data, data concerned with diabetes and additional laboratory tests including risk factors for cardiovascular diseases were assessed. The median age of the examined group was 13.7 years. The body mass deficiency was noted in 0.66%, normal body mass in 71.6%, overweight in 15.3% and obesity in 12.3%. The abdominal obesity was noted in 16.0% of children. The rise in the body weight between 3-6 months from the beginning of the insulin therapy and the present assessment was statistically significant. Children with normal weight had a better metabolic control in comparison to children with overweight/obesity. Girls had a higher rise in body mass index values between the time of diagnosis and the present investigation compared to boys. Higher values of blood pressure or hypertension were noted in 16.6% of children. Altogether in 25.3% of children some dyslipidemia was observed. The metabolic syndrome criteria were noted in: 28.0% - one criterion, 13.0% - two criteria, and 0.3% - three criteria. The population of children with type 1 diabetes is characterized by high frequency of overweight/obesity, abdominal obesity, dyslipidemia and hypertension. The features of metabolic syndrome are less frequent. It is worthwhile to monitor the risk for development of cardiovascular diseases in this group of children.

• Diminished expression of ICOS, GITR and CTLA-4 at the mRNA level in T regulatory cells of children with newly diagnosed type 1 diabetes.

  Authors: Włodzimierz Luczyński, Natalia Wawrusiewicz-Kurylonek, Anna Stasiak-Barmuta, Remigiusz Urban, Elżbieta Iłendo, Mirosława Urban, Marek Hryszko, Adam Krętowski, Maria Górska

  Acta biochimica Polonica. 06/2009;

  Diabetes mellitus is one of the most common chronic diseases in children. T regulatory cells (Tregs) modulate response to autoantigens and probably play a role in pathogenesis of type 1 diabetesDiabetes mellitus is one of the most common chronic diseases in children. T regulatory cells (Tregs) modulate response to autoantigens and probably play a role in pathogenesis of type 1 diabetes (T1DM). The aim of the present study was the assessment of T regulatory cells including their percentages and expression of critical genes in these cells in children with newly diagnosed type 1 diabetes. The examined group consisted of 50 children with T1DM. A flow cytometric analysis of T-cell subpopulations was performed using the following markers: anti-CD4, anti-CD25 and anti-CD127 (=IL-7R). Additionally, T regulatory cells were isolated for assessment of mRNA levels for chosen genes with the real-time RT PCR technique. The percentages of CD4CD25highCD127(dim/-) were very low and did not differ between T1DM and control children. We did not observe any statistically significant differences between healthy and diabetic children in mRNA expression for FoxP3, IL-7R (CD127), IL-8RA, IL-10RA, IL-12A, IL-2RA (CD25), IL-21, STAT1, STAT3, SOCS2, SOCS3, TGF-beta1-R1, TGF-beta-R2 and TBX-21 genes. Interestingly the mRNA level for CTLA-4, ICOS1, IL-23, IL-27, SMAD3 and GITR were lower in Treg cells of children with diabetes compared to the control patients. No disturbances in the percentages of T regulatory cells in patients with diabetes but diminished expression of some elements important in Treg function could be the result of an immunologic imbalance accompanying the onset of the diabetes. The results of our study should be used in future research in the field of immunotherapy in pediatric diabetes.

• Lower percentages of T regulatory cells in children with type 1 diabetes - preliminary report.

  Authors: Włodzimierz Luczyński, Anna Stasiak-Barmuta, Remigiusz Urban, Mirosława Urban, Bożena Florys, Marek Hryszko

  Endokrynologia, diabetologia i choroby przemiany materii wieku rozwojowego : organ Polskiego Towarzystwa Endokrynologow Dzieciecych. 02/2009; 15(1):34-38.

  Introduction:Type 1 diabetes (T1D) isa well-known autoimmune disease, however there are still some processes in its pathogenesis to be elucidated. In the last few years the role of T regulatory cellsIntroduction:Type 1 diabetes (T1D) isa well-known autoimmune disease, however there are still some processes in its pathogenesis to be elucidated. In the last few years the role of T regulatory cells in the pathogenesis of T1D has been investigated. The aim of study was to determine the percentages and numbers of T regulatory cells in the peripheral blood of children with type 1 diabetes. Material and methods: A total of 25 children with newly diagnosed type 1 diabetes were studied, and compared to the control group consisted of 30 healthy children with no signs of autoimmune, chronic, inflammatory, neoplastic disease, and no evidence ofT1D in their families. Flow cytometric analysis ofT-cell subpopulations was performed using thefollowing markers: anti-CD3, anti-CD4, anti-CD25, and anti-CD127 (IL-7R). Results: In the group of children with type 1 diabetes we noted statistically significant lower percentages of T regulatory cells, i.e., CD4+CD25 high and CD4+CD127 low comparing to the control children. There were no differences in other assessed parameters: white blood cell count, lymphocytes, CD4+ and CD4+CD25 high CD127 low cells (percentage and count). We did not find the correlation between patients' age and any of analysed parameters. Conclusions: Our results suggestthe lower percentages ofTregs in children with T1D, however those data need to beconfirmed in a larger cohort of patients and complemented with functional studies, e.g. at mRNA level. In our opinion T regulatory cells could be excellent candidates for cell therapy in newly diagnosed type 1 diabetic children.

• [Do the regulatoryT cells play a role in the pathogenesis and therapy oftype 1 diabetes?]

  Authors: Włodzimierz Luczyński, Remigiusz Urban, Anna Stasiak-Barmuta, Mirosława Urban

  Endokrynologia, diabetologia i choroby przemiany materii wieku rozwojowego : organ Polskiego Towarzystwa Endokrynologów Dziecięcych. 02/2009; 15(1):41-4.

  In the paper the current knowledge concerning the role of T regulatory cells in the pathogenesis and modern therapy of diabetes type 1 is discussed. Type 1 diabetes is one of the most well-knownIn the paper the current knowledge concerning the role of T regulatory cells in the pathogenesis and modern therapy of diabetes type 1 is discussed. Type 1 diabetes is one of the most well-known autoimmunological diseases. In its pathogenesis the destruction of pancreatic beta cells by autoreactive T lymphocytes plays the main role. It is considered that in the autoimmunologic reaction participate the disturbances in the number/function of T regulatory cells - responsiblefor the inhibition of hyperreactive immune response. According to the newestdata the regulatory cells possess CD4+/ CD25++/CD127low phenotype. Results concerning the role of T regulatory cells in the pathogenesis of diabetes are diverse, further both the experimental and clinical studies are required including the use ofthose cells in immunotherapy. Most authors observed the lackof number and/or function of T regulatory cells in type 1 diabetes. Among produced cytokines most valuable is TGF-p. Important disturbances in this field were also observed in the patients' families including the carriers of high-risk group genes. There are also ideas to use T regulatory cells in the prevention and therapy of diabetes. Some reports noted the role of T regulatory cells in the response to novel immunotherapies in diabetes like the use ofTNF-a, anti-CD3 antibodies or dendritic cells inducing pancreatic islet tolerance. It is possible that further research will allow to use the immunotherapy including T regulatory cells in type 1 diabetes.

• [Myeloid-derived suppressor cells - the new mechanism of immunosuppression in cancer]

  Authors: Włodzimierz Luczyński, Maryna Krawczuk-Rybak, Anna Stasiak-Barmuta

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2008; 62:18-22.

  In this paper we summarized the current knowledge about the new mechanism of immunosuppression in cancer i.e. the lately discovered myeloid-derived suppresor cells (MDSC). MDSC are produced in boneIn this paper we summarized the current knowledge about the new mechanism of immunosuppression in cancer i.e. the lately discovered myeloid-derived suppresor cells (MDSC). MDSC are produced in bone marrow under the influence of tumor cell derived substances. MDSC show negative effect on T lymphocytes function through arginine depletion and nitric oxide production. In mice MDSC are characterized by Gr-1 and CD11b expression, however in human there are no definitive markers of this subpopulation. Some laboratory experiments in turning back the negative influence of MDSC on immunological system are provided, mainly through the administration of monoclonal antibodies against MDSC or drugs which block their function. The elucidation of MDSC characteristics can allow us to build new immunotherapeutic protocols in the future.

• Upregulation of antigen-processing machinery components at mRNA level in acute lymphoblastic leukemia cells after CD40 stimulation.

  Authors: Włodzimierz Luczyński, Oksana Kowalczuk, Elzbieta Iłendo, Anna Stasiak-Barmuta, Maryna Krawczuk-Rybak

  Annals of hematology. 06/2007; 86(5):339-45.

  The development of immunotherapy in hematologic malignancies has been observed in the last few years. One of the approaches is the use of cancer vaccines based on leukemia-derived dendritic cellsThe development of immunotherapy in hematologic malignancies has been observed in the last few years. One of the approaches is the use of cancer vaccines based on leukemia-derived dendritic cells (DC). Recent studies from our laboratory and other laboratories have shown that CD40 stimulation improves leukemia cells immunogenicity and generates an antitumor immune response. The design of future cancer vaccines requires the knowledge concerning the function of dendritic cells including antigen processing. The aim of our present study was the assessment of antigen-processing machinery (APM) components in acute lymphoblastic leukemia (ALL) cells before and after CD40 stimulation at messenger RNA (mRNA) level. Twenty-five children with ALL were enrolled into the study. Leukemic cells were stimulated (or not) with CD40L and IL-4. Elements of the antigen-processing machinery (MB1, LMP2, LMP7, LMP10, TAP1, TAP2, calnexin, calreticulin, tapasin, ERp57, zeta, delta) were determined by real-time PCR technique. The expression of important costimulatory and adhesion molecules considered as DC markers (CD40, CD54, CD80, CD83, CD86) were determined at the mRNA (PCR) and protein (flow cytometry) levels. The following are the results of our study: (1) We noted an upregulation of all costimulatory and adhesion molecules at the mRNA and protein levels in ALL cells after the culture; (2) the significant rise in expression of nearly all APM components after CD40 stimulation was observed. This confirms specific stimulation of the antigen-processing system in ALL cells by CD40L. Future work should focus on the clinical significance of these findings for immunotherapy in leukemias.

• [Experimental and selected clinical aspects of active immunotherapy in leukemia]

  Authors: Włodzimierz Luczyński, Maryna Krawczuk-Rybak, Anna Stasiak-Barmuta

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2006; 60:379-86.

  The aim of the review is to summarize current knowledge concerning active immunotherapy in leukemia. The molecular mechanisms and selected clinical implications of different cancer vaccines used inThe aim of the review is to summarize current knowledge concerning active immunotherapy in leukemia. The molecular mechanisms and selected clinical implications of different cancer vaccines used in pediatric and adult leukemias are discussed. Escape of neoplasmatic cells from elimination by host cells can be caused by immunological disturbances, such as the production of immunosuppressive cytokines and downregulation of costimulatory and adhesion molecules. Cells of acute lymphoblastic leukemia and chronic lymphocytic leukemia can be induced into antigen-presenting cells with the CD40 ligation system. After CD40 stimulation, leukemic cells achieve the phenotypic and functional characteristics of dendritic cells. In many studies it was confimed that these cells stimulate auto- and/or allogeneic T-cell response. Immunological response can be of cellular and humoral origin and is extensively examined. Similar effects using different cytokines such as GM-CSF, TNF-alpha, and IL-4 can be observed in acute myeloid leukemia and myelodysplastic syndromes. Clinical experience with such vaccines is limited, but the results of some preliminary reports are quite promising. Cancer vaccines are safe, result in host response, and probably prolong patients survival.

• CD40 stimulation induces differentiation of acute lymphoblastic leukemia cells into dendritic cells.

  Authors: Włodzimierz Luczyński, Anna Stasiak-Barmuta, Elzbieta Iłendo, Maryna Krawczuk-Rybak, Iwona Malinowska, Małgorzata Mitura-Lesiuk, Adam Parfieńczyk, Marcin Szymański

  Acta biochimica Polonica. 02/2006; 53(2):377-82.

  Despite the very high percentage of long-term remissions in acute lymphoblastic leukemia (ALL) in children, some of them suffer from recurrence of the disease. New treatment modalities, e.g.Despite the very high percentage of long-term remissions in acute lymphoblastic leukemia (ALL) in children, some of them suffer from recurrence of the disease. New treatment modalities, e.g. effective geno- and immunotherapy are needed. The use of neoplasmatic cells to present tumor antigens is one of the approaches in cancer vaccines. ALL cells lack the expression of costimulatory molecules and are poor antigen presenting cells (APCs) for T-cell activation. CD40/40L interaction stimulates B-cells to proliferate, differentiate, upregulate costimulatory molecules and increase antigen presentation. The aim of the study was to test the hypothesis that ALL cells can be turned into professional APCs by CD40L activation. Children with B-cell precursor ALL were enrolled into the study. Mononuclear cells from bone marrow or peripheral blood were stimulated with CD40L and interleukin 4. Results: 1) after culture we noted upregulation of all assessed costimulatory, adhesion and activatory molecules i.e. CD1a, CD11c, CD40, CD54, CD80, CD83, CD86, CD123, HLA class I and II; 2) CD40L activated ALL cells induced proliferation of allogeneic T-cells (measured by [(3)H]thymidine incorporation). These results confirm the possibility of enhancing the immunogenicity of ALL cells with the CD40L system and indicate that this approach can be used in immunotherapeutic trials.

• Lower expression of mRNA for interferon-gamma in T helper cells of children with newly diagnosed lymphomas.

  Authors: Włodzimierz Luczyński, Oksana Kovalchuk, Maryna Krawczuk-Rybak, Małgorzata Mitura-Lesiuk, Iwona Malinowska, Lech Chyczewski, Jerzy Kowalczyk, Michał Matysiak

  Folia histochemica et cytobiologica / Polish Academy of Sciences, Polish Histochemical and Cytochemical Society. 02/2005; 43(3):169-71.

  The complex interactions between cancer and host cells are far from being fully elucidated. Assessment of Th1/Th2/Th3/Tr1 balance is an interesting approach to explain immunological disturbances inThe complex interactions between cancer and host cells are far from being fully elucidated. Assessment of Th1/Th2/Th3/Tr1 balance is an interesting approach to explain immunological disturbances in lymphomas. The aim of our study was to assess mRNA for pro- and anti-inflammatory cytokines in T-cells in 20 children with Hodgkin- and non-Hodgkin lymphomas. CD4+ and CD8+ cells were isolated from whole peripheral blood and four different cytokine mRNA levels (IFN-gamma, IL-10, IL-4, TGF-beta) were determined by real-time PCR technique. Comparing to the control group, we found lower expression of mRNA for IFN-gamma in CD4+ cells at the time of lymphoma diagnosis. It may be one of the pathogenetic mechanisms of impaired immunity in these patients.

• [Mechanisms of immunosuppression in leukemia]

  Authors: Włodzimierz Luczyński, Maryna Krawczuk-Rybak

  Postȩpy higieny i medycyny doświadczalnej (Online). 02/2005; 59:28-33.

  The mechanisms of immunosuppression and immunotherapy in vitro in leukemia are reviewed. A predominance of Th2 lymphocytes, diminished percentages of T lymphocyte with costimulatory moleculeThe mechanisms of immunosuppression and immunotherapy in vitro in leukemia are reviewed. A predominance of Th2 lymphocytes, diminished percentages of T lymphocyte with costimulatory molecule expression and impaired monocyte function play major roles in impaired immunity in this group of diseases. Neoplastic cells can produce immunosuppressive cytokines and have low expressions of costimulatory molecules. In immunotherapy in vitro, blasts are converted to antigen-presenting cells by cytokines (acute myeloid leukemia blasts) or CD40L stimulation (acute lymphoblastic leukemia blasts). Manipulation of the immune system to stimulate anticancer response can be a promising tool in hematology and oncology.

• [Immunotherapy in aplastic anaemia as a cause of reactivation of hepatitis B virus-immunologic aspects]

  Authors: Włodzimierz Luczyński, Katarzyna Muszyńska-Rosłan, Maryna Krawczuk-Rybak, Dariusz M Lebensztejn

  Wiadomości lekarskie (Warsaw, Poland : 1960). 02/2005; 58(11-12):697-9.

  We present history of 16-year-old boy, HBsAg carrier, treated with interferon alpha at the age of 6 because of hepatitis B (HBeAg/antyHBe seroconversion). In August 2002--admitted to Department ofWe present history of 16-year-old boy, HBsAg carrier, treated with interferon alpha at the age of 6 because of hepatitis B (HBeAg/antyHBe seroconversion). In August 2002--admitted to Department of Pediatric Oncology due to pancytopenia--diagnosis of severe aplastic anaemia was made (bone marrow cellularity--10%). We found no relative donor for hematopoietic cells transplantation and started immunosuppresive therapy (ATG, G-CSF, methyloprednisolon, cyclosporin). Haematologic parameters were improving. At day +60 he was admitted to our Department due to the increase in aminotransferases and cyclosporin activity. He was treated with cefuroxim, acyclovir and drugs improving liver cell function, cyclosporin was stopped. Presence of HBV DNA in serum confirmed HBV reactivation--a boy received lamivudine and cyclosporin again (as a maintenance therapy of aplastic anaemia). Aminotransferase activity and haematological parameters returned to normal. This case indicates the possibility of HBV reactivation in the course of immunosuppressive therapy (e.g. after antithymocytic globulin and cyclosporin) for aplastic anaemia.

• [Costimulatory and activation molecules of lymphocytes T in acute lymphoblastic leukemia in children]

  Authors: Włodzimierz Luczyński, Anna Stasiak-Barmuta, Maryna Krawczuk-Rybak, Iwona Malinowska, Michał Matysiak, Małgorzata Mitura-Lesiuk, Jerzy Kowalczyk

  Przegla̧d lekarski. 02/2004; 61(9):914-8.

  In the last years one rises importance of costimulatory molecules in immune response in leukemias. Aim of the study was to assess lymphocytes T function in children with acute lymphoblastic leukemiaIn the last years one rises importance of costimulatory molecules in immune response in leukemias. Aim of the study was to assess lymphocytes T function in children with acute lymphoblastic leukemia during remission induction on the grounds of chosen costimulatory and activatory molecules expression. To assess percentages of lymphocytes subpopulations we used tricolor flow cytometry. Results: 1. In the moment of diagnosis and remission induction we noted higher percentages of lymphocytes T with adhesion molecule ICAM-1; 2. During remission induction we observed lower percentage values of lymphocytes T with CD38 coexpression; 3. In the end of remission induction rised the percentage values of lymphocytes T helper with IL-2 receptor expression; 4. In the group of patients with fever/infection we observed higher percentage of activated lymphocytes T (CD3-HLA-DR) comparing to non-infected patients. Summarizing, we suggest lymphocytes T activation during appearance and remission induction of acute lymphoblastic leukemia in children. This confirms participation of cellular immunity in leukemic process.