Reservoir of human immunodeficiency virus in the brain:New insights into the role of T cells

Human immunodeficiency virus(HIV)infection of the central nervous system(CNS)has attracted significant attention because it contributes to severe complications of acquired immunodeficiency syndrome(AIDS)and seriously impairs the life quality of infected patients.In this review,we briefly describe the latent infection of HIV in CNS and focus on the role of the important immune cells,such as T cells,in the formation and maintenance of the HIV reservoir in CNS.This review explores the mechanisms by which T cells enter CNS and establish latent infection of HIV in the CNS.In conclusion,we summarize the role of these cells in the interaction between HIV and CNS.With our better understanding of the underlying mechanisms,we propose future directions for the development of novel strategies to eliminate HIV reservoirs in the CNS based on cellular components.

CD4+ T cells and natural killer cells: Biomarkers for hepatic fibrosis in human immunodeficiency virus/hepatitis C virus-coinfected patients

AIM To characterize peripheral blood natural killer(NK) cells phenotypes by flow cytometry as potential biomarker of liver fibrosis in human immunodeficiency virus(HIV)/hepatitis C virus(HCV) coinfected patients.METHODS Peripheral mononuclear cells from 24 HIV/HCV(HBVnegative) coinfected and 5 HIV/HCV/HBV seronegative individuals were evaluated. HIV/HCV coinfected patients were divided in to groups: G1, patients with METAVIR F0-F2 and G2, patients with METAVIR F3-F4. NK surface cell staining was performed with: AntiCD3(APC/Cy7), anti-CD56(PE/Cy5), anti-CD57(APC), anti-CD25(PE), anti-CD69(FITC), anti-NKp30(PE), antiNKp46(PE/Cy7), anti-NKG2D(APC), anti-DNAM(FITC); anti-CD62L(PE/Cy7), anti-CCR7(PE), anti-TRAIL(PE), anti-Fas L(PE), anti CD94(FITC). Flow cytometry data acquisition was performed on BD FACSCanto, analyzed using Flow Jo software. Frequency of fluorescence was analyzed for all single markers. Clinical records were reviewed, and epidemiological and clinical data were obtained.RESULTS Samples from 11 patients were included in G1 and from 13 in G2. All patients were on ARV, with undetectable HIV viral load. Liver fibrosis was evaluated by transient elastography in 90% of the patients and with biopsy in 10% of the patients. Mean HCV viral load was(6.18 ± 0.7 log10). Even though, no major significant differences were observed between G1 and G2 regarding NK surface markers, it was found that patients with higher liver fibrosis presented statistically lower percentage of NK cells than individual with low to mild fibrosis and healthy controls(G2: 5.4% ± 2.3%, G1: 12.6% ± 8.2%, P = 0.002 and healthy controls 12.2% ± 2.7%, P = 0.008). It was also found that individuals with higher liver fibrosis presented lower CD4 LT count than those from G1(G2: 521 ± 312 cells/μL, G1: 770 ± 205 cells/μL; P = 0.035).CONCLUSION Higher levels of liver fibrosis were associated with lower percentage of NK cells and LTCD4+ count; and they may serve as noninvasive biomarkers of liver damage.

Immune therapy for human papillomaviruses-related cancers

Human papillomaviruses(HPVs) are a large family of double strand DNA viruses comprising more than 180 types. Infection with HPV is very common and it is associated with benign and malignant proliferation of skin and squamous mucosae. Many HPVs, considered lowrisk such as HPV 6 and 11, produce warts; while highrisk viruses, such as HPVs 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, and 58, induce tumors. About 5% of all cancers in men and women are associated with HPV infection. Because there are not antiviral drugs for HPV infection, current therapies for low-risk HPV infections involve physical removal of the lesion by cryotherapy, trichloracetic acid, laser, or surgical removal. Surgical procedures are effective in the treatment of precancerous lesions, however after these procedures, many recurrences appear due to new re-infections, or to failure of the procedure to eliminate the HPV. In addition, HPV can inhibit recognition of malignant cellsby the immune system, leading to the development of cancer lesions. When this occurs, radiotherapy and chemotherapy are then used. Unfortunately, about 50% of the HPV-cancer patients still die. In the past decade, a better knowledge of the natural history of the virushost interaction and of the immune response against this viral infection has brought new therapeutic strategies geared to modulate the immune system to generate an efficient virus-specific cytotoxic response. Novel HPV protein-expressing vaccines have shown some significant clinical efficacy and systemic HPV-specific cytotoxic T cell responses. This review will describe the current status of the several therapeutic strategies used to treat HPV-induced lesions, and discuss the various new therapies now being tested.

Lymphocyte subsets predictive value and possible involvement of human papilloma virus infection on breast cancer molecular subtypes

AIM To detect human papilloma virus(HPV) presence and to characterize cellular immune response in breast cancer patients. METHODS A total of 74 women were included, of which 48 samples were from patients diagnosed with breast cancer and 26 patients with benign pathology of the breast. Molecular subtype classification was performed based on the immunohistochemical reports of the tumor piece. HPV genome detection and genotyping from fresh breast biopsies was performed using the INNO-LIPA HPV Genotyping Extra test(Innogenetics, Ghent, Belgium). CD3+, CD4+, CD8+ and natural killer(NK)+ cells levels from peripheral blood samples from patients with breast cancer and benign pathology were measured by flow cytometry. RESULTS Luminal A was the most frequent breast cancer molecular subtype(33.33%). HPV was detected in 25% of the breast cancer patients, and genotype 18 was the most frequent in the studied population. The mean of CD3+, CD4+ and CD8+ subpopulations were decreased in patients with breast cancer, in relation to those with benign pathology, with a statistically significant difference in CD8+ values(P = 0.048). The mean of NK+ cells was increased in the benign pathology group. The average level of CD3+, CD4+, CD8+ and NK+ cells decreased as the disease progressed. HER2+ and Luminal B HER2+ tumors had the lowest counts of cell subsets. HPV breast cancer patients had elevated counts of cellular subsets. CONCLUSION Determining level changes in cellular subsets in breast cancer patients is a useful tool to evaluate treatment response.

Direct effects of hepatitis C virus on the lymphoid cells

It has been reported that the direct binding of hepatitis C virus(HCV)and/or the replication of HCV in the extrahepatic organs and,especially,lymphoid cells,might affect the pathogenesis of extrahepatic diseases with HCV infection.More than one decade ago,several reports described the existence of HCV-RNA in peripheral blood mononuclear cells.Moreover,many reports describing the existence of HCV in B lymphocytes and B cell lymphoma have been published.In addition to B lymphocytes,it was reported that HCV replication could be detected in T lymphocytes and T cell lines.Among the extrahepatic diseases with HCV infection,mixed cryoglobulinemia-related diseases and autoimmunerelated diseases are important for understanding the immunopathogensis of HCV persistent infection.Moreover,HCV persistent infection can cause malignant lymphoma.The biological significance of lymphotropic HCV has not yet become clear.However,several candidates have been considered for a long time.One is that lymphotropic HCV is an HCV reservoir that might contribute to the recurrence of HCV infection and difficultto-treat disease status.The other important issue is the carcinogenesis of the lymphoid cells and disturbances of the immune responses.Therefore,the extrahepatic diseases might be induced by direct interaction between HCV and lymphoid cells.In this article,we summarize various studies showing the direct effect of HCV on lymphoid cells and discuss the biological significance of lymphotropic HCV.

Host and viral factors contributing to CD8+ T cell failure in hepatitis C virus infection

Virus-specific CD8+ T cells are thought to be the major anti-viral effector cells in hepatitis C virus (HCV) infection. Indeed, viral clearance is associated with vigorous CD8+ T cell responses targeting multiple epitopes. In the chronic phase of infection, HCV-specific CD8+ T cell responses are usually weak, narrowly focused and display often functional defects regarding cytotoxicity, cytokine production, and proliferative capacity. In the last few years, different mechanisms which might contribute to the failure of HCV-specific CD8+ T cells in chronic infection have been identified, including insufficient CD4+ help, deficient CD8+ T cell differentiation, viral escape mutations, suppression by viral factors, inhibitory cytokines, inhibitory ligands, and regulatory T cells. In addition, host genetic factors such as the host’s human leukocyte antigen (HLA) background may play an important role in the efficiency of the HCV- specific CD8+ T cell response and thus outcome of infection. The growing understanding of the mechanisms contributing to T cell failure and persistence of HCV infection will contribute to the development of successful immunotherapeutical and -prophylactical strategies.

Human T-lymphotropic virus type 1-associated myelopathy/tropical spastic paraparesis: Clinical presentation and pathophysiology

Human T-cell lymphotropic virus type 1(HTLV-1)-associated myelopathy/tropical spastic paraparesis(HAM/TSP) is a slowly progressive neurodegenerative disorder in which lesions of the central nervous system cause progressive weakness, stiffness, and a lower limb spastic paraparesis. In some cases, polymyositis, inclusion bodymyositis, or amyotrophic lateral sclerosis-like syndromes are associated with HTLV-1. TSP was first described in Jamaica in 1888 and known as Jamaican peripheral neuritis before TSP was related to HTLV-1 virus, the first retrovirus being identified, and the disease is since named HAM/TSP. There is no established treatment program for HAM/TSP. Prevention is difficult in lowincome patients(i.e., HTLV-1 infected breast feeding mothers in rural areas, sex workers). Thus, there is a need for new therapeutic avenues. Therapeutic approaches must be based on a better understanding, not only of clinical and clinicopathological data, but also of the pathophysiology of the affection. Consequently, a better understanding of existing or newly developed animal models of HAM/TSP is a prerequisite step in the development of new treatments.

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。

Activation-induced pyroptosis contributes to the loss of MAIT cells in chronic HIV-1 infected patients

Background: Mucosal-associated invariant T(MAIT) cells are systemically depleted in human immunodeficiency virus type 1(HIV-1) infected patients and are not replenished even after successful combined antiretroviral therapy(cART).This study aimed to identify the mechanism underlying MAIT cell depletion.Methods: In the present study, we applied flow cytometry, single-cell RNA sequencing and immunohistochemical staining to evaluate the characteristics of pyroptotic MAIT cells in a total of 127 HIV-1 infected individuals, including 69 treatment-naive patients, 28 complete responders, 15 immunological non-responders, and 15 elite controllers, at the Fifth Medical Center of Chinese PLA General Hospital, Beijing, China.Results: Single-cell transcriptomic profiles revealed that circulating MAIT cells from HIV-1 infected subjects were highly activated, with upregulation of pyroptosis-related genes. Further analysis revealed that increased frequencies of pyroptotic MAIT cells correlated with markers of systemic T-cell activation, microbial translocation, and intestinal damage in cART-naive patients and poor CD4+ T-cell recovery in long-term cART patients. Immunohistochemical staining revealed that MAIT cells in the gut mucosa of HIV-1 infected patients exhibited a strong active gasdermin-D(GSDMD, marker of pyroptosis) signal near the cavity side, suggesting that these MAIT cells underwent active pyroptosis in the colorectal mucosa. Increased levels of the proinflammatory cytokines interleukin-12(IL-12) and IL-18 were observed in HIV-1 infected patients. In addition, activated MAIT cells exhibited an increased pyroptotic phenotype after being triggered by HIV-1 virions, T-cell receptor signals, IL-12 plus IL-18, and combinations of these factors, in vitro.Conclusions: Activation-induced MAIT cell pyroptosis contributes to the loss of MAIT cells in HIV-1 infected patients,which could potentiate disease progression and poor immune reconstitution.

An atlas of immune cell transcriptomes in human immunodeficiency virus-infected immunological non-responders identified marker genes that control viral replication

Background:Previous studies have examined the bulk transcriptome of peripheral blood immune cells in acquired immunodeficiency syndrome patients experiencing immunological non-responsiveness.This study aimed to investigate the characteristics of specific immune cell subtypes in acquired immunodeficiency syndrome patients who exhibit immunological non-responsiveness.Methods:A single-cell transcriptome sequencing of peripheral blood mononuclear cells obtained from both immunological responders(IRs)(CD4^(+)T-cell count>500)and immunological non-responders(INRs)(CD4^(+)T-cell count<300)was conducted.The transcriptomic profiles were used to identify distinct cell subpopulations,marker genes,and differentially expressed genes aiming to uncover potential genetic factors associated with immunological non-responsiveness.Results:Among the cellular subpopulations analyzed,the ratios of monocytes,CD16^(+)monocytes,and exhausted B cells demonstrated the most substantial differences between INRs and IRs,with fold changes of 39.79,11.08,and 2.71,respectively.In contrast,the CD4^(+)T cell ratio was significantly decreased(0.39-fold change)in INRs compared with that in IRs.Similarly,the ratios of natural killer cells and terminal effector CD8^(+)T cells were also lower(0.37-fold and 0.27-fold,respectively)in the INRs group.In addition to several well-characterized immune cell-specific markers,we identified a set of 181 marker genes that were enriched in biological pathways associated with human immunodeficiency virus(HIV)replication.Notably,ISG15,IFITM3,PLSCR1,HLA-DQB1,CCL3L1,and DDX5,which have been demonstrated to influence HIV replication through their interaction with viral proteins,emerged as significant monocyte marker genes.Furthermore,the differentially expressed genes in natural killer cells were also enriched in biological pathways associated with HIV replication.Conclusions:We generated an atlas of immune cell transcriptomes in HIV-infected IRs and INRs.Host genes associated with HIV replication were identified as markers of,and were found to be differentially expressed in,different types of immune cells.

宫颈鳞状上皮内病变和宫颈癌Th17、HIF-1α水平变化与HPV感染的关系

目的探讨宫颈鳞状上皮内病变(SIL)和宫颈鳞状细胞癌(SCC)辅助性T细胞17(Th17)、缺氧诱导因子-1α(HIF-1α)水平变化与人乳头瘤病毒(HPV)感染的关系。方法收集2020年1月至2023年1月于金寨县人民医院病理科就诊的40例宫颈SIL患者(SIL组)、40例宫颈SCC患者(SCC组)的病理组织和静脉血指标,并收集同期同一医院20例行全子宫切除的患者(对照组)的正常宫颈上皮组织和静脉血指标。对三组Th17、HIF-1α水平及HPV阳性感染率进行检测并比较。结果SCC组和SIL组Th17、HIF-1α水平显著高于对照组(P<0.05),且SCC组高于SIL组(P<0.05)。SCC组和SIL组HPV单一、多重感染率比较无统计学差异(P>0.05);HPV感染患者的Th17、HIF-1α水平均显著高于未感染患者(P<0.05),且多重感染患者的Th17和HIF-1α水平显著高于单一感染患者(P<0.05)。Th17、HIF-1α水平与HPV感染呈显著相关(P<0.05),且两种指标之间亦存在显著相关(P<0.05)。结论Th17、HIF-1α水平可作为SCC患者预后的客观预测指标及指导临床治疗的参考。

三种人乳头瘤病毒疫苗接种后T、 B细胞应答差异的研究进展

宫颈癌是全球女性常见癌症之一,高危型人乳头瘤病毒16(HPV16)、 HPV18的长期感染是宫颈癌的主要病因,目前广泛应用的HPV疫苗主要有2价Cervarix疫苗、 4价Gardasil疫苗及9价Gardasil-9疫苗,三种疫苗接种后在T细胞效应分子变化,B细胞产生的抗体水平、持续时间、年龄及接种针剂等方面存在差异。

整合酶抑制剂治疗人免疫缺陷病毒1型的临床疗效及对患者病毒载量和CD4^(+)T细胞的影响

目的探讨整合酶抑制剂治疗人免疫缺陷病毒1型(HIV-1)患者的临床疗效及对患者病毒载量和CD4^(+)T细胞的影响。方法选取2020年8月至2022年2月九江市第三人民医院收治的88例HIV-1患者作为研究对象,按照随机抽签方式分为观察组与对照组,每组44例。对照组采用常规HIV-1治疗方案,观察组采用整合酶抑制剂治疗。比较两组临床疗效、CD4^(+)T细胞计数、病毒载量水平及治疗安全性。结果观察组治疗总有效率高于对照组,差异有统计学意义(P<0.05)。治疗后1个月、3个月、6个月、1年、1.5年,观察组CD4^(+)T细胞计数水平均高于对照组,病毒载量水平均低于对照组,差异有统计学意义(P<0.05)。两组不良事件发生率比较差异无统计学意义。结论整合酶抑制剂治疗HIV-1患者的临床疗效显著,能更大程度地降低病毒载量和提升CD4^(+)T细胞水平,且安全性较高,值得推广应用。

A novel mouse model of adult T-cell leukemia cell invasion into the spinal cord

Adult T-cell leukemia( ATL) is a mature T-cell malignancy caused by human T-cell leukemia virus type I infection, and 10%-25% of patients show central nervous system( CNS) involvement. CNS involvement significantly reduces survival and there are no effective treatments for CNS involvement. Therefore, an appropriate animal model is required to evaluate the inhibitory effects of novel drugs on the progression of ATL with CNS involvement. Here, we established a mouse model of ATL with CNS involvement using NOD.Cg-Prkdc~ (scid) Il2 rg ^(tm1Wjl)/SzJ mice inoculated with ATL cells intramuscularly in the postauricular region, and these mice showed paraparesis. Of the 10 mice inoculated with ATL cells intramuscularly(I.M.) at 5 weeks of age, 8(80%) showed paraparesis, whereas none of the 10 mice inoculated with ATL cells subcutaneously(S.C.) showed paraparesis. In the I.M. group, PCR detected HTLV-1-specific genes in the thoracic and lumbar vertebrae; however, in the S.C. group, the vertebrae were negative for HTLV-1 genes. Histological analysis revealed a particularly high incidence of tumors, characterized by accumulation of the injected cells, in the thoracic vertebrae of mice in the I.M. group. Tumor cell infiltration was relatively high in the bone marrow. Spinal cord compression caused by invasion of the tumor mass outside the pia mater was observed in the thoracic vertebrae of the spinal cord. In conclusion, we have reported a mouse model of tumor growth with paraparesis that may be used to assess novel therapeutic agents for ATL with CNS involvement.

尖锐湿疣患者人乳头瘤病毒基因型分布及Th1/Th2亚群失衡与复发的关系

目的探讨感染不同基因型人乳头瘤病毒(HPV)及T淋巴细胞亚群状态与尖锐湿疣(CA)复发的关系。方法选取2019年1月至2020年12月新疆维吾尔自治区人民医院诊治的142例CA患者作为研究对象(CA组),采集疣体组织及外周血,根据是否复发分为CA复发组与CA非复发组。另选取同期20例健康人作为对照组,采集外周血。采用聚合酶链反应-反向杂交法检测疣体组织中HPV型别;采用流式细胞术检测外周血中T淋巴细胞亚群;采用流式微球分析法检测细胞因子的表达。结果CA组检出HPV感染亚型主要为HPV 11(47.18%)、HPV 6(38.73%)及HPV 16(19.01%),CA复发率为76.76%;CA患者CD4^(+)/CD8^(+)比值显著低于对照组(P<0.05),CA复发组Treg比率显著低于CA非复发组(P<0.05);CA复发组辅助性T(Th)1型细胞因子白介素-2(IL-2)及γ干扰素(IFN-γ)均显著低于CA非复发组(P<0.05),而Th2型细胞因子白介素-4(IL-4)及白介素-10(IL-10)均显著高于CA非复发组(P<0.05)。结论新疆地区CA患者HPV感染型别以HPV 11、6、16为主,同时均出现了T淋巴细胞亚群的紊乱及Th1/Th2细胞亚群的失衡,这种变化可能与CA发病期的免疫应答及CA复发期的免疫耐受存在关联。

AIDS患者CD4^(+)T细胞水平与并发结核病风险关系的研究

目的:探讨艾滋病(AIDS)患者CD4^(+)T细胞水平与并发结核病风险的关系。方法:选择2021年4月至2022年12月定期在贵州省安顺市人民医院进行CD4^(+)T细胞水平检测及结核病筛查的50例AIDS患者作为研究对象。根据患者在研究过程中是否并发结核病将其进一步分为对照组和患病组,分析患者并发结核病前后的CD4^(+)T细胞水平,总结AIDS患者CD4^(+)T细胞水平与并发结核病风险的关系。结果:分别于2021年4月、2021年10月、2022年4月、2022年10月4次检测50例AIDS患者的CD4^(+)T细胞水平及结核分枝杆菌感染情况,结果显示,2021年4月无患者并发结核病,2021年10月、2022年4月、2022年10月分别有9例、18例、23例患者并发结核病;2021年10月、2022年4月、2022年10月3次检测时,患病组的CD4^(+)T细胞水平均显著低于对照组,差异有统计学意义(P<0.05)。结论:AIDS患者的CD4^(+)T细胞水平对并发结核病的风险具有一定的提示作用,CD4^(+)T细胞水平越低者并发结核病的风险越高。因此,临床上应定期检测AIDS患者的CD4^(+)T细胞水平,以评估其并发结核病的风险,争取做到早发现、早诊断、早治疗,从而提高患者的生存质量。

HIV感染者接受长期抗病毒治疗后CD4^(+)T平台期水平及影响因素分析

目的 了解人类免疫缺陷病毒(human immunodeficiency virus, HIV)感染者长期接受抗逆转录病毒治疗(antiretroviral therapy, ART)后CD4^(+)T进入平台期水平及影响因素。方法 以重庆市公共卫生医疗救治中心感染科诊治,截止2021年12月连续ART超过8年且HIV RNA<200拷贝/mL患者为研究对象,回顾性收集年龄、ART方案、各时间点CD4^(+)T计数等,统计CD4^(+)T增长,分析影响平台期CD4^(+)T的因素。结果 纳入766例ART超过8年患者,平台期CD4^(+)T中位数394.0(292.0,507.0)个/μL,其中66例(8.6%)为重度免疫重建不良(suboptimal immune responders, SIR),CD4^(+)T<200个/μL;458例(59.8%)≥350个/μL,198例(25.8%)≥500个/μL。以基线CD4^(+)T分组(CD4^(+)T<200,200~349,350~499,≥500个/μL),各组平台期CD4^(+)T及较基线增长值均存在统计学差异(P<0.05)。以年龄分组,青少年平台期CD4^(+)T 403(306,519)个/μL,显著高于中老年组(P<0.05)。单因素分析显示重度SIR组年龄大于44岁患者占比显著高于非重度SIR组(P<0.01),基线CD4^(+)T<100个/μL患者占比重度SIR组(54.5%)显著高于非重度SIR组(17.6%,P<0.05)。多因素分析显示年龄>44岁(OR=2.061,95%CI:1.157~3.672)、基线CD4^(+)T<100个/μL(OR=4.803,95%CI:2.767~8.340)与平台期重度SIR相关(P<0.05)。结论 本组HIV感染者平台期CD4^(+)T普遍较低,中年与老年患者启动ART时基线CD4^(+)T低于100个/μL发生平台期重度SIR的风险更大;早筛查、早治疗是防控重度SIR发生的关键。

BMI-1抑制剂PTC-596对人乳腺癌MCF-7细胞增殖、周期和凋亡的影响

目的 分析BMI-1抑制剂PTC-596对人乳腺癌MCF-7细胞增殖、周期和凋亡的影响。方法以人乳腺癌MCF-7细胞为肿瘤细胞模型,正常培养为阴性对照组,25、50 nmol/L PTC-596处理SGC-7901细胞48 h为低、高药物浓度实验组。利用CCK8法分析细胞增殖能力;流式细胞术分别结合PI单染、DCFHDA探针、JC-1探针以及PI/FITC-Annexin V双染分析细胞周期、活性氧(reactive oxygen species, ROS)累积、线粒体膜电位和凋亡细胞比例;Western blot法检测细胞BIM-1蛋白和周期相关蛋白CyclinD1、CyclinB1、P21以及凋亡相关蛋白Bax、Bcl-2、c-PARP蛋白相对表达水平。结果 与对照组相比,PTC-596高效抑制人乳腺癌MCF-7细胞的增殖,处理48 h后IC_(50)为(49.33±7.02)nmol/L。低、高药物浓度实验组细胞中BMI-1表达显著减少(P <0.01)。实验组细胞中CyclinB1和P21相对表达增加,CyclinD1表达减少,细胞有丝分裂被抑制,出现G_2/M期周期阻滞;同时活性氧累积增多,线粒体膜电位下降,Bax表达上调,Bcl-2表达下调,c-PARP增加,凋亡细胞比例从2.04%显著上升为10.56%、26.74%。与对照组相比较,以上结果差异均有统计学意义(P <0.05)。结论 PTC-596高效杀伤人乳腺癌MCF-7细胞,其机制可能与抑制BMI-1、诱导细胞周期阻滞和内源性线粒体途径细胞凋亡密切相关。

徐长卿中的天然产物XCQ-9对Jurkat细胞增殖和凋亡的影响及机制研究

目的探讨徐长卿中天然产物XCQ-9抑制人急性T淋巴细胞白血病Jurkat细胞增殖和凋亡的作用及可能机制。方法以Jurkat细胞作为白血病细胞模型,采用MTT法测定0(空白对照)、2.5、5、10、20、40μmol/L XCQ-9作用24、48、72 h后对Jurkat细胞增殖的抑制作用。用0(空白对照)、2.5、5、10μmol/L XCQ-9作用于Jurkat细胞24、48 h后,利用流式细胞术分析XCQ-9对细胞周期和细胞凋亡的影响,并通过Western blot实验检测上述药物作用24 h后细胞中胱天蛋白酶9(Caspase-9)、活化的Caspase-9(Cleaved Caspase-9)、Caspase-3、活化的Caspase-3(Cleaved Caspase-3)、聚腺苷二磷酸-核糖聚合酶(PARP)、活化的PARP(Cleaved PARP)、细胞周期蛋白依赖性激酶1(CDK1)和细胞周期蛋白B1(Cyclin B1)的表达情况。结果与空白对照比较,不同浓度XCQ-9均可显著降低Jurkat细胞的存活率(P<0.01),并呈时间和浓度依赖性趋势。5、10μmol/L XCQ-9作用48 h后均可显著诱导Jurkat细胞凋亡(P<0.05或P<0.01),将细胞周期阻滞在G2期(P<0.01)。10μmol/L XCQ-9作用24 h后,可显著下调细胞中CDK1、Caspase-9蛋白的表达(P<0.01),上调细胞中Cyclin B1、Cleaved Caspase-9、Cleaved Caspase-3和Cleaved PARP蛋白的表达(P<0.05或P<0.01)。结论XCQ-9通过诱导G2期阻滞抑制Jurkat细胞增殖,并激活Caspase通路促进细胞凋亡,从而发挥其抗肿瘤作用。

人类免疫缺陷病毒/乙型肝炎病毒合并感染者CD4^(+)T细胞计数和乙型肝炎病毒复制、中性粒细胞与淋巴细胞比值水平的相关性

目的探讨人类免疫缺陷病毒(HIV)/乙型肝炎病毒(HBV)合并感染者CD4^(+)T细胞计数和HBV复制、中性粒细胞与淋巴细胞比值(NLR)的相关性。方法回顾性分析2018年2月至2022年1月扬州大学附属医院收治的59例HBV单纯感染者(HBV组)与76例HIV/HBV合并感染者(HIV/HBV组)的临床资料。HIV/HBV组根据CD4^(+)T细胞计数分为A组(CD4^(+)T细胞计数>200个/μl,31例)和B组(CD4^(+)T细胞计数≤200个/μl,45例)。比较HBV组与HIV/HBV组CD4^(+)T细胞计数、HBV DNA水平及NLR,进一步比较A组与B组的HBV DNA水平及NLR,分析HIV/HBV合并感染者CD4^(+)T细胞计数与HBV DNA水平及NLR的相关性。结果HIV/HBV组CD4^(+)T细胞计数、HBV DNA水平低于HBV组,NLR高于HBV组,差异有统计学意义(P<0.05)。A组HBV DNA水平及NLR均低于B组,差异有统计学意义(P<0.05)。HIV/HBV组CD4^(+)T细胞计数与HBV DNA水平及NLR均呈负相关(r<0,P<0.05)。结论HIV/HBV合并感染者CD4^(+)T细胞计数降低,HBV DNA复制更加活跃,对临床指导用药具有重要意义。