Fluoxetine treatment to rats modifies serotonin transporter and cAMP in lymphocytes,CD4+ and CD8+ subpopulations and interleukins 2 and 4

Several lines of evidences indicate that antidepressants produce various immunomodulatory effects. Fluoxetine, an antidepressant and selective serotonin reuptake inhibitor, modulates immune cells in vitro. To explore the in vivo influence of fluoxetine on lymphocytes, male Sprague–Dawley rats were treated daily, 10 mg/kg, or with saline solution for 1, 2 and 3 weeks. The presence of serotonin transporter in CD3+, CD4+ and CD8+ subpopulations of T lymphocytes was determined by immunofluorescence. Serotonin transporter was also labeled with [³H]paroxetine, specific binding defined with imipramine. Plasma levels of pro-inflammatory interleukin 2 (IL-2), and anti-inflammatory interleukin 4 (IL-4), were measured by ELISA; and cAMP concentration by radioimmunoassay. Fluoxetine significantly increased the number of lymphocytes expressing serotonin transporter and elevated the binding of [³H]paroxetine. The percentage of CD4+ cells decreased, that of CD8+ increased, and CD3+ did not change. The ratio CD4+/CD8+ was significantly lowered. Fluoxetine administration elevated the levels of IL-4 at 1, 2 and 3 weeks; and of IL-2, at 2 and 3 weeks. IL-4/IL-2 ratio was significantly increased in fluoxetine group respecting the controls and was similar during the 3 weeks of treatment. Fluoxetine produced a significant decrease in cAMP concentrations in lymphocytes, probably by secondary activation of serotonin receptors. Treatment with fluoxetine modified immune parameters in plasma and lymphocytes of rats, which might be relevant for its systemic therapeutic action as an antidepressant.     

Follicular regulatory T cells infiltrated the ovarian carcinoma and resulted in CD8 T cell dysfunction dependent on IL-10 pathway

A high Treg/CD8 T cell ratio in ovarian carcinoma was negatively associated with the prognosis of the patients. The human follicular regulatory T (Tfr) cells are a newly characterized subset of Treg cells with features of both follicular helper T (Tfh) cells (CXCR5⁺) and canonical Treg cells (CD25⁺Foxp3⁺). The role of Tfr cells in ovarian cancer is yet unclear. We found that in peripheral blood, the ovarian cancer patients presented significantly higher levels of both CD4⁺CD25⁺CD127⁻CXCR5⁺ T cells and CD4⁺CD25⁺CD127⁻CXCR5⁺Foxp3⁺ T cells than the healthy controls. In resected tumor samples, Tfr cells represented a much greater percentage of lymphocytes than in peripheral blood. Interestingly, the circulating Tfr cells from ovarian cancer patients presented significantly higher TGFB1 and IL10 expression than their counterparts in healthy controls directly ex vivo, and significantly higher IL10 after stimulation. The tumor-infiltrating Tfr cells presented further upregulated expression of TGFB1 and IL10. In addition, the levels of TGFB1 and IL10 expression by Tfr cells negatively associated with the expression of IFNG in tumor-infiltrating CD8 T cells. In an in vitro CD8 T cell/Tfr cell coculture system, we found that Tfr cells could significantly suppress the activation of CD8 T cells, in a manner that was dependent on IL-10 and probably on TGF-β. Overall, our study found that Tfr cells could suppress CD8 T cells, and in ovarian cancer patients, the Tfr cells were increased in both frequency and function.     

CXCR5+ CD8+ T cells present elevated capacity in mediating cytotoxicity toward autologous tumor cells through interleukin 10 in diffuse large B-cell lymphoma

Diffuse large B-cell lymphoma (DLBCL) is a common and aggressive subtype of non-Hodgkin's lymphomas, with limited treatment options in refractory and relapsed patients. Growing evidence supports the notion that CD8⁺ T cell immunity could be utilized to eliminate B cell lymphomas. CXCR5⁺ CD8⁺ T cell is a novel cell subtype and share CXCR5 expression with CD19⁺ tumor cells. In this study, we investigated the frequency and function of existing CXCR5⁺ CD8⁺ T cells in DLBCL patients. We found that DLBCL patients as a group demonstrated significantly higher level of CXCR5⁺ CD8⁺ T cells than healthy individuals, with huge variability in each patient. Using anti-CD3/CD28-stimulated CD8⁺ T cells as effector (E) cells and autologous CD19⁺ tumor cells as target (T) cells, at high E:T ratio, no difference between the intensities of CXCR5⁺ CD8⁺ T cell- and CXCR5⁻ CD8⁺ T cell-mediated cytotoxicity were observed. However, at intermediate and low E:T ratios, the CXCR5⁺ CD8⁺ T cells presented stronger cytotoxicity than CXCR5⁻ CD8⁺ T cells. The expressions of granzyme A, granzyme B, and perforin were significantly higher in CXCR5⁺ CD8⁺ T cells than in CXCR5⁻ CD8⁺ T cells, with no significant difference in the level of degranulation. Tumor cells in DLBCL were known to secrete high level of interleukin 10 (IL-10). We therefore blocked the IL-10/IL-10R pathway, and found that the expressions of granzyme A, granzyme B, and perforin by CXCR5⁺ CD8⁺ T cells were significantly elevated. Together, these results suggest that CXCR5⁺ CD8⁺ T cells are potential candidates of CD8⁺ T cell-based immunotherapies, could mediate elimination of autologous tumor cells in DLBCL patients, but are also susceptible to IL-10-mediated suppression.     

PD-1 regulates CXCR5+ CD4 T cell-mediated proinflammatory functions in non-small cell lung cancer patients

PD-1 inhibitors have been used to revive exhausted T cell responses in non-small cell lung cancer (NSCLC) and other malignancies. CXCR5⁺ T follicular helper (Tfh) cells are characterized by constitutive high PD-1 expression and have been associated with the formation of tertiary lymphoid structures and implicated in antitumor immunity. In this study, we investigated the effect of PD-1 and PD-1 inhibition on CXCR5⁺ CD4 T cells. Data showed that CXCR5⁺ CD4 T cells in both healthy subjects and NSCLC patients presented markedly higher PD-1 expression than CXCR5⁻ CD4 T cells. Both CXCR5⁻ and CXCR5⁺ CD4 T cells from NSCLC patients presented higher PD-1 expression than their counterparts in healthy subjects. PD-1⁺ CXCR5⁺ CD4 T cells were functional, could express IL-21, IL-10, and CXCL13 upon stimulation, demonstrated auxiliary effects toward CD8 T cell-mediated IFN-γ production and proliferation, and promoted IgM and IgG production. However, the potency of PD-1⁺ CXCR5⁺ CD4 T cells was lower than the potency of PD-1⁻ CXCR5⁺ CD4 T cells. PD-1 blocking could significantly enhance the effector functions of PD-1⁺ CXCR5⁺ CD4 T cells. Overall, this study demonstrated that PD-1⁺ CXCR5⁺ CD4 T cells could promote CD8 T cell and B cell inflammation and could be modulated by PD-1 inhibition.     

The CD4/CD8 ratio is associated with coronary artery disease (CAD) in elderly Chinese patients

Objective: The aim of this study was to investigate the relationship between number of circulating T cells and coronary artery disease (CAD) in an elderly Chinese population.Methods: A total of 295 elderly inpatients (age ≥ 60) were included in this cross-sectional study. Their clinical and biochemical characteristics were recorded. Patients were divided to two groups: control patients and CAD patients. The risk factors of CAD were explored by binary logistic regression analysis.Results: Compared with control patients, the ratio of CD4 to CD8 T cells was significantly increased in CAD patients. There was no difference in the number of CD3, CD4, and CD8 T cells between the two groups. Multiple logistic regression analysis showed that CAD was independently associated with age, gender, body mass index (BMI), systolic blood pressure (SBP), chronic heart failure (CHF) and the CD4/CD8 ratio. In addition, after adjusting for different clinical parameters (including gender, age, CHF, hypertension, arrhythmia, SBP, and BMI), the risk of CAD was significantly increased in patients with a CD4/CD8 ratio > 1.5.Conclusions: There was a strong and independent association between the ratio of CD4/CD8 and CAD in elderly Chinese population.     

CD28 in thymocyte development and peripheral T cell activation in mice exposed to suspended particulate matter

The CD28:B7 signaling pathway is very important for the activity of mature peripheral T lymphocytes and thymocyte development. The proper development of thymocytes into mature single positive CD4⁺and CD8⁺ T cells is crucial for almost all immune functions. In naturally occurring conditions, T cells maturation in the thymus is influenced by environmental agents. The expression of CD28 and the distribution of CD28^low/high thymocytes have been examined at various stages of thymocyte development in BALB/c mice exposed to air-suspended particulate matter (ASM). Acute exposure to ASM resulted in the decrease of CD28 expression in the total thymocyte population. The increase of the percentage of CD28^low and the decrease of CD28^high thymocytes were observed, which may account for the acceleration of thymocyte development under the conditions of elevated risk resulting from the exposure of animals to environmental xenobiotics. ASM exposure resulted in the increase of the level of proliferation of lymph node T cells induced by anti-CD3 and anti-CD28 monoclonal antibodies activation despite normal expression of CD28 molecule. In contrast, the level of proliferation of spleen T cells was lowered or normal dependently of the concentration of stimuli used for activation. Results of these studies demonstrate that acute exposure of mice to ASM can result in the progression of two contrasting processes in the immune system: upregulation of thymocyte development, which contributes to the maintenance of peripheral T cell pool, and over-activation of lymph node lymphocytes, which may lead to uncontrolled immunostimulation.     

Measurement of circulating T cell subsets positive for Fas antigen (CD95) in patients with alcoholic hepatitis.

T cell subsets positive for Fas antigen in peripheral blood of patients with alcoholic hepatitis were measured, using monoclonal antibodies in two colour immunofluorescence assay with flowcytometry. 1) In the patients with alcoholic hepatitis, the ratio of mean +/- standard deviation (M +/- SD) of CD4+ cells positive for CD95 (Fas antigen) in peripheral blood lymphocytes of patients with alcoholic hepatitis tended to increase, but the ratio of CD95-positive cells in CD4+ cells of peripheral blood was almost the same, compared with those of healthy controls. In the alcoholics (overdrink) who did not show alcoholic hepatitis with or without apparent alcoholic damage, the ratio of CD95-positive CD4+ cells in peripheral blood lymphocytes was within normal range, while CD95-positive cells in CD4+ cells of peripheral blood tended to decrease. 2) In the alcoholic hepatitis, the ratios of CD8+ cells positive for CD95 and CD95-positive cells in CD8+ cells of peripheral blood decreased significantly, and in the alcoholics (overdrink) they also tended to decrease. 3) The fluorescence intensity of CD95 on CD4+ cells in peripheral blood of the alcoholics (overdrink) decreased apparently, although the one on CD8+ cells did not. 4) The ratios of T cell subsets, that is, CD4+ and CD8+ cells, positive for HLADR in peripheral blood of the patients with alcoholic hepatitis increased significantly, respectively. These results showed that the ratio of CD8+ cells positive for Fas antigen in peripheral blood of the patients with alcoholic hepatitis decreased. It was suspected that this finding might be due to the direct effect of intaken alcohol to T cell subsets rather than hepatitis, and/or the result of immunological homeostasis in alcoholic hepatitis.     

Erythropoietin exerts direct immunomodulatory effects on the cytokine production by activated human T-lymphocytes

The effect of erythropoietin-β (Epo-β) on the functional profile of activated human T-lymphocytes remains largely unknown, which hinders clinical application of Epo as an immunomodulatory agent. We studied the direct impact of Epo on the activation status of human T lymphocytes following activation by particles loaded with antibodies (Abs) against human CD2, CD3, and CD28. T cell activation was assessed by the surface expression of CD38 activation marker. Epo did not significantly affect activation status of both CD4⁺ and CD4⁻ T cells, as well as of naive (CD45RA⁺ CD197⁺), central memory (CD45RA⁻ CD197⁺), effector memory (CD45RA⁻ CD197⁻), and terminally-differentiated (CD45RA⁺ CD197⁻) T cells. However, Epo markedly augmented production of IL-2, IL-4 and IL10 by activated T cells with concomitant reduction in IFN-γ secretion. Taken together, our data showed that Epo could directly down-regulate pro-inflammatory T cell responses without affecting T cell activation status.     

Th9 cells are subjected to PD-1/PD-L1-mediated inhibition and are capable of promoting CD8 T cell expansion through IL-9R in colorectal cancer

Th9 cells are named after their expression of IL-9. Studies in recent years demonstrated that Th9 cells could contribute to antitumor immunity by enhancing the recruitment and activation of mast cells, natural killer cells, CD8 T cells, and dendritic cells in the tumor microenvironment. To determine whether Th9 cells participate in colorectal cancer (CRC), we collected resected tumor samples from 20 CRC patients. In the tumor-infiltrating lymphocytes (TILs), IL-9⁺IL-4⁻ CD4⁺ T cells could be observed and were present at higher frequencies than the IL-9⁺IL-4⁺ and the IL-9⁻IL-4⁺ cells, suggesting that the majority of IL-9-producing TILs were bona fide Th9 cells. IL-9-secreting TILs presented particularly high PD-1 expression directly ex vivo. The expression of IL-9 was significantly reduced with PD-L1-mediated inhibition, which in turn was suppressed by anti-PD-1 blocking. Interestingly, the circulating CD4⁺ T cell compartment in CRC patients also presented Th9 enrichment, characterized by higher IL-9⁺IL-4⁻ and IL-9⁺IL-4⁺ cell frequencies in the CXCR3⁻CCR6⁻ compartment as compared to that in non-cancer controls. Using exogenous TGF-β and IL-4, we were capable of enriching Th9 cells without concurrent enrichment of Th2 cells. Th9-enriched CD4⁺ T cells, but not Th9-non-enriched cells, significantly increased the expansion of activated CD8⁺ T cells, in a manner that was dependent on the expression of IL-9R. In addition, the frequencies of Th9 cells in the tumor were positively correlated with the frequencies of CD8⁺ TILs. Together, we demonstrated that Th9 cells infiltrated CRC tumor, could be regulated via the PD-1/PD-L1 pathway, and could contribute the CD8⁺ T cell expansion.     

虫草多糖调节小鼠T淋巴细胞及其亚群数量的机制研究

目的 研究冬虫夏草菌丝体多糖（虫草多糖）对正常小鼠T淋巴细胞及其亚群数量的影响及可能的机制。方法 ICR小鼠,随机分成空白对照组、阳性组（香菇多糖1 mg·kg^-1）和虫草多糖高、中、低剂量（200,100,50 mg·kg^-1）组,连续腹腔注射10 d后,流式细胞仪检测外周血T淋巴细胞（CD³⁺细胞）及其亚群细胞CD4⁺CD8^-和CD4-CD8+数量、CD³⁺细胞凋亡率;MTT法测定小鼠脾淋巴细胞转化功能;实时荧光定量PCR检测脾脏组织Bcl-2 mRNA、Bax mRNA的转录水平,并测定胸腺和脾脏指数。结果 与空白对照组比较,虫草多糖能提高小鼠脾脏质量、促进未经ConA诱导的脾淋巴细胞增殖转化能力,并呈现剂量依赖性;虫草多糖高剂量组能明显增加外周血CD³⁺细胞和CD4⁺CD8^-细胞亚群的数量,降低CD4^-CD8⁺细胞亚群数量,显著提高CD4⁺CD8^-/CD4-CD8⁺的比值;高剂量虫草多糖能明显降低小鼠外周血CD³⁺细胞凋亡率,显著升高脾组织Bcl-2 mRNA转录水平和Bcl-2/Bax比值,而对Bax mRNA转录水平没有明显影响。结论 虫草多糖可刺激小鼠主要免疫器官（胸腺和脾脏）增生,提高T淋巴细胞及CD4^-CD8⁺亚群的数量,其作用机制可能与上调抑制凋亡基因Bcl-2 mRNA的转录,提高Bcl-2/Bax比值,从而抑制淋巴细胞凋亡有关。     

Changes of CD4+CD25+FOXP3 + and CD8+CD28 − regulatory T cells in non-small cell lung cancer patients undergoing surgery

Little is known about the regulatory T cells (Tregs) in the peripheral blood after surgery of non-small cell lung cancer (NSCLC) patients. In this study, we investigated whether CD4+CD25+FOXP3 + and CD8+CD28 − regulatory T cells are decreased in the peripheral blood of NSCLC patients undergoing surgery. The study group (n = 49) comprised NSCLC, and the control group (n = 24) consisted of age- and sex-matched nonmalignant diseases. The prevalence of CD4+CD25+FOXP3 + and CD8+CD28 − Tregs was analyzed using flow cytometry. The study group showed significantly higher percentage of CD4+CD25+FOXP3 + and CD8+CD28 − Tregs than control. The percentage of CD4+CD25+FOXP3 + and CD8+CD28 − Tregs increased with tumor stage. One way ANOVA test shows the significant differences between all subgroups. LSD test shows that there was a statistical significance between each of the two subgroups except stage II in CD4+CD25+FOXP3 + Tregs and control vs. each stage, stage I vs. stage III, and stage IV in CD8+CD28 − Tregs. There is no significant difference among stages II, III, and IV in CD8+CD28 − Tregs. No differences were found between squamous carcinoma and adenocarcinoma. These levels were dropped significantly after operation. Furthermore postoperative Treg percentage in the early stages (stage I and stage II) was not statistically different from that of controls. Postoperative Treg percentage in advanced stage (III + IV) remained above the values shown by controls. Our findings indicate that the percentage of CD4+CD25+FOXP3 + and CD8+CD28 − Tregs correlated with the pathological stage in NSCLC and tumor burden.     

卡介菌多糖核酸对尖锐湿疣病人外周血CD8~+T细胞细胞因子表达影响

目的:研究卡介菌多糖核酸(BCG-PSN)对尖锐湿疣(CA)病人外周血CD8q+T细胞内白细胞介素类(IL-2、IL-4、IL-12)和干扰素γ(IFN-γ)表达的影响。方法:60例CA病人分为2组,均采用CO_2激光去除疣体。BCG-PSN组40例,激光术后开始加用BCG-PSN 1.0mg,im,qod,连续18次,36d为一个疗程。病例对照组20例,激光术后未再用任何药物治疗。另设正常对照组20例。采用流式细胞仪测定3组外周血CD8+T细胞IL-2、IL-4、IL-12、IFN-γ的水平。结果:治疗前CA病人外周血IL-2、IL-12和IFN-γ阳性CD8+T细胞百分率显著低于正常对照组,Tcl/Tc2亦显著低于正常对照组(P<0.01)。治疗后BCG-PSN组外周血IL-2、IL-12和IFN-γ阳性CD8+T细胞百分率显著升高(P<0.01),病例对照组无显著变化(P>0.05),2组变化差异非常显著(P<0.01)。BCG-PSN组治愈率82％,显著高于病例对照组(45％,P<0.01);复发率18％,低于病例对照组(55％,P<0.01)。结论:CA病人存在Tcl/Tc2失调,BCG-PSN可通过上调CA病人外周血CD8+T细胞IL-2、IL-12、IFN-γ的表达,纠正机体细胞因子失调而提高CA病人临床疗效、减少其复发。     

CXCR5+ CD8+ T cells could induce the death of tumor cells in HBV-related hepatocellular carcinoma

The follicular CXCR5⁺ CD8⁺ T cells have recently emerged as a critical cell type in mediating peripheral tolerance as well as antiviral immune responses during chronic infections. In this study, we investigated the function of CXCR5⁺ CD8⁺ T cells in HBV-related hepatocellular carcinoma patients. Compared to CXCR5⁻ CD8⁺ T cells, CXCR5⁺ CD8⁺ T cells presented elevated PD-1 expression but reduced Tim-3 and CTLA-4 expression. Upon anti-CD3/CD28 stimulation, CXCR5⁺ CD8⁺ T cells demonstrated higher proliferation potency than CXCR5⁻ CD8⁺ T cells, especially after PD-1 blockade. CXCR5⁺ CD8⁺ T cells also demonstrated significantly higher granzyme B synthesis and release, as well as higher level of degranulation. Tumor cells were more readily eliminated by CXCR5⁺ CD8⁺ T cells than by CXCR5⁻ CD8⁺ T cells. Interestingly, we found that B cells were more resistant to CXCR5⁺ CD8⁺ T cell-mediated killing than tumor cells, possibly through IL-10-mediated protection. In addition, the CXCR5⁺ CD8⁺ T cell-mediated cytotoxic effects on tumor cells could be significantly enhanced by PD-L1 blockade. Together, we presented that in patients with in HBV-related hepatocellular carcinoma, CXCR5⁺ CD8⁺ T cells could mediate tumor cell death more potently than the CXCR5⁻ CD8⁺ T cells in vitro while the autologous B cells were protected.     

APL-1, an altered peptide ligand derived from human heat-shock protein 60, selectively induces apoptosis in activated CD4 + CD25 + T cells from peripheral blood of rheumatoid arthritis patients

Rheumatoid arthritis (RA) is a chronic T-cell mediated autoimmune disease that affects primarily the joints. The induction of immune tolerance through antigen-specific therapies for the blockade of pathogenic CD4 + T cells constitutes a current focus of research. In this focus it is attempted to simultaneously activate multiple regulatory mechanisms, such as: apoptosis and regulatory T cells (Tregs). APL-1 is an altered peptide ligand derived from a novel CD4 + T-cell epitope of human heat-shock protein of 60 kDa, an autoantigen involved in the pathogenesis of RA. Previously, we have reported that APL-1 induces CD4 + CD25^highFoxp3 + Tregs in several systems. Here, we investigated the ability of APL-1 in inducing apoptosis in PBMCs from RA patients, who were classified as active or inactive according to their DAS28 score. APL-1 decreased the viability of PBMCs from active but not from inactive patients. DNA fragmentation assays and typical morphological features clearly demonstrated that APL-1 induced apoptosis in these cells. Activated CD4 + CD25 + T cells but not resting CD4 + CD25 − T cells were identified as targets of APL-1. Furthermore, CD4 + T-cell responses to APL-1 were found to be dependent on antigen presentation via the HLA-DR molecule. Thus, APL-1 is a regulatory CD4 + T cell epitope which might modulate inflammatory immune responses in PBMCs from RA patients by inducing CD4 + CD25^highFoxp3 + Tregs and apoptosis in activated CD4 + T cells. These results support further investigation of this candidate drug for the treatment of RA.     

Proton pump inhibitor therapy in gastro-oesophageal reflux disease decreases the oesophageal immune response but does not reduce the formation of DNA adducts

Background  Chronic oesophageal inflammation and related oxidative stress are important in the pathogenesis of erosive oesophagitis (EO) and its malignant progression.
Aim  To study the effect of proton pump inhibitors (PPIs) on oesophageal cellular immune response and oxidative damage in EO patients.
Methods  Forty gastro-oesophageal reflux disease (GERD) patients [non-erosive reflux disease (NERD): 15, EO: 25] were included, after 7 days off antisuppressive drugs. EO patients were randomized to 20-mg rabeprazole once daily for either 4 or 8 weeks with baseline and follow-up endoscopy with distal oesophageal biopsies. T lymphocytes, macrophages and mast cells were quantified by immunohistochemistry. DNA adducts were measured by analysis of 8-oxo-deoxyguanosine levels.
Results  Erosive oesophagitis patients had more T lymphocytes and CD8⁺ T lymphocytes in squamous epithelium than NERD patients ( P  = 0.001, P  = 0.002, respectively). Levels of DNA adducts between both groups were, however, not different ( P  = 0.99). Four- and eight-week rabeprazole treatment in EO patients resulted in a significant decrease in number of T lymphocytes and CD8⁺ T lymphocytes (all P  < 0.05). PPIs did not, however, affect levels of DNA adducts.
Conclusions  Short-term PPI therapy in EO patients reduces the oesophageal cellular immune response, but does not change oxidative damage. PPI therapy may therefore not be effective in reducing the risk of oesophageal cancer in GERD patients.     

Cocaine administration increases CD4/CD8 lymphocyte ratio in peripheral blood despite lymphopenia and elevated corticosterone

The CD4/CD8 lymphocyte ratio in peripheral blood is used in the diagnosis of HIV infection, autoimmune disorders or susceptibility to infections. The present experiment aimed to evaluate the lymphocyte subsets, their distribution and CD4/CD8 ratio in blood after repeated, intravenous administration of cocaine. Adult male Wistar rats received three daily, in 30 min intervals, intravenous infusions of cocaine hydrochloride (5 mg/kg) or saline for 14 consecutive days. After each infusion the locomotor-activating effects of cocaine were assessed. Blood samples were collected 30 min after the last daily infusion on the 1st, 7th and 14th day of treatment. Total leukocyte numbers, percentages of leukocyte subpopulations, and T, B, NK, T CD4⁺, and T CD8⁺ lymphocyte subsets, IFN-γ, and plasma corticosterone were determined. Repeated cocaine treatment resulted in an increase in neutrophil numbers and a significant decrease in total leukocyte and lymphocyte numbers involving a significant reduction in numbers of T, B, and NK lymphocyte subsets. T CD4⁺ and T CD8⁺ lymphocyte numbers were reduced but with a considerably smaller decrease in T CD4⁺ number. Cocaine treatment altered proportions between the lymphocyte subsets by decreasing the percentages of T CD8⁺, B, and NK cells but increasing a percentage of T CD4⁺ cells. Destabilization in proportions between T CD4⁺ and T CD8⁺ was manifested as an elevated CD4/CD8 ratio that occurred despite increased plasma corticosterone and the lymphocytopenia. Cocaine did not affect the concentration of IFN-γ. The results suggest that although cocaine induced lymphopenia, it did not suppress the overall immune activity in terms of the CD4/CD8 ratio.     

Crucial CD8+ T-lymphocyte cytotoxic role in amphotericin B nanospheres efficacy against experimental visceral leishmaniasis

This work aims to develop poly(d,l-lactide-co-glycolide) (PLGA)-nanospheres containing amphotericin B (AmB) with suitable physicochemical properties and anti-parasitic activity for visceral leishmaniasis (VL) therapy. When compared with unloaded-PLGA-nanospheres, the AmB-loaded PLGA-nanospheres displayed an increased particle size without affecting the polydispersity and its negative surface charge. AmB stability in the PLGA-nanospheres was > 90% over 60-days at 30 °C. The AmB-PLGA-nanospheres demonstrated significant in vitro and in vivo efficacy and preferential accumulation in the visceral organs. In addition, an immune-modulatory effect was observed in mice treated with AmB-PLGA-nanospheres, correlating with improved treatment efficacy. The in vitro cytotoxic response of the T-lymphocytes revealed that AmB-PLGA-nanospheres efficacy against VL infection was strictly due to the action of CD8⁺- but not CD4⁺-T lymphocytes. Overall, we demonstrate a crucial role for CD8⁺ cytotoxic T lymphocytes in the efficacy of AmB-PLGA nanospheres, which could represent a potent and affordable alternative for VL therapy.From the Clinical EditorThis study demonstrates a crucial role for CD8+ T lymphocytes in eliminating visceral leishmaniasis in a murine model by enhancing the cytotoxic efficacy of CD8+ T-cells via amphotericin-B-PLGA nanospheres, paving a way to a unique, potentially more potent and cost-effective therapeutic strategy.     

Peripheral blood CD4+CRTH2+ cells in advanced stage non small cell lung cancer

To investigate CD4+CRTH2+ cells in peripheral blood in advanced stage non small cell lung cancer (NSCLC) patients. Forty-six patients with advanced stage NSCLC, who are chemotherapy or radiotherapy naïve, and 17 healthy volunteers, were enrolled in this study. The study was performed using flow cytometry and a complete blood cell counter analyser. CD4+ T cell percentage, CD4/CD8 ratio, CRTH2+CD4+ cell percentages, counts, and mean fluorescein intensity (MFI) and hematological parameters were evaluated in both groups. A survival analysis was performed to compare the patients with high CD4+CRTH2+ cell percentage and those with low CD4+CRTH2+ percentage. CD4+ T cell percentage in total lymphocytes and the CD4/CD8 ratio were lower in the patient group than in the control group. The absolute CD8 T cell count was higher in the patient group than in the control group, whereas the total T cells was not different. The CRTH2+ cell percentage in CD4+ T cells (7.96% ± 6.21% vs 3.37% ± 3.55%; respectively; p: 0,001) and the absolute count of CRTH2+CD4+ cells ( 97 mm-3 ± 109 mm-3 vs 37 mm-3 ± 38 mm-3, respectively; p: 0,033) in the patient group were higher than in the control group, but CRTH2-PE MFI values were not different between groups. Cox regression analysis did not show that CRTH2+CD4+ cell count or percentage is an independent prognostic factor. The study found that CRTH2 expression of CD4+ T cells and CRTH2+CD4+ cell number are higher in the peripheral blood of NSCLC patients than in that of healthy subjects. Further studies that explore the biological significance of high CD4+CRTH2+ cells in lung cancer patients, should be pursued.     

消癌平注射液联合化疗对中晚期肺癌患者CD4~+CD25~+FOXP3~+调节型T细胞的影响

目的探讨消癌平注射液联合化疗对中晚期肺癌患者CD4+CD25+FOXP3+调节型T细胞的影响。方法 71例Ⅲ～Ⅳ期非小细胞肺癌患者随机分为治疗组与对照组,治疗组36例采用化疗联合消癌平注射液;对照组35例单独采用化疗,两组化疗均采用GP方案。完成2周期后,通过流式细胞仪测定两组患者治疗前后外周血中CD4+CD25+FOXP3+调节型T细胞比例。结果治疗后治疗组患者CD4+FOXP3+T细胞占CD4+T细胞及CD4+CD25+T细胞的比例分别为(6.2±2.4)%和(27.6±6.0)%,而对照组分别为(8.2±0.5)%和(32.1±7.6)%,两组比较差异有统计学意义(P<0.05)。CD4+T细胞占总淋巴细胞的比例,治疗组较对照组明显提高(P<0.05)。结论消癌平注射液是一种较为理想的免疫增强剂,联合化疗可明显改善非小细胞肺癌患者的细胞免疫功能。