Correlation between radiculalgia and counts of T lymphocyte subsets in the peripheral blood of patients with lumbar disc herniation

Objective: To determine the correlation between the degree of radiculalgia and counts of T lymphocyte subsets in the peripheral blood of patients with lumbar disc herniation. Methods: Forty‐nine patients with lumbar disc herniation (group A) were divided into three subgroups according to Visual Analogue Scale (VAS) pain scores (group A₁: n= 12, VAS 0–4.0; A₂: n= 24, VAS 4.1–7.0; A₃: n= 13, VAS 7.1–10.0. Twenty health blood donors who volunteered to be involved in the study comprised the control group (group B). Peripheral blood counts of various T lymphocyte subsets were measured in each group. Results: (i) The counts of CD4+ T and CD4+/CD8+ lymphocytes were higher in group A than in group B, and the difference between the two groups was statistically significant (P < 0.05). There were also statistically significant differences between group A and group B in the counts of CD3+ and CD8+ T lymphocytes (P < 0.05); (ii) There was no correlation between the VAS scores and the counts of CD3+ T lymphocytes (r= 0.194, P > 0.05). A strong significant correlation was observed between the VAS scores and counts of CD4+ T lymphocytes (r= 0.542, P < 0.05), CD4+/CD8+ (r= 0.468, P < 0.05), which increased with increasing VAS scores in the three subgroups of group A (P < 0.05). However there was a significant negative linear correlation between CD8+ T lymphocyte counts and pain scores (r=?0.462, P < 0.05). Conclusion: Our results suggest that changes in T lymphocyte subsets in peripheral blood take place after prolapse of lumbar intervertebral discs. The current results may provide support for involvement of immunologic mechanisms in low back pain secondary to herniation of the lumbar disc. T lymphocytes may play an important role in the development of symptoms in patients with lumbar intervertebral disc herniation.     

Changes in T Cells After ABO-Incompatible Liver Transplantation

Purpose: T lymphocytes are an essential component of allograft rejection and tolerance. The aims of the present study are to analyze the characteristics of T-cell subsets between ABO-incompatible living donor liver transplantation (ABO-I LDLT) and ABO-compatible LDLT (ABO-C LDLT). Materials and Methods: Between April 2013 and June 2014, 61 patients underwent adult LDLT. ABO-I LDLT patients received rituximab and all patients received basiliximab as induction therapy and tacrolimus as maintenance therapy. The distribution of peripheral blood T lymphocyte subsets pretransplant and 4, 8, 12, and 24 weeks post-transplant were serially monitored. Results: Eight patients underwent ABO-I LDLT. Patient characteristics did not vary between the ABO-I and ABO-C groups. Absolute lymphocyte counts and CD4+ T cells in the ABO-I group were lower than those in ABO-C group after LDLT (p =.034 and p =.039, respectively). However, the comparison between the ABO-I and ABO-C groups revealed that the CD8+ T cells, CD4/CD8 ratio, Vδ1 cells, Vδ2 cells, γδ T cells, Vδ1/Vδ2 ratio, CD3-CD56+ cells, and CD4+Foxp3+ T cells did not change significantly over time. Conclusions: Absolute lymphocyte counts and CD4+ T cell levels are different between ABO-I and ABO-C groups after LDLT. The present study suggests that T-cell lymphocyte changes in peripheral blood in ABO-I LDLT patients were similar to those in ABO-C LDLT patients.     

Morbidly obese human subjects have increased peripheral blood CD4+ T cells with skewing toward a Treg- and Th2-dominated phenotype

Obesity is associated with local T-cell abnormalities in adipose tissue. Systemic obesity-related abnormalities in the peripheral blood T-cell compartment are not well defined. In this study, we investigated the peripheral blood T-cell compartment of morbidly obese and lean subjects. We determined all major T-cell subpopulations via six-color flow cytometry, including CD8+ and CD4+ T cells, CD4+ T-helper (Th) subpopulations, and natural CD4+CD25+FoxP3+ T-regulatory (Treg) cells. Moreover, molecular analyses to assess thymic output, T-cell proliferation (T-cell receptor excision circle analysis), and T-cell receptor-β (TCRB) repertoire (GeneScan analysis) were performed. In addition, we determined plasma levels of proinflammatory cytokines and cytokines associated with Th subpopulations and T-cell proliferation. Morbidly obese subjects had a selective increase in peripheral blood CD4+ naive, memory, natural CD4+CD25+FoxP3+ Treg, and Th2 T cells, whereas CD8+ T cells were normal. CD4+ and CD8+ T-cell proliferation was increased, whereas the TCRB repertoire was not significantly altered. Plasma levels of cytokines CCL5 and IL-7 were elevated. CD4+ T-cell numbers correlated positively with fasting insulin levels. The peripheral blood T-cell compartment of morbidly obese subjects is characterized by increased homeostatic T-cell proliferation to which cytokines IL-7 and CCL5, among others, might contribute. This is associated with increased CD4+ T cells, with skewing toward a Treg- and Th2-dominated phenotype, suggesting a more anti-inflammatory set point.     

Phenotypic changes in lymphocyte subpopulations in pediatric renal-transplant patients after T-cell depletion

BACKGROUND: T-cell depletion causes a novel homeostasis in lymphocyte subsets in adult transplant recipients. Little is known about long-term changes in pediatric patients. METHODS: Twenty-one pediatric renal-transplant patients (mean age 11.8 years) were selected according to their initial postoperative immunosuppressive therapy: (1) baseline immunosuppression (BI) with cyclosporine, azathioprine, and steroids, n=11; and (2) BI plus polyclonal antibodies, n=10. Lymphocyte surface markers were measured in the mean 2.3 years after transplantation and analyzed between the patient groups and in regard to 46 age-matched healthy controls. RESULTS: The patient groups did not differ with respect to age, sex, renal function, and previous infections. Total lymphocyte counts, CD4+ T-cell numbers, and distribution of naive to memory CD4+ T cells were not different between transplant groups and controls. However, patients with postoperative T-cell depletion showed significantly lower ratios of CD4+ to CD8+ T cells, elevated CD8+ T-cell numbers, increased counts of CD8+ T cells coexpressing CD57, and higher numbers of CD8+ cells with a naive phenotype. In addition, the numbers of double-positive T cells and lymphocytes bearing both natural killer (NK) and T-cell markers were elevated in the patients with postoperative depletion. NK and B-cell counts were lower in the transplant patient groups compared with the healthy controls. CONCLUSIONS: Pediatric transplant patients show characteristic long-term changes in lymphocyte subsets after T-cell depletion. In contrast with adult patients, these perturbations are less pronounced and predominant in the CD8+ T-cell compartment.     

Functional defects and the influence of age on the frequency of CD4+ CD25+ T-cells in type 1 diabetes

CD4+ CD25+ T-cells appear to play a crucial role in regulating the immune response. Therefore, we evaluated the peripheral blood frequency and function of CD4+ CD25+ T-cells in 70 type 1 diabetic patients and 37 healthy individuals. Interestingly, a positive correlation was observed between increasing age and CD4+ CD25+ T-cell frequency in both subject groups. In contrast to previous studies of nonobese diabetic mice and type 1 diabetic patients, similar frequencies of CD4+ CD25+ and CD4+ CD25(+Bright) T-cells were observed in healthy control subjects and type 1 diabetic patients of similar age. There was no difference between type 1 diabetic subjects of recent-onset versus those with established disease in terms of their CD4+ CD25+ or CD4+ CD25(+Bright) T-cell frequency. However, type 1 diabetic patients were markedly defective in their ability to suppress the proliferation of autologous effector T-cells in vitro. This type 1 diabetes-associated defect in suppression was associated with reduced production of interleukin (IL)-2, gamma-interferon, and transforming growth factor-beta, whereas other cytokines including those of adaptive and innate immunity (IL-10, IL-1beta, IL-6, IL-8, IL-12p70, and tumor necrosis factor-alpha) were similar in control subjects and type 1 diabetic patients. These data suggest that age strongly influences the frequency of CD4+ CD25+ T-cells and that function, rather than frequency, may represent the means by which these cells associate with type 1 diabetes in humans.     

乳腺癌患者外周血中T细胞亚群与淋巴结转移和组织学分级的关系

目的:探讨乳腺癌患者外周血中T细胞亚群的变化及其与淋巴结转移和组织学分级的关系。方法:用流式细胞术检测86例乳腺癌患者以及20例乳腺腺病患者外周血T细胞亚群的百分率。结果:乳腺癌患者外周血总T细胞与CD4~+T细胞百分数与腺病患者无统计学差异(均P0.05),但CD8~+T细胞百分数低于腺病患者(P0.05)。乳腺癌患者中,淋巴结转移者CD4~+T细胞百分数高于无淋巴结转移者(P0.05);CD8~+T细胞百分数随组织学分级增加而升高(P0.05)。结论:乳腺癌患者存在细胞免疫功能紊乱,外周血中CD4~+T细胞、CD8~+T细胞比例的变化分别与淋巴结转移、组织学分级密切相关。监测外周血T细胞亚群的变化,有助于病情及预后的判断。     

冷冻消融治疗软组织肉瘤后外周血T细胞亚群变化与患者生存期的相关性

目的 观察冷冻消融治疗软组织肉瘤(STS)后外周血T细胞亚群变化及其与患者生存期的相关性。方法 纳入22例接受冷冻消融治疗的晚期STS患者,比较治疗前、后外周血T细胞亚群变化,分析影响患者生存期的相关因素,以及总生存期(OS)和无进展生存期(PFS)与外周血T细胞亚群变化的相关性。结果 共对22个病灶实施冷冻消融。治疗后疾病客观缓解率为81.82%(18/22),患者中位OS为15个月,中位PFS为7个月。OS与肿瘤病理分级和消融效果相关(P均<0.01)。治疗后外周血CD4⁺T、Treg细胞较治疗前降低、自然杀伤(NK)细胞较前升高(P均<0.05);患者PFS与NK细胞水平呈正相关(r=0.539,P=0.010),OS与Treg细胞水平呈负相关(r=-0.463,P=0.030),PFS则与治疗前、后外周血CD4⁺T细胞差值呈正相关(r_s=0.424,P=0.049)。结论 冷冻消融治疗STS可在一定程度上改善机体免疫功能;治疗后血清NK、Treg细胞水平可用于评估患者生存期。     

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目的 研究乳腺癌腋窝淋巴结发生癌转移和未发生癌转移时的免疫功能。方法2004年8月至2005年7月采用流式细胞技术检测乳腺癌病人乳腺癌前哨淋巴结（SLN）与乳腺癌非前哨淋巴结（NSLN）中免疫细胞CD3^＋T、CD4^＋T、CD8^＋T、CD16^＋NK、CD^19^＋B的比例，并相互比较。结果淋巴结未发生癌转移时，SLN与NSLN的免疫细胞CD3^＋T、CD4^＋T、CD8^＋T、CD16^＋NK、CD^19^＋B的比例差异无统计学意义（P〉0．05）；而且当SLN与NSLN发生癌转移后，它们的免疫细胞的比例也无差异。但SLN发生癌转移与未发生癌转移时相比，其CD4^＋T、CD8^＋T、CD16^＋NK的比例发生显著改变（P〈0．05），CD3^＋T、CD19^＋B的比例改变无统计学意义（P〉0．05）。结论 当腋窝淋巴结未发生癌转移时，它仍有正常的免疫功能。当腋窝淋巴结发生癌转移后，它的免疫微环境发生了改变，免疫功能呈现一种抑制状态。     

Detection of alloreactive T cells by flow cytometry: a new test compared with limiting dilution assay

BACKGROUND: Frequencies of alloreactive T cells determined by limiting dilution assays (LDA) may not adequately reflect the donor-reactive immune status in transplant recipients. To reevaluate LDA frequencies, we developed a flow cytometry test for direct determination of alloreactive T-cell frequencies and compared these frequencies with classical LDA estimates of frequencies. METHODS: For determination of frequencies by flow cytometry, peripheral blood lymphocytes (or lymphocytes taken from primary mixed lymphocyte culture) were stimulated with either Epstein-Barr virus-transformed lymphoblastoid cell lines or T cell-depleted spleen cells and stained for intracellular interferon (IFN)-gamma production and CD69. In lung transplant recipients, frequencies of IFN+ alloreactive T cells were compared with LDA frequencies, that is, cytotoxic T lymphocyte precursors and helper T lymphocyte precursors. RESULTS: With flow cytometry, alloreactive T cells were detected after overnight allostimulation as IFN-gamma CD69bright cells (range, 0.1-0.58% and 0.1-0.66% of total CD4 and CD8 cells, respectively). Frequencies increased 25-fold or more when lymphocytes were prestimulated in primary mixed lymphocyte culture before testing. After lung transplantation, mean donor-specific IFN+ CD8 T-cell frequencies did not decrease as mean donor-specific LDA cytotoxic T lymphocyte precursor frequencies, whereas no difference was seen in pretransplantation samples or third-party-specific frequencies at both time points. Mean frequencies of IFN+ CD4 did not differ from helper T lymphocyte precursors at both time points, but frequencies did not correlate. CONCLUSIONS: The flow cytometry test allows a direct measurement of alloreactive T-cell frequencies and demonstrates a discrepancy between donor-specific IFN+ CD8 T-cell frequencies and LDA CLTp after transplantation. This may be a result of the existence of "functional diverse" alloreactive T cells or of activation-induced cell death of donor-reactive T cells during long (LDA) culturing, which is avoided in the flow cytometry test.     

大肠癌患者术后NDV-ATV治疗对细胞免疫功能的影响

目的　探讨大肠癌患者术后NDV -ATV治疗对细胞免疫功能的影响。方法　采用流式细胞仪分析 70例术后接受NDV -ATV治疗的大肠癌患者外周静脉血T淋巴细胞亚群及NK细胞活性的变化情况 ,并与 45例术后未接受NDV -ATV治疗的大肠癌患者作为对照。结果　NDV -ATV治疗组 ,术后外周血CD+3、CD+4、NK细胞和CD+4 CD+8均较治疗前显著增高 (P <0 0 1 ) ,且明显高于未接受NDV -ATV治疗的术后常规治疗组 (P <0 0 5 )。结论　NDV -ATV可改善大肠癌患者术后机体的免疫功能     

腹膜透析患者腹主动脉钙化与外周血淋巴细胞亚群的相关研究

目的探讨腹膜透析(peritoneal dialysis,PD)患者腹主动脉钙化(abdominal aortic calcification,AAC)与外周血淋巴细胞亚群之间的关系。方法收集86例PD患者临床资料,腹部侧位平片判断患者发生AAC的程度,并计算腹主动脉钙化积分(AAC score,AACs),依据患者的AACs分为3组:无和轻度钙化组(0≤AACs≤4分)41例、中度钙化组(5≤AACs≤15分)28例及重度钙化组(16≤AACs≤24)17例。利用流式细胞技术分别检测各组患者外周血中淋巴细胞亚群,其中包括CD3⁺、CD3⁺CD4⁺、CD3⁺CD8⁺、CD4⁺/CD8⁺、Treg、CD19⁺、CD16⁺CD56⁺等淋巴细胞的百分率及CD8⁺DR⁺淋巴细胞计数,计算CD4⁺/CD8⁺比值。采用Spearman相关性分析方法分析AAC与外周血淋巴细胞亚群及临床指标的相关性,用Logistic回归分析PD患者发生AAC的危险因素。结果3组患者年龄、原发病为糖尿病肾病、慢性肾小球肾炎以及白蛋白、血磷、碱性磷酸酶水平比较,差异有统计学意义(P<0.05)。Spearman相关性分析结果显示:AAC程度与患者的年龄、透析龄、甲状旁腺素、血磷、碱性磷酸酶、超敏C反应蛋白、CD4⁺/CD8⁺、CD16⁺CD56⁺、CD8⁺DR⁺呈正相关(r=0.352、0.284、0.226、0.299、0.265、0.285、0.236、0.228、0.292,P<0.05),与白蛋白、Treg呈负相关(r=-0.222、-0.354,P<0.05)。Logistic回归模型结果显示:年龄(OR=1.060,95%CI 1.008~1.116,P<0.05),CD4⁺/CD8⁺(OR=25.441,95%CI 1.561~414.659,P<0.05),Treg(OR=0.427,95%CI 0.255~0.715,P<0.05)是PD患者发生腹主动脉中重度钙化的独立预测因素。结论PD患者的AAC程度与外周血中淋巴细胞亚群改变具有相关性。Treg细胞百分率减少、CD4⁺/CD8⁺比值增加可能会加速AAC。PD患者外周血淋巴细胞亚群的改变可能会为AAC的防治提供新方向。     

Lymphocytes Subsets in the Course of Continuous Ambulatory Peritoneal Dialysis (CAPD)

Background: We studied lymphocyte subset counts in comparison with normal subjects in order to clarify the abnormalities of cellular immune responses in uremic patients undergoing continuous ambulatory peritoneal dialysis (CAPD). Methods: The study included 37 CAPD patients and 45 normal individuals, as the control group. For the study, CAPD patients were divided into four groups depending on duration of replacement therapy. Group I consisted of patients treated for 0–6 months (n = 6), group II for 6–12 months (n = 6), group III for 13–24 months (n = 16), and group IV for more than 25 months (n = 9). Flow cytometry was used for estimation of lymphocyte subsets (determination of CD2, CD3, CD3 +/CD4 +, CD3 +/CD8 +, CD3 ?/16 + 56 +, CD19, CD4/CD8). Results: Our patients started CAPD with decreased lymphocyte subset counts, slightly above the normal range (excluding CD3 ?/16 + 56 +, CD2). After 6 months of CAPD therapy, an increase in CD4/CD8 ratio was observed and all examined lymphocyte subset counts decreased (excluding CD2). In patients on CAPD for more than 25 months, CD3 +/CD4 +, CD19 counts were below the normal range, CD3 ?/16 + 56 + exceeded the upper limit of normal range and at the same time mean total lymphocyte count (TLC) was maintained in the normal range. Conclusions: We recommend lymphocyte subset determinations for detection of immune abnormalities in the course of CAPD treatment.     

微创食管癌根治术对老年患者肺部感染与T淋巴细胞亚群及MMP－9的影响

目的探讨微创食管癌根治术对老年患者术后肺部感染、T淋巴细胞亚群及MMP－9的影响。 方法选取2015年1月到2017年12月98例老年食管癌患者作为研究对象,采用随机数字表法分为微创组(实施微创胸腹腔镜食管癌根治术)和传统组(实施传统食管癌根治术)。应用SPSS 19.0统计软件进行数据分析,术中术后指标、T细胞亚群、MMP－9水平指标以均值±标准差表示,组间比较采用独立t检验;术后肺部感染及死亡发生率指标,组间比较采用χ²检验。以P<0.05认为差异具有统计学意义。 结果微创组患者手术各项指标(手术切口平均总长度、平均手术时间、手术出血量、术后住院天数)均优于传统组患者(P<0.05);微创组术后肺部感染率6.1%(3/49)明显低于传统组26.5%(13/49),P<0.05;微创组患者术后第1天、3天T细胞亚群的各项指标的水平(CD3^＋、CD4^＋、CD4^＋/CD8^＋)均高于传统组(P<0.05);微创组术后第1天、3天MMP－9水平均低于传统组,差异均有统计学意义(P<0.05)。 结论老年食管癌患者行微创胸腹腔镜食管癌根治术效果更好,可明显减轻手术创伤,改善外周血T淋巴细胞亚群及血清MMP－9水平,值得临床推广。     

Th1/Th2 balance and CD45-positive T cell subsets in primary nephrotic syndrome

T cells are involved in the pathogenesis of nephrotic syndrome (NS). The aim of the study was to determine whether the activity of T-helper-1 (Th1) and T-helper-2 (Th2) cells and the distribution of the lymphocyte subsets, namely CD45RA+CD4+ (”naive” helper T cells, suppressor-inducer), CD45RA+CD8+ (”naive” suppressor T cells, suppressor-effector), CD45RO+CD4+ (”memory” helper T cells), are predictive for steroid sensitivity in children with primary NS. These parameters were assessed at the onset of disease, before initiation of steroid therapy. Two groups of NS children were retrospectively formed according to steroid sensitivity (SS) or resistance (SR). The activity of Th1 and Th2 cells was defined by the production of interleukin-2 (IL-2), interferon-γ, IL-4, and IL-10 in the supernatants of CD4+ T cell cultures activated with autologous monocytes presenting tetanus toxoid (TT). Peripheral lymphocyte subsets were determined using double- or triple-color flow cytometry. In SS children with NS we found a decreased proliferative response of CD4+ T cells to TT stimulation, cytokine synthesis indicating the predominance of Th2 activity, and an increased percentage of activated suppressor-inducer (CD45RA+ CD4+CD25+, 5.18±0.8, P<0.001) and suppressor-effector (CD45RA+CD8+CD25+, 2.05±0.6, P<0.01) cells, with the concomitant reduction of activated memory cells (CD45RO+CD4+CD25+, 0.2±0.1, P<0.001). In children with SRNS we found an increased proliferative response of CD4+ T cells to TT, a rise in activated memory (CD45RO+CD4+CD25+, 3.82±0.7, P<0.01) and suppressor-inducer peripheral T cells (CD45RA+ CD4+CD25+, 3.85±0.6, P<0.01), but a low percentage of activated suppressor-effector (CD45RA+CD8+ CD25+, 0.5±0.2, P<0.05) T cells. We conclude that prior to treatment the distribution of lymphocyte subpopulations in peripheral blood together with Th1 and Th2 cell activity provides a useful tool for evaluating the likelihood of steroid sensitivity in patients with primary NS. Received: 3 November 1998 / Revised: 1 September 1999 / Accepted: 8 September 1999     

他克莫司对肝移植受者外周血T淋巴细胞亚群及共刺激分子表达的影响

目的 探讨他克莫司（FK506）对肝移植受者外周血T淋巴细胞亚群及其表面共刺激分子表达的影响。方法 采用荧光标记单克隆抗体和流式细胞技术，测定术后采用FK506治疗的肝移植受者（FK506治疗组）在用FK506后1、2、3、4周时的外周血T淋巴细胞亚群及其表面共刺激分子CD28、CD152和ICOS分子的表达情况，以健康志愿者（健康对照组）和患终末期肝脏疾病拟行肝移植者（肝病对照组）为对照。结果 CD3^＋T淋巴细胞在各组间的差异均无统计学意义（P〉0．05）。经FK506治疗后，肝移植患者的CD4^＋T淋巴细胞逐渐减少，CD8^＋T淋巴细胞逐渐增加，并恢复至健康对照组水平（P〉0．05）。FK506治疗组T淋巴细胞亚群表面CD28分子和ICOS分子表达逐渐下降，并明显低于健康对照组（P〈0．05），而CD152分子表达增加，且明显高于健康对照组（P〈0．05）；其ICOS分子表达水平的下降晚于CD28分子，CD4^＋CD28^＋T淋巴细胞、CD8^＋CD28^＋T淋巴细胞和CD4^＋ICOS^＋T淋巴细胞均呈现相近的变化规律。结论 FK506能迅速纠正移植受者T淋巴细胞亚群紊乱，并抑制正性共刺激分子CD28和ICOS的表达，促进负性共刺激分子CD152的表达。     

T-cell subsets,interleukin-2 receptor expression and production of interleukin-2 in minimal change nephrotic syndrome

This study was designed to investigate T-lymphocyte subsets interleukin-2 receptor (IL-2R) expression and IL-2 production in minimal change nephrotic syndrome (MCNS). Peripheral blood T-lymphocytes and IL-2R expression were analysed using fluorescein isothiocyanatelabelled CD3, CD4, CD8 and CD25 monoclonal antibodies with flow cytometry. IL-2 production was determined by enzyme immunoassay. Ten children with MCNS in relapse and in remission were evaluated. Thirteen healthy children served as controls. The patients in relapse demonstrated a moderate decrease in the total absolute lymphocyte counts and CD8(+) T-lymphocytes compared with controls (P<0.05) and had a greatly increased IL-2R expression in frashly isolated, unstimulated peripheral lymphocytes compared with patients in remission and controls. While this was not statistically significant, IL-2R expression on cultured lymphocytes stimulated with phytohaemagglutinin was significantly elevated in relapse compared with those in remission and controls (P<0.05). IL-2 production did not correlate well with IL-2R expression and there was no significant difference between the groups. Our results suggest that T-cell subset changes and high IL-2R expression on peripheral lymphocytes may indicate the presence of stimulated T-cell populations in MCNS which could contribute to the immunopathogenesis.     

The prognostic value of peripheral blood lymphocyte subsets in patients with bladder carcinoma treated using neoadjuvant M-VEC chemotherapy

OBJECTIVES: To assess the prognostic value of peripheral blood lymphocyte subsets in patients with bladder cancer who were treated with neoadjuvant chemotherapy. PATIENTS, SUBJECTS AND METHODS: Thirty patients with a histological diagnosis of invasive bladder transitional cell carcinoma and 30 age-matched controls with no evidence of cancer and immunological disorders were evaluated. Peripheral blood samples were assessed in both groups using monoclonal antibodies. Patients with bladder cancer who achieved complete or partial responses and those who had progression of the disease after systemic chemotherapy with methotrexate, vinblastine, epirubicin and cisplatin were compared according to the pretreatment values of the peripheral blood lymphocyte subsets. RESULTS: There were no significant differences in B lymphocyte levels between the groups. In patients with bladder cancer, the percentages of T lymphocytes (P<0.01), natural killer (NK) cells (P<0.05) and the CD4+/CD8+ ratio (P<0.05) were significantly lower than in the control group. In patients who responded to the chemotherapy regimen, the pretreatment values of T lymphocytes (P<0.001), the CD4+/CD8+ ratio (P<0.01) and NK cell levels (P<0.01) were significantly higher than in the patients who did not. CONCLUSION: In patients with invasive bladder carcinoma, cell-mediated immunity may have a role in the resistance to this malignancy and in these patients the pretreatment levels of T lymphocyte subsets may be an indicator of the potential response to chemotherapy.     

2型糖尿病肾病患者外周血CD28/CTLA4共刺激分子的表达及其意义

目的:探讨2型糖尿病(T2DM)肾病患者外周血T细胞亚群、共刺激分子CD 28及细胞毒T淋巴细胞抗原(CTLA)4的表达及其临床意义。方法:选取2015年07月~2018年06月至我科室和内分泌科治疗的患者,根据尿微量清蛋白排泄率(UAER)将其分为DM组(T2DM无肾病,n=42)和DN组(T2DM合并肾病,n=40),同时选取健康人群40例作为对照(NC组)。采用流式细胞技术测定两组患者外周血T淋巴细胞上CD4、CD8的表达,并用流式细胞技术测定CD4^+、CD8^+T淋巴细胞表面CD28、CTLA4分子的表达。结果:三组外周血CD4^+和CD4^+/CD8^+按NC组-DM组-DN组顺序均呈显著递增趋势(P<0.01),CD3^+、CD8^+按照顺序呈显著递减趋势(P<0.01),且各组间差异明显(P<0.01);三组外周血CD4^+CD28^+、CD8^+CD28^+ T细胞按NC组-DM组-DN组顺序均呈显著递增趋势(P<0.01),CD4^+CTLA4+及CD8^+CTLA4+T细胞按照顺序呈显著递减趋势(P<0.01),且各组间差异明显(P<0.01)。结论:T2DM患者T细胞亚群失衡,其中合并肾病患者表达失衡更为严重,提示糖尿病肾病患者存在自身免疫调节异常。同时T2DM合并肾病患者外周血CD28和CTLA4表达也显著异于正常对照,提示共刺激分子可能参与了T2DM合并肾病的免疫功能紊乱。     

Effect of Surgically-Induced Weight Loss on Leukocyte Indicators of Chronic Inflammation in Morbid Obesity

Background: Recent evidence suggests that morbid obesity is a chronic inflammatory condition that may be associated with immune dysfunction.To test this hypothesis, we investigated several leukocyte cell surface markers of chronic inflammation and followed their response to surgically-induced weight loss. Methods: 26 patients having Roux-en-Y gastric bypass (RYGBP) for morbid obesity (BMI>40) were compared to 10 normal controls (BMI<25). Relative monocyte and neutrophil frequencies and expression of the activation antigens CD11b (adhesion molecule), CD16 (Fc receptor), and CD62L (Lselectin), were evaluated by flow cytometry preoperatively and at 1, 3, 6 and 12 months after RYGBP. Cases served as their own controls but were also compared to non-obese controls. The results were statistically analyzed using Student's t-test and ANOVA for parametric values and Mann-Whitney along with Kruskal-Wallis ANOVA for nonparametric values Results: The control group had mean age 37 ± 7.6 with mean 23 ± 2.5 and no comorbidities. The mean age of the sample group was 40.36 ± 13.7 with mean BMI 52 ± 8.2. The neutrophil and monocyte relative frequencies of CD11b (monocytes and neutrophils), and CD16 (neutrophils only) were comparable to controls at baseline and did not change significantly with weight loss throughout the study period. However, a significant reduction of CD62L (Lselectin) expression was noted in monocytes and neutrophils at baseline (neutrophils 103 vs 240 gmf, p<0.001) (monocytes104 vs 246 gmf, P<0.001) when compared to normal controls. Levels of L-selectin normalized by 6 months in both monocytes and neutrophils, and by 12 months had become abnormally elevated in monocytes (monocytes 391 gmf, P=0.007); in neutrophils, there was an upward trend that did not reach significance.The expression of the LPS receptor CD14 in the study group was elevated significantly compared to controls at baseline (1129 vs 719 gmf, P=0.004); this marker appeared to return to normal by 3 months. Monocyte CD14+/CD16+ subset percentage were also elevated significantly at baseline (14.3% vs 5.25%, P <0.001), declined throughout the time period but was still significant at 1 year (8.8%, P<0.001). Eosinophil percentages were elevated at baseline (3.3% obese vs 1.8% controls, P=0.003) and remained so throughout the time period. Conclusion: Deficiencies in the immune system of morbidly obese individuals include elevated levels of eosinophils, monocyte CD14, and monocyte CD14+/CD16+ subsets, with depression of monocyte and neutrophil CD62L. These abnormal levels reverse rapidly with surgically-induced weight loss. RYGBP is not only a weight loss operation but also appears to be an immune restorative procedure.     

Lymphocyte Subset Ratio Cannot Diagnose Immune Failure of a TKA

BackgroundUp to 20% of patients are dissatisfied following total knee arthroplasty (TKA), most often due to pain and/or stiffness. The differential diagnosis includes immune reaction to the prosthesis. However, there is no consensus on diagnostic criteria for immune failure, an allergic reaction, to a TKA.Histologic evaluation could provide evidence as to whether an allergic reaction caused TKA failure. A recent study showed an increase in CD4+ lymphocytes compared to CD8+ lymphocytes in patients lymphocyte transformation testing (LTT) + for Ni. This finding is consistent with Ni sensitization, but can lymphocyte subsets be used to diagnose immune failure on a case-by-case basis?MethodsPeriprosthetic tissues from 18 revision cases of well-fixed, aseptic, but painful and/or stiff primary TKAs were analyzed. Six patients LTT? for Ni were matched as a cohort for age, sex, and body mass index (BMI), to 12 patients LTT + for Ni. Periprosthetic tissue biopsies underwent immunohistochemical IHC staining for CD4+ and CD8+ lymphocyte subsets and were compared by LTT status. The immunohistochemicalIHC results were also compared with periprosthetic histology.ResultsThere was no relationship between LTT status and mean CD4+ cells/hpf or CD4+:CD8+ lymphocyte ratio. No relationship was found between LTT stimulation index (continuous or categorical) and CD4+:CD8+ ratio or aseptic lymphocyte-dominant vasculitis-associated lesion ALVAL score.ConclusionLymphocytes in periprosthetic tissue are highly variable in number, subtype ratio, and location, and have no relationship to LTT result or ALVAL score on a case-by-case basis. Based on these results, lymphocyte subsets cannot diagnose immune failure. Further work is needed to determine criteria for the diagnosis of immune failure of a TKA.