Significance of CMV-seropositivity in the alterations of CD4+ T-cell subsets and expression of their phenotyping markers in B-cell chronic lymphocytic leukemia

زمینه و هدف: لوسمی لنفوسیتی مزمن سلول B (B-CLL) یک بیماری بدخیم لنفوسیت های B است که همراه با تغییرات متعددی در لنفوسیت های T غیر بدخیم می باشد. با توجه به اینکه آنتی ژن های ویروسی می توانند تغییرات قابل توجهی در لنفوسیت های T ایجاد نمایند و سیتومگالوویروس (CMV) در افراد پیر سالم باعث تغییرات گوناگون در لنفوسیت های T بوده است، لذا این مطالعه با هدف بررسی ارتباط بین تغییرات این سلول ها و CMV در بیماران CLL انجام شده است. روش بررسی: در این مطالعۀ توصیفی -تحلیلی 79 بیمار مبتلا به B-CLL وارد مطالعه شدند (41 بیمار CMV+ و 38 بیمار CMV-). بررسی زیر گروه های لنفوسیت های T و تعیین فنوتیپ سلول ها با استفاده از آنتی بادی های منوکلونال و فلوسایتومتری انجام شد. ترشح سیتوکسین ها پس از تحریک آنتی ژنیک و کشت کوتاه مدت و سپس با روش رنگ آمیزی داخل سلولی سیتوکین ها تعیین گردید. از آزمون آماری من ویتنی جهت مقایسه دو گروه استفاده شد. یافته ها: در بیماران CMV+، زیر گروه لنفوسیت های CD8+ (Cluster of differentiation) و CD4+ CD8dim به صورت معنی داری نسبت به بیماران CMV- بیشتر بود (001/0

CD4+ immune response to cytomegalovirus (CMV) in healthy carriers and hematological malignancies

Background and aim: Cytomegalovirus (CMV) infects the majority of human population in their life time and triggers strong immune responses from all arms of the immune system. However the cellular immune response is the major mechanism by which CMV replication is controlled. CMV-specific CD4+ T cells have a substantial role in maintenance of CMV-specific CD8+ T cell response. The aim of the study was an overview of CD4+T cell response to CMV in healthy donors and patients with hematological malignancies. Methods: In this review, abstract or full text articles related to CMV-specific CD4+ T cell response, published during 1990 until 2010, were collected from the Medline. The Persian articles were searched through the IranMedex database and used if they were appropriate. Results: In chronic infection, the CMV-specific CD4+ T cells secrete interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α) and interleukin-2 (IL-2). CMV-specific CD4+ response increases with age and the response has been up to 32% with intracellular cytokine detection technique. The cells have also lower activation threshold. The CMV-specific CD4+ T cell response increases and comprises up to 47% of whole CD4+ compartment in patients who received hematopoietic stem cells transplants. It was up to 44% in chronic lymphocytic leukemia patients and broad phenotyping alterations have also been observed in relation to CMV-seropositivity. Conclusion: Considering the high level of CMV-specific CD4+ T cell response, the viral replication can be controlled and the reactivation can be prevented. Because of the possibility of intracellular cytokine, it is now possible to determine the phenotype of the cells. Therefore, CMV has served as an excellent model for effectors-memory phenotype studies and could be a possible tool in the way to achieve immunotherapy

Evaluation of T cell phenotype and function in 10-30 years old individuals and the correlation of its alteration with age

زمینه و هدف: ایمنی همراه با افزایش سن تغییر می کند که در نهایت به پیری ایمنی منجر می شود. این پدیده در لنفوسیت های T با تغییرات فتوتیپی و عملکردی همراه است. هدف از مطالعه حاضر بررسی فتوتیپ لنفوسیت های T و تعیین داخل سلولی IL2 و IFNγ در یک گروه از افراد سالم و همبستگی تغییرات آن ها با سن بود. روش بررسی: در این مطالعه توصیفی،خون از 26 فرد سالم گرفته شد تعیین IL2 و IFNγ داخل سلولی به شیوه تعیین سایتوکین ها انجام شد. فتوتیپ ایمنی لنفوسیت های با فلوسایتومتری بررسی گردید. داده های به دست آمده از شمارش کامل خونی و سایر داده های موجود با آمار توصیفی و آزمون همبستگی اسپیرمن تجزیه و تحلیل گردید. یافته ها: نسبت لنفوسیتT 87/14±81/54، تعداد لنفوسیت CD4 5/294±15/644 و تعداد لنفوسیتTCD8 ، 44/155±04/417 در میلی متر مکعب بود. میزان لنفوسیت های CD4 بیان کننده هر یک از شاخص های CD27, CD28, CD57, CCR7 و نسبت های سلول های تولید کننده IL2 و IFNγ بیش از میزان آن ها در زیر گروه CD8 بود. از شاخص های بررسی شده تنها بین میزان سلول های CD8+CCR7+ و سن همبستگی منفی معنی دار دیده شد. نتیجه گیری: یافته های مطالعه نشان می دهد که تغییرات فتوتیپی مرتبط با کاهش لنفوسیت بکر و خاطره ای مرکزی و افزایش لنفوسیت های خاطره ای اجرایی (که همراه با پیری ایمنی است) ظاهراً در دوران جوانی با سن همبستگی معنی دار ندارد. برای داشتن اطلاعات فراگیرتر، مطالعه فتوتیپ و عمل سیستم ایمنی در گروه های مختلف جوان و پیر پیشنهاد می گردد

Association of homozygous thrombophilia polymorphisms and venous thromboembolism in Shahrekord, Iran

Background: Venous thromboembolism (VTE) is a major cause of mortality. Factor V Leiden (FVL), methylenetetrahydrofolate reductase (MTHFR) C677T, and prothrombin (FII) G20210A polymorphisms are the main inherited risk factors for VTE. Since evidence is limited on homozygotes, the aim of this study was to investigate the association between homozygous variants of these polymorphisms and VTE in Shahrekord, southwest Iran. Materials and Methods: In this case-control study, blood samples of 72 VTE patients admitted to Hajar Hospital, Shahrekord and 306 sex- and age-matched healthy volunteers as controls were taken in EDTA Vacutainers. The polymorphisms of FVL, MTHFR C677T, and FIIG20210A were investigated by PCR-RFLP. The data were analyzed by descriptive statistics and independent t-test. Results: The frequency of all homozygous polymorphisms was found to be 16.77% in patients and 4.90% in controls with a significant difference (P=0.004). Homozygous FVL mutation was more frequent in patients than in controls with no significant difference. Regarding the frequency of homozygous MTHFR C677T, a significant difference was noted between patients and controls (P=0.03). There was no significant difference in homozygous FIIG20210A and heterozygous variants of the above-mentioned polymorphisms between the patients and controls. Conclusion: Homozygous MTHFR C677T polymorphism is associated with VTE in Shahrekord. Control of the acquired risk factors may be necessary in homozygous form of this polymorphism. VTE patients with this polymorphism may need to be managed differently. © 2016 NRITLD

Alteration of T cell subtypes in beta-thalassaemia major: Impact of ferritin level

Introduction: Oxidative damage and regular antigenic stimulation are main factors in accelerating immunosenescence. The present study was conducted to investigate new concepts of early immunosenescence in thalassaemia patients. aterials and Methods: Twenty seven beta-thalassaemia major patients and a group of matched healthy volunteers aged 10-30 years in Shahrekord, Iran were recruited into the study. Ferritin level was determined and CD4 or CD8 T cells were analysed versus phenotyping markers, CD27, CD28, CD57 and CCR7, by flowcytometry. Data were analysed by Mann-Whitney and Spearman’s correlation coefficient test in SPSS 11.5. Results: Absolute lymphocytosis and partial decrease in T cells were observed in the patients. CD4+CD57+ and CD4+CCR7- T cells were significantly higher, whereas CD8+CD27+ and CD8+CCR7+ T cells were partially higher in patients. A negative correlation was observed between ferritin level and number of CD8+CD27+ and CD8+CCR7+ T cells, whereas the correlation was positive between ferritin level and number of CD57+ T cells. Conclusion: Moderate alteration of T cell repertoire and increase in CCR27-, CCR7-, and CD57+ T cells could reflect antigenic stimulation, decline in naïve T cells, and being closer to terminally differentiated cells. Effect of iron overload is potentially explained by positive correlation of blood transfusion and ferritin level with frequency of CD3+CD27- and that of ferritin with frequency of CD57+ T cells

Association between fibrinogen receptor (Glycoprotein IIb) polymorphism and the risk of venous thromboembolism: a systematic review

Background: The fibrinogen receptor is an integrin on the platelet surface and is shaped from two types of glycoprotein (GP) subunits, GPIIb and GPIIIa. Membrane glycoprotein IIb/IIIa plays an important role in platelet function. The gene encoding the glycoprotein IIIa shows a common polymorphism, PLA2 that increases the binding of the receptor to fibrinogen and enhances the platelet aggregation. The clinical impact of PLA2 polymorphism has been studied in some diseases, but the definition of its exact role on venous thromboembolism complications has been challenging. The present systematic review aimed to clarify the association of PLA2 polymorphism and venous thromboembolism. Main text: In this study, Electronic databases including PubMed, Embase, Scopus, Web of Science, and Cochrane Library were searched. All the assessed studies focused on the relationship between PLA2 polymorphism and venous thromboembolism. Five studies were eligible for systematic review. One study revealed a significant correlation between PLA2 polymorphism and venous thromboembolism. PLA2 polymorphism was associated with deep vein thrombosis in one study and pulmonary thromboembolism in another one. Conclusion: The published data supported the hypothesis that having the PLA2 polymorphism of GPIIIa may be a risk factor for venous thromboembolism, but the association cannot be concluded; it needs more clinical investigation Keywords: Venous thromboembolism, Deep vein thrombosis, Pulmonary embolism, PLA2 polymorphis

Association between homozygous Methylene Tetrahydrofolate Reductase and Platelet PLA2 Antigen Polymorphisms with Venous Thromboembolism (VTE) in Shahrekord

Materials and Methods: In this cross-sectional study, EDTA venous blood was taken from 72 patients with venous thromboembolism referred to Shahrekord Hajar Hospital, and 306 age and sex matched healthy volunteers. Genotyping of factor V Leiden, MTHFR C677T and PLA 2 polymorphisms was done by PCR-RFLP. Statistical analysis was done using descriptive statistics and χ 2 test. Results: There was a significant difference between patients and controls in the number of homozygous polymorphisms with the frequency of ..

Antiproliferative and Apoptosis-Inducing Activities of Thymoquinone in Lymphoblastic Leukemia Cell Line

Acute lymphoblastic leukemia is one of the malignant proliferations of lymphoid cells in the early stages of differentiation and accounts for about 80% of all cases of childhood leukemia. Side effects of available treatment are still main concern. Thymoquinone (TQ), a natural compound isolated from Nigella sativa, induces growth inhibition and apoptosis in several cancer cell lines. The aim of the present study was to investigate the effect of TQ alone and in combination with doxorubicine on the proliferation inhibition and apoptosis induction of TQ in a lymphoblastic leukemia cell line. Jurkat cell line was cultured in standard condition and with concentrations of TQ (0–30 μm) and doxorubicine for 24, 48 and 72 h. Cell viability was measured by MTS assay. Apoptosis induction by TQ was assessed by annexin V-FITC/PI and flow cytometry analysis. TQ and DOX decreased cell viability with a time and dose dependent manner. The IC50 values were 19.461 ± 1.141, 17.342 ± 1.949 and 14.123 ± 1.874 μM in 24, 48 and 72 h, respectively for TQ. IC50 values for DOX were. 075 ± .0124, .028 ± .007 and.007 ± .001 μM in 24, 48 and 72 h, respectively. The level of cell apoptosis in all used concentrations of TQ (4, 8, 12, 16 and 20 μm) was higher than control group (10.2, 14.1, 36.6, 87.5 and 93.3% respectively after 24 h; 10.7, 13.9, 64.6, 92.2 and 93.1 respectively after 48 h; 2.83, 5.83, 41.4, 71.6 and 86.6% respectively after 72 h) and reached to a significant level at 12, 16 and 20 μm concentration for 24 and 48 h and 16 and 20 μm for 72 h incubation. Combination of doxorubicine and TQ lead to a synergistic cytotoxicity as compared to any of them alone. The study indicated that TQ is effective on proliferation inhibition and is a strong apoptotic inducer in Jurkat lymphoblastic cell line and has synergistic effect in combination with DOX. This combination strategy can be an alternative way for more powerful anticancer effects. Therefore, the study of the mechanism of apoptosis induction of TQ can be a step forward to in target therapy which might be considered in the future studies. © 2016 Indian Society of Haematology & Transfusion Medicin

Gallic Acid Inhibits Proliferation and Induces Apoptosis in Lymphoblastic Leukemia Cell Line (C121)

Leukemia is known as the world's fifth most prevalent cancer. New cytotoxic drugs have created considerable progress in the treatment, but side effects are still the important cause of mortality. Plant derivatives have been recently considered as important sources for the treatment of various diseases, including cancer. Gallic acid (GA) is a polyhydroxyphenolic compound with a wide range of biological functions. The aim of the present study was to evaluate the effect of GA on proliferation inhibition and apoptosis induction of a lymphoblastic leukemia cell line. Jurkat cell (C121) line was cultured in RPMI 1640 supplemented with 10% heat-inactivated fetal bovine serum (FBS) with different concentrations of GA (10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 mu M) for 24, 48 and 72 hours. The effect of GA on cell viability was measured using MTS assay. Induction of apoptosis was evaluated with Annexin V-FITC/PI kit and flow cytometry. Data were analyzed by SPSS version 20 using Kruskal-Wallis and Dunn's multiple comparison tests. Decline of cell viability to less than 50% was observed at 60.3+/-1.6, 50.9+/-1.5, and 30.9+/-2.8 mu M concentration after 24, 48, and 72 hours incubation, respectively. All concentrations of GA (10, 30, 50 and 80 mu M) enhanced apoptosis compared to the control (P<0.05). The results demonstrate that the polyphenolic compound, GA, is effective in inhibition of proliferation and induction of apoptosis in Jurkat cell line. It is recommended to study the mechanism of apoptosis induction in future investigations

The Inhibitory Effect of Epigallocatechin Gallate on the Viability of T Lymphoblastic Leukemia Cells is Associated with Increase of Caspase-3 Level and Fas Expression

Acute lymphoblastic leukemia is the most prevalent cancer in children. Novel components to help struggle aggressive malignancies and overcome some side effects of conventional treatments could be a promising strategy. Epigallocatechingallate (EGCG), have attracted the attention of scientists for prevention or treatment of some cancers. Jurkat cells were incubated with the different concentrations of EGCG (30–100 µm) for 24, 48, and 72 h and cell viability was investigated using MTS test. Apoptosis and the level of caspase 3 alterations were evaluated using flowcytometry and expression of Fas by Real Time PCR. EGCG decreased viability of cells with an inhibition concentration (IC50) of 82.8 ± 3.1, 68.8 ± 4 and 59.7 ± 4.8 μM in 24,48 and 72 h. 50, 70 and 100 µM concentrations of EGCG induced apoptosis in about 31, 40 and 71% of the cells, respectively. The mean value of caspase 3 positive cells in the presence of 50, 70 and 100 µm concentrations of EGCG was 19.3 ± 2.9, 29.5 ± 3.1 and 61.2 ± 3.4 respectively compared to 7.8 ± 1.1 in control with a significant difference at 100 µm concentration. Treatment with EGCG for 48 h enhanced the expression of Fas reaching to a significant level at 100 µM concentration. EGCG is effective in decrease cell viability, apoptosis induction and enhancement of caspase 3 and Fas expression level in jurkat cells. A comprehensive understanding of molecular events and pharmacokinetics of the component and experiments in animal models are required for dose determination and its interaction with other components of combination chemotherapy