Increased frequency of CD39+CD73+ regulatory T cells and Deltex-1 gene expression level in kidney transplant recipients with excellent long-term graft function

BackgroundThe ability of regulatory T cells (Tregs) to limit inflammatory responses has been demonstrated. However, different subpopulations of this cell have varying abilities to suppress alloreactive immune responses. The primary goal of this study was to assess the frequency of CD4⁺FOXP3⁺CD39⁺CD73⁺ Tregs and Deltex-1 gene expression on long-term renal transplant function.MethodsA total of 49 subjects were divided into 3 groups: (i) the excellent long-term graft function (ELTGF) group, (ii) the chronic graft dysfunction (CGD) group, and (iii) the healthy control (HC) group. Following sample collection, peripheral blood mononuclear cells (PBMCs) were isolated, and the Deltex-1 gene expression level and the frequency of CD4⁺FOXP3⁺CD39⁺CD73⁺ Tregs were evaluated.ResultsThe ELTGF group had more CD4⁺FOXP3⁺ Tregs than the CGD group, but the difference was not statistically significant (P = 0.07). However, the frequency of CD4⁺FOXP3⁺CD39⁺CD73⁺ Tregs and the ratio of these cells to total CD4⁺ lymphocytes significantly increased in the ELTGF group than in the CGD group (P = 0.04 and P = 0.02 respectively). In addition, the expression level of the Deltex-1 gene was significantly lower in the CGD group than in the other 2 groups (P = 0.01 and P = 0.04 respectively).ConclusionsGiven the increased frequency of CD4⁺FOXP3⁺CD39⁺CD73⁺ Tregs and the expression level of the Deltex-1 gene in the ELTGF group, it appears that these factors probably improved function and long-term survival of the transplanted organ through the suppression of alloreactive responses and reduction of inflammation. In other words, one of the immunological mechanisms involved in the CGD group may be a deficiency in Tregs.     

Absence of CD4CD25 regulatory T cell expansion in renal transplanted patients treated in vivo with Belatacept mediated CD28–CD80/86 blockade

AimsBelatacept is a new recombinant molecule (CTLA4-Ig) that interferes with the second activation signal of T lymphocytes. CTLA4-Ig induced T cell allograft tolerance in rodents but not in primates. We examined the changes in peripheral lymphocyte subsets, including regulatory T cells, in renal transplant patients treated with Belatacept.MethodsA cross-sectional immunological study was carried out 6 months after transplantation in 28 patients enrolled in the Belatacept phase II study. Eighteen patients received Belatacept, mycophenolate mofetil and steroids (Belatacept group), while the control group of 10 patients received cyclosporine, mycophenolate mofetil and steroids (CsA group). Lymphocyte subsets were examined by flow cytometry. Foxp3 mRNA expression was measured by quantitative PCR.ResultsThe number of T lymphocytes and the percentage of CD3+ T cells were similar in both groups. However, the percentage of CD3+ CD4+ T cells was lower in the Belatacept group than in the control CsA group (B = 42.5% ± 13.7 vs CsA = 52.9% ± 9, p < 0.005), and the percentage of CD3+ CD8+ cells was higher in the Belatacept group than in the control (B = 32.9% ± 6.7 vs CsA = 19.5% ± 8.2, p < 0.0002). The percentage of CD19+ cells was similar in both groups. Among CD56+cells, only the percentage of CD16+ cells was significantly higher in the Belatacept group than in the control (B = 82% ± 12 vs CsA = 59.7% ± 25, p = 0.01). Among CD4 and CD8 T cells the percentage of activated lymphocytes expressing CTLA4, HLA-DR or CD40L was similar in both groups. The percentage of CD4+CD25+ T cells was higher in the CsA group. The percentage of regulatory CD4+CD25+ cells with bright CD25 staining was similar in both groups (B = 3.6 ± 2.3% vs CsA = 4.7 ± 1.9%, ns) as was the expression of FoxP3.ConclusionOur results indicated that Belatacept did not induce regulatory T cell expansion in vivo. We suggest that Belatacept treatment should be maintained after transplantation to allow graft acceptance.     

Ibrutinib suppresses alloantibody responses in a mouse model of allosensitization

BackgroundIbrutinib is a Bruton's tyrosine Kinase (BTK) antagonist that inhibits B cell receptor (BCR) signaling. Complete BTK deficiency is associated with absence of B-cells. Ibrutinb is currently approved by FDA for treatment of B-cell malignancies, including Waldenström macroglobulinaemia. We recently carried out studies to determine if ibrutinib could modify alloantibody responses.Materials and methodsA mouse model of allogenic sensitization using a C57BL/6 mouse as the recipient of a skin allograft from an HLA-A2 transgenic mouse was utilized to examine the effects of ibrutinib on alloantibody responses and B cell effector functions. Donor-specific antibody (DSA) levels were measured in a flow-cytometric antibody binding assay. Splenic T and B cell subsets and plasma cells were analyzed in flow cytometry.ResultsControl mice developed peak levels of DSA IgM at day 14 PTx while the ibrutinib treated mice had significantly lower levels of DSA IgM (p = 0.0047). Control mice developed HLA.A2-specific IgG antibodies at day 14 (230 ± 60 MFI) and reached peak levels at day 21 (426 ± 61 MFI). In contrast, mice in the treatment group had low levels of HLA.A2-specific IgG at day 14 (109 ± 59 MFI, p = 0.004) and day 21 (241 ± 86 MFI, p = 0.003). FACS analysis found a reduction of B220⁺ or CD19⁺ B cell population (p < 0.05). In addition, ibrutinib attenuated recall DSA IgG responses to re-sensitization (p < 0.05) and reduced CD38⁺ CD138⁺ plasma cells (p < 0.05) in the spleens.ConclusionsIbrutinib is effective in suppressing alloantibody responses through blocking BTK-mediated BCR signaling, leading to reduction of B cells and short-lived plasma cells in the spleens. Use of ibrutinib may provide benefits to HLA-sensitized transplant patients for alloantibody suppression.     

Effect of everolimus initiation and early calcineurin inhibitor withdrawal on myocardial FOXP3 + regulatory T cells in heart transplantation

BackgroundThrough immunosuppression CD4 + FoxP3 + regulatory T-cells (Tregs) play an indispensable role in allograft rejection. Post-HTx treatment with everolimus is associated with slower progression of cardiac allograft vasculopathy (CAV) – chronic rejection – than CNI based therapy. We hypothesized treatment with everolimus reduced the risk of CAV by modulating myocardial FoxP3 levels.Methods15 patients from the Schedule trial comparing everolimus, MMF, steroid and early CNI (Everolimus, n = 8) withdrawal to conventional CNI based immunosuppression (Controls, n = 7) after de novo HTx were included and FoxP3 + cells were quantified in 56 endomyocardial biopsies, and compared in the two patient groups. CAV was evaluated invasively using coronary intravascular ultrasound (IVUS).ResultsBaseline FoxP3 biopsy levels were similar in the two groups. The Everolimus group showed a significant increase in Foxp3 densities from baseline to time of one-year follow-up (median (IQR) = 4.8 × 10^− 7(20.4) Tregs/μm², P = 0.046) while Controls showed no significant change (median (IQR) = 3.1 × 10^− 7(6.5) Tregs/μm², P = 0.116). At 1-month follow-up FoxP3 densities correlated with the observed change in TAV from baseline to time of 1-year follow-up (r = 0.641, P = 0.034). FoxP3 densities at 1-week predicted acute cellular rejection (ACR) levels at 1 month (P = 0.026). No other correlations with ACR were found.ConclusionEverolimus treatment combined with early CNI elimination is associated with increased densities of Tregs 12-months post-HTx compared to patients receiving CNI based regimen. Furthermore, the density of myocardial FoxP3+ cells early after transplantation appears to predict at least one measure of CAV burden after one year.     

Effects of B cell depletion on T cell allogeneic Immune responses: A strategy to reduce allogeneic sensitization

BackgroundB cell depletion has been employed to treat antibody-mediated organ transplantation rejection, although the effects on cellular immune responses have not been extensively investigated.MethodsA model of B cell depletion used SCID/beige mice reconstituted with BALB/c splenocytes either depleted of B cells (BD) or not (BN). BD and B/N mice received C57BL/6 skin grafts and were sacrificed after 6 weeks (BD-S6 and BN-S6).ResultsRecall proliferative responses of BD-S6 splenocytes to C57BL/6 were significantly reduced compared to BN-S6, and central memory T cells' proportions (CD4⁺CD44⁺CD62L⁺ or CD8⁺CD44⁺CD62L⁺) were significantly decreased in BD-S6 spleens. Recall IFN-γ production by BD-S6 splenocytes was significantly reduced compared to BN-S6 splenocytes (p = 0.0028). Survival times of C57BL/6 heart grafts were significantly longer in SCID/beige mice reconstituted with BD-S6 splenocytes (8.5 ± 1.1 days) than for SCID/beige reconstituted with BN-S6 splenocytes (6.0 ± 1.1 days; p = 0.0006). Under cyclosporine therapy, C57BL/6 heart survival was significantly longer for SCID/beige reconstituted with BD-S6 splenocytes (17.5 ± 6.4 days) than those reconstituted with BN-S6 splenocytes (6.2 ± 1.5 days; p < 0.0001).ConclusionB cell depletion during allogeneic sensitization decreased memory T cells and recalls IFN-γ production and reduced second-set allograft rejection.     

High proportion of CD95+ and CD38+ in cultured CD8+ T cells predicts acute rejection and infection,respectively, in kidney recipients

The aim of this study was to find noninvasive T-cell markers able to predict rejection or infection risk after kidney transplantation.We prospectively examined T-lymphocyte subsets after cell culture stimulation (according to CD38, CD69, CD95, CD40L, and CD25 expression) in 79 first graft recipients from four centers, before and after transplantation. Patients were followed up for one year.Patients who rejected within month-1 (n = 10) showed high pre-transplantation and week-1 post-transplantation percentages of CD95⁺, in CD4⁺ and CD8⁺ T-cells (P < 0.001 for all comparisons). These biomarkers conferred independent risk for early rejection (HR:5.05, P = 0.061 and HR:75.31, P = 0.004; respectively). The cut-off values were able to accurately discriminate between rejectors and non-rejectors and Kaplan–Meier curves showed significantly different free-of-rejection time rates (P < 0.005). Patients who rejected after the month-1 (n = 4) had a higher percentage of post-transplantation CD69⁺ in CD8⁺ T-cells than non-rejectors (P = 0.002). Finally, patients with infection (n = 41) previously showed higher percentage of CD38⁺ in CD8⁺ T-cells at all post-transplantation times evaluated, being this increase more marked in viral infections. A cut-off of 59% CD38⁺ in CD8⁺ T-cells at week-1, week-2 and month-2 reached 100% sensitivity for the detection of subsequent viral infections.In conclusion, predictive biomarkers of rejection and infection risk after transplantation were detected that could be useful for the personalized care of kidney recipients.     

Pre-transplant infusion of donor-derived dendritic cells maintained at the immature stage by sinomenine increases splenic Foxp3+ Tregs in recipient rats after renal allotransplantation

ObjectiveThe immunosuppressive mechanism of sinomenine in organ allotransplantation was investigated, especially its effect of blocking dendritic cell (DC) maturation, which might influence the frequency of regulatory T cells (Tregs).MethodsBone marrow cells from male donor Wistar rats were induced to differentiate into DCs in vitro in the presence or absence of sinomenine, and characterized by flow cytometry. These two groups of DCs were respectively injected into male recipient Sprague-Dawley rats via the tail vein, at both high and low doses. Sprague-Dawley rats receiving saline injection were used as controls. Seven days later, renal transplantation was performed from donor Wistar rats to the recipient Sprague-Dawley rats. Seven days after transplantation, spleens were collected from the recipients. The proportions of Tregs and Foxp3⁺ Tregs to CD4⁺ T cells were determined using flow cytometry.ResultsWith sinomenine treatment, the frequency of mature DCs was reduced, as indicated by lower expression of the surface markers CD80, CD86, and RT1B. In recipient Sprague-Dawley rats that received sinomenine-treated DCs before renal allotransplantation, the proportions of splenic Tregs and Foxp3⁺ Tregs were significantly higher than in control recipients receiving saline or DCs without sinomenine treatment (all p < 0.05). A high dose of sinomenine-treated DCs (10⁶ cells) had a more obvious effect in increasing Tregs than the low dose (10⁵ cells) (p < 0.05).ConclusionPre-transplant infusion of donor-derived sinomenine-induced maturation arrested DCs could result in the increase of Foxp3⁺ Tregs in the spleens of recipients after renal allotransplantation.     

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.     

Donor age negatively affects the immunoregulatory properties of both adipose and bone marrow derived mesenchymal stem cells

PurposeAge negatively impacts the biologic features of mesenchymal stem cells (MSCs), including decreased expansion kinetics and differentiation potential. Clinically, donor-age may be within a wide spectrum; therefore, investigation of the role of donor's age on immunoregulatory potential is of critical importance to translate stem cell therapies from bench to bedside.MethodsAdipose and bone marrow derived MSCs (ASCs and BMSCs) were isolated in parallel from Lewis and Brown Norway rats of young (less than 4-week old) and senior groups (older than 15-month). The presentation of cells and time required for growth to 90% confluence was recorded. FACS sorting based on the expression of CD90 and CD29 double positive and CD45 CD11 double negative quantified the proportions of MSCs. After expansion, ASCs and BMSCs from different age groups were co-cultured in mixed lymphocyte reaction (MLR; Lewis vs. Brown Norway) assays. The suppression of CD3⁺CD4⁺ and CD3⁺CD8⁺ T cell populations by different sources of MSCs were compared.ResultsThe kinetics of cell growth was slower in old animals (17.3 ± 2 days) compared with young animals (8.8 ± 3 days), and cell morphology was irregular and enlarged in the senior groups. The yield of MSCs by FACS sorting was significantly higher in young groups compared to senior groups (p < 0.02). With regard to immunoregulatory potential, senior ASCs failed to induce any CD3⁺CD4⁺ T cell suppression (p > 0.05). In addition, young BMSCs-induced suppression was more prominent than seniors (p < 0.05).ConclusionsDonor age should be taken into consideration when using recipient MSC of either bone marrow or adipose origin in clinical applications.     

Increased levels of CD4+ and CD8+ T cells expressing CCR1 in patients developing allograft dysfunction; a cohort study

BackgroundLeukocyte infiltration into the graft has pivotal effects on kidney transplantation outcome. The present study sought to determine whether the expression of sequential chemokine receptors on CD4⁺ and CD8⁺ T cells in human renal allograft can predict clinical episodes.MethodsBlood samples from 52 consecutive renal transplant patients were evaluated at the time of transplantation and at three times (2, 90 and 180 days) after transplantation to analyze the expression of CCR1 and CXCR3 on CD4⁺ and CD8⁺ T cells by flowcytometry. A total of 30 biopsies, including protocol biopsy (n = 24) and cause biopsy (n = 6), were investigated according to the Banff criteria.ResultsThe mean percentage of CD4⁺ and CD8⁺ T cells expressing CCR1 was significantly increased in patients with allograft dysfunction (n = 25) (p = 0.006, p = 0.004). The mean fluorescence intensity of CXCR3 on CD4⁺ and CD8⁺ T cells were found to be significantly higher in graft dysfunction than that in well-functioning grafts. (p < 0.001, p = 0.007). Receiver Operating Characteristic (ROC) Curve Analysis showed that the calculated AUC was 0.86 at the third month for CD4⁺ CCR1⁺ and CD8⁺ CCR1⁺ (p < 0.001). Multiple logistic regression analysis showed that an increase in CD4⁺ expressing CXCR3 leads to a lower risk of graft dysfunction (OR = 0.37), while an increase in CD8⁺ expressing CCR1 results in a higher risk of graft dysfunction (OR = 3.66).ConclusionDuring renal transplantation, CD4⁺ and CD8⁺ T cells expressing CCR1 were increased in patients who developed graft dysfunction. These findings may prospectively predict allograft dysfunction, and help elucidate the underlying pathogenic mechanisms.     

ASP0028 in combination with suboptimal-dose of tacrolimus in Cynomolgus monkey renal transplantation model

FTY720, a S1P-receptor modulator, has shown to be effective in several transplant and autoimmune disease models, via modulating lymphocyte homing into secondary lymphoid organs (SLOs), and thereby reducing these cells in peripheral blood. ASP0028, a newly developed S1P₁/S1P₅-selective agonist, presented comparable efficacy to FTY720 and wider safety margins than FTY720. In this study, we assessed the efficacy and safety of ASP0028 co-administered with suboptimal-dose of tacrolimus in the Cynomolgus monkey renal transplantation model. Seven animals in group-1 or group-2 received mono-tacrolimus 1.0 mg/kg once a day (QD), or ASP0028 0.6 mg/kg plus tacrolimus 1.0 mg/kg QD, respectively. Eight animals in group-3 received ASP0028 1.2 mg/kg plus tacrolimus 1.0 mg/kg QD. The allograft median survival time (MST) in group-2 and group-3 were significantly extended to 41 and 61.5 days, versus that of 28 days in group-1 (p = 0.036 and 0.001, respectively). ASP0028 administration remarkably reduced absolute numbers of peripheral lymphocytes, particularly subsets of CD4⁺/ or CD8⁺/naive and central memory cells, CD4⁺/Treg cells, and to a lesser extent on B cells, but not CD4⁺/ or CD8⁺/effector memory cells and NK cells. These data show ASP0028 combined with suboptimal-dose of tacrolimus effectively prolongs renal allograft survival in nonhuman primates (NHPs) with well tolerated safety, supporting its further investigation to optimize CNI-sparing regimens.     

Cytokines affecting CD4+ T regulatory cells in transplant tolerance. III. Interleukin-5 (IL-5) promotes survival of alloantigen-specific CD4+ T regulatory cells

CD4⁺ T cells mediate antigen-specific allograft tolerance, but die in culture without activated lymphocyte derived cytokines. Supplementation of the media with cytokine rich supernatant, from ConA activated spleen cells, preserves the capacity of tolerant cells to transfer tolerance and suppress rejection. rIL-2 or rIL-4 alone are insufficient to maintain these cells, however. We observed that activation of naïve CD4⁺ CD25⁺ FOXP3⁺ Treg with alloantigen and the Th2 cytokine rIL-4 induces them to express interleukin-5 specific receptor alpha (IL-5Rα) suggesting that IL-5, a Th2 cytokine that is produced later in the immune response may promote tolerance mediating Treg.This study examined if recombinant IL-5(rIL-5) promoted survival of tolerant CD4⁺, especially CD4⁺ CD25⁺ T cells. CD4⁺ T cells, from DA rats tolerant to fully allogeneic PVG heart allografts surviving over 100 days without on-going immunosuppression, were cultured with PVG alloantigen and rIL-5. The ability of these cells to adoptively transfer tolerance to specific-donor allograft and suppress normal CD4⁺ T cell mediated rejection in adoptive DA hosts was examined. Tolerant CD4⁺ CD25⁺ T cells' response to rIL-5 and expression of IL-5Rα was also assessed.rIL-5 was sufficient to promote transplant tolerance mediating CD4⁺ T cells' survival in culture with specific-donor alloantigen. Tolerant CD4⁺ T cells cultured with rIL-5 retained the capacity to transfer alloantigen-specific tolerance and inhibited naïve CD4⁺ T cells' capacity to effect specific-donor graft rejection. rIL-5 promoted tolerant CD4⁺ CD25⁺ T cells' proliferation in vitro when stimulated with specific-donor but not third-party stimulator cells. Tolerant CD4⁺ CD25⁺ T cells expressed IL-5Rα.This study demonstrated that IL-5 promoted the survival of alloantigen-specific CD4⁺ CD25⁺ T cells that mediate transplant tolerance.     

Correlação entre saturação venosa central de oxigênio perioperatória e mortalidade em cirurgia cardíaca: estudo prospectivo observacional

BackgroundCardiac surgery can produce persistent deficit in the ratio of Oxygen Delivery (DO₂) to Oxygen Consumption (VO₂). Central venous oxygen Saturation (ScvO₂) is an accessible and indirect measure of DO₂/VO₂ ratio.ObjectiveTo monitor perioperative ScvO₂ and assess its correlation with mortality during cardiac surgery.MethodsThis prospective observational study evaluated 273 patients undergoing cardiac surgery. Blood gas samples were collected to measure ScvO₂ at three time points: T0 (after anesthetic induction), T1 (end of surgery), and T2 (24 hours after surgery). The patients were divided into two groups (survivors and nonsurvivors). The following outcomes were analyzed: intrahospital mortality, length of Intensive Care Unit (ICU) and hospital stay (LOS), and variation in ScvO₂.ResultsOf the 273 patients, 251 (92%) survived and 22 (8%) did not. There was a significant perioperative reduction of ScvO₂ in both survivors (T0 = 78% ± 8.1%, T1 = 75.4% ± 7.5%, and T2 = 68.5% ± 9%; p < 0.001) and nonsurvivors (T0 = 74.4% ± 8.7%, T1 = 75.4% ± 7.7%, and T2 = 66.7% ± 13.1%; p < 0.001). At T0, the percentage of patients with ScvO₂ < 70% was greater in the nonsurvivor group (31.8% vs. 13.1%; p = 0.046) and the multiple logistic regression showed that ScvO₂ is an independent risk factor associated with death, OR = 2.94 (95% CI 1.10 − 7.89) (p = 0.032). The length of ICU and LOS were 3.6 ± 3.1 and 7.4 ± 6.0 days respectively and was not significantly associated with ScvO₂.ConclusionsEarly intraoperative ScvO₂ < 70% indicated a higher risk of death. A perioperative reduction of ScvO₂ was observed in patients undergoing cardiac surgery, with high intraoperative and lower postoperative levels.     

Active Comparator Trial of Teriparatide vs Alendronate for Treating Glucocorticoid-Induced Osteoporosis: Results From the Hispanic and Non-Hispanic Cohorts

Glucocorticoid use is a leading cause of secondary osteoporosis. This post hoc analysis compared teriparatide vs alendronate on bone mineral density (BMD) in Hispanic and non-Hispanic patients with glucocorticoid-induced osteoporosis. The 18-mo results from all patients (N = 428) in a double-blind trial of teriparatide (20 μg/d) and alendronate (10 mg/d) who had taken glucocorticoids for ≥3 mo were reported (Saag et al. N Engl J Med 2007). The present study analyzed results from the Hispanic (n = 61) and non-Hispanic (n = 367) cohorts. The BMD was measured by dual-energy X-ray absorptiometry (DXA). In the Hispanic cohort at 18 mo, there were significantly greater increases from baseline in the teriparatide vs alendronate group in lumbar spine BMD (9.8% ± 1.7% vs 4.2% ± 1.4%; p < 0.001; mean ± SE) and total hip BMD (5.9% ± 1.6% vs 1.3% ± 1.3%, p < 0.001), with no significant difference between groups at the femoral neck (4.3% ± 2.2% vs 2.0% ± 1.8%, p = 0.228). Within each treatment group, the BMD responses were not significantly different in the Hispanic vs non-Hispanic cohort. The number of patients reporting ≥1 adverse event was not significantly different between treatments in either cohort, with more patients reporting nausea in the teriparatide group. In summary, teriparatide was more efficacious than alendronate in increasing BMD in Hispanic and non-Hispanic patients with glucocorticoid-induced osteoporosis. Both treatments were generally well tolerated.     

Patterns of Physical Activity Progression in Patients With COPD

IntroductionAlthough mean physical activity in COPD patients declines by 400?500 steps/day annually, it is unknown whether the natural progression is the same for all patients. We aimed to identify distinct physical activity progression patterns using a hypothesis-free approach and to assess their determinants.MethodsWe pooled data from two cohorts (usual care arm of Urban Training [NCT01897298] and PROactive initial validation [NCT01388218] studies) measuring physical activity at baseline and 12 months (Dynaport MoveMonitor). We identified clusters (patterns) of physical activity progression (based on levels and changes of steps/day) using k-means, and compared baseline sociodemographic, interpersonal, environmental, clinical and psychological characteristics across patterns.ResultsIn 291 COPD patients (mean ± SD 68 ± 8 years, 81% male, FEV₁ 59 ± 19%_pred) we identified three distinct physical activity progression patterns: Inactive (n = 173 [59%], baseline: 4621 ± 1757 steps/day, 12-month change (?): ∧487 ± 1201 steps/day), Active Improvers (n = 49 [17%], baseline: 7727 ± 3275 steps/day, ?: + 3378 ± 2203 steps/day) and Active Decliners (n = 69 [24%], baseline: 11 267 ± 3009 steps/day, ?: ∧2217 ± 2085 steps/day). After adjustment in a mixed multinomial logistic regression model using Active Decliners as reference pattern, a lower 6-min walking distance (RRR [95% CI] 0.94 [0.90?0.98] per 10 m, P = .001) and a higher mMRC dyspnea score (1.71 [1.12?2.60] per 1 point, P = .012) were independently related with being Inactive. No baseline variable was independently associated with being an Active Improver.ConclusionsThe natural progression in physical activity over time in COPD patients is heterogeneous. While Inactive patients relate to worse scores for clinical COPD characteristics, Active Improvers and Decliners cannot be predicted at baseline.     

Impact of pillow height on double-lumen endotracheal tube intubation with McGRATH MAC: a prospective randomized clinical trial

Study objectiveThis study aimed to compare the impact of pillow height on double-lumen tracheal tube (DLT) intubation with McGRATH MAC (McG) in patients undergoing elective surgery.DesignRandomized clinical trial.SettingOperating room.PatientsFifty adult patients scheduled for elective surgery under 1-lung ventilation with an American Society of Anesthesiologists physical status of 1 to 3.InterventionsDLT intubation with McG was performed with a high pillow (HP group; 25 patients) or low pillow (LP group; 25 patients) by anesthesiologists.MeasurementsIntubation time, number of laryngoscopy, number of tracheal intubation attempts to successful intubation, percentage of glottic opening score, and subjective difficulty of laryngoscopy and tube passage through the glottis were assessed.Main ResultsIntubation time was significantly shorter in the HP group compared with the LP group (HP: 32.1 ± 14.9 seconds vs LP: 49.4 ± 11.2 seconds, P < .001). The number of laryngoscopy were 1 (HP group, 22 patients; LP group, 17 patients), 2 (HP group, 3 patient; LP group, 7 patients), and 3 (HP group, 0 patient; LP group, 1 patient), with no significant difference between the 2 groups (P = .197). Although the percentage of glottic opening score did not significantly differ between HP and LP groups (HP: 95.6% ± 6.7% vs LP: 96.0% ± 12.3%, P = .08), the number of tracheal intubation attempts was significantly lower in the HP group compared with the LP group (P = .009). The visual analog scale score for laryngoscopy did not significantly differ between the 2 groups (P = .54). However, the visual analog scale for tube passage through the glottis was significantly higher in the LP group than in the HP group (P < .001).ConclusionsIntubation with an HP was associated with a better DLT intubation profile than with an LP with McG, possibly due to smoother tracheal tube progression through the glottis.     

Clinical utility of CD4 + function assessment (ViraCor-IBT ImmuKnow test) in lung recipients

The ImmuKnow assay measures cell-mediated immunity, quantifying ATP production from peripheral blood CD4 + T-cells in solid-organ transplant patients who undergo immunosuppressive therapy. We aimed to measure functional immunity in lung transplant recipients and correlate Immuknow values with immunosuppression levels, presence of chronic lung allograft dysfunction (CLAD) and infections. We evaluated 61 lung recipients who underwent follow-up for lung transplantation between 2010 and 2014. Rejection and infection were retrospectively analyzed. The association between over-immunosuppression and a number of predictors was assessed by means of univariate and multivariate logistic regression models. 71 out of 127 samples (56%) showed an over-immunosuppression with an ImmuKnow assay mean level of 112.92 ng/ml (SD ± 58.2), vs. 406.14 ng/ml (SD ± 167.7) of the rest of our cohort. In the over-immunosuppression group we found 51 episodes of infection (71%) (OR 2.754, 95% CI 1.40–5.39; P-value 0.003). In the other group, only 25 samples (44%) were taken during an infectious episode. The mean absolute ATP level was significantly different between patients with or without infection (202.38 ± 139.06 ng/ml vs. 315.51 ± 221.60 ng/ml; P < 0.001). RAS (Restrictive allograft syndrome) was associated to low ImmuKnow level (P < 0.001). These results were confirmed by the multivariate analysis. The ImmuKnow assay levels were significantly lower in infected lung transplant recipients compared with non-infected recipients and in RAS patients.     

Alloantigen-specific CD4+ regulatory T cells induced in vivo by ultraviolet irradiation after alloantigen immunization require interleukin-10 for their induction and activation,and flexibly mediate bystander immunosuppression of allograft rejection

Ultraviolet (UV) irradiation prior to antigen immunization is employed to induce antigen-specific regulatory T cells (Tregs). UV-induced Tregs demonstrate unique bystander suppression, although antigen-specific activation is required initially. We previously reported the phenotype of alloantigen-specific transferable Tregs induced by UV-B irradiation after immunization was the same as T regulatory type 1-like CD4⁺ T cells, with antigen-specific interleukin (IL)-10 production. Here, by using semi-allogeneic transplantation models in vivo, we investigated the role of IL-10 in the induction and activation of these Tregs, and the possibility of bystander suppression of third-party allograft rejection. Naïve mice (H-2^b) were immunized with alloantigen (H-2^b/d), and received UV-B irradiation (40 kJ/m²) 1 week later. Four weeks afterwards, splenic CD4⁺ T cells were purified from the UV-irradiated immunized mice, and were transferred into naïve mice (H-2^b). Allografts expressing the same alloantigen as T-cell donors were immunized against (H-2^b/d) or an irrelevant alloantigen (H-2^b/k) were transplanted to CD4⁺ T-cell-transferred mice, and an alloantigen-specific prolongation of allograft survival observed. Experiments where IL-10 was neutralized by monoclonal antibody in the induction or effector phase revealed that IL-10 is critical, not only for induction but also for immunosuppressive function of CD4⁺ Tregs induced by UV irradiation after alloantigen immunization. Third-party allografts (H-2^d/k) were transplanted to CD4⁺ T-cell-transferred mice, and graft survival was also prolonged. Even a graft only partially compatible with immunized alloantigen worked well in vivo to activate CD4⁺ Tregs induced by UV irradiation after alloantigen immunization, which resulted in the bystander suppression of third-party allograft rejection.     

Combined Superoxide Dismutase Mimetic and Peroxynitrite Scavenger Protects Against Neointima Formation After Endarterectomy in Association with Decreased Proliferation and Nitro-oxidative Stress

ObjectiveReactive oxygen and nitrogen species (e.g., peroxynitrite) may trigger neointima formation leading to restenosis. In a rat carotid endarterectomy (CEA) model, we investigated the effects of the manganese(III)tetrakis(4-benzoic acid)porphyrin (MnTBAP), a superoxide dismutase (SOD) mimetic and peroxynitrite scavenger on neointima formation.MethodsCEA was performed in male Sprague–Dawley rats. Animals received either vehicle (control group; n = 15) or 15 mg kg^?1 day^?1 MnTBAP intraperitoneally for 3 weeks (treatment group; n = 13). Four groups of carotids were analysed: the left, uninjured carotids (sham) and the right, injured carotids (control CEA) from the control group, the right, injured carotids from the treatment group (CEA + MnTBAP) and an additional group of carotids that were harvested 1 h following endarterectomy. The analysis of carotid arteries was performed by histology, immunohistochemistry and real-time polymerase chain reaction (PCR). Plasma malondialdehyde (MDA) levels were measured by lipid hydroperoxidase assay.ResultsStenosis rate (10.5 ± 8.1% vs. 45.4 ± 28.3%), the percentage of proliferating cell nuclear antigen-positive cells (13.4 ± 7.1% vs. 23.3 ± 11.0%) and nitrotyrosine immunoreactivity (5.8 ± 1.9 vs. 8.0 ± 2.0) were significantly reduced in the vascular wall of the CEA + MnTBAP group compared with control CEA group. Ratio of Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL)-positive nuclei was significantly lower after antioxidant therapy (41.7 ± 26.7% vs. 64.9 ± 18.5%). Plasma MDA levels increased after endarterectomy (11.7 ± 4.8 vs. 4.1 ± 2.0 μmol l^?1) and reduced in the treatment group (3.2 ± 2.1 μmol l^?1). No significant gene regulation after MnTBAP treatment could be noted.ConclusionsMnTBAP decreased neointima formation, which was associated with reduced vascular smooth muscle cell proliferation and attenuated local and systemic nitro-oxidative stress.