Significance of mast cell renal infiltration in patients with anti-GBM nephritis

To investigate the role of mast cells (MCs) renal infiltration in the progression of human anti-GBM nephritis, 38 patients diagnosed with anti-GBM nephritis were enrolled. Renal biopsies were performed. Immunohistochemistry was conducted to detect MCs in renal tissues. Patients were divided into group 1 (MCs?<50?mm^?2, n?=?18) and group 2 (MCs?≥50?mm^?2, n?=?20) according to the infiltrating renal MC count. The clinical–pathological indices were compared. And, correlation between MCs and the clinical–pathological indices was analyzed. Patients of group 2 had more severe renal dysfunctions, expressed as higher levels of serum creatinine (SCr 8.95?±?3.66 vs. 4.75?±?2.73?mg/dL, p?<?0.001), urine retinol-binding protein (RBP 29.8?±?13.9 vs. 15.7?±?11.5?mg/dL, p?=?0.005), and lower urinary osmotic pressure. Pathologically, patients of group 2 had a higher percentage of fibrous/fibrocellular crescents (66.7?±?21.9 vs. 47.0?±?33.6%, p?=?0.037) but a lower percentage of cellular crescents. More CD8 (268?mm^?2 vs. 180?mm^?2, p?=?0.045) and CD68 (268?mm^?2 vs. 180?mm^?2, p?=?0.045) positive cells infiltrating the interstitium were observed in group 2. Furthermore, renal MCs correlated significantly with the total number of crescents and the tubular interstitial CD8 and CD68 positive cells. And, the number of MCs was associated with the histological types. The renal function was significantly different between the two groups at presentation. However, at 3 and 6 month follow-up, the patient outcome was associated with the histological types. Our study showed that MC infiltrations were associated with chronic lesions in anti-GBM nephritis and may be involved in the loss of renal function with pathological changes.     

Renal functional reserve and renal hemodynamics in hypertensive patients

The renal functional reserve (RFR) is the ability of the kidneys to increase renal plasma flow and glomerular filtration rate (GFR) in response to protein intake. It is a measure of functional and anatomic integrity of nephrons. It is not known what relation between RFR and kidney Doppler parameters. We aimed to study the relation between the RFR and renal hemodynamic parameters in hypertensive patients with and without nephropathy who had normal kidney function. Twenty-four hypertensive subjects with nephropathy (HTN-n, n?=?10) and hypertension without nephropathy (HTN, n?=?14) were included in the study. Control group included 11 healthy subjects. Baseline GFR (GFR1) and GFR after intake of egg protein 1?mg/kg of body weight were determined (GFR2). RFR was calculated by the following formula: (GFR2-GFR1)/GFR1?×?100%. Doppler ultrasonography was performed. Arterial blood pressure (BP), body mass index (BMI), and estimated GFR were also recorded. HTN and HTN-n groups had impaired levels of RFR compared with controls (p?<?0.05), significantly decreased value of flow velocity parameters (Vmax, Vmin), and increased RRI compared with controls. There was significant negative correlation of RFR with blood pressure levels (sBP, r?=??0.435, p?=?0.009; dBP, r?=??0.504, p?=?0.002), RRI (r?=??0.456, p?=?0.008), micro albuminuria (MAU, r?=??0.366, p?=?0.031) and positive correlation with Vmax and Vmin (r?=?0.556, p?=?0.001 and r?=?0.643, respectively, p?<?0.001). Linear regression showed that RRI and MAU were independent predictors of decreased RFR. RFR is lower in hypertensive patients despite near-normal level of kidney function and is related to particular level of BP. RRI and MAU were independent predictors of decreased RFR.     

Alloreactive natural killer cells promote haploidentical hematopoietic stem cell transplantation by expansion of recipient‐derived CD4+CD25+ regulatory T cells

Alloreactive NK cells (Allo‐NKs) have been shown to exert advantageous effects on the outcomes of haploidentical hematopoietic stem cell transplantation (Haplo‐HSCT) for cancer treatment. However, the mechanisms of action of Allo‐NKs remain unclear. We established a novel Haplo‐HSCT conditioning regimen composed of Allo‐NKs and a low dose of immunosuppressive drugs (Allo‐NKs + Chemo) to investigate alternative mechanisms besides direct cytotoxicity. The inhibitory effects of different cell subsets on the donor–recipient mixed lymphocyte reactions (MLRs) were evaluated after Haplo‐HSCT. The quantities and functions of CD4⁺CD25⁺ regulatory T cells (Tregs) and dendritic cells (DCs) in the spleen and the thymus were examined. Our results showed that the Allo‐NKs + Chemo regimen induced systemic tolerance, and that CD4⁺CD25⁺ Tregs played a significant role in inducing and maintaining systemic tolerance after Haplo‐HSCT. Alloreactive NK cells promoted the expansion of recipient‐derived CD4⁺CD25⁺CD127^? Tregs in the thymus and the spleen which could be amplified in vitro by the immature donor‐derived DC subset isolated from the thymus of Allo‐NKs + Chemo‐treated mice. Our findings suggested that Allo‐NKs are capable of inducing systemic tolerance after Haplo‐HSCT by assembling donor‐derived immature DCs to expand recipient‐derived Treg cells in the thymus.     

T cell receptor beta chain (TCR-Vβ) repertoire of circulating CD4+ CD25−, CD4+ CD25low and CD4+ CD25high T cells in patients with long-term renal allograft survival

The mechanisms underlying maintenance of renal allografts in humans under minimal or conventional immunosuppression are poorly understood. There is evidence that CD4⁺ CD25⁺ regulatory T cells and clonal deletion, among other mechanisms of tolerance, could play a key role in clinical allograft survival. Twenty‐four TCR‐Vβ families were assessed in CD4⁺ CD25^?, CD4⁺ CD25^low and CD4⁺ CD25^high T cells from patients with long‐term renal allograft survival (LTS), patients exhibiting chronic rejection (ChrRx), patients on dialysis (Dial) and healthy controls (HC) by flow cytometry. LTS patients presented a higher variability in their TCR‐Vβ repertoire, such decreased percentage of Vβ2⁺, Vβ8a⁺ and Vβ13⁺ in CD4⁺ CD25^low and ^high compared with CD4⁺ CD25^? subset and increased Vβ4 and Vβ7 families in CD4⁺ CD25^high T cells exclusively. Additionally, LTS patients, particularly those that were not receiving calcineurin inhibitors (CNI), had increased percentages of CD4⁺ CD25^high T cells when compared with Dial (P < 0.05) and ChrRx (P < 0.05) patients. Our results suggest that a differential expression of particular TCR‐Vβ families and high levels of circulating CD4⁺ CD25^high T cells in long‐term surviving renal transplant patients could contribute to an active and specific state of immunologic suppression. However, the increase in this T cell subset with regulatory phenotype can be affected by CNI.     

The characterization of peritoneal and pleural exudate cells from malignant effusions

The immune function of peripheral blood cells and cells from the pleural and abdominal effusions of patients with advanced cancer was compared to that of peripheral blood cells from controls. The parameters examined included lymphocyte subsets, natural killer (NK) cell activity, and anti-Daudi and lymphokine-activated killer (LAK) cell activity. The percentage of CD4⁺ pleural and peritoneal exudate cells (PEC) was significantly higher than the percentage of peripheral blood mononuclear cells (PBMC) in the patients. The percentage of CD8⁺CD11⁺ PEC and PBMC, being the suppressor T-cells, of the patients was increased compared with controls, while the percentage of CD8⁺CD11^– PEC, being the cytotoxic T-cells, was identical to the PBMC of both patients and controls. The NK activity of PEC was significantly lower than that of PBMC in both patients and controls, and there was no correlation between the NK activity of PBMC and PEC. Although the anti-Daudi activity of PEC was markedly low, LAK cells with high activity could be induced by culture with interleukin-2 for 4 days. These results suggest that the immune function of cells in malignant effusions may be depressed due to a low population of cytotoxic T cells, low NK activity and increased suppressor T cells, while the local administration of interleukin-2 may induce LAK cells. Therefore, effective local immunotherapy for malignant effusions should not only augment effector cells, but also inhibit supprssor cells.     

A Single Dose of Rituximab Does Not Deplete B Cells in Secondary Lymphoid Organs but Alters Phenotype and Function

A single dose of the anti‐CD20 monoclonal antibody rituximab induces a nearly complete B cell depletion in peripheral blood, but not in secondary lymphoid organs. Modulation of this remaining B cell population due to rituximab treatment may contribute to the therapeutic effects of rituximab. To assess the in vivo effects of rituximab we used lymph nodes (LNs) collected during renal transplant surgery in patients who had received rituximab 4 weeks earlier in preparation for an ABO‐incompatible transplantation. Rituximab treatment resulted in a lower percentage of naïve (IgD⁺CD27^?) and a higher percentage of switched memory (IgD^?CD27⁺) B cells. Remarkably, transitional (CD24⁺⁺CD38⁺⁺) B cells were virtually lacking in the LNs of rituximab‐treated patients. Moreover, LN‐derived B cells from rituximab‐treated patients produced different amounts of various Ig‐subclasses after anti‐CD40/IL‐21 stimulation ex vivo. Finally, after stimulation of allogeneic T cells with LN‐derived B cells from rituximab‐treated patients, the proliferated T cells showed a decreased production of IL‐17. In conclusion, after treatment with rituximab there remains a B cell population with different functional capacities. Consequently, the effect of rituximab on the immune response will not only be determined by the extent of B cell depletion, but also by the functional properties of the remaining B cells.

     

Analysis of mucosal bladder leucocyte subpopulations in patients treated with intravesical Bacillus Calmette-Guerin

Summary Immunohistochemical techniques were used to investigate leucocyte subpopulations in the bladders of patients with superficial transitional cell carcinoma treated with BCG Pasteur. Leucocyte subsets were enumerated with a panel of monoclonal antibodies which included CD3, CD4, CD8, TQ1, Leu7, CD15, HLA-DR, CD25, CD22. We demonstrated in the bladders of patients treated with BCG a particular lymphocyte population; the major subset was an inducer (CD4⁺, TQ1^-) which was activated (CD25⁺, HLA-DR⁺) and associated with polymorphonuclear eosinophils. There was neither inducer of suppression nor major cytotoxic/suppressive subsets. CD8⁺ and NK cells could not be the primary mediators of BCG activity. These data supported the hypothesis of a helper T lymphocyte activity associated with lymphokine production and activation of effector killer cells.     

Salt sensitivity of blood pressure in non-dialysis patients with chronic kidney disease

Background: Chronic kidney disease (CKD) is a world-wide public health problem. Hypertension is both a cause and a complication of CKD, and a risk factor for progression of kidney disease. The effect of salt intake on blood pressure (BP) and the salt sensitivity in non-dialysis patients with CKD were studied. Methods: One hundred and thirty non-dialysis patients with CKD were enrolled in the present study. Daily urinary excretion of sodium (representative of daily sodium intake) and BP was monitored in conditions of original eating habits. Estimated glomerular filtration rate (eGFR) was measured by the creatinine clearance (Ccr). Results: There was a linear positive relationship between the salt intake and systolic blood pressure (SBP) (β?=?0.250, p?=?0.004). It had been found that the log of BP/24-h urinary sodium (salt sensitivity index) had linear relationship with the log of eGFR (β_syst?=??0.364, p?=?0.000, β_diast?=??0.345, p?=?0.000, respectively). Multi-stepwise regression analysis showed SBP was mainly influenced by salt intake and eGFR. There was a negative correlation between diastolic blood pressure (DBP) and age. Conclusion: These results demonstrated a linear relationship between the salt intake and SBP in non-dialysis patients with CKD. The salt sensitivity of BP rose with the decline of renal function.     

Febuxostat for treating allopurinol-resistant hyperuricemia in patients with chronic kidney disease

Background: Availability of the novel xanthine oxidase inhibitor febuxostat, which has multiple excretion pathways, enables investigation of the significance of serum uric acid control on renal function in patients with chronic kidney disease (CKD). Methods: This was an exploratory, retrospective, observational study conducted at a single Japanese center. Serum uric acid concentrations and serum creatinine levels in the 6 months before and after the start of febuxostat treatment were collected for CKD patients switched from allopurinol after failing to achieve serum uric acid concentrations ≤6.0?mg/dL. Results: Evaluable data were available for 60 patients, 67% of whom had advanced CKD (eGFR <30?mL/min/1.73?m²). Mean dose of febuxostat was 15.9 (±?8)?mg/day. Mean serum uric acid concentration decreased from 8.4 (±1.4) mg/dL at baseline to 6.2 (±1.2)?mg/dL at 6 months; 47.5% of patients achieved a level ≤6.0?mg/dL. The change from baseline in eGFR was positive at all time points during febuxostat treatment and the increase of 2.3 (±5.6)?mL/min/1.73?m² at 6 months was significant (p?=?0.0027). Whereas the eGFR slope was negative during allopurinol treatment, it became positive after the switch to febuxostat. The change in eGFR slope before and after febuxostat treatment was significant for all patients (p?p?2 (p?Conclusions: In patients with CKD, febuxostat reduces serum uric acid concentrations effectively and may suppress the progressive decline in renal function.     

Low utility of serum 25–hydroxyvitamin D3 and 1, 25-dihydroxyvitamin D3 in predicting peripheral Treg and Th17 cell counts in ESRD and renal transplant patients

BackgroundVitamin D has shown an immune-modulatory effect in different studies. Vitamin D stimulates Tregs and inhibits Th17 cells. The immune-modulatory role of vitamin D in chronic kidney disease (CKD) and renal transplant patients is unclear. We measured whether different serum levels of vitamin D were associated with an increased or decreased presence of lymphocyte subsets including Treg and Th17 cells in end-stage renal disease (ESRD) and renal transplant recipients.MethodsEighty-seven renal transplant recipients and 53 end-stage renal disease (ESRD) patients were enrolled in this study. The absolute counts of CD4 + and CD8 + T, CD16 + CD56 + NK, CD19 + B, CD4 + CD25 + CD127- Foxp3 + (Tregs), Helios + Tregs, CD38 + Tregs, and CD4 + CD17 + (Th17) cells were analyzed in peripheral blood in both patient groups. In addition, serum 25 (OH) D₃, 1, 25 (OH)₂ D₃, IL-6, IL-17, IL-23, and TGF-β₁ were measured. The association between lymphocyte subset counts and 1, 25 (OH)₂ D₃ or 25 (OH) D₃ was studied, as was the association between serum IL-6, IL-17, IL-23, or TGF-β₁ and 1,25 (OH)₂ D₃ or 25 (OH) D₃.ResultsSerum 25 (OH) D₃ and 1,25 (OH)₂ D₃ levels were not independently associated with peripheral CD4 + T, CD19 + B, CD16 + CD56 + NK, Treg, or Th17 cell counts. In contrast to serum 25 (OH) D₃, serum1, 25 (OH)₂ D₃ was positively associated with CD8 + T cells counts in renal transplant recipients.ConclusionOur findings indicate low utility of serum 25 (OH) D₃ and 1, 25 (OH)₂ D₃ levels in predicting a change in lymphocyte subset counts in ESRD and renal transplant patients.     

Impact of radiofrequency ablation on PBMC subpopulation in patients with renal cell carcinoma

Purpose

With the development of diagnostic techniques, renal cell carcinoma (RCC) is currently diagnosed in earlier stages, allowing the introduction of less invasive techniques in its management. One of the most promising new treatment methods is based on the utilization of high temperature created by radiofrequency current circulating around the needle probe introduced into the tumor. Besides the direct destruction of the cancer tissue, the treatment may induce immunologic reaction to tumor antigens released from destroyed tumor cell. This paper describes changes observed in the peripheral blood lymphocyte population after radiofrequency ablation (RFA) of RCC.

Methods

Blood was tested before, and 2, 4, and 6 weeks after the RFA in 6 patients with RCC for the proportions and numbers of CD3⁺, CD3⁺HLA-DR⁺, CD3⁺CD4⁺, CD3⁺CD8⁺, and CD56⁺CD16⁺ cells. The blood was stained with fluorochrome-conjugated monoclonal antibodies and percentages of cells expressing various markers were determined by flow cytometry.

Results

In all patients, the changes were most pronounced 2 weeks after the procedure. The proportion of CD4⁺ and CD8⁺ lymphocytes were changed. In 1 patient, an increase in both CD4⁺ and CD8⁺ cells was observed. In 5 out of 6 patients, the proportion of activated (DR⁺) cells was increased over the whole follow-up period with the highest values in the second week after RFA. The percentage of the CD56⁺CD16⁺ was decreased in most of the patients.

Conclusions

Our study confirms that in the majority of patients, RFA of the renal tumors causes significant changes in the proportion of the peripheral immune cells. We suggest that the results presented in this article shows the necessity for further studies.     

Effect of polyflux membranes on the improvement of hemodialysis-associated eosinophilia: a case series

Hemodialysis-associated eosinophilia (HAE) is believed to be associated with allergic reactions to dialyzer materials. This study aimed to investigate the use of Polyflux membranes to improve HAE. Thirty-one patients suffering from HAE were included. Patients were dialyzed with polysulfone membranes when they developed HAE. After that, patients were dialyzed with Polyflux membranes three times every week, 4?h every time without changing the dialysis parameters and medication. Levels of peripheral eosinophils, hsCRP, IgE, C3a, IL-5 and peripheral CD4+ lymphocytes and CD8+ lymphocytes were assessed before Polyflux treatment, and at 4th, 8th and 12th weeks of treatment. Any symptoms including chest tightness and skin itching were observed during the study period. After 12 weeks of Polyflux membrane dialysis and compared with polysulfone membrane dialysis, levels of peripheral eosinophils were significantly decreased (1.26?±?0.61 vs. 0.71?±?0.29?×?10⁹/L, p?<?0.001); serum IL-5 levels were significantly decreased (24.43?±?10.21 vs. 9.11?±?4.21?pg/mL, p?<?0.001); and chest tightness and skin itching were significantly improved (45.2% vs. 19.4%, p?=?0.028). After 12 weeks, there was no significant change in serum levels of hsCRP (2.00?±?0.94 vs. 1.81?±?0.79?mg/L, p?=?0.352), IgE (104.61?±?98.79 vs. 114.95?±?101.07?IU/mL, p?=?0.422) and C3a (121.61?±?34.04 vs. 120.29?±?32.81?µg/L, p?=?0.316), and in peripheral levels of CD4+ (589?±?181 vs. 569?±?171?cells/mm³, p?=?0.672) and CD8+ (443?±?123 vs. 414?±?140 cells/mm³, p?=?0.395) cells. Eosinophil count was correlated with serum IL-5 levels (r?=?0.873, p?<?0.001). Changing to a Polyflux membrane may alleviate HAE and reduce serum IL-5 levels. Therefore, this could be a strategy to manage HAE in the clinical practice.     

Characterization of baboon NK cells and their xenogeneic activity

Kennett SB, Porter CM, Horvath‐Arcidiacono JA, Bloom ET. Characterization of baboon NK cells and their xenogeneic activity. Xenotransplantation 2010; 17: 288–299. © 2010 John Wiley & Sons A/S. Abstract: Background: Baboons are commonly used as models for transplantation and preclinical testing of various types of therapeutic agents. For proper assessment of information gathered from these models, differences between the baboon and human immune systems need to be characterized. Natural killer (NK) cells are the first line of defense against many infectious agents and cancer and are important mediators of transplantation rejection reactions, particularly during xenotransplantation. In this study, we examined baboon NK cell function and developed methods for purifying and expanding these cells. Methods: Baboon NK cells were analyzed using a combination of extracellular and intracellular cell staining, cell sorting, interleukin (IL)‐2 mediated stimulation and expansion, and 4 h cytotoxicity assays with human and pig target cell lines. Results: Baboon peripheral blood mononuclear cell (PBMC) exert very low but detectable cytolytic activity against both human (K562) and pig (PAEC, J2) target cells, and this activity is enhanced within 4 h of treatment with IL‐2. Like human NK cells, many baboon PBMC express the lytic enzymes granzyme A, granzyme B, and perforin. Based on these markers, we identified a subpopulation of CD3^? baboon lymphocytes that are CD8^dim and CD16^bright that likely represents the baboon NK cells. These cells also are characterized by expression of the natural cytotoxicity receptor NKp46. Baboon CD3^?NKp46⁺ cells purified by flow cytometric cell sorting have high cytolytic capacity that can be further enhanced by IL‐2 stimulation. These baboon NK cells can be expanded in vitro and retain extremely high cytolytic capacity. While fresh baboon lymphocytes express very little CD56, the expanded baboon NK cells are predominantly CD56⁺; approximately 10% of the expanded NK cells are CD56^dim, and the remainder are CD56^bright. Conclusions: Baboon NK cells that are IL‐2 responsive can be identified on the basis of a CD3^?NKp46⁺CD8^dimCD16^+/? or CD3^?CD8^dimCD16^bright phenotype and can be isolated and expanded in culture. These results may allow for a more accurate representation of the human innate immune system in baboon models and more accurate analyses of the role of the baboon innate immune system cells in preclinical models.     

Liver Transplantation in a Patient With CD40 Ligand Deficiency and Hyper‐IgM Syndrome: Clinical and Immunological Assessments

Monoclonal antibodies that disrupt CD40–CD40 ligand (CD40L) interactions are likely to have use in human transplantation. However, the extent of the immunosuppressive effects of CD40–CD40L blockade in humans is unknown. Hyper‐IgM syndrome (HIGM) is a rare primary immunodeficiency syndrome characterized by defects in the CD40–CD40L pathway, severe immune deficiency (IgG), and high or normal IgM levels. However, the effects of CD40L deficiency on T‐ and natural killer (NK)‐cell function is not established. Here, we present a patient with HIGM syndrome who underwent liver transplantation for hepatitis C virus infection. Posttransplantation, NK‐cell antibody‐dependent cytokine release (γ‐interferon) to alloantigens and T cell responses to viral antigens and mitogens were assessed and showed normal CD4⁺, CD8⁺, and NK‐cell responses. We also examined antibody‐dependent cellular cytotoxicity against a CD40⁺ and HLA‐expressing cell line. These experiments confirmed that the patient's NK cells were equivalent to those of normal subjects in mediating antibody‐dependent cellular cytotoxicity despite the absence of CD40–CD40L interactions. Mitogenic stimulation of the patient's peripheral blood mononuclear cells showed no expression of CD40L on T and NK cells compared with increased expression in normal subjects. Taken together, these data suggest that absence of CD40L expression is responsible for aberrant B cell immunity but had little impact on NK‐ and T cell immune responses in vitro.     

A comparison of quality of sleep between patients with chronic kidney disease not on hemodialysis and end-stage renal disease on hemodialysis in a developing country

Few studies have compared quality of sleep between pre-dialysis chronic kidney disease (pre-dialysis CKD) patients and end-stage renal disease patients on dialysis (ESRD) and have found inconsistent results. Objective of this study is to compare quality of sleep between patients with pre-dialysis CKD and ESRD in a developing country. This study was conducted in an out-patient department and hemodialysis unit of a tertiary care facility. Patients included had either pre-dialysis CKD or ESRD. Assessment of quality of sleep was done using Pittsburgh sleep quality index (PSQI). A total of 152 patients were included in the study. Out of these patients, 79 (52%) had ESRD and 73 (48%) had pre-dialysis CKD. Median PSQI score was 6 (IQR 3–8.8). Poor sleep quality (PSQI ≥5) was present in 100 (65.8%) patients. Only hemoglobin (β?=??0.39, p?β?=?0.56, p?β?=?0.22, p?r?=??0.34, p value .80) in pre-dialysis CKD patients. Poor sleep quality is common in patients with CKD including hemodialysis patients in a developing country, which is independent of kidney function in non-dialysis patients. There is no difference in quality of sleep between pre-dialysis CKD and ESRD patients.     

High serum chemerin level in CKD patients is related to kidney function,but not to its adipose tissue overproduction

Abstract

Chemerin is an adipokine modulating inflammatory response and affecting glucose and lipid metabolism. These disturbances are common in CKD. The aim of the study was: (a) to evaluate circulating chemerin level at different stages of CKD; (b) to measure subcutaneous adipose tissue chemerin gene expression; (c) to estimate the efficiency of renal replacement therapy in serum chemerin removal. 187 patients were included into the study: a) 58 patients with CKD; (b) 29 patients on hemodialysis; (c) 20 patients after kidney transplantation. 80 subjects constituted control group. Serum chemerin concentration was estimated by ELISA. The adipose tissue chemerin mRNA level was measured by RT-qPCR. The mean serum chemerin concentration in CKD patients was 70% higher than in the control group (122.9?±?33.7 vs. 72.6?±?20.7?ng/mL; p?<?0.001) and it negatively correlated with eGFR (r?=??0.71, p?<?0.001). The equally high plasma chemerin level was found in HD patients and a HD session decreased it markedly (115.7?±?17.6 vs. 101.5?±?16.4?ng/mL; p?<?0.001). Only successful kidney transplantation allowed it to get down to the values noted in controls (74.8?±?16.0 vs. 72.6?±?20.7?ng/mL; n.s.). The level of subcutaneous adipose tissue chemerin mRNA in CKD patients was not different than in patients of the control group. The study demonstrates that elevated serum chemerin concentration in CKD patients: (a) is related to kidney function, but not to increased chemerin production by subcutaneous adipose tissue, and (b) it can be efficiently corrected by hemodialysis treatment and normalized by kidney transplantation.     

Foxp3+ T cells in peripheral blood of renal transplant recipients and clinical correlations

Aim: Immunophenotype peripheral blood T cells from renal transplant recipients (RTR) using cellular markers of regulatory T cells (Tregs) and flow cytometry, including Foxp3, and correlate these findings with clinical parameters. Methods: Expression of phenotypic markers of Tregs was assessed by flow cytometric analysis of peripheral blood lymphocytes (PBL) from (i) RTR (n = 95); (ii) patients with end‐stage renal failure (ESRF) awaiting transplantation (n = 17); and (iii) normal healthy controls (n = 18). Results: The percentage of CD4⁺CD25⁺Foxp3⁺ cells within the CD4⁺ cell population did not significantly alter at different time points post‐transplant. However, the percentage of CD4⁺CD25⁺Foxp3⁺ cells within the CD4⁺ population was significantly lower in RTR compared with patients with ESRF. In contrast, RTR and ESRF had a similar percentage of CD4⁺CD25⁺ cells expressing Foxp3. Multivariate analysis of PBL and clinical parameters demonstrated (i) a positive linear relationship between the percentage CD4⁺CD25⁺ cells expressing Foxp3 and estimated glomerular filtration rate and (ii) a higher percentage of CD4⁺CD25⁺ cells in the CD4⁺ cell population in patients with malignancy (the majority were skin cancers). Malignancy also correlated strongly with time post‐transplant and age of the RTR. Conclusion: Immune monitoring of the PBL phenotype in RTR using CD4, CD25 and Foxp3 may stratify RTR and predict graft outcome and function, and risk of complications from immunosuppression. Longitudinal and functional studies of Tregs are essential to extend the findings of the present study.     

Generation of an effective anti-lung cancer vaccine by DTPP-mediated photodynamic therapy and mechanistic studies

The objective of this study was to generate an effective vaccine against lung cancer using photosensitizing drug-mediated photodynamic therapy (PDT) and to study the mechanism. The efficiency of a photosensitizing drug (DTPP) was investigated by singlet oxygen yield determination, killing effect analysis, and cell apoptosis induction effect assessment. DTPP-based PDT tumor cell lysates and cell surface antigens obtained from acid-eluted adherent cells were then used as vaccines to prevent lung cancer using LA795 murine lung cells. The optimal protocol for PDT vaccine preparation was selected based on the tumor growth retardation effect of the vaccines, DTPP concentration, illumination dose, and numbers of DTPP-based PDT cells. To study the mechanism of the anti-tumor effect of vaccines, host anti-tumor immune responses were studied, including CD4⁺/CD8⁺ ratios and percentage of NK cells and serum cytokine levels. A comparison of cytokine (IFN-γ and IL-1) secretion from splenocytes and tumor pathologic features from mice immunized with vaccines were compared with controls and showed that the optimal protocol for PDT vaccine preparation was LA795 cells exposed to 10 μg/ml DTPP photosensitizer for 24 h, illuminated with 7.2 J/cm² at 20 mW/cm² (630 nm) and 2?×?10⁷ PDT cell lysates injected per mouse. DTPP-based PDT cell lysate vaccination had a significant inhibitory effect on tumor growth based on increased CD4⁺/CD8⁺ ratios, NK cell percentages, elevated serum IFN-γ and IL-1 levels, and lymphocyte aggregation at the edge of tumors. Thus, DTPP-based PDT can induce LA795 cell apoptosis that can generate anti-tumor effects without use of an adjuvant.     

The correlation of inflammatory markers and plasma vaspin levels in patients with diabetic nephropathy

Vaspin, a recently identified adipokine, is a visceral adipose tissue-derived serine protease inhibitor that may have insulin sensitizing effect on adipose tissue. Herein, we measured vaspin level in patients with different stages of diabetic nephropathy (DNP), and investigated the correlation of the vaspin level with other inflammatory parameters. 106 adult type 2 diabetic patients with no known chronic inflammatory disease were included and grouped according to the stage of DNP: Albuminuria <30?mg/day and estimated glomerular filtration rate (eGFR)?>?60?mL/min/1.73m² (Group-1); albuminuria 30–300?mg/day and eGFR >60?mL/min/1.73m² (Group-2); albuminuria >300?mL/min and eGFR <60?mL/min/1.73m² (Group-3). Demographic, clinical and laboratory data were recorded as well as vaspin, high sensitivity C-reactive protein (hsCRP), interleukin (IL)-1 and tumor necrosis factor (TNF)-α levels. There were 38, 35 and 33 patients in Group 1, 2 and 3, respectively. Groups were similar regarding age and gender. Vaspin level did not differ between groups. When all the groups were considered, vaspin was positively correlated with IL-6 level (r?=?0.215, p?=?0.041). No correlation of vaspin was found with IL-1, TNF-α and hsCRP levels (p?=?0.580, r?=?0.054; p?=?0.463, r?=?0.072; p?=?0.812, r?=?0.025, respectively). Vaspin levels of the patients with GFR ≥60?mL/min/1.73m² was less than that of patients with GFR <60?mL/min/1.73m² (p?=?0.03). Age and IL-6 were found to be the major determinants of vaspin level with linear regression analysis. In patients with DNP, vaspin level does not change within the early stages of DNP; while it is higher in patients with decreased GFR, which may be related with increasing inflammation regardless of the stage of the kidney disease.     

CD4+ CD25+ regulatory T cells suppressed the indirect xenogeneic immune response mediated by porcine epithelial cell pulsed dendritic cells

Nishimura T, Onda M, Takao S. CD4⁺ CD25⁺ regulatory T cells suppressed the indirect xenogeneic immune response mediated by porcine epithelial cell pulsed dendritic cells. Xenotransplantation 2010; 17: 313–323. © 2010 John Wiley & Sons A/S. Abstract: Background: CD4⁺ CD25⁺ regulatory T cells have been reported to suppress T cell‐mediated xenogeneic immune responses. Although the direct T cell response to xenogeneic cells is important, the indirect xenogeneic immune response mediated by dendritic cells (DCs) is also likely involved in rejection. We have generated an in vitro indirect immune reaction model and evaluated the effect of CD4⁺ CD25⁺ regulatory T cells on this system. Methods: Human DCs were generated from peripheral blood and cultured with X‐ray‐irradiated porcine kidney epithelial cells. Porcine cell‐pulsed DCs were mixed with autologous CD4⁺ T cells, CD4⁺ CD25^? T cells and/or CD4⁺ CD25⁺ T cells. After 7 days of culture, T cell proliferation was measured. Results: The co‐culture of human DCs and X‐ray‐irradiated porcine epithelial cells resulted in observable DC phagocytic activity within 2 days. These porcine cell–pulsed DCs stimulated CD4⁺ T cell proliferation much more potently than unpulsed DCs or porcine cells. This proliferation was blocked by CTLA4‐Ig or an anti‐HLA‐DR antibody. CD4⁺ CD25⁺ regulatory T cells also suppressed CD4⁺ CD25^? T cell proliferation in response to porcine cell‐pulsed DCs. Conclusions: An in vitro model of the indirect xenogeneic immune response was established. Porcine cell‐pulsed DCs stimulated CD4⁺ T cells, and CD4⁺ CD25⁺ regulatory T cells suppressed this response.