Evaluation of specific protective immune response in tuberculosis pleurisy

Dünyada önemli bir problem olmaya devam eden tüberküloz (TB)’un kontrolü ve gelecekte eradikasyonu için etkili aşılara, spesifik tanı reaktiflerine; tedavisi için de alternatif immün modülatör yaklaşımların belirlenmesine ihtiyaç vardır. Çalışmamızda, TB aşısı için aday peptidler olarak kabul edilen CFP-10 ve ESAT- 6’nın koruyucu immün cevaptaki rolü ile TB’da lokal koruyucu immün cevapta yer alan sitokin ve mononükleer hücrelerin etkinliği araştırıldı. Bu amaçla TB plörezi (hasta grubu) ve malign plörezili (kontrol grubu) hastalardan elde edilen plevral sıvı mononükleer hücreleri PHA, PPD, CFP-10 ve ESAT-6 ile stimüle edilerek intrasellüler ve ekstrasellüler sitokin düzeyleri (IFN-γ, IL-10, IL-13, IL-4) değerlendirildi. Ayrıca hasta ve kontrol gruplarında plevra sıvısı IFN-γ, IL-12p40, IL-18, IL-8, TNF-α, IL-4, IL-10, IL-13, sFas, sFasL düzeyleri ile mononükleer hücre oranları araştırıldı. Plevra sıvısı mononükleer hücrelerinin CFP-10 ve ESAT-6 ile stimülasyonu sonucu, TB plörezi olgularında ekstrasellüler ve intrasellüler IFN-γ düzeylerinde, kontrol grubuna göre belirgin artış olduğu, antienflamatuvar bir sitokin olan IL-10 düzeylerinin de arttığı saptandı. TB plörezili hastalarda plevra sıvısı IFN-γ, IL-12p40, IL-18, IL-8, TNF-α düzeyleri, kontrol gruplarına göre yüksek bulundu. Elde edilen veriler ışığında TB’daki koruyucu immünitede Th1 tipi spesifik hücresel immünitenin primer yanıt olduğu; ESAT-6 ve CFP-10’un TB plörezili olgularda bu yanıtı endükledikleri; bu peptitlerin TB aşı preparasyonlarında yer almaları sonucu koruyucu etkinliği daha güçlü aşıların geliştirilebileceği; ESAT-6 ve CFP-10’un yer aldığı testlerden BCG’ye bağlı Tüberkülin pozitifliği ile doğal enfeksiyonun ayrımında ve özellikle TB’un endemik olmadığı bölgelerde tanıda yararlanılabileceği; ayrıca plevra sıvısındaki IFN-γ düzeylerinin TB plörezi ve malign plörezi ayırımda kullanılabileceği sonucuna varıldı.There are requirements for effective vaccines and specific diagnostic reagents to control and eradicate Tuberculosis (TB) which remains an important health problem. Alternative immunomodulatory approaches also needs to be developed for better treatment of the disease. In our study, the roles of ESAT and CFP-10, which are candidate peptides for new TB vaccine preparations, and effectiveness of cytokines and mononuclear cells were investigated to determine their involvement in local protective immune response. For this aim, pleural fluid mononuclear cells obtained from TB pleurisy (study group) and malign pleurisy or non-TB non-malign pleurisy (control groups) patients were stimulated with PHA, PPD, ESAT-6 and CFP-10 prior to the measurement of intracellular and extracellular cytokines (IFN-γ, IL-4, IL-10 and IL-13). Additionally, the levels of IFN-γ, IL-4, IL-8, IL-10, IL-12p40, IL-13, TNF-α, sFas and sFasL, and the proportional profiles of mononuclear cell subsets in pleural fluids were evaluated. As a result of stimulation of pleural fluid mononuclear cells with ESAT-6 and CFP-10, extracellular and intracellular levels of IFN-γ dramatically increased in TB pleurisy cases when compared to control groups, and the levels of anti-inflammatory cytokine IL-10 were also determined to be elevated. It was also shown that levels of pleural fluid IFN-γ, IL-8, IL-12p40, IL-18, TNF-α and sFasL were significantly higher in the patients with TB pleurisy in comparison to control groups. In the light of the data obtained in our study it can be suggested that Th1 type specific cellular immunity is the primary response responsible for protective immunity in TB; that more potent vaccines may be developed by including ESAT-6 and CFP-10 in new vaccine preparations since these peptides induce protective response providing selfcontrol of TB pleurisy; that diagnostic tests using ESAT-6 and CFP-10 could be used to distinguish natural infection from Tuberculin test positivity due to previous BCG vaccination and to diagnose TB in the regions where TB is not endemic; and additionally, that the measurements of pleural fluid IFN-γ levels might be used to differ TB pleurisy and malign pleurisy.Uludağ Üniversitesi Araştırma Fonu T-2002/70 numaralı proj

The Senescence Program is Reduced in Proteasome Inhibitor Bortezomib-Resistant PC3 Prostate Cancer Cell Line

Objective: Senescence may act as an antitumor mechanism by preventing the proliferation of cancer cells. Here we investigated the hypothesis that PC3 prostate cancer cells resistant to bortezomib respond differently to proteasomal inhibition with respect to induction of the senescence program as compared to the parental cells. Materials and Methods: The degree of senescence was measured by β-galactosidase activity and the level of senescence-associated p16 INK4a by Western blotting after treatment of cells with varying concentrations of bortezomib. In addition, the senescence-associated secretory phenotype was analyzed by Human Cytokine Antibody Array. Results: It is reported that the basal level of senescence was lower in resistant cells compared to non-resistant cells. It was found that the basal level of the senescence marker p16 INK4a was lower in bortezomib-resistant cells than in parent non-resistant cells. Moreover, p16 INK4a was significantly reduced in both cells under conditions of 100 nM bortezomib treatment, a finding suggesting that the reduced senescence after proteasomal inhibition was likely due to the reduced levels of p16 INK4a. Finally, it is reported here for the first time that basal levels of the proteins NAP2, FGF-6, MIP-3α, and PARC are significantly increased in the resistant cells compared to the parental cells. Conclusion: Overall, the results suggest that inhibition of senescence may play an important function in the development of resistance to bortezomib

The effects of Spirulina platensis (Arthrospira platensis) and Saccharomyces cerevisiae on the distribution and cytokine production of CD4+and CD8+T-lymphocytes in rabbits

Natural additives have become one of the most alternative immune enhancer nowadays. In particular, Spirulina platensis (Arthrospira platensis) (SP) and Saccharomyces cerevisiae (SC) have been used for improving the immune system and quality of life. The aim of this study was to regulate the immune effect of S. cerevisiae and S. platensis (A. platensis) combination. Forty male New Zealand white rabbits, aged 5-6 weeks, were analysed in 4 groups: Control (basal diet); SC (added 3 g/kg diet), SP (added 5% of the diet); SC and SP (added 3 g/kg SC and 5% SP of the diet). The entire experiment lasted 90 days. Blood samples were obtained by ear venipuncture on the 90th day. The CD4+ and CD8+ T lymphocyte values were determined by flow cytometry and cytokines (IFN-gamma and IL-4) were determined by ELISA. According to the results, there were no significant differences in the expression of cytokines, but serum CD4+/CD8+ increased in the animals fed SP and SC+SP supplemented diets (3 g/kg and 5% of the diet, respectively). It was concluded that S. platensis (A. platensis) may be used as an immune enhancer, although further studies are needed to clarify the effects of spirulina supplement on immunity.Research Foundations of Uludag UniversityUludag University [2011/19]This study was supported by a grant from the Research Foundations of Uludag University (Project number: 2011/19). Also, this manuscript was originated from the PhD thesis of Nilay Seyidoglu

Cytokine signal suppressor (SOCS) 1-1478 CA/del gene polymorphism in Turkish patients with polycystic ovary syndrome

Eighty-four subjects, premenopausal female patients (n = 42, mean (SD) age: 26.4 (4.2) years) diagnosed with polycystic ovary syndrome (PCOS) and age-matched healthy volunteers (n = 42, mean (SD) age: 27.6(3.4) years), were included in this study. Data on physical examination, anthropometric measurements and blood biochemistry analysis were recorded for each subject along with analysis for SOCS1-1478 CA/del polymorphism by polymerase chain reaction-restriction fragment length polymorphism. The relation of SOCS1-1478 CA/del polymorphism to PCOS status and insulin resistance was analysed via logistic regression analysis. Mean (SD) levels for BMI (28.5(6.5) vs.22.5 (4.9) kg/m2, p < .001), HOMA-IR (3.1(1.8) vs.1.5 (1.0), p < .001), LDL-cholesterol (115.9(32.7) vs.100.7 (27.3)mg/dL, p = .03) and triglyceride (113.8(64.9) vs.83.3(36.3)mg/dL, p = .017) were significantly higher in patients. Groups were similar in terms of SOCS1-1478 CA/del polymorphism. No significant relation of this polymorphism was noted to PCOS and HOMA-IR. Our findings revealed no difference between groups in terms of the rate of SOCS1-1478 CA/del polymorphism, and no significant relation of this polymorphism to insulin resistance and PCOS status.Impact statement Polycystic ovary syndrome (PCOS), the most common cause of anovulation and the most commonly encountered form of female endocrine disease. SOCS proteins have been suggested to play a fundamental role in the negative feedback regulation of the JAK-STAT pathway, which is the major signalling pathway involved in a wide range of physiologic and pathologic processes, including inflammatory diseases, malignancies and immune disorders. Pathways involving the induction of suppression of SOCS proteins were also shown likely to be involved in mediating cytokine-induced insulin resistance. The present study was designed to determine the frequency of SOCS1-1478 CA/del gene polymorphism in patients with PCOS in relation to healthy controls and insulin resistance. Our findings revealed significantly higher rates of insulin resistance, obesity and dyslipidaemia in Turkish patients with PCOS compared with age-matched healthy controls, while no difference between study groups in terms of the rate of SOCS1-1478 CA/del polymorphism along with no significant relation of SOCS1-1478 CA/del polymorphism to insulin resistance and PCOS status. Future larger scale studies with the application of standardised diagnostic methods and criteria, and of state-of-the-art modern techniques including genomics, proteomics and pharmacogenetics would provide better understanding of the association between PCOS and genomic variants

Altered Expressions of miR-1238-3p, miR-494, miR-6069, and miR-139-3p in the Formation of Chronic Brucellosis

Brucellosis is a zoonotic disease that is still endemic in developing countries. Despite early diagnosis and treatment of patients, chronic infections are seen in 10–30% of patients. In this study, we aimed to investigate the immunological factors that play roles in the transition of brucellosis from acute infection into chronic infection. Here, more than 2000 miRNAs were screened in peripheral blood mononuclear cells (PBMCs) of patients with acute or chronic brucellosis and healthy controls by using miRNA array, and the results of the miRNA array were validated through qRT-PCR. Findings were evaluated using GeneSpring GX (Agilent) 13.0 software and KEGG pathway analysis. Four miRNAs were expressed in the chronic group but were not expressed in acute and control groups. Among these miRNAs, the expression level of miR-1238-3p was increased while miR-494, miR-6069, and miR-139-3p were decreased (p<0.05, fold change > 2). These miRNAs have the potential to be markers for chronic cases. The differentially expressed miRNAs and their predicted target genes involved in endocytosis, regulation of actin cytoskeleton, MAPK signaling pathway, and cytokine-cytokine receptor interaction and its chemokine signaling pathway indicate their potential roles in chronic brucellosis and its progression. It is the first study of miRNA expression analysis of human PBMC to clarify the mechanism of inveteracy in brucellosis