Imbalance of Circulating Follicular Regulatory and Follicular Helper T Cell Subpopulations Is Associated with Disease Progression and Serum CYFRA 21-1 Levels in Patients with Non-small Cell Lung Cancer

Objective This study aimed to investigate the changes of follicular helper T(TFH)and follicular regulatory T(TFR)cell subpopulations in patients with non-small cell lung cancer(NSCLC)and their significance.Methods Peripheral blood was collected from 58 NSCLC patients at different stages and 38 healthy controls.Flow cytometry was used to detect TFH cell subpopulation based on programmed death 1(PD-1)and inducible co-stimulator(ICOS),and TFR cell subpopulation based on cluster determinant 45RA(CD45RA)and forkhead box protein P3(FoxP3).The levels of interleukin-10(IL-10),interleukin-17a(IL-17a),interleukin-21(IL-21),and transforming growth factor-β(TGF-β)in the plasma were measured,and changes in circulating B cell subsets and plasma IgG levels were also analyzed.The correlation between serum cytokeratin fragment antigen 21-1(CYFRA 21-1)levels and TFH,TFR,or B cell subpopulations was further explored.Results The TFR/TFH ratio increased significantly in NSCLC patients.The CD45RA^(+)FoxP3^(int) TFR subsets were increased,with their proportions increasing in stages Ⅱ to Ⅲ and decreasing in stage IV.PD-1^(+)ICOS+TFH cells showed a downward trend with increasing stages.Plasma IL-21 and TGF-β concentrations were increased in NSCLC patients compared with healthy controls.Plasmablasts,plasma IgG levels,and CD45RA^(+)FoxP3^(int) TFR cells showed similar trends.TFH numbers and plasmablasts were positively correlated with CYFRA 21-1 in stages Ⅰ-Ⅲ and negatively correlated with CYFRA 21-1 in stage IV.Conclusion Circulating TFH and TFR cell subpopulations and plasmablasts dynamically change in different stages of NSCLC,which is associated with serum CYFRA 21-1 levels and reflects disease progression.

Analysis of the Peripheral Blood Helper T-Cell 17- Cell Level and Monocyte/Lymphocyte Ratio for Colorectal Cancer Prognosis Prediction

Objective: To investigate the value of peripheral blood helper T cell 17 cell level and monocyte/lymphocyte ratio to predict the prognosis of colorectal cancer patients. Methods: 74 colorectal cancer patients who attended Hospital 960 from January 2021 to January 2022 were retrospectively analyzed. Clinical data of the patients were collected, including gender, age, and histologic type. Immunohistochemical indexes such as Th17 cell level and monocyte/ lymphocyte ratio in the peripheral blood of patients were also collected. The prognosis of patients after treatment, as well as peripheral blood Th17 and MLR levels, were observed and analyzed. Results: After follow-up after treatment, in the final 74 patients, the prognosis was good in 32 patients, accounting for 43.24%, and the prognosis was bad in 42 patients, accounting for 56.76%. There were no significant differences between the average age and tumor diameters of the good prognosis and poor prognosis groups (P > 0.05). However, the TNM staging, intervention taken, differentiation degree, presence of distant metastasis, presence of lymph node metastasis, Th17 level, and MLR level are significantly different between the two groups (P < 0.05). Conclusion: Peripheral blood Th17 and MLR have predictive value for the prognosis of colorectal cancer patients, and high levels of peripheral blood Th17 and MLR imply poor prognosis. The detection of peripheral blood Th17 and MLR levels is simple and convenient and can be used as indicators to provide a reference for the prognostic assessment of colorectal cancer patients.

Impaired balance of T helper 17/T regulatory cells in carbon tetrachloride-induced liver fibrosis in mice

AIM: To investigate the effect of T helper (Th) 17/T regulatory (Treg) cells on hepatic fibrosis in mice and its possible mechanism.

Ribavirin contributes to eradicate hepatitis C virus through polarization of T helper 1/2 cell balance into T helper 1 dominance

The mechanism of action of ribavirin(RBV) as an immunomodulatory and antiviral agent and its clinical significance in the future treatment of patients with hepatitis C virus(HCV) infection are reviewed.RBV up-regulates type 1 and/or 2 cytokines to modulate the T helper(Th) 1/2 cell balance to Th1 dominance.Examination of co-stimulatory signaling indicated that RBV down-modulates inducible co-stimulator on Th cells,which contributes to differentiating na?ve Th cells into Th2 cells while reducing their interleukin-10 production.The effects on T-regulatory(Treg) cells were also investigated,and RBV inhibited the differentiation of na?ve Th cells into adaptive Treg cells by downmodulating forkhead box-P3.These findings indicate that RBV mainly down-regulates the activity of Th2 cells,resulting in the maintenance of Th1 activity that contributes to abrogating HCV-infected hepatocytes.Although an interferon-free treatment regimen exhibits almost the same efficacy without serious complications,regimens with RBV will be still be used because of their ability to facilitate the cellular immune response,which may contribute to reducing the development of hepatocellular carcinogenesis in patients infected with HCV.

Probiotic Mixture VSL#3 Alleviates Dextran Sulfate Sodium-induced Colitis in Mice by Downregulating T Follicular Helper Cells

Clinical trials have shown beneficial effects of probiotics on inflammatory bowel diseases (IBD), although the exact mechanism remains unknown. VSL#3, a mixture of 8 probiotic bacteria, has been confirmed to have adjunctive therapeutic effects on colitis. T follicular helper (Tfh) cells, a new separate subset of CD4+ T helper cells, have been proved to play a vital role in autoimmunity. The present study aimed to identify the beneficial effect of the probiotic mixture VSL#3 on the mouse model of colitis by regulating Tfh cells. Dextran sulfate sodium (DSS) was used to induce chronic colitis in C57BL/6 mice. VSL#3 (3x109 live bacteria) was given to C57BL/6 mice every other day for 60 days by gavage. The disease activity index (DAI), histological activity index (HAI), colon length and myeloperoxidase (MPO) activity were detected. Immunofluorescence was used to visualize the location of Tfh cells. Immunoglobulins, Tfh cells and plasma cells were quantified by enzyme-linked immunosorbent assay (ELISA), flow cytometry, real-time PCR or Western blotting. The results showed that after DSS treatment, the humoral immunity was disordered in C57BL/6 mice, with increased IgM, IgG and IgA levels in colonic mucus and increased Tfh cells in mesenteric lymph nodes (MLN). VSL#3 treatment showed anti-inflammatory effects as evidenced by reduced DAI score, HAI score and MPO activity. IgM, IgG and IgA levels were significantly reduced in colon mucus, and the number of Tfh cells was markedly decreased in MLN after VSL#3 treatment. It was concluded that VSL#3 alleviates DSS-induced colitis by downregulating Tfh cells, and Tfh cells may become a potential therapeutic target for IBD.

Immunomodulation of adipose-derived mesenchymal stem cells on peripheral blood mononuclear cells in colorectal cancer patients with COVID-19

BACKGROUND Accumulating evidence has shown that adipose tissue-derived mesenchymal stem cells(ADSCs)are an effective therapeutic approach for managing coronavirus disease 2019(COVID-19);however,further elucidation is required to determine their underlying immunomodulatory effect on the mRNA expression of T helper cell-related transcription factors(TFs)and cytokine release in peripheral blood mononuclear cells(PBMCs).AIM To investigate the impact of ADSCs on the mRNA expression of TFs and cytokine release in PBMCs from colorectal cancer(CRC)patients with severe COVID-19(CRC^(+)patients).METHODS PBMCs from CRC^(+)patients(PBMCs-C+)and age-matched CRC patients(PBMCs-C)were stimulated and cultured in the presence/absence of ADSCs.The mRNA levels of T-box TF TBX21(T-bet),GATA binding protein 3(GATA-3),RAR-related orphan receptor C(RORC),and forkhead box P3(FoxP3)in the PBMCs were determined by reverse transcriptase-polymerase chain reaction.Culture supernatants were evaluated for levels of interferon gamma(IFN-γ),interleukin 4(IL-4),IL-17A,and transforming growth factor beta 1(TGF-β1)using an enzyme-linked immunosorbent assay.RESULTS Compared with PBMCs-C,PBMCs-C+exhibited higher mRNA levels of T-bet and RORC,and increased levels of IFN-γ and IL-17A.Additionally,a significant decrease in FoxP3 mRNA and TGF-β1,as well as an increase in Tbet/GATA-3,RORC/FoxP3,IFN-γ/IL-4,and IL-17A/TGF-β1 ratios were observed in PBMCs-C+.Furthermore,ADSCs significantly induced a functional regulatory T cell(Treg)subset,as evidenced by an increase in FoxP3 mRNA and TGF-β1 release levels.This was accompanied by a significant decrease in the mRNA levels of T-bet and RORC,release of IFN-γ and IL-17A,and T-bet/GATA-3,RORC/FoxP3,IFN-γ/IL-4,and IL-17A/TGF-β1 ratios,compared with the PBMCs-C+alone.CONCLUSION The present in vitro studies showed that ADSCs contributed to the immunosuppressive effects on PBMCs-C+,favoring Treg responses.Thus,ADSC-based cell therapy could be a beneficial approach for patients with severe COVID-19 who fail to respond to conventional therapies.

T helper cell dysregulation with hepatitis B and rebalance with glucocorticoids

AIM: To investigate T helper 17/regulatory T cell alterations in early severe hepatitis B and the effect of glucocorticoids.

Upregulated adenosine 2A receptor accelerates post-infectious irritable bowel syndrome by promoting CD4+T cells’T helper 17 polarization

BACKGROUND Post-infectious irritable bowel syndrome(PI-IBS)is generally regarded as a functional disease.Several recent studies have reported the involvement of lowgrade inflammation and immunological dysfunction in PI-IBS.T helper 17(Th17)polarization occurs in IBS.Adenosine and its receptors participate in intestinal inflammation and immune regulation.AIM To investigate the role of Th17 polarization of CD4+T cells regulated by adenosine 2A receptor(A2AR)in PI-IBS.METHODS A PI-IBS model was established by infecting mice with Trichinella spiralis.The intestinal A2AR and CD4+T lymphocytes were detected by immunohistochemistry,and the inflammatory cytokines were detected by enzyme-linked immunoassay.CD4+T lymphocytes present in the animal’s spleen were separated and cultured with or without A2AR agonist and antagonist.Western blotting and real-time quantitative polymerase chain reaction were performed to determine the effect of A2AR on the cells and intestinal tissue.Cytokine production was determined.The protein and mRNA levels of A2AR associated signaling pathway molecules were also evaluated.Furthermore,A2AR agonist and antagonist were injected into the mouse model and the clinical features were observed.RESULTS The PI-IBS mouse model showed increased expression of ATP and A2AR(P<0.05),and inhibition of A2AR improved the clinical features in PI-IBS,including the abdominal withdrawal reflex and colon transportation test(P<0.05).The number of intestinal CD4+T cells and interleukin-17(IL-17)protein levels increased during PI-IBS,which was reversed by administration of the A2AR antagonist(P<0.05).CD4+T cells expressed A2AR and produced IL-17 in vitro,which was regulated by the A2AR agonist and antagonist.The A2AR antagonist increased the production of IL-17 by CD4+T cells via the Janus kinase-signal transducer and activator of transcriptionreceptor-related orphan receptorγsignaling pathway.CONCLUSION The results of the present study suggested that the upregulation of A2AR increases PI-IBS by promoting the Th17 polarization of CD4+T cells.

Effect of pretreatment with different concertrations of morphine, fentanyl and tramadol on the differentiation of human helper T cells in vitro

Objective： To investigate and compare the .effects of different concentrations of morphine, fentanyl and tramadol on the differentiation of human adult helper T cells in vitro. Methods： Twenty out-patients without immune disease were selected and their peripheral blood was collected. Then the Whole blood of peripheral blood mononuclear cells （PBMCs） were pretreated with different concentration of morphine, fentanyl and tramadol for 24 h. The level of CD4^＋ IFN-γ^＋ IL-2^＋/CD4^＋ IL-4^＋ IL-10^＋ was analyzed by three-color flow cytometry, and the CD4^＋ CCR5^＋ and CD4^＋ CCR3 ^＋ cells were counted to observe the imbalance of Th2/Th2. Results： The number of Th2 increased significantly and the ratio of Th2/Th2 decreased dramatically compared with the control group, and there was a dose-dependent fashion in all drugs. Conclusion： Morphine, fentanyl and tramadol can direct Th0 cells toward Th2 differentiation, especially morphine and fentanyl.

Involvement of glycated albumin in adipose-derived-stem cellmediated interleukin 17 secreting T helper cell activation

BACKGROUND Advanced glycation end products(AGE)are a marker of various diseases including diabetes,in which they participate to vascular damages such as retinopathy,nephropathy and coronaropathy.Besides those vascular complications,AGE are involved in altered metabolism in many tissues,including adipose tissue(AT)where they contribute to reduced glucose uptake and attenuation of insulin sensitivity.AGE are known to contribute to type 1 diabetes(T1D)through promotion of interleukin(IL)-17 secreting T helper(Th17)cells.AIM To investigate whether lean adipose-derived stem cells(ASC)could be able to induce IL-17A secretion,with the help of AGE.METHODS As we have recently demonstrated that ASC are involved in Th17 cell promotion when they are harvested from obese AT,we used the same co-culture model to measure the impact of glycated human serum albumin(G-HSA)on human lean ASC interacting with blood mononuclear cells.IL-17A and pro-inflammatory cytokine secretion were measured by ELISA.Receptor of AGE(RAGE)together with intercellular adhesion molecule 1(ICAM-1),human leukocyte Antigen(HLA)-DR,cluster of differentiation(CD)41,and CD62P surface expressions were measured by cytofluorometry.Anti-RAGE specific monoclonal antibody was added to co-cultures in order to evaluate the role of RAGE in IL-17A production.RESULTS Results showed that whereas 1%G-HSA only weakly potentiated the production of IL-17A by T cells interacting with ASC harvested from obese subjects,it markedly increased IL-17A,but also interferon gamma and tumor necrosis factor alpha production in the presence of ASC harvested from lean individuals.This was associated with increased expression of RAGE and HLA-DR molecule by cocultured cells.Moreover,RAGE blockade experiments demonstrated RAGE specific involvement in lean ASC-mediated Th-17 cell activation.Finally,platelet aggregation and ICAM-1,which are known to be induced by AGE,were not involved in these processes.CONCLUSION Thus,our results demonstrated that G-HSA potentiated lean ASC-mediated IL-17A production in AT,suggesting a new mechanism by which AGE could contribute to T1D pathophysiology.

Dynamic interplay of T helpercell subsets in experimental autoimmune encephalomyelitis

AIM： To investigate the temporal onset and dynamic interplay of CD4^＋ T helper cell subsets in experimental autoimmune encephalomyelitis （EAE）.METHODS： EAE was induced in C57BL/6 mice by im-munization with myelin oligodendrocyte glycoprotein peptide p35-55. The clinical signs were scored and the tissue samples and immune cells isolated for analysis at different phases of EAE. The expression levels of inflammatory cytokines and related transcription fac-tors were detected by quantitative reverse transcription polymerase chain reaction （PCR） and enzyme linked immunosorbant assay （ELISA）. The percentages of Th1, Th17, Th2, Treg and memory T cell subsets in EAE were analyzed by immunostaining and fow cytometry. The data were analyzed by statistical techniques.RESULTS： Quantitative real-time PCR analysis showed that EAE mice express elevated levels of Th1 [interferon gamma （ IFNγ ）, interleukin （ IL ） -12p40 ], Th17 [ IL-17 , related orphan receptor gamma （RORγ ）, IL-12p40] and Treg [ Foxp3, Epstein-Barr virus induced gene 3 （EBI3）, IL-10] genes in the central nervous system at the peak of the disease. Whereas, the expression of Th1 （ IFNγ , T-bet, IL-12p35, IL-12p40 ）, Th17 （RORγ, IL-12p40 ）, Th2 （ IL-4） and Treg （ Foxp3, EBI3） response genes was reduced in the spleen during pre-disease but gradually recovered at the later phases of EAE. ELISA and fow cytometry analyses showed an increase in Th17 re-sponse in the periphery, while Th1 response remained unchanged at the peak of disease. The mRNA levels of IFNγ, IL-17 and IL-12p40 in the brain were increased by 23 （P 〈 0.001）, 9 （P 〈 0.05） and 14 （P 〈 0.01） fold, respectively, on day 21 of EAE. Conversely, the mRNA expression of IL-10 was increased by 2 fold （P 〈 0.05） in the spleen on day 21. CD4^＋CD25^＋Foxp3＋Treg response was reduced at pre-disease but recovered to na？ve levels by disease onset. The percentage of CD25 Foxp3 regulatory T cells decreased from 7.7% in the na？ve to 3.2% （P 〈 0.05） on day 7 of EAE, which then increased to 8.4% by day 28. Moreover, the CD4＋CD127＋CD44high memory T cell response was increased during the onset and recovery phases of EAE. The memory and effector cells showed an in-verse relationship in EAE, where the memory T cells increased from 12.3% in nave to 20% by day 21, and the effector cells decreased from 32% in na？ve to 21% （P 〈 0.01） by day 21. The wild type C57BL/6 mice with EAE showed elevated levels of effector-memory T cells （TEM） with concomitant reduction in central-memory T cells （TCM）, but the EAE-resistant IL-7R defcient mice showed elevated TCM with no effect on TEM cells in EAE.CONCLUSION： Our fndings highlight the temporal on-set and dynamic interplay of effector, memory and regu-latory CD4^＋ T cell subsets and its signifcance to clinical outcome in EAE and other autoimmune diseases.

Function of Helper T Cells in the Memory CTL-mediated Anti-tumor Immunity

To investigate the role of CD4 + helper T (Th) cells in the memory CTL-mediated anti-tumor immunity, the RAG-1 gene knock out mice were adoptively transferred with OT-1 cells to generate the memory CTL, the C57BL/6 mice immunized with the epitope peptide of OVA specific Th cells and with different adjuvants were adoptively transferred with these memory-CTLs, and then the animals were challenged with tumor cells EG7. It was found that although the simple immunization of mice with the epitope peptide of the OVA specific Th cells could generate more effect CTL, but this effect was not so strong enough to resist completely the challenges with tumor cells. Nevertheless, the memory CTL-mediated anti-tumor immune effect required the helps of Th1 and Th2 cells. The cross-regulation between Th1 and Th2 cells seemed to be beneficial for the host to generate more effector CTL for mounting an efficient anti-tumor response. It concluded that the interaction between Th1 and Th2 cells might be more important than the single subset of Th cells in the memory CTL-mediated anti-tumor immune response. More attention should be paid in this regard for the future studies.

Expression of Helper T Cell Type 17 and CD4^(+)CD25^(+)Tregs in AMA-M2 Positive PBC Patients

Objective:To investigate the expression and impact of helper T cell type 17 and CD4^(+)CD25^(+)regulatory T(Treg)cells in anti-mitochondrial M2 antibody(AMA-M2)positive primary biliary cholangitis(PBC)patients.Methods:Thirty PBC patients with positive AMA(M2 type)(antibody titer above 1:320)by indirect immunofluorescence assay under the Affiliated Hospital of Hebei University from November 2021 to August 2022 were selected as the experimental group,while 30 healthy individuals were selected as controls.The subjects were observed and analyzed for AFP-L3 and immunoglobulin expression.Results:The levels of Th17,Treg,Th17/Treg,interleukin(IL)-17A,IL-2,IL-10,and transforming growth factor(TGF)-β1 cytokines of the experimental group were 2.61±0.48,1.15±0.54,2.41±0.47,310.94±21.14,276.36±36.12,317.89±28.97,and 197.48±31.04,respectively,while those of the control group were 1.14±0.58,0.88±0.29,1.47±0.25,9.69±1.26,57.69±2.45,154.01±19.87,and 514.36±36.12,respectively,wherein P<0.05;the CD4^(+),CD8^(+),and CD4^(+)/CD8^(+)of the experimental group were 39.48±4.19,20.12±4.41,and 1.76±0.14,respectively,while those of the control group were 35.78±4.21,22.01±4.16,and 1.51±0.13,respectively,wherein P<0.05.Conclusion:In patients with PBC,there is a significant imbalance in Th17/Treg cells.Il-17A,IL-2,IL-10,and TGF-β1 cytokines play important roles in the differentiation and functional expression of both Th17 and Treg cells.

益胃解毒方治疗慢性萎缩性胃炎的效果及对Cyclin E、Th17/Treg比值的影响分析

目的分析益胃解毒方治疗慢性萎缩性胃炎中的效果及对细胞周期蛋白E(Cyclin E)水平、辅助性T细胞(Th17)/调节性T细胞(Treg)比值的影响。方法选取2019年6月至2022年6月在河北省石家庄市中医院确诊的150例慢性萎缩性胃炎患者作为研究对象,按照随机数字表法分为益胃解毒方组、六君子汤组、益胃解毒方+六君子汤组,每组50例。益胃解毒方组使用益胃解毒方治疗,六君子汤组使用六君子汤治疗,益胃解毒方+六君子汤组使用益胃解毒方联合六君子汤治疗,均连续治疗24周。比较3组治疗效果、中医证候积分、Cyclin E水平、Th17/Treg比值、不良反应发生情况。结果益胃解毒方组治疗总有效率高于六君子汤组和益胃解毒方+六君子汤组,且益胃解毒方+六君子汤组高于六君子汤组,差异均有统计学意义(P<0.05)。治疗后3组各项中医证候积分低于治疗前,差异均有统计学意义(P<0.05)。治疗后益胃解毒方组各项中医证候积分低于六君子汤组和益胃解毒方+六君子汤组,且益胃解毒方+六君子汤组低于六君子汤组,差异均有统计学意义(P<0.05)。治疗后3组Cyclin E水平、Th17/Treg比值均低于治疗前,差异均有统计学意义(P<0.05)。治疗后益胃解毒方组Cyclin E水平、Th17/Treg比值低于六君子汤组和益胃解毒方+六君子汤组,且益胃解毒方+六君子汤组低于六君子汤组,差异均有统计学意义(P<0.05)。益胃解毒方组总不良反应发生率低于六君子汤组和益胃解毒方+六君子汤组,且益胃解毒方+六君子汤组低于六君子汤组,差异均有统计学意义(P<0.05)。结论益胃解毒方治疗慢性萎缩性胃炎效果显著,不仅能够改善患者临床症状,调节Cyclin E、Th17/Treg表达,还能减少不良反应发生,值得临床推广应用。

帕金森病患者免疫球蛋白、Th9亚群水平变化及其与IGF-1、S-100B蛋白的相关性

目的:研究帕金森病(PD)患者免疫球蛋白(IgG、IgA和IgM)、辅助性T细胞亚群Th9水平变化及其与胰岛素生长因子-1(IGF-1)、S-100B蛋白的相关性。方法:选取2020年12月至2022年12月期间在河北省中医院确诊的108例PD患者,将其作为研究组,并根据患者病变程度分为轻度组(35例)、中度组(44例)和重度组(29例);另选108例健康成人作为对照组。对比研究组和对照组免疫球蛋白和Th9亚群水平,以及轻度组、中度组及重度组免疫球蛋白、Th9亚群、IGF-1和S-100B蛋白水平,采用Pearson相关性分析免疫球蛋白、Th9亚群和IGF-1、S-100B蛋白的相关性。采用Spearman相关性分析PD患者疾病程度和所有差异指标间的相关性。结果:研究组IgM水平较对照组低,且重度组低于中度组,中度组低于轻度组(P<0.05);研究组IgG、IgA、IL-9和Th9亚群水平较对照组高,且重度组高于中度组,中度组高于轻度组(P<0.05)。重度组IGF-1水平低于中度组,中度组低于轻度组;重度组S-100B蛋白水平高于中度组,中度组高于轻度组(P<0.05)。Pearson相关性分析结果显示,PD患者IgM水平与IGF-1水平呈正相关,与S-100B蛋白水平呈负相关;IgG、IgA、IL-9和Th9亚群水平均与IGF-1水平呈负相关,与S-100B蛋白水平呈正相关(P<0.05)。Spearman相关性分析结果显示,PD患者疾病程度与IgM、IGF-1水平呈负相关,与S-100B蛋白、IgG、IgA、IL-9和Th9亚群水平呈正相关(P<0.05)。结论:PD患者IgM水平降低,IgG、IgA、Th9亚群水平升高,且其水平变化与IGF-1、S-100B明显相关,可以用于评估病情的严重程度。

结直肠癌组织高表达IL-35对Th1和Th17可塑性的作用

目的 探讨结直肠癌组织中高表达的抑制性细胞因子白细胞介素(IL)-35对辅助性T细胞1(Th1)和辅助性T细胞17(Th17)可塑性的作用。方法 选取2019年3月—2020年2月上海交通大学医学院附属新华医院结直肠癌患者90例(结直肠癌组)和健康体检者28名(正常对照组)。采用流式细胞术检测结直肠癌组和正常对照组外周血Th1百分比(Th1%)和Th17百分比(Th17%)。基于Kaplan-Meier Plotter数据库分析γ-干扰素(IFN-γ)和IL-17A高表达对结直肠癌患者总生存期和预后不良的影响。采用免疫组化法检测结直肠癌患者癌组织和癌旁组织(距肿瘤边缘≥2 cm)中IL-35的表达。采用酶联免疫吸附试验检测结直肠癌患者和正常对照者血清IL-35水平。采用体外诱导Th1分化实验观察IL-35对Th1分泌IL-17A和IFN-γ的影响。结果与正常对照组比较,结直肠癌组Th1%显著降低(P=0.019),Th17%显著升高(P<0.001)。结直肠癌组Th1%与Th17%的r2值(0.393 4)与正常对照组(0.041 0)比较差异有统计学意义(P=0.007)。基于KaplanMeierPlotter数据库的分析结果显示,IFN-γmRNA低表达是结直肠癌患者总生存期缩短的危险因素[风险比(HR)=0.74,95%可信区间(CI)为0.58~0.94,P=0.013],IL-17A mRNA高表达是结直肠癌患者总生存期缩短的危险因素(HR=1.26,95%CI为1.02~1.59,P=0.020)。结直肠癌患者癌组织IL-35表达显著高于癌旁组织(P<0.001)。Ⅰ~Ⅱ期(早期)和Ⅲ~Ⅳ期(晚期)结直肠癌患者血清IL-35水平均显著高于正常对照者(P<0.05)。体外诱导Th1分化实验结果显示,IL-35可降低Th1中IFN-γ表达,提高IL-17A表达。结论 结直肠癌患者外周血Th1和Th17呈异常变化,且IL-35表达增加。高表达的IL-35可降低Th1中IFN-γ表达,提高IL-17A表达。

Th17和调节性T细胞在人类免疫缺陷病毒疾病进展中的作用及其调控机制

目的本研究通过观察人类免疫缺陷病毒(HIV)感染者外周血Th17、Treg细胞及其相关转录因子、细胞因子的表达,进一步揭示Th17、Treg细胞的调控机制及其在HIV感染者疾病进展中的作用。方法选取2019年1月—2019年10月河南省上蔡县HIV感染者80例作为研究组,选取同地区健康人群20例作为对照组。采用流式细胞术检测外周血CD4^(+)T、CD8^(+)T及外周血单个核细胞Th17、Treg,酶联免疫吸附试验检测血浆中细胞因子白细胞介素-17(IL-17)、IL-23水平,实时聚合酶链反应检测转录因子ROR-γt及Foxp3 mRNA表达,Pearson法分析Th17、Treg与CD4^(+)T的相关性。结果研究组CD4^(+)T、CD4^(+)T/CD3+T、CD4^(+)T/CD8^(+)T低于对照组(P<0.05),CD8^(+)T和CD8^(+)T/CD3+T高于对照组(P<0.05)。研究组Th17/CD4^(+)T、Th17/Treg低于对照组(P<0.05),Treg/CD4^(+)T高于对照组(P<0.05)。研究组ROR-γtmRNA高于对照组(P<0.05),Foxp3mRNA低于对照组(P<0.05)。研究组IL-17、IL-23水平低于对照组(P<0.05)。Pearson相关性分析结果表示,Th17细胞百分比与CD4^(+)T细胞计数呈正相关(r=0.293,P<0.05),Treg细胞百分比与CD4^(+)T细胞计数呈负相关(r=-0.198,P<0.05)。结论HIV感染能降低Th17细胞、升高Treg细胞,破坏机体免疫平衡,其机制与调控转录因子ROR-γt、Foxp3及细胞因子IL-17、IL-23表达有关,Th17细胞、Treg细胞与HIV感染者疾病进展密切相关。

核孔蛋白160基因对肾病综合征小鼠肾脏辅助性T细胞17/调节性T细胞免疫平衡的影响

目的探讨核孔蛋白160(NUP160)基因对肾病综合征小鼠肾脏辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡的影响。方法将30只小鼠随机分为空白组(n=10)和造模组(n=20)。造模组小鼠经尾静脉注射阿霉素溶液(7.5 mg·kg-1)制备原发性肾病综合征(PNS)模型,空白组小鼠经尾静脉注射等容积的生理盐水。造模成功后将造模组小鼠随机分为PNS组和NUP160组,每组10只。NUP160组小鼠经尾静脉注射NUP160(10 mg·kg-1),每日1次;PNS组和空白组小鼠经尾静脉注射等量生理盐水,每日1次;3组小鼠均连续干预4周。给药0、2、4周时,应用生化自动检测仪检测小鼠24 h尿蛋白定量(24 h HUPQ);给药结束后,采用酶联免疫吸附试验法检测各组小鼠血清白细胞介素(IL)-17和IL-10水平;苏木精-伊红(HE)染色、过碘酸希夫(PAS)染色、Masson染色法观察各组小鼠肾组织病理学变化;流式细胞术检测各组小鼠肾组织中Th17、Treg细胞比例。结果给药0、2、4周时,PNS组和NUP160组小鼠24 h HUPQ显著高于空白组(P<0.05)。给药0周时,PNS组与NUP160组小鼠24 h HUPQ比较差异无统计学意义(P>0.05);给药2、4周时,NUP160组小鼠24 h HUPQ显著低于PNS组(P<0.05)。与空白组比较,PNS组和NUP160组小鼠血清IL-17水平显著升高,IL-10水平显著降低(P<0.05);与PNS组比较,NUP160组小鼠血清IL-17水平显著降低,IL-10水平显著升高(P<0.05)。空白组小鼠肾组织无明显病理学变化。与空白组相比,PNS组小鼠HE染色发现肾小球细胞排列紊乱,肾小球轻度萎缩,少许炎症细胞浸润,球囊壁轻度增厚,小球囊腔轻度扩张;PAS染色发现系膜出现增生,肾间质可见炎症细胞浸润;Masson染色发现小鼠肾小球基底膜和间质蓝色胶原沉积明显增多,组织纤维化程度显著升高。与PNS组相比,NUP160组小鼠肾组织以上症状均得到明显改善。与空白组比较,PNS组和NUP160组小鼠肾组织中Th17占比、Th17/Treg比值显著增加,Treg占比显著降低(P<0.05);与PNS组比较,NUP160组小鼠肾组织中Th17占比、Th17/Treg比值显著降低,Treg占比显著增加(P<0.05)。结论NUP160可调节PNS小鼠的Th17/Treg免疫平衡,进而发挥改善PNS的作用。

苦参碱调节RhoA-ROCK信号通路对冠心病模型大鼠Th17/Treg细胞平衡的影响

目的探讨苦参碱(Matrine)对冠心病(coronary heart disease,CHD)大鼠辅助T细胞17(helper T cell 17,Th17)/调节性T细胞(regulatory T cells,Treg)细胞平衡及Ras同源基因家族成员A(RhoA)-Rho相关的卷曲螺旋激酶(ROCK)信号通路的影响。方法建立冠心病模型,将实验大鼠分为对照组、模型组、苦参碱低剂量(50 mg·kg^(-1))组、苦参碱高剂量(200 mg·kg^(-1))组及苦参碱高剂量(200 mg·kg^(-1))+LPA组(10 mg·kg^(-1))。超声心动图进行大鼠心功能检测;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)法进行白细胞介素17(IL-17)、转化生长因子β(TGF-β)水平检测;流式细胞术检测Th17、Treg数量及Th17/Treg比值;免疫组化进行内皮型一氧化氮合酶(eNOS)、内皮素1(ET-1)蛋白表达水平检测;Masson染色进行大鼠心肌组织的病理形态变化观察;TTC染色检测各组大鼠心肌梗死情况;TUNEL染色进行心肌组织中细胞凋亡情况检测;试剂盒检测RhoA活性;Western Blot法进行半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、B细胞淋巴瘤因子2(Bcl-2)、Bcl-2相关X蛋白(Bax)、RhoA、ROCK1、ROCK2蛋白表达水平检测。结果与对照组比较,模型组心肌组织有大量蓝色胶原纤维沉积,左室舒张末期容积(left ventricular end-diastolic volume,LVEDV)、左室收缩末期容积(left ventricular end-systolic volume,LVESV)、IL-17、Th17、Th17/Treg、ET-1、心肌梗死面积、细胞凋亡率、TUNEL阳性率、Bax、Caspase-3、RhoA活性、RhoA、ROCK1、ROCK2表达水平明显升高,左室射血分数(left ventricular ejection fraction,LVEF)、左室缩短分数(left ventricular shortening fraction,LVFS)、TGF-β、Treg、eNOS、Bcl-2表达水平明显降低(P<0.05)。与模型组比较,Matrine-L组、苦参碱高剂量组心肌组织蓝色胶原纤维逐渐减少,LVEDV、LVESV、IL-17、Th17、Th17/Treg、ET-1、心肌梗死面积、细胞凋亡率、TUNEL阳性率、Bax、Caspase-3、RhoA活性、RhoA、ROCK1、ROCK2表达水平依次明显降低,LVEF、LVFS、TGF-β、Treg、eNOS、Bcl-2表达水平依次明显升高(P<0.05)。与苦参碱高剂量组比较,苦参碱高剂量+LPA组心肌组织蓝色胶原纤维增多,LVEDV、LVESV、IL-17、Th17、Th17/Treg、ET-1、心肌梗死面积、细胞凋亡率、TUNEL阳性率、Bax、Caspase-3、RhoA活性、RhoA、ROCK1、ROCK2表达水平明显升高,LVEF、LVFS、TGF-β、Treg、eNOS、Bcl-2表达水平明显降低(P<0.05)。结论苦参碱通过抑制RhoAROCK信号通路调节Th17/Treg细胞平衡,改善冠心病大鼠心肌损伤。

缺血性脑卒中后抑郁患者外周血CDC42、Th17细胞表达及与抑郁程度的相关性分析

目的观察缺血性脑卒中患者外周血细胞分裂周期蛋白42(CDC42)、辅助性T细胞17(Th17)表达并探究其表达与患者抑郁程度的相关性。方法选取2022年1月—2023年1月河北北方学院附属第一医院收治的200例缺血性脑卒中患者为观察组,另选取同期该院200例体检健康者为对照组。患者入院后均接受常规抗血小板聚集、降脂等对症支持治疗,并采用多感官刺激方案对患者进行干预。采用蒙哥马利抑郁评定量表评估患者的抑郁状态,并将患者分为无抑郁组116例、轻度抑郁组38例、中度抑郁组31例、重度抑郁组15例。比较各组医院焦虑抑郁量表(HADS)和简易精神状态评价量表(MMSE)评分,比较观察组干预前后日常生活活动能力(ADL)评分;检测各组外周血CDC42 mRNA、Th17细胞、调节性T细胞(Treg)、Th17/Treg表达。以Pearson相关系数分析HADS评分与患者外周血CDC42 mRNA、Th17细胞、Treg细胞、Th17/Treg表达水平的相关性。结果观察组HADS-A评分、HADS-D评分均高于对照组(P<0.05),MMSE评分低于对照组(P<0.05)。重度抑郁组HADS-A评分、HADS-D评分高于其他组(P<0.05),MMSE评分低于其他组(P<0.05);中度抑郁组HADS-A评分、HADS-D评分高于轻度抑郁组和无抑郁组(P<0.05),MMSE评分低于轻度抑郁组和无抑郁组(P<0.05);轻度抑郁组HADS-A评分、HADS-D评分高于无抑郁组(P<0.05),MMSE评分低于无抑郁组(P<0.05)。观察组外周血CDC42 mRNA、Treg细胞的表达水平低于对照组(P<0.05),外周血Th17细胞、Th17/Treg的表达水平高于对照组(P<0.05)。重度抑郁组Th17细胞、Th17/Treg表达水平高于其他组(P<0.05),Treg细胞表达水平低于另外组(P<0.05),中度抑郁组Th17细胞、Th17/Treg表达水平高于轻度抑郁组、无抑郁组(P<0.05),Treg细胞表达水平低于轻度抑郁组、无抑郁组(P<0.05),轻度抑郁组Th17细胞、Th17/Treg表达水平高于无抑郁组(P<0.05),Treg细胞表达水平低于无抑郁组(P<0.05)。观察组干预后HADS-A评分、HADS-D评分低于干预前(P<0.05),MMSE评分、ADL评分高于干预前(P<0.05)。观察组干预前后外周血CDC42 mRNA表达水平比较,差异无统计学意义(P>0.05)。观察组干预后外周血Th17细胞、Th17/Treg表达水平低于干预前(P<0.05),外周血Treg细胞表达水平高于干预前(P<0.05)。观察组患者外周血Th17细胞、Th17/Treg表达与HADS评分呈正相关(r=0.673、0.603,均P<0.05),外周血Treg细胞表达与HADS评分呈负相关(r=-0.586,P<0.05),外周血CDC42 mRNA与HADS评分无相关性(r=-0.375,P>0.05)。结论缺血性脑卒中患者外周血CDC42mRNA表达明显降低、Th17细胞表达明显升高,随着患者抑郁程度加重,Th17细胞表达逐渐升高,而CDC42mRNA表达与患者的抑郁程度无相关性,可通过合理的干预措施减轻抑郁情绪,防止病情加重。