t-复合多肽1基因在原发性肾病综合征患者外周血单个核细胞中的表达

目的:初步探讨t-复合多肽1(TCP-1)基因在原发性肾病综合征(PNS)患者外周血单个核细胞(PBMC)中的表达及临床意义。方法:收集体检20例正常人(A组)和30例PNS患者(B+C组,其中B组为初发PNS患者,未用激素;C组为复诊PNS患者,使用激素2～3个月。各15例)的新鲜外周血液。用RT-PCR检测其外周血PBMC中TCP-1基因的表达。结果:A组分别与B+C组、B组、C组相比,TCP-1基因的表达差异均有显著性(P<0.01);B组与C组中的表达差异无显著性(P>0.05)。结论:TCP-1可能参与了PNS的发生发展过程。     

Modulation of the Proteome of Peripheral Blood Mononuclear Cells from HIV-1-Infected Patients by Drugs of Abuse

Introduction  We used proteomic analyses to assess how drug abuse modulates immunologic responses to infections with the human immunodeficiency virus type 1 (HIV-1). Methods  Two-dimensional difference gel electrophoresis was utilized to determine changes in the proteome of peripheral blood mononuclear cells (PBMC) isolated from HIV-1-positive donors that occurred after treatment with cocaine or methamphetamine. Both drugs differentially regulated the expression of several functional classes of proteins. We further isolated specific subpopulations of PBMC to determine which subpopulations were selectively affected by treatment with drugs of abuse. Monocytes, B cells, and T cells were positively or negatively selected from PBMC isolated from HIV-1-positive donors. Results  Our results demonstrate that cocaine and methamphetamine modulate gene expression primarily in monocytes and T cells, the primary targets of HIV-1 infection. Proteomic data were validated with quantitative, real-time polymerase chain reaction. These studies elucidate the molecular mechanisms underlying the effects of drugs of abuse on HIV-1 infections. Several functionally relevant classes of proteins were identified as potential mediators of HIV-1 pathogenesis and disease progression associated with drug abuse.     

Increased secretion of TGF-beta1 by peripheral blood mononuclear cells from patients with Type 1 diabetes mellitus with diabetic nephropathy.

AIMS: To study transforming growth factor (TGF)-beta1 secretion by peripheral blood mononuclear cells (PBMC) from Type 1 diabetic patients with and without nephropathy. METHODS: Thirty normoalbuminuric Type 1 diabetic patients (urinary albumin excretion rate (AER) < 20 microg/min), 12 microalbuminuric (AER 20-200 microg/min), 10 nephropathic (AER > 200 microg/min), and 13 non-diabetic individuals were recruited. TGF-beta1 secretion by PBMC was measured by enzyme immunoassay (EIA) after 48 h culture with and without phytohaemagglutinin (PHA) (5 microg/ml). RESULTS: After 48 h culture, the highest TGF-beta1 levels secreted by unstimulated PBMC were found in patients with nephropathy (median 6.2 (range 0.9-20.0) ng/ml) when compared to patients with normal albumin excretion (4.1 (0.2-11.3) ng/ml), microalbuminuria (1.8 (0.2-6.4) ng/ml) and healthy controls (1.0 (0.2-7.0) ng/ml); P = 0.02 for the three diabetic groups and P = 0.006 for all groups. At 48 h, the PHA-stimulated TGF-beta1 levels were 12.4 (2.9-30.0) ng/ml in nephropathic, 7.3 (0.5-21.2) ng/ml in normoalbuminuric, and 5.5 (0.5-27.6) ng/ml in microalbuminuric patients (P = 0.05). A correlation was observed between TGF-beta1 and diastolic blood pressure in the subgroup of patients with incipient and overt nephropathy (r = 0.45, P = 0.04). CONCLUSIONS: Type 1 diabetic patients with overt nephropathy show increased TGF-beta1 secretion by PBMC. Diastolic blood pressure levels correlated with TGF-beta1 secretion in diabetic patients with nephropathy. Increased TGF-beta1 secretion by PBMC may be associated with renal and vascular disease in Type 1 diabetes mellitus.     

肿瘤患者外周血单个核细胞诱导的CD_3AK细胞体外抗肿瘤作用的初步观察

采用抗ＣＤ３单抗（１０μｇ／ｍｌ）联合ＩＬ－２（５０Ｕ／ｍｌ）诱导肿瘤病人外周血单个核细胞成为ＣＤ３ＡＫ细胞，并观察其体外增殖及抗肿瘤作用，结果显示：肿瘤病人的ＮＫ活性显著低于正常人，但其ＣＤ３ＡＫ细胞体外杀伤Ｋ５６２细胞的细胞毒活性与正常人比较无明显差异（Ｐ＞００５）。本研究初步表明：由肿瘤病人外周血单个核细胞诱导的ＣＤ３ＡＫ细胞，可作为过继免疫治疗的效应细胞进行自身淋巴细胞治疗     

High permeability haemofiltration improves peripheral blood mononuclear cell proliferation in septic patients with acute renal failure.

BACKGROUND: Continuous veno-venous haemofiltration (HF) with high permeability (HP) haemofilters is a novel approach in the adjuvant therapy of septic patients. HP haemofilters are characterized by an increased pore size which facilitates the filtration of inflammatory mediators. The present study examines whether HP-HF has an impact on peripheral blood mononuclear cell (PBMC) proliferation and whether ultrafiltrate can alter PBMC function in isolates from healthy volunteers. METHODS: Twenty-eight septic patients with acute renal failure were randomly allocated to either HP-HF or conventional HF (C-HF). HP-HF was performed with a newly developed high-flux polyamide membrane (P2SH) with a nominal cut-off point of 60 kDa. For C-HF, a high-flux polyamide haemofilter (Polyflux 11S; cut-off, 30 kDa) was used. RESULTS: Septic patients demonstrated a significantly reduced proliferation of anti-CD3-stimulated PBMCs compared to healthy controls (P = 0.016). Initiating HF led to a restoration of the PBMC proliferation in HP-HF but not in C-HF. Exposing PBMCs isolated from healthy donors to ultrafiltrates from patients with sepsis demonstrated a significant suppressive effect of HP ultrafiltrates on the anti-CD3-stimulated PBMC proliferation (P = 0.011). Ultrafiltrate from patients with sepsis who received C-HF had no impact on PBMC proliferation. CONCLUSION: HP-HF restores PBMC proliferation in septic patients probably by eliminating immunomodulatory mediators. HP-HF may represent a new renal replacement therapy able to modulate PBMC function in sepsis.     

脆弱类杆菌脂多糖对正常人外周血单个核细胞分泌白细胞介素的影响

目的　通过研究脆弱类杆菌脂多糖 (LPS)对正常人外周血单个核细胞 (PBMC)分泌白细胞介素 2 (IL 2 )和白细胞介 4素 (IL 4 )的影响 ,探讨脆弱类杆菌感染的机制。　方法　采用脆弱类杆菌临床分离菌和标准菌株 (NCTC9343)提取的LPS ,以不同浓度作用于PBMC ,2 4h后收集培养细胞上清 ,运用ELISA法检测其IL 2和IL 4的含量变化。　结果　脆弱类杆菌LPS对正常人PBMC分泌IL 2有显著抑制作用 ,并呈明显的浓度依赖关系 (r=0 .80 2 4 ,P <0 .0 1)。脆弱类杆菌LPS对正常人PB MC分泌IL 4有显著刺激作用 (P <0 .0 5 ) ,但无浓度依赖关系。脆弱类杆菌临床分离菌与标准菌株LPS对正常人PBMC分泌IL 2和IL 4具有相同效应 ,两组之间无明显差异 (r =0 .10 95 ,P >0 .0 5 )。　结论　脆弱类杆菌LPS对正常人PBMC分泌IL 2有抑制作用 ,而对IL 4的分泌有刺激作用     

In vitro effects of monophthalates on cytokine expression in the monocytic cell line THP-1 and in peripheral blood mononuclear cells from allergic and non-allergic donors

It has recently been shown that plasticizers are present in indoor air dust, which may lead to human exposure via the inhalation route. Moreover, studies have indicated that plasticizers may possess adjuvant effects increasing the health damaging potential of allergens. The aim of this study was to investigate the in vitro effect of metabolites of phthalate plastisizers, such as whether an adjuvant effect is paralleled by changes of the cytokine expression in the monocytic cell line THP-1 and in peripheral blood mononuclear cells (PBMCs) from allergics and non-allergics. The toxicity monitored by cell viability was determined by incubating THP-1 cells with a 10-fold dilution series of monophthalates for 24 h. At different points in time cytokine expression (IL-1β, IL-6, IL-12 (p35)) in THP-1 cells incubated with non-toxic concentrations of monophthalate (2–20 μg/ml)±LPS (1 μg/ml) were determined using Quantitative Competitive RT-PCR. PBMCs from allergics and non-allergics were incubated with monophthalate 220 μg/ml) for up to 48 h and cytokine expression (IL-4, IL-5, IFN-γ) was measured using real-time PCR. The cytotoxic level of monophthalates is 20–200 μg/ml, depending on the individual monophthalate. There seems to be a correlation between increasing side-chain length and toxicity. Monophthalates did not induce changes in cytokine expression in THP-1 cells, though there is an increase when co-incubating with LPS. Cytokine expression in PBMC seems virtually unchanged when co-incubated with monophthalate, though mono-n-butyl phthalate (MBUP) tends to increase the level of IL-4 in PBMCs from allergic individuals. The two cellular models demonstrated the dynamics of regulated cytokine mRNA and are applicable for in vitro immunotoxicological investigations. The results regarding monophthalates suggest these to have a limited effect on cytokine expression in the monocytic cell line THP-1 and weak effect on cytokine expression in PBMCs from allergic and non-allergic individuals.     

Influence of immunosuppressive drugs on the CD30 molecule in kidney transplanted patients

Background

Soluble CD30 (sCD30) is a suggested marker for kidney transplantation outcomes. We investigated whether sCD30 serum levels are influenced by immunosuppression and whether they correlate with findings in protocol biopsies and with CD30 gene expression in peripheral blood mononuclear cells (PBMC).

Biomarkers of immune tolerance in liver transplantation

The liver exhibits intrinsic immune tolerogenic properties that contribute to a unique propensity toward spontaneous acceptance when transplanted, both in animal models and in humans. Thus, in contrast to what happens after transplantation of other solid organs, several years following liver transplantation a significant subset of patients are capable of maintaining normal allograft function with histological integrity in the absence of immunosuppressive drug treatment. Significant efforts have been put into identifying sensitive and specific biomarkers of tolerance in order to stratify liver transplant recipients according to their need for immunosuppressive medication and their likelihood of being able to completely discontinue it. These biomarkers are currently being validated in prospective clinical trials of immunosuppression withdrawal both in Europe and in the United States. These studies have the potential to transform the clinical management of liver transplant recipients by mitigating, at least in part, the burden of lifelong immunosuppression.     

Increased T cell immunosenescence and accelerated maturation phenotypes in older kidney transplant recipients

Older kidney transplant recipients experience increased rates of infection and death, and less rejection, compared with younger patients. However, little is known about immune dysfunction in older compared with younger kidney transplant recipients and whether it is associated with infection. We evaluated T cell phenotypes including maturation, immune senescence, and exhaustion in a novel investigation into differences in older compared with younger patients receiving identical immune suppression regimens.We evaluated PBMC from 60 kidney transplant recipients (23 older and 37 matched younger patients) by multiparameter immune phenotyping. Older kidney transplant recipients demonstrated decreased frequency of naïve CD4+ and CD8+ T cells, and increased frequency of terminally differentiated, immune senescent, and NK T cells expressing KLRG1. There was a trend towards increased frequency of T cell immune senescence in patients experiencing infection in the first year after transplantation, which reached statistical significance in a multivariate analysis.This pilot study reveals immune dysfunction in older compared with younger transplant recipients, and suggests a likely mechanism for increased vulnerability to infection. The ability to assess T cell maturation and immune senescence in transplant recipients offers the potential for risk stratification and customization of immune suppression to prevent infection and rejection after transplantation.