狂犬病疫苗对人外周血B淋巴细胞分泌特异性抗体的体外诱导作用

目的探索在体外用狂犬病疫苗诱导人外周血B淋巴细胞(PBL)分泌特异性抗体,并使B淋巴细胞保持良好增殖和分化状态的方法。方法以狂犬病疫苗免疫志愿者的外周血淋巴细胞进行体外诱导,并对诱导抗原量、诱导时间及IL-2、IL-6和胞壁酰二肽(MDP)等生物因素对诱导产生特异性抗体的影响进行检测,并观察PBL的活化与增殖状态。结果在特异抗原及其他生物因子的共同作用下,经免疫的人PBL体外抗原诱导5d后产生特异性抗体的量达到最大值。MDP、IL-2、IL-6对免疫志愿者的PBL体外特异性抗体的分泌有显著影响,但对未经免疫志愿者无明显影响。显微镜观察显示PBL生长与分化正常。结论已建立了体外诱导人外周血B淋巴细胞分泌特异性抗体的方法,并保持了B淋巴细胞良好的生长增殖和分化状态。     

猪弓形虫重组卡介苗的构建及其免疫保护性

目的构建弓形虫Rhomboid基因和猪IL-2基因的重组卡介苗(BCG),并分析其免疫保护效果。方法应用RT-PCR技术分别扩增弓形虫Rhomboid基因和猪IL-2基因,克隆至p MD18-T载体中,双酶切鉴定及测序正确后,将弓形虫Rhomboid基因和猪IL-2基因串联后分别连接至穿梭表达载体pMV261和整合表达载体pMV361中,电转化BCG,制备重组BCG pMV261-IL-2-Rho和pMV361-IL-2-Rho。分别经颈部肌肉注射免疫猪,共3次,每次间隔2周,免疫剂量均为108 CFU/只,采用ELISA法检测免疫前后猪体内抗体(IgG)滴度、Th1型及Th2型细胞因子的变化情况;第3次免疫后2周通过腹腔注射的方式接种弓形虫China I株速殖子1×108个/只进行攻虫试验,观察重组BCG对猪的免疫保护效果。结果成功构建了重组BCG p MV261-IL-2-Rho和p MV361-IL-2-Rho。重组BCG免疫猪后,随着免疫次数的增加,血清中抗体滴度也逐渐升高,且与PBS对照组相比,差异有统计学意义(P0.01)。3次免疫后,pMV261-IL-2-Rho组血清Th1型细胞因子IL-2、IFNγ和IL-12的表达水平均明显提高,与PBS对照组相比,差异有统计学意义(P0.05),而pMV361-IL-2-Rho组血清IL-2、IFNγ和IL-12的水平虽略有升高,但与PBS对照组相比,差异无统计学意义(P0.05);各组在免疫过程中Th2型细胞因子IL-4水平均未发生明显变化。攻虫后各组猪出现精神沉郁,食欲下降,体温升高等症状,耐过发病期后的猪,食欲逐渐恢复,体温渐趋正常,各种症状减轻,且各组猪在攻虫后20 d均无死亡。结论构建的基因重组BCG pMV261-IL-2-Rho对猪弓形虫感染具有一定的免疫保护作用。     

细胞因子诱导的杀伤细胞免疫治疗的研究进展

过继免疫细胞治疗作为一种新的方法已用于常规治疗无效的实体肿瘤,在肿瘤的治疗中有着巨大的应用潜力。细胞因子诱导的杀伤细胞(cytokine-induced killer,CIK)是人外周血单个核细胞经多种细胞因子体外扩增所得的一群异质细胞。CIK兼有T淋巴细胞的强大抗瘤活性和自然杀伤细胞(natural killer cells,NK)非主要组织相容性复合体(major histochompatibility complex,MHC)限制杀瘤特点,对多种实体肿瘤有较好的疗效。本文就CIK细胞的生物学特点、杀瘤机制及其临床疗效的最新进展进行综述。     

白细胞介素-15在感染性疾病中作用的研究进展

白细胞介素-15(interleukin,IL-15)是一种具有多种生物学活性的细胞因子,可活化NK、CD8+T淋巴细胞等免疫细胞,参与机体的先天性免疫和适应性免疫;还可作为佐剂,提高免疫细胞对抗原性物质的应答能力。因此,IL-15在细菌、病毒、寄生虫等微生物感染性疾病的致病及预后过程中发挥重要作用。     

大规模扩增人外周血来源自然杀伤细胞体系的筛选

目的：探讨人外周血来源的单个核细胞经三种体外培养体系活化扩增后的自然杀伤细胞（NK细胞）在扩增倍数、生物学表型和细胞毒性上的异同。方法：收集健康供者捐赠的外周血,经淋巴细胞分离液介导的密度梯度离心法富集单个核细胞。使用含自体血浆、重组人细胞因子的无血清培养液,各体系均添加IL-2和IL-15,体系B额外添加抗人CD3单抗,体系C额外添加IL-18,在相同条件下培养扩增14 d,比较不同培养体系的NK细胞在细胞表型、细胞活力及细胞毒性的异同。结果：体系A、B、C中NK细胞扩增倍数分别为811.43±30.07、2 800.79±36.42和1 047.97±85.21,表明抗人CD3单抗可以增强NK细胞的扩增。三种体系培养NK细胞在细胞表型、细胞周期、细胞活力和细胞毒性方面无统计学差异。结论：使用含自体血浆、重组人IL-2、IL-15细胞因子和抗人CD3单抗的无血清培养液,在37℃、5%CO2条件下可以实现NK细胞大规模培养,获取高纯度的NK细胞。     

融合表达IL-2和EGFP逆转录病毒载体的构建

目的构建融合表达IL-2和EGFP报告基因的逆转录病毒载体。方法设计载体pRevTet-On和pEGFP-C1的接头序列,经酶切连接重组为过渡载体pRevEGFP-C1,同时对质粒pBV220-IL-2和pEGFP-C1进行酶切,连接重组为过渡载体pEGFP-C1-IL-2。分别酶切质粒pRevEGFP-C1和pEGFP-C1-IL-2,琼脂糖凝胶电泳回收目的片段IL-2,并将其连接到pRevEGFP-C1的多克隆位点(MCS)中,转化至E.coliDH5α进行扩增,提取质粒DNA获得重组体pRevEGFP-C1-IL-2。结果经分析鉴定,所构建的重组载体pRevEGFP-C1-IL-2完全正确。结论已成功地构建了融合表达IL-2和EGFP的逆转录病毒载体pRevEG-FP-C1-IL-2。     

人细胞内Ca2+对CD4+T细胞免疫功能调控作用的研究进展

CD4⁺T淋巴细胞是人体免疫系统中重要的免疫细胞之一,主要分为Th1、Th2、Th17、滤泡辅助T细胞(T f ollicular helper cells,Tfh)及调节性T细胞(regulatory T cells,Tregs)。CD4⁺T淋巴细胞功能取决于T细胞受体(T cell receptor,TCR),TCR的激活触发了内质网释放Ca²⁺,Ca²⁺作为细胞的第二信使,通过改变其在细胞内外的浓度来参与细胞周期进展和增殖等多个生物活动过程,以维持免疫细胞的正常功能。CD4⁺T淋巴细胞与脂肪肝、肝脂沉积、肝炎、肿瘤等多种疾病的发生发展有关。因此,本文就细胞内Ca²⁺对CD4⁺T细胞免疫功能调控作用的研究进展作一综述,以期为某些疾病新临床治疗方案的研发提供参考。     

MF59佐剂对脊髓灰质炎灭活疫苗诱导恒河猴细胞免疫应答的影响

目的分析以MF59为佐剂的脊髓灰质炎灭活疫苗(IPV)免疫恒河猴后诱导的细胞免疫应答。方法将恒河猴分为4组:无佐剂低剂量组(IPVL-PBS组)、低剂量MF59佐剂组(IPVL-MF59组)、无佐剂高剂量组(IPVH-PBS组)和低剂量铝佐剂组(IPVL-AL组)。高剂量组IPV每剂中Ⅰ、Ⅱ、Ⅲ型抗原含量分别为30、32、42 DU,低剂量组IPV每剂中Ⅰ、Ⅱ、Ⅲ型抗原含量分别为3、3.2、4.2 DU,0.5 ml/剂,其中MF59为0.25 ml/剂,氢氧化铝为0.5 mg/剂,均于第1、28、56天经恒河猴后腿肌肉注射,分别于免疫后第28、56、84天采血,分离血清,微量中和试验法检测血清中和抗体效价;分别于免疫后第56和84天,取外周血,分离淋巴细胞,上流式细胞仪检测特异性抗原刺激下的Th1型细胞因子(IL-2、IFNγ、TNF)和Th2型细胞因子(IL-4、IL-5、IL-6)分泌水平,以及CD3+、CD4+、CD8+T淋巴细胞比例的分布情况。结果恒河猴经3次免疫后,IPVL-MF59组血清中和抗体阳转率明显高于其他各组,达75%以上。恒河猴免疫后第56天,IPVL-MF59组IFNγ和TNF分泌水平明显高于IPVL-AL组(P均0.05),IL-6分泌水平明显高于IPVL-PBS和IPVH-PBS组(P均0.05);免疫后第84天,各组IL-2、IFNγ、TNF均稳定在同一水平,IL-4、IL-5分泌水平均明显高于第56天(P均0.05)。免疫后第84天,IPVL-MF59组CD8+T淋巴细胞百分比均明显高于IPVH-PBS和IPVL-PBS组(P均0.05)。结论 MF59佐剂能增强IPV诱导的TNF和IL-6的分泌水平,增强Th1和Th2类反应。     

重组弓形虫磷酸甘油酸变位酶2联合白细胞介素-2或干扰素γ诱导小鼠的脾淋巴细胞免疫应答

目的观察重组弓形虫磷酸甘油酸变位酶2(Recombinant Toxoplasma gondii phosphoglycerate mutase 2,rTgPGAM2)联合白细胞介素-2(Interleukin-2,IL-2)或干扰素γ(Interferonγ,IFNγ)滴鼻免疫小鼠诱导的脾淋巴细胞免疫应答,探讨IL-2和IFNγ的佐剂效应。方法将48只BALB/c小鼠随机均分为6组:rTgPGAM2组(30μg)、IL-2组(500 IU)、IFNγ组(1 000 IU)、rTgPGAM2(30μg)+IL-2(500 IU)组和rTgPGAM2(30μg)+IFNγ(1 000 IU)组和对照组(20μl PBS),免疫途径均为滴鼻免疫,共免疫3次。末次免疫后第14天,颈椎脱臼处死小鼠,CCK-8法检测各组小鼠脾淋巴细胞增殖活性,ELISA法检测脾淋巴细胞培养上清中IL-2、IFNγ、IL-4和IL-10水平。结果经rTgPGAM2刺激后,与对照组比较,各免疫组小鼠脾淋巴细胞刺激指数(Stimulation index,SI)均显著高于对照组(P<0.05或P<0.01),rTgPGAM2联合IL-2或IFNγ组显著高于rTgPGAM2组(P<0.01或P<0.05);经ConA刺激后,仅IL-2组SI显著高于对照组(P<0.01)。各免疫组小鼠脾淋巴细胞培养上清中IL-2和IFNγ含量均显著高于对照组(P<0.05或P<0.01),rTgPGAM2联合IL-2或IFNγ组显著高于rTgPGAM2组(P<0.01或P<0.05);rTgPGAM2组及其联合IL-2或IFNγ组IL-4含量显著高于对照组(P<0.05);而IL-10的含量,只有IFNγ组和rTgPGAM2+IFNγ组显著高于对照组(P<0.05)。结论 rTgPGAM2联合IL-2或IFNγ鼻内免疫小鼠诱导的脾淋巴细胞免疫应答优于rTgPGAM2单独免疫,表明IL-2和IFNγ具有良好的佐剂效应,IL-2的佐剂效应更佳。     

Immune Profile Analysis in Peripheral Blood and Tumor in Patients with Malignant Melanoma

Melanoma is a severe and life-threatening malignancy derived from melanocytes. The traditional treatment for melanoma could not sustain satisfactory outcomes long term; however, the recent immune checkpoint treatment has made a breakthrough in these problems. Nivolumab is a representative immune checkpoint treatment, and this PD-1-targeted therapy has evolutionally developed and improved the clinical outcome in a recent decade. On the other hand, the clinical application of immune checkpoint treatment presents clinicians with novel questions, especially how to obtain additional efficacy and overcome the disadvantage by using this treatment. To answer these problems, we first investigated the distribution of PD-L1 in various organs to clarify the organs most affected by anti-PD-1 antibody treatment. Among various organs, lung, placenta, spleen, heart, and thyroid highly expressed PD-L1, while skin, thalamus, hippocampus, ovary, stomach, testis, and prostate showed lower expressions of PD-L1. Furthermore, the immune profiles were also examined in tumors and peripheral blood in patients with melanoma. PD-1 was highly expressed in CD8 and CD4 cells, and B cells also highly expressed PD-1 compared with NK cells. However, there was no significant difference in Th1/Th2/Th17 cytokines and inhibitory cytokine IL-10. Although nevus showed a low expression of PD-L1 compared with healthy skin, PD-L1 expression was increased in growth-phase melanoma. Finally, we analyzed the peripheral blood profiles in patients treated with nivolumab. PD-1-bearing dendritic cells (DCs) were increased during nivolumab treatment and Lin-CD11c+HLA-DR+ cells were highly increased during nivolumab treatment. These findings indicate a clue to answering the problems during nivolumab treatment and suggest to us the importance of multiple aspect observation during immune checkpoint treatment.     

IL-2、15、22基因对CVB3 VP1 DNA疫苗免疫效果的影响

目的观察白细胞介素2(Interleukin2,IL2)、白细胞介素15(Interleukin15,IL15)和白细胞介素22(Interleukin22,IL22)基因对柯萨奇病毒(CoxasckievirusB3,CVB3)VP1DNA疫苗诱导小鼠免疫应答及保护作用的影响。方法取雄性BALBc小鼠,随机分为6组,每组20只,分别肌内注射盐水、pcDNA3、pcDNA3VP1、pcDNA3IL2+pcDNA3VP1、pcDNA3IL15+pcDNA3VP1和pcDNA3IL22+pcDNA3VP1,每周1次,共3次,每次免疫后6d取血清用微量中和试验检测CVB3中和抗体,3次免疫后用800TCID50CVB3感染小鼠,观察小鼠的生存时间和生存率。结果pcDNA3VP1、pcDNA3IL2+pcDNA3VP1和pcDNA3IL15+pcDNA3VP1组均能诱导小鼠产生中和抗体,抗体滴度随免疫次数增加而提高。病毒攻击后,各组的生存率差异无显著意义,pcDNA3IL2+pcDNA3VP1和pcDNA3IL15+pcDNA3VP1组较其他4组生存时间明显延长。pcDNA3IL22+pcDNA3VP1组虽也能诱导小鼠产生中和抗体,但抗体滴度和生存情况无明显变化。结论IL2、IL15基因对CVB3VP1DNA疫苗诱导小鼠产生免疫应答和免疫保护有一定的增强作用,而IL22基因无明显作用。     

What Do We Have to Know about PD-L1 Expression in Prostate Cancer? A Systematic Literature Review. Part 3: PD-L1, Intracellular Signaling Pathways and Tumor Microenvironment

The tumor microenvironment (TME) includes immune (T, B, NK, dendritic), stromal, mesenchymal, endothelial, adipocytic cells, extracellular matrix, and cytokines/chemokines/soluble factors regulating various intracellular signaling pathways (ISP) in tumor cells. TME influences the survival/progression of prostate cancer (PC), enabling tumor cell immune-evasion also through the activation of the PD-1/PD-L1 axis. We have performed a systematic literature review according to the PRISMA guidelines, to investigate how the PD-1/PD-L1 pathway is influenced by TME and ISPs. Tumor immune-escape mechanisms include suppression/exhaustion of tumor infiltrating cytotoxic T lymphocytes, inhibition of tumor suppressive NK cells, increase in immune-suppressive immune cells (regulatory T, M2 macrophagic, myeloid-derived suppressor, dendritic, stromal, and adipocytic cells). IFN-γ (the most investigated factor), TGF-β, TNF-α, IL-6, IL-17, IL-15, IL-27, complement factor C5a, and other soluble molecules secreted by TME components (and sometimes increased in patients’ serum), as well as and hypoxia, influenced the regulation of PD-L1. Experimental studies using human and mouse PC cell lines (derived from either androgen-sensitive or androgen-resistant tumors) revealed that the intracellular ERK/MEK, Akt-mTOR, NF-kB, WNT and JAK/STAT pathways were involved in PD-L1 upregulation in PC. Blocking the PD-1/PD-L1 signaling by using immunotherapy drugs can prevent tumor immune-escape, increasing the anti-tumor activity of immune cells.     

Activation of Peripheral Blood Mononuclear Cells and Leptin Secretion: New Potential Role of Interleukin-2 and High Mobility Group Box (HMGB)1

Activation of innate immunity and low-grade inflammation contributes to hyperglycemia and an onset of Type 2 Diabetes Mellitus (T2DM). Interleukin-2 (IL-2), leptin, High Mobility Group Box-1 (HMGB-1), and increased glucose concentrations are mediators of these processes also by modulating peripheral blood mononuclear cells (PBMCs) response. The aim of this study was to investigate if HMGB-1 and IL-2 turn on PBMCs and their leptin secretion. In isolated human PBMCs and their subpopulations from healthy individuals and naïve T2DM patients, leptin release, pro-inflammatory response and Toll-like Receptors (TLRs) activation was measured. After treatment with IL-2 and HMGB1, NK (Natural Killer) have the highest amount of leptin secretion, whilst NK-T have the maximal release in basal conditions. TLR4 (TAK242) and/or TLR2 (TLR2-IgA) inhibitors decreased leptin secretion after IL-2 and HMGB1 treatment. A further non-significant increase in leptin secretion was reported in PBMCs of naive T2DM patients in response to IL-2 and HMGB-1 stimulation. Finally, hyperglycemia or hyperinsulinemia might stimulate leptin secretion from PBMCs. The amount of leptin released from PBMCs after the different treatments was enough to stimulate the secretion of IL-1β from monocytes. Targeting leptin sera levels and secretion from PBMCs could represent a new therapeutic strategy to counteract metabolic diseases such as T2DM.     

Secreted Ligands of the NK Cell Receptor NKp30: B7-H6 Is in Contrast to BAG6 Only Marginally Released via Extracellular Vesicles

NKp30 (Natural Cytotoxicity Receptor 1, NCR1) is a powerful cytotoxicity receptor expressed on natural killer (NK) cells which is involved in tumor cell killing and the regulation of antitumor immune responses. Ligands for NKp30, including BAG6 and B7-H6, are upregulated in virus-infected and tumor cells but rarely detectable on healthy cells. These ligands are released by tumor cells as part of the cellular secretome and interfere with NK cell activity. BAG6 is secreted via the exosomal pathway, and BAG6-positive extracellular vesicles (EV-BAG6) trigger NK cell cytotoxicity and cytokine release, whereas the soluble protein diminishes NK cell activity. However, the extracellular format and activity of B7-H6 remain elusive. Here, we used HEK293 as a model cell line to produce recombinant ligands and to study their impact on NK cell activity. Using this system, we demonstrate that soluble B7-H6 (sB7-H6), like soluble BAG6 (sBAG6), inhibits NK cell-mediated target cell killing. This was associated with a diminished cell surface expression of NKG2D and NCRs (NKp30, NKp40, and NKp46). Strikingly, a reduced NKp30 mRNA expression was observed exclusively in response to sBAG6. Of note, B7-H6 was marginally released in association with EVs, and EVs collected from B7-H6 expressing cells did not stimulate NK cell-mediated killing. The molecular analysis of EVs on a single EV level using nano flow cytometry (NanoFCM) revealed a similar distribution of vesicle-associated tetraspanins within EVs purified from wildtype, BAG6, or B7-H6 overexpressing cells. NKp30 is a promising therapeutic target to overcome NK cell immune evasion in cancer patients, and it is important to unravel how extracellular NKp30 ligands inhibit NK cell functions.     

Diphtheria toxin receptor-binding domain substitution with interleukin 6: genetic construction and interleukin 6 receptor-specific action of a diphtheria toxin-related interleukin 6 fusion protein

We have genetically replaced that portion of the diphtheriatoxin structural gene which encodes the native receptor-bindingdomain with a synthetic gene encoding the cytokine interleukin6 (IL-6/IFN-ß2/BSF-2). The resulting gene fusion encodesthe chimeric toxin DAB₃₈₉-IL-6. Following expression and purification,we demonstrate that DAB₃₈₉-IL-6 is selectively cytotoxic foreukaryotic cells bearing the interleukin 6 receptor. In addition,the cytotoxic action of DAB₃₈₉-IL-6 is shown to require bindingto the IL-6 receptor, internalization by receptor-mediated endocytosisand passage through an acidic compartment. Following the deliveryof the catalytically active fragment A to the cytosol of targetcells, cellular protein synthesis is inhibited by the ADP-ribosylationof elongation factor 2. While eukaryotic cells which are devoidof the IL-6 receptor are uniformly resistant to the action ofthis fusion toxin, the data presented suggest that a minimalnumber of IL-6 receptors may be necessary to mediate the internalizationof sufficient levels of DAB₃₈₉-IL-6 to result in the intoxicationof target cells.     

A Chimeric IL-15/IL-15Rα Molecule Expressed on NFκB-Activated Dendritic Cells Supports Their Capability to Activate Natural Killer Cells

Natural killer (NK) cells, members of the innate immune system, play an important role in the rejection of HLA class I negative tumor cells. Hence, a therapeutic vaccine, which can activate NK cells in addition to cells of the adaptive immune system might induce a more comprehensive cellular response, which could lead to increased tumor elimination. Dendritic cells (DCs) are capable of activating and expanding NK cells, especially when the NFκB pathway is activated in the DCs thereby leading to the secretion of the cytokine IL-12. Another prominent NK cell activator is IL-15, which can be bound by the IL-15 receptor alpha-chain (IL-15Rα) to be transpresented to the NK cells. However, monocyte-derived DCs do neither secrete IL-15, nor express the IL-15Rα. Hence, we designed a chimeric protein consisting of IL-15 and the IL-15Rα. Upon mRNA electroporation, the fusion protein was detectable on the surface of the DCs, and increased the potential of NFκB-activated, IL-12-producing DC to activate NK cells in an autologous cell culture system with ex vivo-generated cells from healthy donors. These data show that a chimeric IL-15/IL-15Rα molecule can be expressed by monocyte-derived DCs, is trafficked to the cell surface, and is functional regarding the activation of NK cells. These data represent an initial proof-of-concept for an additional possibility of further improving cellular DC-based immunotherapies of cancer.     

肠道病毒71型感染患儿外周血自然杀伤细胞比例、亚群、表面受体及免疫效应分子的变化

目的探讨肠道病毒71型(Enterovirus 71,EV71)感染患儿外周血中自然杀伤(Natural killer,NK)细胞比例、亚群、表面受体及免疫效应分子的变化。方法将EV71阳性的急性期患儿按疾病严重程度分为重症病例组和普通病例组,同时设健康对照组。提取各组患儿外周血,采用流式细胞术检测外周血中NK细胞比例、NK细胞亚群、自然细胞毒受体NKp30和NKp46、激活性受体NKG2D、抑制性受体CD94和NKG2A、脱颗粒标记CD107a及免疫效应分子(PF、GrB和GNLY)阳性细胞所占NK细胞的比例。结果重症病例组患儿外周血NK细胞比例较健康对照组明显减少(P<0.01),普通病例组CD16+NK细胞亚群比例较健康对照组升高(P<0.05),重症病例组较普通病例组低,但差异无统计学意义(P>0.05);重症病例组和普通病例组NKG2D阳性细胞百分率均较健康对照组低(P<0.01),而CD94和NKG2A阳性细胞百分率均较健康对照组明显升高(P<0.01),NKp30和NKp46阳性细胞率3组之间差异无统计学意义(P>0.05);重症病例组和普通病例组CD107a、PF、GrB、GNLY的阳性细胞百分率均较健康对照组明显降低(P<0.01或P<0.05)。结论 EV71感染可能抑制宿主NK细胞的增殖、激活及其杀伤功能。重症病例组与普通病例组比较,NK细胞比例和CD16+NK细胞亚群降低,抑制性受体CD94表达增高,胞浆内CD107a、GrB和PF表达显著降低,可能与重症病例的发生有关。     

Mechanisms,Characteristics, and Treatment of Neuropathic Pain and Peripheral Neuropathy Associated with Dinutuximab in Neuroblastoma Patients

Prognosis of metastatic neuroblastoma is very poor. Its treatment includes induction chemotherapy, surgery, high-dose chemotherapy, radiotherapy, and maintenance with retinoic acid, associated with the anti-GD2 monoclonal antibody (ch14.18) dinutuximab. Immunotherapy determined a significant improvement in survival rate and is also utilized in relapsed and resistant neuroblastoma patients. Five courses of dinutuximab 100 mg/m² are usually administered as a 10-day continuous infusion or over 5 consecutive days every 5 weeks. Dinutuximab targets the disialoganglioside GD2, which is highly expressed on neuroblastoma cells and minimally present on the surface of normal human neurons, peripheral pain fibers, and skin melanocytes. Anti GD2 antibodies bind to surface GD2 and determine the lysis of neuroblastoma cells induced by immune response via the antibody-dependent cellular cytotoxicity and the complement-dependent cytotoxicity. Dinutuximab has significant side effects, including neuropathic pain, peripheral neuropathy, hypersensitivity reactions, capillary leak syndrome, photophobia, and hypotension. The most important side effect is neuropathic pain, which is triggered by the same antibody–antigen immune response, but generates ectopic activity in axons, which results in hyperalgesia and spontaneous pain. Pain can be severe especially in the first courses of dinutuximab infusion, and requires the administration of gabapentin and continuous morphine infusion. This paper will focus on the incidence, mechanisms, characteristics, and treatment of neuropathic pain and peripheral neuropathy due to dinutuximab administration in neuroblastoma patients.     

Peritoneal Fluid from Patients with Ovarian Endometriosis Displays Immunosuppressive Potential and Stimulates Th2 Response

Endometriosis is a common gynaecological disorder characterized by the ectopic growth of endometrial tissue outside the uterine cavity. It is associated with chronic pelvic inflammation and autoimmune reactivity manifesting by autoantibody production and abrogated cellular immune responses. Endometriotic peritoneal fluid contains various infiltrating leucocyte populations and a bulk of proinflammatory and immunoregulatory cytokines. However, the nature and significance of the peritoneal milieu in women with endometriosis still remains obscure. Therefore, the aim of the present study was to investigate the immunoregulatory activity of the peritoneal fluid (PF) from women with endometriosis. The peritoneal fluid samples were collected during laparoscopic surgery from 30 women with and without endometriosis. Immunoregulatory cytokines (IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ and TNF) and chemokines (CCL2, CCL5, CXCL8 and CXCL9) were evaluated in PF and culture supernatants generated by unstimulated and CD3/CD28/IL-2-stimulated CD4⁺ T cells cultured in the presence of PF. The effect of PF on the generation of Treg and Th17 cells in CD4⁺ T cell cultures, as well as the natural cytotoxic activity of peripheral blood mononuclear cells, was also investigated. Concentrations of IL-6, IL-10, CCL2, CXCL8 and CXCL9 were significantly upregulated in the PF from women with endometriosis when compared to control women, whereas concentrations of other cytokines and chemokines were unaffected. The culturing of unstimulated and CD3/CD28/IL-2-stimulated CD4⁺ T cells in the presence of endometriotic PF resulted in the downregulation of their IL-2, IFN-γ, IL-17A and TNF production as compared to culture medium alone. On the other side, endometriotic PF significantly stimulated the production of IL-4 and IL-10. Endometriotic PF also stimulated the release of CCL2 and CXCL8, whereas the production of CCL5 and CXCL9 was downregulated. Endometriotic PF stimulated the generation of Treg cells and had an inhibitory effect on the generation of Th17 cells in cultures of CD4⁺ T cells. It also inhibited the NK cell cytotoxic activity of the peripheral blood lymphocytes. These results strongly imply that the PF from patients with endometriosis has immunoregulatory/immunosuppressive activity and shifts the Th1/Th2 cytokine balance toward the Th2 response, which may account for deviation of local and systemic immune responses. However, a similar trend, albeit not a statistically significant one, was also observed in case of PF from women without endometriosis, thus suggesting that peritoneal milieu may in general display some immunoregulatory/immunosuppressive properties. It should be stressed, however, that our present observations were made on a relatively small number of PF samples and further studies are needed to reveal possible mechanism(s) responsible for this phenomenon.