Marine peptides as immunomodulators: Californiconus californicus-derived synthetic conotoxins induce IL-10 production by regulatory T cells (CD4+Foxp3+)

Abstract

Context: CD4⁺ T lymphocytes are able to differentiate into distinct subtypes according to several immunological scenarios, including T helper (Th)1, Th2, Th17 and regulatory T (Treg) cells. CD4⁺ T cells are phenotypically flexible and have specific ion channels, such as the nicotinic acetylcholine receptors (nAChR) that could be modulated by peptides produced by marine snails, known as conotoxins. Their effect on T lymphocytes has not been explored and emerging evidence suggests that these peptides may have immunomodulatory activities.

Objective: This study investigated the effect of two Californiconus californicus-derived synthetic conotoxins on the proliferation and differentiation of T lymphocyte subpopulations Th1, Th2, Th17 and Treg.

Methods: Cells from lymph nodes of BALB/c mice were cultured in the presence of conotoxins cal14.1b and cal14.2c (5.5?μM), during 96?h. Cell proliferation and intracellular cytokine production (IFN-γ, IL-4, IL-17 and IL-10) were analyzed by flow cytometry.

Results and Discussion: cal14.1b and cal14.2c increased intracellular IL-10 production in Treg (CD3⁺CD4⁺Foxp3⁺) cells and decreased intracellular IL-17 production (CD3⁺CD4⁺) after 72?h of culture. Conotoxins did not show any effect on T cell proliferation nor Th1/Th2 balance.

Conclusion: These results suggest that synthetic conotoxins exert immunomodulatory activity, especially by regulating specific functions on T lymphocytes.     

Predictors of progression from moderate to severe coronavirus disease 2019: a retrospective cohort

ObjectiveMost cases of coronavirus disease 2019 (COVID-19) are identified as moderate, which is defined as having a fever or dry cough and lung imaging with ground-glass opacities. The risk factors and predictors of prognosis in such cohorts remain uncertain.MethodsAll adults with COVID-19 of moderate severity diagnosed using quantitative RT-PCR and hospitalized at the Central Hospital of Wuhan, China, from 1 January to 20 March 2020 were enrolled in this retrospective study. The main outcomes were progression from moderate to severe or critical condition or death.ResultsAmong the 456 enrolled patients with moderate COVID-19, 251/456 (55.0%) had poor prognosis. Multivariate logistic regression analysis identified higher neutrophil count: lymphocyte count ratio (NLR) on admission (OR 1.032, 95% CI 1.042–1.230, p 0.004) and higher C-reactive protein (CRP) on admission (OR 3.017, 95% CI 1.941–4.690, p < 0.001) were associated with increased OR of poor prognosis. The area under the receiver operating characteristic curve (AUC) for NLR and CRP in predicting progression to critical condition was 0.77 (95% CI 0.694–0.846, p < 0.001) and 0.84 (95% CI 0.780–0.905, p < 0.001), with a cut-off value of 2.79 and 25.95 mg/L, respectively. The AUC of NLR and CRP in predicting death was 0.81 (95% CI 0.732–0.878, p < 0.001) and 0.89 (95% CI 0.825–0.946, p < 0.001), with a cut-off value of 3.19 and 33.4 mg/L, respectively.ConclusionsHigher levels of NLR and CRP at admission were associated with poor prognosis of individuals with moderate COVID-19. NLR and CRP were good predictors of progression to critical condition and death.     

The association of γδ-T cells with bronchopulmonary dysplasia in premature infants

BackgroundAs the survival rate of premature infants increases, the incidence of bronchopulmonary dysplasia (BPD), a chronic complication of premature infants, is also higher than before. The pathogenesis of BPD is complicated, and immune imbalance and inflammatory response may play important roles in it.ObjectiveTo investigate the correlation between lymphocyte subsets in peripheral blood, especially γδ-T cells, and BPD of preterm infants.Materials and methodThe study was carried out with the peripheral blood of premature infants (GA < 32 weeks, BW < 1500 g), which were collected at 24 h or 3–4 weeks after birth. The infants were divided into non-BPD groups and BPD groups that were classified as mild or moderate and severe in preterm infants based on the magnitude of respiratory support at 28 days age and 36 weeks postmenstrual age. The γδ-T, CD3+, CD4+, CD8+ and total lymphocyte subsets in peripheral blood were detected by flow cytometry.ResultsThe percentages of T lymphocyte subsets in peripheral blood were not different between BPD and non-BPD within 24 h after birth. And no significant difference was found in T lymphocyte subsets among neonates with BPD of different severities. However, the infants who developed BPD had a significant increase in γδ-T cells compared to non-BPD ones within 3–4 weeks after birth.ConclusionsIt seems that γδ-T cells in peripheral blood are correlated with BPD. However, the causality of BPD and various lymphocytes remains unclear, which need to be further studied.     

Neutrophil-lymphocyte ratio: a controversial marker in predicting Crohn’s disease severity

Peripheral blood-derived inflammation-based scores such as the neutrophil-lymphocyte ratio (NLR) have recently been proposed as prognostic markers in ulcerative colitis. In some previous serological markers are commonly used to detect the severity of the Crohn’s disease (CD), but their sensitivity and specificity are relatively low. So we want to use simple indicators which are easy to obtain to predict disease severity. Now, we investigated and compared the capacity of NLR and other inflammatory markers in detecting CD activity and differentiating CD patients from healthy controls. These CD patients had not received corticosteroid or immunosuppressive drugs within a defined period of time. Data from our hospital between 2010 and 2012 was used. Neutrophil-lymphocyte ratio (NLR), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), white blood cells (WBC), platelet count and albumin were measured in 44 patients with active CD, 66 patients with inactive CD, and 55 healthy blood donors. Disease activity was assessed by the Crohn’s Disease Activity Index. In the active CD group, NLR values were found to be elevated compared to inactive CD patients and controls (6.00±7.38, 5.53±6.18 and 1.84±0.85, respectively), but statistical difference was not found between active and inactive CD groups. The overall accuracy of NLR (cutoff: 2.13 fl), CRP (cutoff: 10.5 mg/dl), ESR (cutoff: 19.5 mm/hour) and WBC (cutoff: 9.2 × 10⁹/l) in differentiating CD patients from healthy controls was 80.9%, 67.3%, 71% and 60% respectively. NLR values were found to be correlated with WBC and CRP levels. NLR increased in CD patients compared with healthy subjects. NLR had the best accuracy in determination of CD patients and healthy controls. NLR did not show a discriminative value in disease activity.     

Combined effects of very short “all out” efforts during sprint and resistance training on physical and physiological adaptations after 2 weeks of training

Purpose

The aim of this study was to compare the combined effects of resistance and sprint training, with very short efforts (5 s), on aerobic and anaerobic performances, and cardiometabolic health-related parameters in young healthy adults.

Methods

Thirty young physically active individuals were randomly allocated into four groups: resistance training (RTG), sprint interval training (SITG), concurrent training (CTG), and control (CONG). Participants trained 3 days/week for 2 weeks in the high-intensity interventions that consisted of 6–12 “all out” efforts of 5 s separated by 24 s of recovery, totalizing ~ 13 min per session, with 48–72 h of recovery between sessions. Body composition, vertical jump, lower body strength, aerobic and anaerobic performances, heart rate variability (HRV), and redox status were evaluated before and after training. Total work (TW), rating of perceived exertion (CR-10 RPE) and mean HR (HR_mean) were monitored during sessions. Incidental physical activity (PA), dietary intake and perceived stress were also controlled.

Results

Maximum oxygen consumption (VO_2max) significantly increased in SITG and CTG (P < 0.05). Lower body strength improved in RTG and CTG (P < 0.05), while countermovement jump (CMJ) was improved in RTG (P = 0.04) only. Redox status improved after all interventions (P < 0.05). No differences were found in TW, PA, dietary intake, and psychological stress between groups (P > 0.05).

Conclusions

RT and SIT protocols with very short “all out” efforts, either performed in isolation, or combined, demonstrated improvement in several physical fitness- and health-related parameters. However, CT was the most efficient exercise intervention with improvement observed in the majority of the parameters.

     

Neutrophil-to-Lymphocyte Ratio at Emergency Room Predicts Mechanical Complications of ST-segment Elevation Myocardial Infarction

BackgroundThe neutrophil-to-lymphocyte ratio (NLR) has been proven to be a reliable inflammatory marker. A recent study reported that elevated NLR is associated with adverse cardiovascular events in patients with ST-segment elevation myocardial infarction (STEMI). We investigated whether NLR at emergency room (ER) is associated with mechanical complications of STEMI undergoing primary percutaneous coronary intervention (PCI).MethodsA total of 744 patients with STEMI who underwent successful primary PCI from 2009 to 2018 were enrolled in this study. Total and differential leukocyte counts were measured at ER. The NLR was calculated as the ratio of neutrophil count to lymphocyte count. Patients were divided into tertiles according to NLR. Mechanical complications of STEMI were defined by STEMI combined with sudden cardiac arrest, stent thrombosis, pericardial effusion, post myocardial infarction (MI) pericarditis, and post MI ventricular septal rupture, free-wall rupture, left ventricular thrombus, and acute mitral regurgitation during hospitalization.ResultsPatients in the high NLR group (> 4.90) had higher risk of mechanical complications of STEMI (P = 0.001) compared with those in the low and intermediate groups (13% vs. 13% vs. 23%). On multivariable analysis, NLR remained an independent predictor for mechanical complications of STEMI (RR = 1.947, 95% CI = 1.136–3.339, P = 0.015) along with symptom-to balloon time (P = 0.002) and left ventricular dysfunction (P < 0.001).ConclusionNLR at ER is an independent predictor of mechanical complications of STEMI undergoing primary PCI. STEMI patients with high NLR are at increased risk for complications during hospitalization, therefore, needs more intensive treatment after PCI.     

Arctigenin from <Emphasis Type="Italic">Arctium lappa</Emphasis> inhibits interleukin-2 and interferon gene expression in primary human T lymphocytes

Background  

Arctium lappa (Niubang), a Chinese herbal medicine, is used to treat tissue inflammation. This study investigates the effects of arctigenin (AC), isolated from A. lappa, on anti-CD3/CD28 Ab-stimulated cell proliferation and cytokine gene expression in primary human T lymphocytes.     

Sarcopenia and high NLR are associated with the development of hyperprogressive disease after second-line pembrolizumab in patients with non-small-cell lung cancer

The aim of this multi-center retrospective study was to evaluate the incidence of hyperprogressive disease (HPD) after second-line treatment with pembrolizumab in patients (n = 167) with metastatic non-small-cell lung cancer (NSCLC) whose tumors expressed programmed cell death ligand 1 (PD-L1) in ≥ 1% and to search for hematological and imaging biomarkers associated with its development. Prior to chemotherapy, neutrophil : lymphocyte ratio (NLR1) and platelet : lymphocyte ratio (PLR1), and prior to immunotherapy, NLR2 and PLR2 were retrospectively analyzed. The psoas major muscle area (PMMA) was calculated at the L3 position on computed tomography before chemotherapy (PMMA1) and before immunotherapy (PMMA2) (n = 112). Patients with ∆PMMA (1-PMMA2/PMMA1) × 100 ≥ 10% were considered to have sarcopenia (low muscle mass). After treatment with pembrolizumab on the first computerized tomography (CT) scan evaluation, patients were subdivided as follows as: hyperprogressors (HPs), progressors (Ps), non-progressors (NPs) and pseudoprogressors (PPs). HPs had significantly higher ∆PMMA levels, NLR2 and PLR2 than the other patients. Moreover, in multinomial logistic regression analysis, higher levels of ∆PMMA were associated with a decreased likelihood of being a P [odds ratio (OR) = 0·81; 95% confidence interval (CI) = 0·65–0·99; P = 0·047] or an NP (OR = 0·76; 95% CI = 0·62–0·94; P = 0·012) versus an HP. Higher NLRs tended to decrease the likelihood of being a P versus an HP (OR = 0·66; 95% CI = 0·42–1·06; P = 0·09) and significantly decreased the likelihood of being an NP versus an HP (OR = 0·44; 95% CI = 0·28–0·69; P < 0·0001). Our data suggest that a high pre-immunotherapy NLR2 and the presence of sarcopenia are potential risk factors for the development of HPD.     

Prognostic impact of neutrophils-to-lymphocytes ratio (NLR), PD-L1 expression,and tumor immune microenvironment in laryngeal cancer

PurposeIn laryngeal carcinoma (LSCC), tumor immune microenvironment is attracting increasing interest, given the recent progresses in immunotherapy. Immune cells migrate to tumors as a result of a tumor antigen-induced immune reaction and cancer cells recruit immune regulatory cells to induce an immunosuppressive network, resulting in the escape from host immunity. This interaction reflects both on tumor microenvironment and systemic inflammatory status. Blood neutrophil-to-lymphocyte ratio (NLR), reflecting a highly pro-inflammatory status, has been related to worse oncological survival outcomes.The aim of this study was to analyze in LSCC the relationship between circulating inflammatory cells (also in terms of NLR) and tumor immune microenvironment histopathological features (programmed cell death ligand 1 [PD-L1] expression, and tumor-infiltrating lymphocytes [TILs]), also investigating their clinical-pathological and prognostic significance.MethodsBlood pre-operative NLR, and, at pathology, PD-L1 (in terms of combined positive score [CPS]) and TILs were assessed on 60 consecutive cases of LSCC.ResultsBlood NLR, neutrophils, and lymphocytes counts showed a significant value in predicting DFS and recurrence risk. Moreover, PD-L1 CPS ≥ 1 and TILs count rate ≥30% were associated with higher disease-free survival (DFS) and reduced recurrence risk. A logistic regression model found a significant positive association between increasing NLR values, and PD-L1 CPS < 1 and TILs count rate <30%.ConclusionsFurther studies are needed to better characterize the role of pre-operative blood NLR in association with PD-L1 expression and tumor immune microenvironment features as prognostic factors and markers of anti-tumor immune response in LSCCs, also with regard to the effectiveness of immunotherapeutic protocols.     

Immunotherapy of Patients with Recurrent Spontaneous Miscarriage and Idiopathic Infertility: Does the Immunization-Dependent Th2 Cytokine Overbalance Really Matter?

Recurrent spontaneous miscarriage (RSM) and idiopathic infertility (IIF) are partially caused by immunologic disturbances. Paternal lymphocyte immunization (PLI) is proposed for restoration of the proper Th1/Th2 balance in these patients, but still there are controversies on PLI mechanism, its efficacy and identification of patients who may benefit from this therapy. The study group consisted of n = 34 RSM and n = 42 IIF women with unexplained miscarriage or IIF. PLI was offered as a treatment in both groups. Peripheral blood lymphocyte (PBL) populations (CD3⁺, CD3⁻/CD19⁺, CD3⁺/CD4⁺, CD3⁺/CD8⁺, CD3⁻/CD16⁺CD56⁺) were studied before immunization, while PBL cytokine secretion (IFN-γ, TNF-α, IL-10, IL-5, IL-4, IL-2), before and after immunization, pre-conceptionally in both groups. The reference PBL ratio and cytokine levels were adopted from previously studied normal fertile women. PBL populations, concentration and ratio of Th1/Th2 cytokines did not differ between RSM and IIF patients. Compared to the results observed in normal fertile women the levels of IFN-γ, TNF-α and IL-2 were higher, while IL-10 lower in both RSM and IIF patients (p < 0.01). After immunization a decrease of IFN-γ (RSM and IIF groups) and IL-4 and IL-10 (RSM group) were observed, as well as an increase in TNF-α/IL-4 ratio (RSM group) (p < 0.05). No differences in Th1/Th2 concentration and ratio between patients with successful and unsuccessful pregnancy were observed. No significant correlations between success and particular cytokine concentration were observed. Concentrations of Th1/Th2 cytokines and PBL populations did not differ between RSM and IIF women. Th1 shift in both RSM and IIF patients was observed in comparison to fertile women. Treatment with PLI-induced pre-conceptionally cytokine changes which neither indicated Th2 shift nor correlated with subsequent pregnancy success.     

外周血中性粒细胞/淋巴细胞比值在慢性阻塞性肺病相关肺动脉高压患者中的预后价值

目的：探讨外周血中性粒细胞/淋巴细胞比值(neutrophil/lymphocyte ratio,NLR)对慢性阻塞性肺病(chronic obstructive pulmonary disease,COPD)相关肺动脉高压患者预后的评判价值。方法：选择2013年1月至2014年3月收治入上海交通大学医学院附属新华医院急诊科的200例COPD相关肺动脉高压(pulmonary hypertension,PH)患者为研究对象,对其进行至少2年的生存随访,随访终点为全因死亡,按照生存情况分为生存组和死亡组;记录各组入院24 h的一般临床资料,血常规[C反应蛋白(C-reactive protein,CRP)、中性粒细胞计数(neutrophils count,NEU)及淋巴细胞计数(lymphocyte count,LYM)并计算两者间比值(NLR)]、肌酐、尿素氮、胆红素、WHO肺动脉高压功能分级、肺动脉收缩压等;绘制受试者工作特征(receiver operating characteristic,ROC)曲线,分析NLR预测患者预后的临床价值;并以Kaplan-Meier法绘制观察指标不同水平下的生存曲线,进行生存分析。COX回归分析各指标提示预后的价值。结果：死亡组患者NLR,CRP,WHO肺动脉高压功能分级、肺动脉收缩压、尿素氮、肌酐、中性粒细胞计数高于生存组,淋巴细胞计数低于生存组,差异均具有统计学意义(P<0.05)。根据ROC曲线分析,NLR的ROC曲线下面积(AUC)为0.720(P<0.01),高于肌酐(AUC=0.716)、中性粒细胞计数(AUC=0.655)、肺动脉收缩压(AUC=0.652)及CRP(AUC=0.643)。当NLR截断值为4.7时,其灵敏度为74.2%,特异度为72.0%。Kaplan-Meier生存曲线分析显示,NLR值水平较高组预后明显差于水平较低组(P<0.01)。单因素Cox回归分析提示NLR是提示患者不良预后的危险因素,多因素Cox回归分析(P>0.05)。结论：NLR水平与COPD相关肺动脉高压患者临床预后呈明显相关;NLR水平越高则提示病情较重,预后较差。     

Evidence of bone marrow downregulation in brain‐dead rats

Experimental findings support the evidence of a persistent leucopenia triggered by brain death (BD). This study aimed to investigate leucocyte behaviour in bone marrow and blood after BD in rats. BD was induced using intracranial balloon catheter inflation. Sham‐operated (SH) rats were trepanned only. Thereafter bone marrow cells were harvested every six hours from the femoral cavity and used for total and differential counts. They were analysed further by flow cytometry to characterize lymphocyte subsets, granulocyte adhesion molecules expression and apoptosis/necrosis [annexin V/propidium iodide (PI) protocol]. BD rats exhibited a reduction in bone marrow cells due to a reduction in lymphocytes (40%) and segmented cells (45%). Bone marrow lymphocyte subsets were similar in BD and SH rats (CD3, P = 0.1; CD4, P = 0.4; CD3/CD4, P = 0.4; CD5, P = 0.4, CD3/CD5, P = 0.2; CD8, P = 0.8). Expression of L‐selectin and beta₂‐integrins on granulocytes did not differ (CD11a, P = 0.9; CD11b/c, P = 0.7; CD62L, P = 0.1). There were no differences in the percentage of apoptosis and necrosis (Annexin V, P = 0.73; PI, P = 0.21; Annexin V/PI, P = 0.29). In conclusion, data presented suggest that the downregulation of the bone marrow is triggered by brain death itself, and it is not related to changes in lymphocyte subsets, granulocyte adhesion molecules expression or apoptosis and necrosis.     

The effects of altered exercise distribution on lymphocyte subpopulations

The effects of exercise distribution on lymphocyte count, lymphocyte subpopulations and plasma cortisol concentration in peripheral blood were assessed in 19 healthy subjects. The subjects were randomly divided into group A (n = 10) or group B (n = 9) according to exercise distribution. Both groups underwent a 10-week programme involving 5 × 2-week blocks: baseline (B), training period 1 (TP1), stabilisation 1 (S1), training period 2 (TP2), and stabilisation 2 (S2). During B, S1 and S2 normal training was undertaken. During TP1 and TP2 the subjects increased the amount of training by 50% in week 1 and by 100% in week 2. During TP1 subjects in group A exercised 6 days·week^–1, while during TP2 these subjects exercised on 3 alternate days·week^–1, but doubled the duration of each training session. The subjects in group B reversed this training order. Blood was collected 36–42 h following exercise period B, and at the end of periods TP1, S1, TP2 and S2, and also 12–18 h following completion of exercise at the end of TP1 and TP2. There were no significant differences (P > 0.05) between the 6 day·week^–1 programme and the 3 alternate day·week^–1 programme in total lymphocyte count, CD3⁺, CD4⁺, CD8+, CD16⁺, or CD19⁺ cells, the CD4:CD8 ratio, HLA-DR⁺ (activated) T cells or plasma cortisol concentrations. Following both TP1 and TP2 there was a nonsignificant decrease in lymphocyte subpopulations. However following both S1 and S2 (baseline training) there was a significant increase in total lymphocyte count, CD3⁺, CD4⁺ and CD8⁺ lymphocytes. The S2 variables statistically significant from B were: total lymphocyte count (P < 0.01), CD3⁺ T-cells and percentage of circulating lymphocytes (^P < 0.01), CD4⁺ cells (P < 0.0001), CD8⁺ cells (P < 0.05), and HLA-DR⁺ (activated) T-cells (P < 0.05). The results indicated that provided the amount of exercise is constant for a given period, then exercise distribution is not a critical variable in the alteration of lymphocyte subpopulations that may occur in response to overload training. However 2 weeks of overload training followed by 2 weeks of active recovery (baseline) training may induce an increase in the lymphocyte count.     

Mobilization without immune depletion fails to restore immunological tolerance or preserve beta cell function in recent onset type 1 diabetes

Granulocyte colony‐stimulating factor (G‐CSF) has been used to restore immune competence following chemoablative cancer therapy and to promote immunological tolerance in certain settings of autoimmunity. Therefore, we tested the potential of G‐CSF to impact type 1 diabetes (T1D) progression in patients with recent‐onset disease [n = 14; n = 7 (placebo)] and assessed safety, efficacy and mechanistic effects on the immune system. We hypothesized that pegylated G‐CSF (6 mg administered subcutaneously every 2 weeks for 12 weeks) would promote regulatory T cell (T_reg) mobilization to a degree capable of restoring immunological tolerance, thus preventing further decline in C‐peptide production. Although treatment was well tolerated, G‐CSF monotherapy did not affect C‐peptide production, glycated haemoglobin (HbA1c) or insulin dose. Mechanistically, G‐CSF treatment increased circulating neutrophils during the 12‐week course of therapy (P < 0·01) but did not alter T_reg frequencies. No effects were observed for CD4⁺ : CD8⁺ T cell ratio or the ratio of naive : memory (CD45RA⁺/CD45RO⁺) CD4⁺ T cells. As expected, manageable bone pain was common in subjects receiving G‐CSF, but notably, no severe adverse events such as splenomegaly occurred. This study supports the continued exploration of G‐CSF and other mobilizing agents in subjects with T1D, but only when combined with immunodepleting agents where synergistic mechanisms of action have previously demonstrated efficacy towards the preservation of C‐peptide.     

Low-Dose Daily Subcutaneous Interleukin-2 in Combination with Highly Active Antiretroviral Therapy in HIV + Patients: A Randomized Controlled Trial

Abstract

Purpose: Previous studies with intermittent interleukin-2 (IL-2) therapy using intermediate and high levels of IL-2 have demonstrated significant increases in the CD4 + T cell count in HIV-infected patients. Intermittent regimens are amenable to outpatient use, but severe adverse events are frequently experienced with intermediate- and high-dose levels of IL-2. Therefore in this study, the effect of daily, subcutaneous low-dose IL-2 therapy on safety and immunological endpoints was investigated to determine whether immunological benefit could be achieved without toxicity in HIV-infected patients also receiving highly active antiretroviral therapy (HAART). Method: A total of 115 patients were enrolled in the trial. Fifty-six asymptomatic HIV-infected patients who had CD4 + T cell counts less than 300 cells/μL at screening and a stable HIV viral load received low-dose IL-2 (1.2 million IU [MIU]/m 2 beginning dose) once daily in conjunction with HAART (IL-2 group). Fifty-nine patients received HAART alone (control group). Results: A dramatic effect of IL-2 on the natural killer (NK) cell population was observed with mean increases of 156 cells/μL in the IL-2 group compared to 19.93 cells/μL in the control group (p < .001). Additionally, IL-2-treated patients experienced a statistically significant increase in the mean percentage of CD4 + T cells (3.52% increase) when compared to control patients (1.33% increase) (p < .001). The expanded CD4 + T cell population was primarily of the naive phenotype, with mean increases of 4.53% for the IL-2 group and 0.31% for the control group (p < .001 for between-group difference). In addition, a higher proportion of IL-2-treated patients (67%) compared to control patients (33%) achieved increases of greater than 50% in the CD4+ T cell count (p = .08). Adverse events of grade 3 or grade 4 toxicity were infrequent in the current study and were substantially lower by comparison to those in studies of intermittent dose IL-2 therapy. Also, negligible changes in the HIV viral load from baseline to final measurement were observed in both groups. A trend toward a reduced number of modifications of antiretroviral therapy was apparent in the IL-2 group when compared to control patients. Conclusion: Daily, low-dose subcutaneous IL-2 therapy in conjunction with HAART is safe and well tolerated and is effective in expanding lymphocyte cell types including NK cells and naive T cells in individuals who have <300 CD4+ T cells.     

Antiretroviral Treatment Regimen Outcomes Among HIV-Infected Prisoners

Abstract

Background: Despite the high prevalence of HIV in correctional settings, the duration of therapy and response to various highly active antiretroviral therapy (HAART) regimens in this setting is unknown. Method: Using a retrospective cohort study (1997-2002) of HIV-infected prisoners in Connecticut that linked demographic, pharmacy, and laboratory data, we compared HIV-1 RNA (VL) and CD4 lymphocyte responses to four treatment strategies at baseline and at the end of incarceration. Results: Using an analysis of 1,044 incarceration periods or 1,099 subjects for whom ≥6 months of continuous data were available, HAART regimens that included a triple NRTI, two NRTIs + either a PI or NNRTI, or a three-class (NRTI+NNRTI+PI) strategy demonstrated no difference in virological and immunological outcomes. The proportion of subjects who were initiated with NRTI, NNRTI, PI, or three-class regimens were 14%, 32%, 46%, and 8%, respectively. For all study groups, the mean change from baseline in CD4 and VL was +74 cells/μL and -0.93 log₁₀ copies/mL (p < .0001), respectively. Overall, 59% of subjects had an HIV-1 RNA level below the level of detection (<400 copies/mL) by the end of their incarceration. Using Kaplan-Meier curves to examine the time to change in the initial HAART strategy over the incarceration period, the three-class strategy was significantly more likely to be changed earlier than all others (p < .05). Conclusion: Although the three-class strategy was less durable, initiating HAART with any strategy resulted in similar and impressive virological and immunological outcomes by the end of incarceration, further supporting prison as an important site for the initiation and provision of effective antiretroviral therapy.     

Lymphocyte Subset Recovery and Outcome after Autologous Hematopoietic Stem Cell Transplantation for Plasma Cell Myeloma

Rapid immune reconstitution—particularly of natural killer cells (NK cells)—after allogeneic hematopoietic stem cell transplantation (HSCT) is associated with protection from relapse. Whether such an association also exists after autologous stem cell transplantation is less clear. We retrospectively assessed lymphocyte subsets after autologous HSCT in 114 patients and correlated lymphocyte recovery with outcome. CD8 T cell and NK cell counts recovered rapidly to pretransplantation levels, whereas B cell and CD4 T cell recovery were delayed. Compared with patients with low NK cells (<100/uL), high NK cell count at 1 month after HSCT was associated with significantly prolonged progression-free survival: for NK cells 100 to 200/uL hazard ratio [HR], .33 (95% confidence interval [CI]; .16 to .80; P = .004); for NK cells > 200/μL HR, .27 (95% CI, .13 to .58; P = .001). No significant protective effects were associated with rapid recovery of any other lymphocyte subset. None influenced overall survival (OS) or time to next treatment. Early NK cell recovery is associated with better progression-free survival after autologous HSCT. The failure to detect an effect on OS might be due to the salvage strategies available to these patients.     

A simple algebraic demonstration of the validity of DeFries-Fulker analysis in unselected samples with multiple kinship levels

DeFries and Fulker's (Behav. Genet. 15, 467–473, 1985) regression procedure (DF analysis) to estimatec ² andh ² was originally applied to selected twin data. Since then, DF analysis has been applied more broadly in unselected data and with multiple (nontwin) kinship levels. Theoretical work based on the matrix algebra of variance-covariance matrices has shown that estimates ofc ² andh ² are unbiased in selected two-group settings. In this article, a simple proof is presented supporting the validity of DF analysis in broader settings. We use scalar algebra to show that parameter estimates ofh ² andc ² are unbiased in unselected settings with multiple (more than two) kinship levels. Caveats are offered, and other DF analysis problems are identified.     

Effect of ingesting carbohydrate only or carbohydrate plus casein protein hydrolysate during a multiday cycling race on left ventricular function,plasma volume expansion and cardiac biomarkers

Purpose

Multiday racing causes mild left ventricular (LV) dysfunction from day 1 that persists on successive days. We evaluated ingesting casein protein hydrolysate–carbohydrate (PRO) compared with carbohydrate-only (CHO) during a 3-day mountain bike race.

Methods

Eighteen male cyclists were randomly assigned to ingest 6.7% carbohydrate without (CHO) or with 1.3% casein hydrolysate (PRO) during racing (~ 4–5 h/day; 68/71/71 km). Conventional LV echocardiography, plasma albumin content, plasma volume (PV) and blood biomarkers were measured before day 1 and post race on day 3.

Results

Fourteen cyclists (n = 7 per group) completed the race. PV increased in CHO (mean increase (95% CI), 10.2% (0.1 to 20.2)%, p = 0.045) but not in PRO (0.4% (− 6.1 to 6.9)%). Early diastolic transmitral blood flow (E) was unchanged but deceleration time from peak E increased post race (CHO: 46.7 (11.8 to 81.6) ms, p = 0.019; PRO: 24.2 (− 0.5 to 48.9) ms, p = 0.054), suggesting impaired LV relaxation. Tissue Doppler mitral annular velocity was unchanged in CHO, but in PRO septal early-to-late diastolic ratio decreased (p = 0.016) and was compensated by increased lateral early (p = 0.034) and late (p = 0.012) velocities. Systolic function was preserved in both groups; with increased systolic lateral wall velocity in PRO (p = 0.002). Effect size increase in serum creatine kinase (CK) activity, CK-MB and C-reactive protein concentrations was less in PRO than CHO (Cohen’s d mean ± SD, PRO: 2.91 ± 2.07; CHO: 7.56 ± 4.81, p = 0.046).

Conclusion

Ingesting casein hydrolysate with carbohydrate during a 3-day race prevented secondary hypervolemia and failed to curb impaired LV relaxation despite reducing tissue damage and inflammatory biomarkers. Without PV expansion, systolic function was preserved by lateral wall compensating for septal wall dysfunction.

     

Increased p70s6k phosphorylation during intake of a protein–carbohydrate drink following resistance exercise in the fasted state

The present study aimed at comparing the responses of myogenic regulatory factors and signaling pathways involved in muscle protein synthesis after a resistance training session performed in either the fasted or fed state. According to a randomized crossover study design, six young male subjects participated in two experimental sessions separated by 3 weeks. In each session, they performed a standardized resistance training. After the sessions, they received during a 4-h recovery period 6 ml/kg b.w. h of a solution containing carbohydrates (50 g/l), protein hydrolysate (33 g/l), and leucine (16.6 g/l). On one occasion, the resistance exercise session was performed after the intake of a carbohydrate-rich breakfast (B), whereas in the other session they remained fasted (F). Needle biopsies from m. vastus lateralis were obtained before (Rest), and 1 h (+1h) and 4 h (+4h) after exercise. Myogenin, MRF4, and MyoD1 mRNA contents were determined by RT-PCR. Phosphorylation of PKB (protein kinase B), GSK3, p70^s6k (p70 ribosomal S6 kinase), eIF2B, eEF2 (eukaryotic elongation factor 2), ERK1/2, and p38 was measured via western blotting. Compared with F, the pre-exercise phosphorylation states of PKB and p70^s6k were higher in B, whereas those of eIF2B and eEF2 were lower. During recovery, the phosphorylation state of p70^s6k was lower in B than in F (p = 0.02). There were no differences in basal mRNA contents between B and F. However, compared with F at +1h, MyoD1 and MRF4 mRNA contents were lower in B (p < 0.05). Our results indicate that prior fasting may stimulate the intramyocellular anabolic response to ingestion of a carbohydrate/protein/leucine mixture following a heavy resistance training session.