Group Of Healthy Individuals Research Articles

TNF-α -308A/G SNP association with kidney allograft rejection in Algerian population: A retrospective case-control study

No consensus has been reached regarding the association beween the -308A/G single nucleotide polymorphism (SNP) in the tumor necrosis factor-α gene (TNFA) and kidney allograft rejection (KAR). Our retrospective case-control study aimed to assess the association of the SNP with KAR in Algerian patients who underwent kidney transplantation. The study enrolled 313 Algerian patients: 58 kidney-transplant recipients without rejection events (PWoR); 58 kidney-transplant recipients with at least one rejection event, with or without graft loss (PWR); and 197 healthy individuals (HI). The TNFA -308A/G SNP was genotyped using a real-time polymerase chain reaction. The results demonstrated that, the frequencies of TNFA -308A allele and AA genotype were higher in the PWR than in the HI groups (p = 0.001, OR = 2.26, CI = 1.33‐3.77 and p = 0.0004, OR = 5.53, CI-1.89-16.6, respectively). Furthermore, the frequencies were higher among the PWR than among the PWoR groups (p = 0.001, OR = 3.29, CI = 1.56‐7.21 and p = 0.0006, OR = 28.26, CI = 1.62‐493.2, respectively), particularly among PWR patients with de novo anti-human leukocyte antigens (HLA) antibodies (PG-a-HLA-Ab). However, the frequency of TNFA -308G allele was lower in the PWR group than in the PWoR group (p = 0.001, OR = 0.3, CI = 0.1‐0.64) and the HI group (p = 0.001, OR = 0.44, CI = 0.27‐0.44). Our results suggest an association of the TNFA -308A/G alleles with KAR in Algerian patients who underwent kidney transplantation. Carriers of TNFA -308A allele who have PG-a-HLA-Ab might have a higher risk, whereas TNFA -308G allele carriers could have a lower risk of KAR. Thus, therapeutic strategies can be adapted to minimize KAR risk in patients who have a genetic proclivity for increased pro-inflammatory TNF-α activity.

Body composition and energy expenditure in anorexia nervosa: preliminary data of outpatients with recovering and active disease

BackgroundIn Anorexia Nervosa (AN) recovery, body mass index (BMI) may not reflect body composition. To investigate recovery, bioelectrical impedance (BIA) parameters and energy expenditure were investigated in patients with active and recovering AN, with emphasis on phase angle (PA), a BIA parameter.MethodsBMI, PA, indirect BIA parameters (fat free mass, fat mass, total body water, fat free mass index, fat mass index) and resting metabolic rate (RMR) were obtained. Data from subjects distributed to active AN (ANact, n = 9), recovered AN (ANrec, n = 9) and healthy individuals (HI) (n = 16) were compared employing univariate methods and ordinal logistic regression.ResultsIn univariate comparison, the BMI would not distinguish recovered individuals; this distinction was observed for the PA (p = < 0,001). PA showed a good capacity to discriminate, between ANrec and HI (AUC = 0.792; CI = 0.564- 1.000; p = 0.017). In 2 models of ordinal logistic regression PA (OR = 0.123; 95% CI 0.030; 0.503 and OR = 0.091; 95% CI 0.016; 0.528) remained as a significant independent variable, indicating that increases in PA are related to higher probabilities of moving from ANact, to ANrec and to HI group. Bivariate regression indicated the presence of a relationship between PA and (R2 = 0.266, p = 0.002).ConclusionsChanges in body composition and energy expenditure were observed in recovered anorexics with normal BMI. PA can play an important role in the assessment of recovering anorexic patients.

FEATURES OF ARTERIAL PRESSURE IN SCHOOLCHILDREN WITH ACUTE OBSTRUCTIVE BRONCHITIS

Objective – to assess blood pressure indicators in children with acute obstructivebronchitis.Materials and methods. The paper presents the results of a survey of 68 childrenhospitalized for acute obstructive bronchitis. Patients were examined for clinical symptomsand severity of the clinical course according to the BSS-ped scale. BP parameters andechocardiography data were determined and compared with the results of a control groupof healthy individuals (20 children).Results. In patients, the severity of bronchitis clinical course on the fi rst day was average –according to the BSS scale – 7.79±0.19 points. BP indicators in patients were probablylower than in healthy children and had an inverse correlative dependence on the severityof the disease. Echocardiography showed a decrease in the end systolic and diastolicvolume of the left ventricle, a decrease of the pressure gradient in the mitral valve and itsincrease in the pulmonary valve.Conclusions. In acute obstructive bronchitis in children, changes in the activity of thecardiovascular system are observed due to a decrease in systemic blood pressure and anincrease of pressure in the pulmonary circulation system.

Clinical Proteomics Profiling for Biomarker Identification Among Patients Suffering With Indian Post Kala Azar Dermal Leishmaniasis.

Background: Post Kala Azar Dermal Leishmaniasis (PKDL) is a non-fatal dermal sequel of Visceral Leishmaniasis (VL), affecting individuals worldwide. Available diagnostic tools lack sensitivity and specificity toward identifying macular (MAC) PKDL patients, due to low parasite load in patients' sample. Confirmatory test like punch biopsy are invasive and painful. Considering the rural nature of this disease and the prevailing situation of diagnostic scenario, PKDL patients mostly remains unattended from receiving proper medical care. They in turn act as “mobile parasite reservoir,” responsible for VL transmission among healthy individuals (HI). This study aims to identify PKDL disease specific glycated protein biomarkers, utilizing the powerful LC-MS/MS technology, which is the tool of choice to efficiently identify and quantify disease specific protein biomarkers. These identified PKDL disease specific novel glycoproteins could be developed in future as immunochromatographic based assay for efficient case detection.Methodology: Previously our lab had identified importance of glycated (Circulating Immune Complexes) CICs, among PKDL patients. This study aims to further characterize disease specific glycated protein biomarkers, among MAC PKDL patients for both diagnostic and prognostic evaluation of the disease. LC-MS/MS based comparative spectral count analysis of MAC PKDL to polymorphic (POLY) PKDL, HI, and Cured (CR) individuals were performed. Proteins level alterations among all study groups were confirmed by Western blot and enzyme-linked immunosorbant Assay (ELISA).Results: Among MAC PKDL patients 43, 60, 90 proteins were altered compared to POLY PKDL, HI, and CR groups, respectively. Filtering for the most significant proteins, Plasminogen (PLG) and Vitronectin (VTN) were identified which promisingly identified MAC PKDL cases. Active surveillance results from endemic districts of West Bengal revealed drastic rise of MAC PKDL cases, alarming the urgency for field adaptive efficient biomarker.Conclusion: This current study aims to establish PLG and VTN as novel diagnostic and prognostic protein biomarker for MAC and POLY PKDL cases management.

Contribution of Leishmania braziliensis antigen-specific CD4+ T, CD8+ T, NK and CD3+CD56+NKT cells in the immunopathogenesis of cutaneous leishmaniasis patients: Cytotoxic, activation and exhaustion profiles.

The pathogenesis of cutaneous leishmaniasis (CL) caused by Leishmania (Viannia) braziliensis is dictated mainly by the immune-mediated-tissue inflammation developed. The understanding of the immunological mechanisms that generate tissue damage or resolution of lesions is the key to the development of effective vaccine protocols and proper therapeutic schemes. It is clear that the specific immune response mediated by T cells is responsible for the beneficial outcome of the disease, however, the roles of CD4+ T, CD8+ T, NK and NKT cell subpopulations in immunopathogenesis of CL need to be elucidated. Peripheral blood cells from patients before, during and after the antimonial therapy, as well as healthy individuals (HI) were cultured with (LbAgS) or without (NS) L. braziliensis antigens (LbAg). Afterwards, the frequencies of LbAg-specific-cytotoxic CD8+ T, CD4+ T, NK and CD3+CD56+ NKT cells, as well as their activation and exhaustion profiles, were defined by flow cytometry. We observed higher frequencies of CD8+ T, NK and CD3+CD56+ NKT cells and lower frequencies of CD4+ T lymphocytes in LbAgS cell cultures from patients before treatment. The specific response to LbAg resulted in an expansion of cytotoxic-activated CD4+ T, CD8+ T, and NK cells, before and during treatment, indicating specificity in the response by these cells against L. braziliensis. Furthermore, comparing the differences of frequencies of cytotoxic-activated CD4+T, CD8+T, and NK cells, among before and during treatment patients and HI groups, we conclude that these cell populations are in charge of immune response elicited by antimonial therapy. Interestingly, we also observed that NK cells were induced by LbAg to an exhaustion profile during all clinical stages of the disease. The increased antigen-specific activation and cytotoxic activity are in line with the strong inflammatory response described in this disease, a likely cause of tissue damage. These findings reinforce the involvement of these distinct cytotoxic-activated cell populations in the immunopathogenesis of CL, showing a character of specificity in this immune response.

Pentraxin-2 is Associated with Renal Fibrosis in Patients Undergoing Renal Biopsy

OBJECTIVES:Progressive renal disease is characterized by histological changes in the kidney and fibrosis is a common outcome. Renal biopsy is the only diagnostic tool to evaluate these histopathological changes. Pentraxin-2 (PTX-2) is an anti-inflammatory constitutive plasma protein associated with the innate immune system. Recently, as a biomarker, the circulating level of PTX-2 is shown to be decreased in chronic fibrotic diseases. In this study, we aimed to investigate the relationship between renal fibrosis severity and serum PTX-2 levels in patients undergoing renal biopsy.METHODS:This cross-sectional study included 45 patients and 16 healthy individuals (HIs). The severity of renal fibrosis was evaluated according to the Banff and Sethi scoring systems by the same pathologist. PTX-2 was measured by an enzyme-linked immunosorbent assay and compared with the demographical, clinical, biochemical, and histopathological data of the patients and HIs.RESULTS:PTX-2 levels were lower in the biopsy group than in the HI group (p=0.12). Patients with moderate renal fibrosis had significantly lower serum PTX-2 levels than those in patients with minimal and mild fibrosis (p=0.017 and p=0.010, respectively). PTX-2 concentrations were correlated with serum albumin (r=0.30, p=0.016), and were negatively correlated with serum creatinine levels (rho=-0.42, p=0.01) and body mass index (r=-0.32, p=0.011).CONCLUSIONS:The results indicated that PTX-2 levels are significantly lower in patients with renal fibrosis than HIs, and declining further in patients with severe fibrosis.

CHARACTERISTIC OF TIM-3 EXPRESSION IN T CELL SUBSETS IN PATIENTS WITH IMMUNE THROMBOCYTOPENIC PURPURA

T cell dysregulation is a common event involved in immune thrombocytopenic purpura (ITP). Recent findings indicated that aberrant expression of immune checkpoints may be related to autoimmune disease pathogenesis as well as in ITP. However, the distribution of Tim-3 and its co-expression with exhausted phenotype in T cell subsets remain unclear. In this study, the frequency of T-cell immunoglobulin mucin 3 (TIM-3) expression and co-expression with CD244 and CD57 in CD3+, CD4+ and CD8+ T cells were evaluated in peripheral blood (PB) from 16 cases with ITP and 21 healthy individuals (HI). Overall, the frequency of Tim-3+CD3+, CD4+ and CD8+T cells in ITP is similar to HI. However, Tim-3+CD244+CD8+ and Tim-3+CD57+CD8+T cells were increased in ITP in compared with HI group. While the percentage of CD244+ and CD57+CD3+, CD4+ or CD8+T cells was similar between ITP and HI groups. Moreover, the Tim-3+CD244+CD3+ and Tim-3+CD244+CD8+ T cells in newly diagnosed ITP group were higher than that in relapsed ITP group, similar trends showed in Tim-3+CD57+CD3+T cells. Obviously, relative higher Tim-3+T cells in most T cell subsets were detected in a case with acute ITP with viral infection. The distribution of Tim-3+ T cells in different subsets was also compared between PB and bone marrow (BM). High trends of Tim-3+T cells was detected in BM group. In conclusion, we firstly characterized T cell immune disorder with higher level of Tim-3+exhausted+CD8+T cells in ITP, the effect and mechanism of such alteration in ITP is needed further investigation.

Diagnostic manifestations of total hemispheric glucose metabolism ratio in neuronal network diaschisis: diagnostic implications in Alzheimer's disease and mild cognitive impairment.

We tested the hypothesis that lateralized hemispheric glucose metabolism may have diagnostic implications in Alzheimer's disease (AD) and mild cognitive impairment (MCI). We performed FDG-PET/CT in 23 patients (mean age 63.7 years, range 50-78, 17 females) diagnosed with AD (n = 15) or MCI (n = 8) during a six-month period in 2014. Ten neurologically healthy individuals (HIs) (mean age 62.5 years, range 43-75, 5 females) served as controls. A neuroimaging expert provided visual assessment of diaschisis. The total hemispheric glucose metabolism ratio (THGr) was calculated, and with area-under the curve of receiver operating characteristics (AUC-ROC) we generated a "Network Diaschisis Test (NDT)". The qualitative detection of cerebral (Ce) and cerebellar (Cb) diaschisis was 7/15 (47%), 0/8 (0%), and 0/10 (0%) in AD, MCI, and HI groups, respectively. Median cerebral THGr was 0.68 (range 0.43-0.99), 0.86 (range 0.64-0.98), and 0.95 (range 0.65-1.00) for AD, MCI, and HI groups, respectively (p =0.04). Median cerebellar THGr was, respectively, 0.70 (range 0.18-0.98), 0.70 (range 0.48-0.81), and 0.84 (range 0.75-0.96) (p = 0.0138). A positive NDT yielded a positive predictive value of 100% for the presence of AD or MCI and a 86% negative predictive value for healthy brain. Moreover, the diagnostic manifestation of THGr between MCI and AD led to a positive predictive value of 100% for AD, but a negative predictive value of 42.9% for MCI. Patients with AD or MCI had more pronounced diaschisis, lateralized hemispheric glucose metabolism and lower THGr compared to healthy controls. The NDT distinguished AD and MCI patients from HIs, and AD from MCI patients with a high positive predictive value and moderate and low negative predictive values. THGr can be a straightforward source of investigating neuronal network diaschisis in AD and MCI and in other cerebral diseases, across institutions.

Plasma levels of soluble NCAM in multiple sclerosis

In multiple sclerosis (MS), several adhesion molecules are involved within the central nervous system in inflammatory and neurodegenerative processes that are associated to progressive disability and increasing brain atrophy. The neural cell adhesion molecule (NCAM) has been suggested to participate in the reparative mechanisms and in the remyelination processes, key issues in MS pathology. We aimed at investigating plasma levels of the seldom investigated soluble (s)NCAM, and as comparison those of intercellular adhesion molecule-1 (sICAM-1) and vascular adhesion molecule-1 (sVCAM-1), and their association with clinical and MRI measures of lesion volumes and of global and regional atrophy. The cross-sectional study was conducted in 85 relapsing-remitting (RR)-MS, 53 progressive (P)-MS patients, and 42 healthy individuals (HI).Correlation of MRI measures with plasma levels of these adhesion molecules were not observed.In the MS and HI groups, sNCAM levels were significantly and positively associated with sVCAM-1 levels. Differently, the correlation between sICAM-1 and sVCAM-1 was observed only in MS patients. sNCAM and sVCAM-1 levels were higher in P-MS compared to HI (P = 0.05 and P = 0.028 respectively). The sVCAM-1 levels differed (P < 0.001) among DMTs groups and HI.The association of sNCAM plasma levels with MS disease, as well as differences in sVCAM-1 levels in patients receiving different DMTs, deserve further investigation.

Association of Cytokines with Alanine Aminotransferase, Hepatitis B Virus Surface Antigen and Hepatitis B Envelope Antigen Levels in Chronic Hepatitis B

Background:Cytokines play an important role in occurrence and recovery of hepatitis B virus (HBV) infection. The aim of this study was to investigate the changes of cytokines concentration and its correlation to alanine aminotransferase (ALT), HBV deoxyribonucleic acid (HBV-DNA), hepatitis B envelope antigen (HBeAg), and HBV surface antigen (HBsAg) in the development of chronic hepatitis B (CHB).Methods:Thirteen healthy individuals (HI), 30 chronic HBV-infected patients in immune tolerant (IT) phase, and 55 CHB patients were enrolled between August 2015 and May 2017. The peripheral blood samples were collected from all individuals. The levels of interferon (IFN)-α2, interleukin (IL)-10, transforming growth factor (TGF)-β1, HBV-DNA, HBsAg, and HBeAg and liver function were measured. The quantitative determinations of cytokines levels, including IFN-α2, IL-10, and TGF-β1 were performed using Luminex multiplex technology. The correlation of cytokines to ALT, HBV-DNA, HBsAg, and HBeAg was analyzed by linear regression analysis.Results:IFN-α2 levels were similar between HI and IT groups (15.35 [5.70, 67.65] pg/ml vs. 15.24 [4.07, 30.73] pg/ml, Z = −0.610, P = 0.542), while it elevated significantly in CHB group (35.29 [15.94, 70.15] pg/ml vs. 15.24 [4.07, 30.73] pg/ml; Z = −2.522, P = 0.012). Compared with HI group (3.73 [2.98, 11.92] pg/ml), IL-10 concentrations in IT group (5.02 [2.98, 10.11] pg/ml), and CHB group (7.48 [3.10, 18.00] pg/ml) slightly increased (χ2= 2.015, P = 0.365), and there was no significant difference between IT and CHB group (Z = −1.419, P = 0.156). The TGF-β1 levels among HI (3.59 ± 0.20 pg/ml), IT (3.62 ± 0.55 pg/ml), and CHB groups (3.64 ± 0.30 pg/ml) were similar (χ2= 2.739, P = 0.254). In all chronic HBV-infected patients (including patients in IT and CHB groups), the elevation of IFN-α2 level was significantly associated with ALT level (β= 0.389, t = 2.423, P = 0.018), and was also negatively correlated to HBV-DNA load (β = −0.358, t = −2.308, P = 0.024), HBsAg (β = −0.359, t = −2.288, P = 0.025), and HBeAg contents (β = −0.355, t = −2.258, P = 0.027). However, when both ALT level and cytokines were included as independent variable, HBV-DNA load, HBsAg, and HBeAg contents were only correlated to ALT level (β = −0.459, t = −4.225, P = 0.000; β = −0.616, t = −6.334, P = 0.000; and β = −0.290, t = −2.433, P = 0.018; respectively).Conclusions:IFN-α2 elevation was associated with ALT level in patients with chronic HBV infection. However, in CHB patients, only ALT level was correlated to HBV-DNA, HBsAg and HBeAg contents.

NEOPTERIN DAN PEROKSIDA SERUM SEBAGAI PETANDA MAKROFAG TERAKTIVASI PADA TUBERKULOSIS PARU AKTIF DAN INDIVIDU BERKEBAHAYAAN TINGGI (Serum Neopterin and Peroxide As Marker of Activated Macrophages on Active Pulmonary Tuberculosis and Individuals At High Risk)

Failure of macrophages to phagocytize Mycobacterium tuberculosis causes the release of hydrogen peroxide/peroxide (H2O2) bythe activated macrophages. Neopterin is one of the most important markers in the activated macrophages. Neopterin is a pteridinederivative produced by the activated macrophages through the stimulation of interferon gamma. Increased levels of Neopterin has beenreported in lung tuberculosis. Activation of macrophages is essential to the development of tuberculosis infection that can lead to activepulmonary tuberculosis or latent tuberculosis, in this case is a high-risk for healthy individuals. To determine the differences in serumlevels of Neopterin and H2O2 between patients with active pulmonary tuberculosis and healthy individuals at high risk of pulmonarytuberculosis. A total of 15 patients with active pulmonary tuberculosis and 15 healthy individuals at high risk examination serumNeopterin levels and peroxide (H2O2). Active pulmonary tuberculosis patients when the results of sputum smear examination chest x-raysis obtained positive results. Healthy individuals at high risk when the results of smear examination and chest x-rays is negative. Thelevel of Neopterin was examined using a double antibody sandwich immunoassay with Human neopterin (NEOP) ELISA Kit ® BioassayTechnology Laboratory. The level of peroxide was examined using quantitative colorimetric peroxidedetermination with QuantiChromTMPeroxide Assay Kit (DIOX-250)®. Neopterin serum levels between patients with active pulmonary tuberculosis and healthy individualsat high-risk were analysed by independent samples t-test. H2O2 serum levels between patients with active pulmonary tuberculosis andhealthy individuals at high-risk were analysed with Mann Whitney Test. The confidence level is p &lt;0.05. The mean Neopterin levelsin patients with active pulmonary tuberculosis was 5.17±4.64 nmol/L, the mean Neopterin levels in group of healthy individuals athigh risk was 3.97±1.79 nmol/L. Statistical analysis by the independent samples t-test found no significant differences between groupsin Neopterin serum levels of patients with active pulmonary tuberculosis and healthy individuals at high risk (p=0.357). The meanserum levels of H2O2 of group of patients with active pulmonary tuberculosis was 26.38±3.00 μM, the mean levels of H2O2of group ofhealthy individuals at high risk of 20.69±4.46 μM. Statistical analysis with non-parametric Mann-Whitney Test was found significantdifference in the peroxide (H2O2) levels between groups of patients with active pulmonary tuberculosis and group of healthy individualat high-risk (p=0.000). The levels of Neopterin in patients with active pulmonary tuberculosis was not significantly higher comparedto the healthy individuals of high risk. Levels of peroxide (H2O2) serum in patients with active pulmonary tuberculosis was significantlyhigher compared to the group of healthy individuals at high risk. This shows that there is increased activity of macrophages in patientswith active pulmonary tuberculosis, but not effective in eliminating of Mycobacterium tuberculosis.

Biochemical Studies of Matrix Metalloproteinase (MMP) and tissue inhibitor of Metalloproteinase (TIMP) as New Biomarkers in HCV - Chronic liver disease

The present study aimed to investigate serum levels of Matrix Metalloproteinase (MMP9) and tissue inhibitor of Metalloproteinase (TIMP-1) as new biomarkers in HCV Chronic liver disease. Patients and methods: The study included 35 patients suffering from Chronic liver disease attending at Gastro Entrology Surgical Center (GEC), Mansoura University from 2016 to 2017 and 25 healthy individuals (HI) used as control group ,the studied groups were categorized into 14 patients (9 men and 5women; mean age 53.34±9.46yrs) suffering from HCC associated with HCV infection, 11patients (9 men and 2 women; mean age 52.34±9.14yrs) suffering from LC associated with HCV infection and 10 patients (7 men and 3women; mean age 48.9 ± 12.4yrs) suffering from Chronic Hepatitis (CH). Sera of all individuals were examined for TNF-α, LDH, AFP, ALT, AST, Albumin, T.Bilirubin, GGT, MMP-9 and TIMP-1 . Results: The obtained results showed a significant increase in serum ALT, AST, total Bilirubin and GGT and significant decrease of Albumin in liver disease groups (I, II and III) compared with Group IV. Significant increase in serum MMP9, TIMP1 in groups (I, II and III) when compared with HI group (IV). Significant correlation was recorded between MMP9 and TIMP1 in one hand and between each one with AST, T. bilirubin and Albumin concentration. Serum concentrations of TNF-α, AFP were elevated significantly in HCC patients compared to LC and CH but the difference between LC and CH was elevated significantly only (p&lt;0.0001) in TNF-α. Significant association was recorded between TNF-α, LDH, AFP, ALT and AST Conclusion: MMP9 and TIMP1 could be used as new biomarker for evaluating disease progression of HCV-chronic liver disease

Clinical significance of anti-dense fine speckled 70 antibody in patients with fibromyalgia.

Background/AimsFibromyalgia (FM) is a common rheumatologic disease characterized by chronic widespread pain, along with various clinical manifestations including atypical autoimmune characteristics. Despite its high prevalence, there remain no approved laboratory tests to identify specific manifestations of FM, or to rule out FM from other rheumatic diseases. Anti-dense fine speckled 70 (anti-DFS70) antibodies were initially identified as a form of anti-nuclear antibodies in a patient with interstitial cystitis. Anti-DFS70 antibodies are found in ≤ 10% of healthy individuals, but have suggestive negative association with autoimmune diseases; however, the clinical significance of these autoantibodies in FM patients remains poorly understood.MethodsWe examined 39 patients with FM, along with 17 patients with systemic lupus erythematosus (SLE), and 19 healthy individuals (HI). Patients were compared based on physical measurements, disease duration, tender point counts, FM Impact Questionnaire (FIQ) scores, visual analog scale (VAS) for pain, somatic symptoms, and anti-DFS70 antibodies.ResultsLevels of anti-DFS70 antibodies were significantly higher in the FM and HI groups than in those with SLE. Both anti-DFS70 antibodies and VAS scores were positively correlated with FM. Within the FM group, patients with arthralgia had higher anti-DFS70 antibody values compared to those without arthralgia (p = 0.024); antibody levels were also higher in patients with sleep disturbances relative to those without sleep issues (p = 0.024). In contrast, there were no correlations between anti-DFS70 antibodies and age, body mass index, disease duration, tender point counts, FIQ, short-form health survey results, or other clinical manifestations.ConclusionsAnti-DFS70 antibodies may represent a useful biomarker for differentiating between FM and other autoimmune diseases. The levels of anti-DFS70 antibodies were also significantly higher among patients with arthralgia and sleep disturbances. Further investigations are necessary to evaluate the relationships between anti-DFS70 antibodies and other cytokines as a predictive marker for pain.

Different aberrant expression pattern of immune checkpoint receptor in patients with T-NHL and NK/T-CL

T-cell immunodeficiency plays a crucial role in the development and progression of tumor. Immune checkpoint (co-inhibitory receptors) and TCR signal pathway regulate T-cell function in opposite way. Our previous study showed the decreasing trend in T cell activation related TCR signal pathway expression in peripheral blood mononuclear cells (PBMCs) from T-cell non-Hodgkin lymphomas (T-NHL) and NK/T-cell lymphoma (NK/T-CL). However, how immunosuppression status regulated by the co-inhibitory receptors, like programmed death 1 (PD-1), cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), B/T lymphocyte attenuator (BTLA) expanded to Lymphocyte-activation gene 3 (LAG-3), T cell immunoglobulin-3 (TIM-3), T cell immunoglobulin and ITIM domain (TIGIT) remains unclear. In this study, we characterized the expression pattern of all these immunosuppressive receptors in PBMCs form T-NHL and NK/T-CL patient by real-time qPCR. Significantly higher PD-1 expression level was detected in both T-NHL (n=14) and NK/T-CL (n=8) when compare to HI (healthy individuals, n=22,), while the expression level of its ligand PD-L1 had no statistic difference in both groups in comparison to HI group. The expression of BTLA, LAG-3, TIM-3, ICOS and TIGIT significantly increased in T-NHL group, but not in NK/T-CL group. While higher expression of CTLA-4 was found only in NK/T-CL group. When disease stage was considered in, BTLA is only up-regulating in advanced stage (stage III and IV), most patients with high ICOS expression level are in stage I and II. Moreover, the positive correlation between the expression level of LAG-3, TIM3 and TIGIT gene in HI group was impacted when it comes to either T-NHL or NK/T-CL patients. In conclusions, aberrant expression pattern of T-lymphocyte inhibitory receptors is a common characteristic in patients with T- NHL or NK/T-CL, which is one of reasons of immunosuppression, moreover, the different upregulating of such inhibitory receptors between T-NHL and NK/T-CL might be as immune biomarkers for evaluation the immunosuppression status and help to establish the precision target of immunotherapy.

The Distribution of T Memory Stem Cells in Cord Blood, Peripheral Blood from Healthy Individuals and Patients with Leukemia/Lymphoma

Human T memory stem cells (TSCM) are a unique memory T cell subset with enhanced self-renewal capacity and the ability to differentiate into potent effectors could be valuable weapons in adoptive T-cell therapy against cancer. The distribution of TSCM cells in umbilical cord blood and leukemia/lymphoma remain unclear. In this study, we investigated the distribution of TSCM and other memory T (TM) subsets in the cord blood (CB) (n = 11), peripheral blood (PB) of lymphoblastic leukemia/lymphoma (LL) (n = 7), Myeloid leukemia (ML) (n = 7) as well as healthy individual (HI) (n = 16) by multicolor flow cytometry.There were high percentage of CD4+ TSCM and CD8+ TSCM in CB group than that in HI-PB, however, fewer CD4+T central memory (TCM), CD4+T effector memory (TEM), CD8+ TEM and CD8+ terminal effector (TE) cells in CB group than in HI-PB (Figure 1). When compared LL or ML group with HI group, there was an increase in the CD4+ TSCM in the ML group, while the frequencies of TCM, TEM, or TE were similar. However, the percentage of memory T cell subsets in patient group was quite disperse, for example, CD4+ TSCM could range from 0.35% to 6.52% (LL) and from 0.67% to 3.71%(ML), the percentage of CD8+ TSCM could even reach to 10.43% and 7.62% in LL and ML group respectively, the same dispersion also exist in other T cell subsets of patient group but not in the blood and cord blood from HI. According to individually analyze distribution of T cell subsets in each patients, a higher percentage of CD4+ TSCM was detected in a B-ALL patient in the LL group comparing with HI, combined with significant lower percentage of CD4+ TCM, TEM and TE (Figure 2 A, L3), similar distribution pattern of CD8 T cell subsets also detected in the same B-ALL patient (Figure 2B, L3). Interesting, a case with BCR-ABL+ B-ALL include in this study showed a relative normal distribution pattern (Figure 2 A and B, L5). There were four patients in LL group showed a relative higher percentage of CD8+ TCM (Figure 2 B, L1, L2, L4 and L6), while two of them followed with quite lower TE in PB (Figure 2 B, L2 and 6). In the ML group, relative higher CD4+ TSCM combined with lower TEM was also detected in a case with AML-M5 (Figure 2 C, ML-3). In terms of CD8 T cell subsets distribution in ML group, we found two patients with AML-M5 possessing a higher CD8+ TSCM proportion compare with HI and other patients in ML group (Figure 2 D, ML-2 and 3).In conclusion, we characterized the distribution of different memory T cell subsets in healthy CB and PB, the reason of high percentage of TSCM in CB is needed further investigation. In addition, the higher CD4+ TSCM detected in ML group and individual cases of LL group may related to the leukemia antigens they encountered, whether such a characteristic would impact their prognostic worth further investigation. Overall, the distribution of TSCM and other memory T cell subsets in blood between healthy and leukemia is not shown obviously difference, while, the large difference among individual patients reflect the various immunosuppression degree and immune reaction ability to tumor antigens. To characterize the relationship between distribution pattern of TSCM and TM cells and immune status of patients will benefit to developing more suitable individually immune treatment strategies.Acknowledgments: The study was supported by grants from the National Natural Science Foundation of China (Nos. 81270604, U1301226, and 81400109), the China Postdoctoral Science Foundation (No. 2013M540685), Guangdong Natural Science Foundation (Nos. S2013040016151 and S2013020012863), the Foundation for High-level Talents in Higher Education of Guangdong, China (No. [2013]246-54), and the Guangzhou Science and Technology Project Foundation (201510010211). [Display omitted] [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Altered expression pattern of miR-29a, miR-29b and the target genes in myeloid leukemia.

ObjectivesThe miR-29 family have been demonstrated acting as vital tumor suppressor in multiple cancers as well as regulators in the adaptive immune system. Little is known about their role in leukemogenesis. The purpose of this study is to analyze the expression pattern of miR-29a/29b and its target genes Mcl-1 (myeloid cell leukemia sequence 1) and B-cell lymphoma 2 (Bcl-2) in myeloid leukemia.MethodsQuantitative real-time PCR was used for detecting genes expression level in peripheral blood mononuclear cells (PBMCs) from 10 cases with newly diagnosed, untreated acute myeloid leukemia (AML) and 14 cases with newly diagnosed, untreated chronic myeloid leukemia (CML) in chronic phase, and 14 healthy individual (HI) served as controls. Correlation between the relative expression levels of different genes have been analyzed.ResultsSignificant lower expression of miR-29a/29b and higher expression level of two potential target genes Bcl-2 and Mcl-1 were found in PBMCs from AML and CML patients compared with HI group. In addtion, miR-29a expression in AML was significantly lower than that in CML. Moreover, negative correlation between miR-29a/29b and its target genes have been found. While, positive correlation between relative expression level of miR-29a and miR-29b or Bcl-2 and Mcl-1 were presented in the total 38 research objects.ConclusionDown-regulated miR-29a and miR-29b, and accompanying up-regulated Bcl-2 and Mcl-1 are the common feature in myeloid leukemias. These data further support the role for miR-29a/29b dysregulation in myeloid leukemogenesis and the therapeutic promise of regulating miR-29a/29b expression for myeloid leukemia in the future.

High-avidity IgG Autoantibodies against DFS70/LEDGF in Atopic Dermatitis

Objective: Dense fine speckles 70 kDa protein (DFS70)/lens epithelium-derived growth factor (LEDGF) autoantibodies have been reported to be present in serum samples from patients with atopic dermatitis (AD) as well as from healthy individuals (HI). We previously revealed that IgE- and IgG4-anti-DFS70 autoantibodies were found in patients with AD and that their presence was associated with high levels of thymus-and activation-regulated chemokine. Our aim was to determine the avidity of IgG-anti-DFS70 antibodies in patients with AD relative to that avidity in HI.Patients and methods: This study was undertaken to measure the avidity of IgG-anti-DFS70 autoantibodies in AD and HI groups, in comparison with the major recall antigen diphtheria toxoid (DT) and to evaluate the relevance of IgG-anti-DFS70 autoantibodies to the disease. We measured the avidity of IgG-anti-DFS70 autoantibodies and anti-DT antibodies in sera that were positive for IgG-anti-DFS70 autoantibodies, obtained from 20 AD patients and 20 HI using enzyme-linked immunosorbent assay.Results: The avidity of anti-DFS70 autoantibodies was significantly higher (p<0.01) in the AD patients than in the HI, whereas there was no difference in the avidity of anti-DT between the two groups. There was also no positive correlation between the avidity and the titer of anti-DFS70 in the both groups. In contrast, positive correlation was shown between the serum levels and the avidities of anti-DT.Conclusion: Our findings indicate that AD patients might have significantly high-avidity IgG-anti-DFS70 autoantibodies and that this avidity might be a novel serological marker for a certain AD subpopulation.

The differential expression pattern of the BMI-1, SALL4 and ABCA3 genes in myeloid leukemia

Background and methodsIn order to characterize the expression pattern of SALL4, BMI-1 and ABCA3 genes in patients with myeloid leukemia and those who achieved complete remission (CR) after chemotherapy. Real-time PCR was used to determine the expression level of these genes in peripheral blood mononuclear cells from 24 patients with AML, eight patients with AML-CR, 13 patients with CML in the chronic phase (CML-CP), 12 patients with CML in blast crisis (CML-BC), 13 patients with CML-CR and 11 healthy individuals (HI).ResultsOverexpression of the BMI-1 gene was found in the AML, CML-CP and CML-BC groups as compared with HI group, while the BMI-1 expression level was lower in patients who achieved CR. In contrast, significantly increased SALL4 expression was only found in AML group, additionally, SALL4 expression was lower in the CML-CP and CML-CR groups compared with the HI group, while the SALL4 expression level in the CML-BC group was higher and significantly greater than that in the CML-CP and CML-CR groups. Moreover, a positive correlation between the expression of SALL4 and BMI-1 genes was found in samples from most groups. There was no significant difference of ABCA3 expression level in AML and CML-BC group in comparison with HI group. Interestingly, the ABCA3 expression level was significantly decreased in the CML-CP, AML-CR and CML-CR in comparison with the HI group. Moreover, the ABCA3 expression level in all of the CR groups was lower than that in their corresponding groups.ConclusionsThese results describe the altered SALL4, ABCA3 and BMI-1 expression pattern in different phases of myeloid leukemia, which may relate to the development and progression to different diseases. SALL4 expression was strongly correlated with BMI-1 in most of the myeloid leukemia patient groups, providing a potential link between SALL4 and BMI-1 in leukemogenesis.

Glutamate dehydrogenase activity in lymphocytes of B-cell chronic lymphocytic leukaemia patients

Objectives: To investigate the pattern of glutamate dehydrogenase (GLDH) activity, GLUD1 and GLUD2 expressions in peripheral blood mononuclear cells (PBMC) of untreated B-chronic lymphocytic leukemia (B-CLL) in healthy individuals (HI) and patients with infectious mononucleosis (IM). Design and methods: GLDH activity was determined in a supernatant obtained from pelleted PBMC. GLUD1 and GLUD2 mRNA expression was determined using a quantitative real-time polymerase chain reaction. CD19 + B cells from PBMC were purified by using positive selection. Results: The highest GLDH activity was found in PBMC of the B-CLL group followed by the HI group and IM group. The PBMC GLDH activity was higher in 60% of the B-CLL patients according to the established reference interval for our HI (2.17–5.70 μkat/g protein). The greater GLDH activity was also found in the CD19 + cell preparation of the B-CLL patients (two of the three) but not in HI ( n = 3). The median value of GLUD1 expression was highest in the IM group ( n = 11), followed by the HI ( n = 14) and B-CLL groups ( n = 59) (median 4.69/3.78, P < 0.005 and 4.69/2.91, P < 0.0005, respectively). GLUD2 expression was not significantly different between groups. Conclusions: The increased GLDH activity is specific for the PBMC of B-CLL patients. The GLUD1 but not the GLUD2 gene expression pattern is different between the PBMC of IM and B-CLL patients.

Effects of FR‐91 on Immune Cells from Healthy Individuals and from Patients with Non‐Hodgkin Lymphoma

The immune system is subject to destruction and dysfunction as a result of attacks by pathogenic and environmental agents. In addition, many clinical situations exist in which it is desirable to stimulate or suppress the immune system. The present study evaluated the screening efficacy of flow cytometric lymphocyte subset typing in peripheral blood mononuclear cells from healthy individuals (HI) and from patients with non-Hodgkin lymphoma (NHL) treated with different concentrations of FR-91, a standardized lysate of microbial cells belonging to the Bacillus genus, and in vitro cytokine production. Increased expression of subset markers (CD3, CD4, CD8) in NHL and CD3 in HI suggests an immunomodulating effect of FR-91. In addition the results of cytokine production also demonstrated a clear effect of FR-91 on both populations. A significant increase of IL-6, IL-12, IFN-γ and TNF-α was observed in the HI group after treatment with FR-91. In a similar manner an increase of IL-2, IL-6, IL-12, IFN-γ and TNF-α was also observed in the NHL group. In conclusion FR-91 seems to affect lymphocyte subpopulations, in vitro cytokine production, as well as mitogen-induced lymphocyte activation in a dose-dependent manner in both healthy individuals and NHL patients.