Immune evasion and therapeutic opportunities based on natural killer cells

Natural killer(NK)cells can elicit an immune response against malignantly transformed cells without recognizing antigens,and they also exhibit cytotoxic effects and immune surveillance functions in tumor immunotherapy.Although several studies have shown the promising antitumor effects of NK cells in immunotherapy,their function is often limited in the tumor microenvironment because tumor cells can easily escape NK cell-induced death.Thus,for efficient tumor immunotherapy,the mechanism by which tumor cells escape NK cell-induced cytotoxicity must be fully understood.Various novel molecules and checkpoint receptors that mediate the disruption of NK cells in the tumor microenvironment have been discovered.In this review,we analyze and detail the major activating and inhibitory receptors on the surface of NK cells to delineate the mechanism by which tumor cells suppress NKG2D ligand expression and increase tumor receptor and inhibitory receptor expression[NKG2A,programmed cell death1(PD-1),and T-cell immunoglobulin and immunoreceptor tyrosine inhibitory motif(TIGIT)]on the NK cell surface,and thus inhibit NK cell activity.We also reviewed the current status of treatments based on these surface molecules.By comparing the therapeutic effects related to the treatment status and bypass mechanisms,we attempt to identify optimal single or combined treatments to suggest new treatment strategies for tumor immunotherapy.

Advancement of chimeric antigen receptor-natural killer cells targeting hepatocellular carcinoma

With the advance of genome engineering technology,chimeric antigen receptors(CARs)-based immunotherapy has become an emerging therapeutic strategy for tumors.Although initially designed for T cells in tumor immunotherapy,CARs have been exploited to modify the function of natural killer(NK)cells against a variety of tumors,including hepatocellular carcinoma(HCC).CAR-NK cells have the potential to sufficiently kill tumor antigen-expressing HCC cells,independent of major histocompatibility complex matching or prior priming.In this review,we summarize the recent advances in genetic engineering of CAR-NK cells against HCC and discuss the current challenges and prospects of CAR-NK cells as a revolutionary cellular immunotherapy against HCC.

HBsAg stimulates NKG2D receptor expression on natural killer cellsand inhibits hepatitis C virus replication

Background： Higher hepatitis B surface antigen （HBsAg） facilitates hepatitis C virus （HCV） clearance inpatients with hepatitis B virus （HBV）/HCV co-infection. We investigated the effect of exogenous HBsAgon the inhibition of HCV replication mediated by natural killer （NK） cells.

Transient Receptor Potential Melastatin 3 and Intracellular Calcium in Natural Killer Cells in Multiple Sclerosis

Background: Natural killer (NK) cell phenotypes have reported to be implicated in the pathomechanism of Multiple Sclerosis (MS). Several investigators have observed reduced peripheral numbers, reduced cytotoxic activity, and altered CD56Dim and CD56Bright NK cell phenotypes. This current project, for the first time, investigates the NK cell cytotoxicity, calcium mobilisation and transient receptor potential melastatin 3 (TRPM3) surface expression. Methods: NK cell cytotoxic activity and calcium signaling were examined in CD56Dim and CD56Bright NK cells before and after stimulation using Ionomycin, Pregnenolone sulphate, 2-Aminoethoxydiphenyl borate and Thapsigargin. Purified NK cells were labelled with antibodies to determine TRPM3, CD69 and CD107a surface expression using flow cytometry. Results: Twenty-two MS patients and 22 healthy controls were recruited for this project. Twelve of the 22 previously received Alemtuzumab (Lemtrada®) and the remaining ten reported nil medication. We report TRPM3 was significantly increased in untreated MS patients compared with healthy controls and treated MS patients (p-value 0.034). There was a significant decrease in CD69 surface expression on CD56Dim NK cell phenotype for untreated MS patients (p-value 0.031) and treated MS patients (p-value 0.036). We report altered calcium mobilisation in CD56Bright NK cells and to a lesser extent CD56Dim NK cells between healthy controls, treated and untreated MS patients. Conclusion: This investigation suggests variations in TRPM3 expression and calcium mobilisation of NK cells may be implicated in the pathogenesis of MS. Further investigation is required to determine the mechanism by which alemtuzumab alters calcium signaling in NK cells.

Targeted Therapy of CEA-CAR-NK Cells Against Colorectal Cancer Cells

Objective:Investigate the cytotoxic effect of CAR-NK cells targeting CEA on colorectal cancer cells with positive CEA expression.Methods:The mRNA and protein levels of CEA in different CRC cell lines were detected by qRT-PCR and Western blot analysis.Lentiviral transduction was used to construct CAR-NK cells and empty vector CON-NK cells targeting CEA.Fluorescence microscopy and WB were used to determine whether the cells successfully constructed and expressed CAR structures.The effector NK cells were co-cultured with target cells,and the levels of LDH,IFN-γ,and GM-CSF were detected.The killing rate of effector cells was calculated,and the release of cytokines during the killing of target cells by different effector cells was compared.Results:The expression level of CEA in colorectal cancer patients was significantly higher than that in normal samples and other tumor samples,and the prognosis survival time of patients with high CEA expression was lower than that of CRC patients with low or no CEA expression(P<0.05).The CEA expression of the HT29 cell line was significantly higher than that of the SW1116 cell line at both the mRNA and protein levels.CEA-CAR-NK92 cells and CON-NK92 cells expressed green fluorescence under a microscope,and WB results showed that CEA-CAR-NK92 cells successfully expressed the CAR structure.Compared with CON-NK92 cells and NK92 cells,CEA-CAR-NK92 cells effectively killed HT29 cells(P<0.05).CEA-CAR-NK92 cells secreted a large amount of IFN-γand GM-CSF during the killing of HT29 cells,while the cytokine secretion of CON-NK92 cells and NK92 cells was not significant(P<0.05).Conclusion:CAR-NK92 cells targeting CEA can effectively kill CEA-positive colorectal cancer cells.

Natural Killer Cell-Based Immunotherapy for Cancer: Advances and Prospects

Natural killer (NK) cells are key innate immune cells that provide the first line of defense against viral infection and cancer. Although NK cells can discriminate between "self" and "non-self," recognize abnormal cells, and eliminate transformed cells and malignancies in real time, tumors develop several strategies to escape from NK cell attack. These strategies include upregulating ligands for the inhibitory receptors of NK cells and producing soluble molecules or immunosuppressive factors. Various types of NK cells are currently being applied in clinical trials, including autologous or allogeneic NK cells, umbilical cord blood (UCB) or induced pluripotent stem cell (iPSC)-derived NK cells, memory-like NK cells, and NK cell line NK-92 cells, for the treatment of different types of tumors. Chimeric antigen receptors (CARs)-NK cells have recently shown great potential due to their redirect specificity and effective antitumor activity. In this review, we summarize the mechanisms of tumor escape from NK cell recognition, the current status and advanced progress of NK cell-based immunotherapy, ways of enhancing the antitumor capacity of NK cells in vivo, and major challenges for clinical practice in this field.

Potential role of killer immunoglobulin receptor genes among individuals vaccinated against hepatitis B virus in Lebanon

AIMTo explore the role of killer immunoglobulin receptor (KIR) genes in responsiveness or non-responsiveness to vaccination against hepatitis B virus. METHODSWe recruited 101 voluntary participants between March 2010 and December 2011. Sera samples from vaccinated and non-vaccinated participants were tested for the presence of anti-HBs antibodies as a measure of protection against hepatitis B, hepatitis B surface antigen and hepatitis B core antibody as indicators of infection by enzyme-linked immunosorbent assay. KIR gene frequencies were determined by polymerase chain reaction. RESULTSSera samples from 99 participants were tested for the levels of anti-HBs as an indicator of protection (&ge; 10 mIU/mL) following vaccination as defined by the World Health Organization international reference standard. Among the vaccinated participants, 47% (35/74) had anti-HBs titers above 100 mIU/mL, 22% (16/74) had anti-HBs ranging between 10-100 mIU/mL, and 20% (15/74) had values of less than 10 mIU/mL. We report the lack of significant association between the number of vaccine dosages and the titer of antibodies among our vaccinated participants. The inhibitory KIR2DL1, KIR2DL4, KIR3DL1, KIR3DL2, and KIR3DL were detected in more than 95%, whereas KIR2DL2, KIR2DL3, KIR2DL5 (KR2DL5A and KIR2DL5B) were expressed in 56%, 84% and 42% (25% and 29%) of participants, respectively. The observed frequency of the activating KIR genes ranged between 35% and 55% except for KIR2DS4, detected in 95% of the study participants (40.6% 2DS4*001/002; 82.2% 2DS4*003/007). KIR2DP1 pseudogene was detected in 99% of our participants, whereas KIR3DP*001/02/04 and KIR3DP1*003 had frequencies of 17% and 100%, respectively. No association between the frequency of KIR genes and anti-HBs antibodies was detected. When we compared the frequency of KIR genes between vaccinated individuals with protective antibodies titers and those who lost their protective antibody levels, we did not detect a significant difference. KIR2DL5B was significantly different among different groups of vaccinated participants (group I > 100 mIU/mL, group II 10-100 mIU/mL, group III CONCLUSIONTo our knowledge, this is the first study screening for the possible role of KIR genes among individuals vaccinated against hepatitis B virus (HBV). Our results can be used to design larger studies to better understand the role of KIR genes in protection against or susceptibility to HBV post vaccination.

黄芪含药血清对NK细胞活性、NKG2A、NKG2D及细胞因子IFN-γ表达的影响

目的探讨黄芪含药血清对自然杀伤(NK)细胞的杀伤活性和NKG2D、NKG2A、细胞因子干扰素-γ(IFN-γ)表达水平的影响。方法20只SPF级SD大鼠,随机分成黄芪低、中、高剂量组(3.1、6.2、12.4 g/kg体重)和正常对照组,灌胃后制备含药血清。用NK细胞专用培养基培养NK-92MI细胞,用1640培养基培养YAC-1细胞。将NK-92MI细胞与20%不同剂量黄芪含药血清及正常对照血清孵育48 h,即分别为正常对照组、黄芪低剂量组、黄芪中剂量组以及黄芪高剂量组。通过乳酸脱氢酶(LDH)释放测定方法检测NK细胞对靶细胞YAC-1的杀伤活性,ELISA法检测IFN-γ的水平,Western blot检测NKG2D以及NKG2A蛋白表达水平。结果与正常对照组和黄芪低剂量组相比,黄芪中、高剂量组的NK细胞活性显著升高(P<0.01)。IFN-γ的表达在黄芪低、中、高剂量组中均升高,但不随浓度升高而增强。与正常对照组相比,黄芪低、中剂量组NKG2D蛋白表达水平明显升高(P<0.01),而NKG2A蛋白表达水平在各组间无明显差异(P>0.05)。结论黄芪含药血清可增强NK细胞杀伤活性、上调IFN-γ表达水平、上调NK细胞活化型受体的表达,通过调控NK细胞实现对机体的保护作用。

自然杀伤细胞家族2成员D受体-配体轴在血液瘤中的作用研究进展

自然杀伤细胞家族2成员D(NKG2D)受体通过表达在自然杀伤(NK)细胞和细胞毒性T淋巴(CTLs)细胞上激活这些免疫细胞对抗肿瘤。肿瘤细胞通过降低自然杀伤细胞家族2成员D配体(NKG2DLs)的表达来逃避免疫系统的监视,从而促进肿瘤的免疫逃逸。最新的研究揭示了NKG2D受体-配体轴在血液瘤的发展、免疫监视和治疗中的关键作用,尤其是在阐明免疫逃逸机制和开发新的治疗策略方面显示出巨大潜力。现从NKG2D受体-配体轴在血液瘤发病机制、免疫监测及治疗中的作用进行阐述,在加深对血液瘤病理机制理解的同时,为提升治疗效果开辟了新的策略。

Why natural killer cells in triple negative breast cancer?

The triple-negative subtype of breast cancer(TNBC)has the bleakest prognosis,owing to its lack of either hormone receptor as well as human epidermal growth factor receptor 2.Henceforth,immunotherapy has emerged as the front-runner for TNBC treatment,which avoids potentially damaging chemotherapeutics.However,despite its documented association with aggressive side effects and developed resistance,immune checkpoint blockade continues to dominate the TNBC immunotherapy scene.These immune checkpoint blockade drawbacks necessitate the exploration of other immunotherapeutic methods that would expand options for TNBC patients.One such method is the exploitation and recruitment of natural killer cells,which by harnessing the innate rather than adaptive immune system could potentially circumvent the downsides of immune checkpoint blockade.In this review,the authors will elucidate the advantageousness of natural killer cell-based immuno-oncology in TNBC as well as demonstrate the need to more extensively research such therapies in the future.

桑黄多糖调控MICB/NKG2D信号通路增强机体对脑胶质瘤细胞的免疫监视作用

目的:探讨桑黄多糖对脑胶质瘤细胞免疫监视的增强作用及其对主要组织相容性复合物Ⅰ链相关基因B(MICB)/自然杀伤细胞活化性受体2D(NKG2D)信号通路的调控机制。方法:取6~8周龄C57BL/6小鼠60只和GL261细胞株培养传代,随机取其中的50只小鼠建立脑胶质瘤模型,其余10只小鼠记为对照组,另将建模成功小鼠采用随机数字表分组,榄香烯组予以100 mg/kg榄香烯与1 ml/100 g生理盐水混匀灌胃,桑黄多糖各剂量组分别予以50、100、200 mg/kg桑黄多糖溶于1 ml/100 g生理盐水中灌胃,对照组和模型组均予以等量生理盐水灌胃,均1次/d,共4周。干预后处死小鼠,比较各组瘤重和抑瘤率;苏木素-伊红(HE)染色观察瘤组织病理变化;以CD57抗体标记检测瘤组织中NK细胞浸润;实时-逆转录聚合酶链反应(RT-qPCR)检测瘤组织主要组织相容性复合物Ⅰ链相关基因A(MICA)、MICB、UL16结合蛋白1(ULBP1)、UL16结合蛋白2(ULBP2)、UL16结合蛋白3(ULBP3)mRNA表达,Western blot检测瘤组织MICA、MICB、ULBP1、ULBP2、ULBP3蛋白表达;采用磁珠亲和细胞分选法分离各组组织中NK细胞,采用流式细胞仪检测其表面NKG2D活性。结果:与模型组比较,榄香烯组和桑黄多糖各剂量组瘤重下降,抑瘤率升高,瘤组织细胞变性、坏死增多,NK细胞浸润增多,MICA、ULBP2、ULBP3 mRNA和蛋白表达下降,MICB、ULBP1 mRNA和蛋白表达升高,且NK细胞表面NKG2D活性增强,上述差异均有统计学意义(P<0.05);桑黄多糖的作用呈剂量依赖性,桑黄多糖低剂量组和榄香烯组所有指标比较差异均无统计学意义(P>0.05)。结论:桑黄多糖可抑制脑胶质瘤增长,促进瘤细胞变性、坏死,增加NK细胞浸润,推测d与调控MICB/NKG2D信号通路,增加MICB、ULBP1表达和NK细胞表面NKG2D活性,下调MICA、ULBP2、ULBP3表达,增强对脑胶质瘤细胞的免疫监视有关。

Hepatitis B virus-persistent infection and innate immunity defect: Cell-related or virus-related?

The outcomes of hepatitis B virus(HBV) infection are closely related to the age at which infection was acquired. Infection acquired in adult life tends to be selflimited, in contrast to perinatal acquirement, for which chronic persistence of the HBV is a general outcome. Innate immunity plays an indispensable role in early virus infection, facilitating virus clearance. However, it has been reported that HBV is under-recognized and poorly eliminated by the innate immune system in the early stages of infection, possibly explaining the long-lasting persistence of viremia afterwards. Furthermore, due to the existence of covalently closed circular DNA, chronic HBV clearance is very difficult, even when patients are given interferon-α and nucleotide/nucleoside analogs for antiviral therapy. The mechanism by which HBV evades innate immune recognition and establishes persistent infection remains a subject of debate. Besides, some researchers are becoming more interested in how to eradicate chronic HBV infection by restoring or boosting innate immunity. This review aimed to summarize the current knowledge on how intrahepatocyte signaling pathways and innate immune cells act after the onset of HBV infection and how these actions are related to the persistence of HBV. We anticipate the insights presented herein to be helpful for future development of novel immune therapeutic strategies to fight HBV infection.

Role of relevant immune-modulators and cytokines in hepatocellular carcinoma and premalignant hepatic lesions

AIM To assess the levels of different immune modulators in patients with hepatocellular carcinoma(HCC),in relation to other hepatic diseases.METHODS Eighty-eight patients were included in the current study and represented patients with HCC(20),liver cirrhosis(28) and chronic hepatitis(CH;25),and normal controls(NC;15).Peripheral blood was isolated for immunophenotyping of active myeloid dendritic cells(m DCs;CD1 c and CD40),mature inactive myeloid cells(CD1 c and HLA),active plasmacytoid cells(p DCs;CD303 and CD40),mature inactive p DCs(CD30 and HLA),active natural killer(NK) cells(CD56 and CD161),active NK cells(CD56 and CD314) and inactive NK cells(CD56 and CD158) was done by flow cytometry.Serum levels of interleukin(IL)-2,IL-10,IL-12,IL-1β,interferon(IFN)-α,IFN-γ and tumor necrosis factor(TNF)-αR2 were assessed by ELISA.RESULTS Active m DCs(CD1 C+/CD40+) and inactive m DCs(CD1 c+/HLA+) were significantly decreased in HCC patients in relation to NC(P < 0.001).CD40+ expression on active p DCs was decreased in HCC patients(P < 0.001),and its level was not significantly changed among other groups.Inactive p DCs(CD303+/HLA+),inactive NKs(CD56+/CD158+) and active NKs(CD56+/CD161+) were not statistically changed among the four groups studied;however,the latter was increased in CH(P < 0.05).NKG2 D was statistically decreased in HCC,CH and cirrhosis(P < 0.001),and it was not expressed in 63%(12/20) of HCC patients.There was significant decrease of IL-2,IFN-α and IFN-γ(P < 0.001),and a significant increase in IL-10,IL-1β,and TNF-αR2(P <0.01,P < 0.001 and P < 0.001;respectively) in HCC patients.There was inverted correlation between IL-12 and IL-1β in HCC(r =-0.565,P < 0.01),with a strong correlation between p DCs(CD303+/CD40+) and NKs(CD56+/CD161+;r = 0.512,P < 0.05) as well as inactive m DCs(CD1 c+/HLA+) and inactive NK cells(CD56+/CD158+;r = 0.945,P < 0.001).CONCLUSION NKG2 D,CD40,IL-2 and IL-10 are important modulators in the development and progression of HCC.

Multiple sclerosis and the role of immune cells

Multiple sclerosis(MS) is a complex disease with many different immune cells involved in its pathogenesis, and in particular T cells as the most recognized cell type. Recently, the innate immune system has also been researched for its effect on the disease. Hence, cells of the immune system play vital roles in either ameliorating or exacerbating the disease. The genetic and environmental factors, as well as the etiology and pathogenesis are of utmost importance for the development of MS. An insight into the roles play by T cells, B cells, natural killer cells, and dendritic cells in MS and the animal model experimental autoimmune encephalomyelitis, will be presented. Understanding the mechanisms of action for current therapeutic modalities should help developing new therapeutic tools to treat this disease and other autoimmune diseases.

Evaluation of antibody-dependent cell-mediatedcy totoxicity activity and cetuximab response in KRAS wildtype metastatic colorectal cancer patients

AIM:To investigate the prognostic role of invariant natural killer T(iNKT) cells and antibody-dependent cell-mediated cytotoxicity(ADCC) in wild type KRAS metastatic colorectal cancer(mC RC) patients treated with cetuximab.METHODS: Forty-one KRAS wt mC RC patients,treated with cetuximab and irinotecan-based chemotherapy in Ⅱ and Ⅲ lines were analyzed. Genotyping of single nucleotide polymorphism(SNP)s in the FCGR2A,FCGR3A and in the 3' untranslated regions of KRAS and mutational analysis for KRAS,BRAF and NRAS genes was determined either by sequencing or allelic discrimination assays. Enriched NK cells were obtained from lymphoprepperipheral blood mononuclear cell and iN KT cells were defined by co-expression of CD3,TCRVα24,TCRVβ11. ADCC was evaluated as ex vivo NK-dependent activity,measuring lactate dehydrogenase release.RESULTS: At basal,mCRC patients performing ADCC activity above the median level(71%) showed an improved overall survival(OS) compared to patients with ADCC below(median 16 vs 8 mo;P=0.026). We did not find any significant correlation of iN KT cells with OS(P=0.19),albeit we observed a trend to a longer survival after 10 mo in patients with iN KT above median basal level(0.382 cells/microliter). Correlation of OS and progression-free survival(PFS) with interesting SNPs involved in ADCC ability revealed not to be significant. Patients carrying alleles both with A in FCGR2 A and TT in FCGR3A presented a trend of longer PFS(median 9 vs 5 mo;P=0.064). Chemotherapy impacted both iN KT cells and ADCC activity. Their prognostic values get lost when we analysed them after 2 and 4 mo of treatment.CONCLUSION: Our results suggest a link between iN KT cells,basal ADCC activity,genotypes in FCGR2A and FCGR3A,and efficacy of cetuximab in KRAS wt mC RC patients.

Engineered human pluripotent stem cell-derived natural killer cells with PD-L1 responsive immunological memory for enhanced immunotherapeutic efficacy

Adoptive chimeric antigen receptor(CAR)-engineered natural killer(NK)cells have shown promise in treating various cancers.However,limited immunological memory and access to sufficient numbers of allogenic donor cells have hindered their broader preclinical and clinical applications.Here,we first assess eight different CAR constructs that use an anti-PD-L1 nanobody and/or universal anti-fluorescein(FITC)single-chain variable fragment(scFv)to enhance antigen-specific proliferation and anti-tumor cytotoxicity of NK-92 cells against heterogenous solid tumors.We next genetically engineer human pluripotent stem cells(hPSCs)with optimized CARs and differentiate them into functional dual CAR-NK cells.The tumor microenvironment responsive anti-PD-L1 CAR effectively promoted hPSC-NK cell proliferation and cytotoxicity through antigen-dependent activation of phosphorylated STAT3(pSTAT3)and pSTAT5 signaling pathways via an intracellular truncated IL-2 receptorβ-chain(ΔIL-2Rβ)and STAT3-binding tyrosine-X-X-glutamine(YXXQ)motif.Anti-tumor activities of PD-L1-induced memory-like hPSC-NK cells were further boosted by administering a FITC-folate bi-specific adapter that bridges between a programmable anti-FITC CAR and folate receptor alpha-expressing breast tumor cells.Collectively,our hPSC CAR-NK engineering platform is modular and could constitute a realistic strategy to manufacture off-the-shelf CAR-NK cells with immunological memory-like phenotype for targeted immunotherapy.

Peripheral blood NKT cell number and function in patients with Graves disease and their correlation with hyperthyroidism

Objective:To assess the peripheral blood NKT cell number and function in patients with Graves disease and study their correlation with hyperthyroidism.Methods: The patients who were diagnosed with Graves disease in our hospital between May 2014 and September 2016 were selected as GD group, and 55 healthy volunteers who received physical examination in our hospital during the same period were selected as control group. Peripheral blood was collected to determine the number of CD3+CD56+NKT cells, and serum was collected to detect the contents of cytokines and thyroid function indexes.Results: Peripheral blood CD3+CD56+NKT cell number in Graves disease group was significantly lower than that in control group;serum IL-2, IFN-γ, TNF-α, IL-10 and TGF-β contents in GD group were significantly lower than those in control group and positively correlated with peripheral blood CD3+CD56+NKT cell number while IL-4, IL-5, IL-17, FT3, FT4, TPOAb, TgAb and TRAb contents were significantly higher than those in control group and negatively correlated with peripheral blood CD3+CD56+NKT cell number.Conclusion:The abnormal reduction of peripheral blood NKT cell number in patients with Graves disease can affect the balance of Th1/Th2 and Th17/Treg and increase the TRAb secretion to cause hyperthyroidism.

ST2、NK细胞和B细胞在T2DM患者中的水平及相关性分析

目的探讨可溶性生长刺激表达基因2蛋白(sST2)、自然杀伤(NK)细胞与B细胞在2型糖尿病(T2DM)患者中的水平及相关性。方法选取2022年10-12月在南京医科大学附属江宁医院住院治疗的50例T2DM患者作为T2DM组,另选取同期该院30例健康体检者作为对照组。采用流式细胞术检测NK细胞(CD45^(+)CD3^(-)CD19-CD56^(+))和B细胞(CD45^(+)CD3^(-)CD56-CD19^(+))比例;采用流式荧光法检测血清IL-6、TNF-α、IFN-γ的水平;采用免疫荧光干式定量法检测所有研究对象血清sST2水平;分析在T2DM患者中血清sST2水平与NK细胞、B细胞的相关性以及在T2DM患者中B细胞与NK细胞的相关性。结果T2DM组患者血清sST2水平高于对照组,B细胞及NK细胞的比例均明显低于对照组,炎症因子IL-6、TNF-α、IFN-γ水平高于对照组,差异均有统计学意义(P<0.05)。T2DM患者血清sST2水平与B细胞比例(r=-0.40,P<0.01)、NK细胞比例(r=-0.29,P<0.05)呈负相关;T2DM患者B细胞比例与NK细胞比例呈正相关(r=0.34,P<0.05)。结论T2DM患者sST2水平、NK细胞与B细胞具有相关性。

莫诺苯宗对肺腺癌细胞自噬和凋亡的分子机制研究

目的:探究莫诺苯宗和自然杀伤细胞激活受体基因(KLRC3)对肺腺癌(LUAD)细胞凋亡和自噬的影响,为LUAD的治疗提供理论基础。方法:分别使用不同浓度莫诺苯宗处理LUAD细胞,并采用CCK-8法检测莫诺苯宗对LUAD细胞活力的影响;随后,使用80μmol/L的莫诺苯宗、50 nmol/L的KLRC3干扰质粒(si-KLRC3)、和100 nmol/L自噬抑制剂3-甲基腺嘌呤(3-MA)1.5μg/ml KLRC3过表达载体(pcDNA-KLRC3)处理细胞后采用流式细胞术和Western blotting检测细胞凋亡以及凋亡相关蛋白和自噬相关蛋白的表达量。结果:莫诺苯宗浓度大于20μmol/L时可有效减弱LUAD细胞活力,且以剂量依赖性地促进了LUAD细胞的凋亡和自噬(均P<0.05)。KLRC3在LUAD细胞中低表达,干扰KLRC3减弱了莫诺苯宗对LUAD细胞自噬和凋亡的促进作用,比较差异有统计学意义(均P<0.05)。此外,过表达KLRC3也促进了LUAD细胞的凋亡和自噬,而KLRC3和莫诺苯宗对LUAD细胞凋亡和自噬的促进作用均可被3-MA所抵消,比较差异有统计学意义(均P<0.05)。结论:莫诺苯宗可能通过促进KLRC3的表达诱导自噬进而促进LUAD细胞的凋亡。

肝细胞癌过继细胞疗法研究进展

肝细胞癌是严重威胁人类生存的一类疾病,中国肝细胞癌死亡率居世界第三。由于肝细胞癌发病早期无特异性症状,晚期的治疗方案又存在复发率高等的种种弊端,因此寻找行之有效的治疗新方法迫在眉睫。过继细胞疗法是一种细胞免疫疗法,在肝脏多样化和复杂的免疫微环境中,通过结合肿瘤特异性抗原或是非特异性的调节免疫平衡,过继细胞疗法在肝细胞癌的治疗中逐渐发挥出优势。在这篇综述中我们阐述了肝细胞癌靶向治疗常用生物标志物的形态结构及信号通路,探讨总结了近年来过继细胞疗法治疗肝细胞癌的研究手段及取得的临床效果,并对接下来的研究方向进行了展望。