Clonal immunoglobulin heavy chain and T-cell receptor γ gene rearrangements in primary gastric lymphoma

AIM:To study the diagnostic value of immunoglobulin heavy chain(IgH)and T-cell receptorγ (TCR-γ)gene monoclonal rearrangements in primary gastric lymphoma(PGL).METHODS:A total of 48 patients with suspected PGL at our hospital were prospectively enrolled in this study from January 2009 to December 2011.The patients were divided into three groups(a PGL group,a gastric linitis plastica group,and a benign gastric ulcer group)based on the pathological results(gastric mucosal specimens obtained by endoscopy or surgery)and follow-up.Endoscopic ultrasonography(EUS)and EUSguided biopsy were performed in all the patients.The tissue specimens were used for histopathological examination and for IgH and TCR-γ gene rearrangement polymerase chain reaction analyses.RESULTS:EUS and EUS-guided biopsy were successfully performed in all 48 patients.In the PGL group(n=21),monoclonal IgH gene rearrangements were detected in 14(66.7%)patients.A positive result for each set of primers was found in 12(57.1%),8(38.1%),and 4(19.0%)cases using FR1/JH,FR2/JH,and FR3/JH primers,respectively.Overall,12(75%)patients with mucosal-associated lymphoid tissue lymphoma(n=16)and 2(40%)patients with diffuse large B-cell lymphoma(n=5)were positive for monoclonal IgH gene rearrangements.No patients in the gastric linitis plastica group(n=17)and only one(10%)patient in the benign gastric ulcer group(n=10)were positive for a monoclonal IgH gene rearrangement.No TCRgene monoclonal rearrangements were detected.The sensitivity of monoclonal IgH gene rearrangements was 66.7%for a PGL diagnosis,and the specificity was96.4%.In the PGL group,8(100%)patients with stage IIE PGL(n=8)and 6(46.1%)patients with stage IE PGL(n=13)were positive for monoclonal IgH gene rearrangements.CONCLUSION:IgH gene rearrangements may be associated with PGL staging and may be useful for the diagnosis of PGL and for differentiating between PGL and gastric linitis plastica.

In vivo anti-tumor activity of murine hematopoietic stem cells expressing a p185HER2-specific chimeric T-cell receptor gene

We have confirmed efficient anti-tumor activities of the peripheral lymphocytes transduced with a p185HEH2-specific chimeric T-cell receptor gene both in murine and in human in our previous studies. To further test the feasibility of chimeric T-cell receptor in a bone marrow transplantation model, we first, made two routine tumor cell lines： MT901 and MCA-205, to express human p185HER2 by retroviral gene transduction. Murine bone marrow cells were retrovirally transduced to express the chimeric T-cell receptor and gene-modified bone marrow cells were transplanted into lethally irradiated mouse. Six months post transplantation, p185HER2-positive tumor ceils： MT-901/HER2 or MCA-205/ HER2 was subcutaneously or intravenously injected to make mouse models simulating primary breast cancer or pulmonary metastasis. The in vivo anti-tumor effects were monitored by the size of the subcutaneous tumor or counting the tumor nodules in the lungs after India ink staining. The size of the subcutaneous tumor was significantly inhibited and the number of pulmonary nodules were significantly decreased in mouse recipients transplanted with chimeric T-cell receptor modified bone marrow cells compared with the control group. Our results suggest the efficient in vivo anti-tumor activities of chimeric T-cell receptor gene modified bone marrow cells.

Tumor Antigen Specific Activation of Primary Human T-Cells Expressing a Virally Encoded Chimeric T-Cell Receptor Specific for p185HER2

We have developed and tested chimeric T-cell receptors (TCR) specific for p185HER2. In these experiments, retroviral vectors expressing the N29γ or N29ζ receptors were constructed in pRET6. Amphotropic viral producer cells were established in the GALV-based PG13 packaging cell line. Ficoll purified human peripheral blood lymphocytes (PBL) were virally transduced using an optimized protocol incorporating activation with immobilized anti-CD3/anti-CD28 monoclonal anti- bodies, followed by viral infection in the presence of fibronectin fragment CH296. Transduced cells were co-cultured with human tumor cell lines that overexpress (SK-OV-3) or underexpress (MCF7) p185HER2 to assay for antigen specific im- mune responses. Both CM+ and CD8+ T-cells transduced with the N29γ or N29ζ chTCR demonstrated HER2-specific anti- gen responses, as determined by release of Th1 like cytokines, and cellular cytotoxicity assays. Our results support the fea- sibility of adoptive immunotherapy with genetically modified T-cells expressing a chTCR specific for p185HER2.

Primary anaplastic lymphoma kinase-positive large B-cell lymphoma of the left bulbar conjunctiva: A case report

BACKGROUND Anaplastic lymphoma kinase(ALK)-positive large B-cell lymphoma(LBCL)is an aggressive and rare variant of diffuse LBCL.Herein,we report an uncommon case of stage IE extranodal ALK-positive LBCL initially originating in the bulbar con-junctiva.CASE SUMMARY A 63-year-old woman presented with a mass in the left bulbar conjunctiva that had persisted for six months,accompanied by swelling and pain that had per-sisted for 3 d.Eye examination revealed an 8 mm slightly elevated pink mass in the lower conjunctival sac of the left eye.Microscopically,the tumor was com-posed of large immunoblastic and plasmablastic large lymphoid cells with scattered anaplastic or multinucleated large cells.Immunophenotypically,the neoplastic cells were positive for ALK,CD10,CD138,Kappa,MUM1,BOB.1,OCT-2,CD4,CD45,EMA,CD79a,CD38,and AE1/AE3,and negative for CD20,PAX5,Lambda,BCL6,CD30 and all other T-cell antigens.The results of gene rearrangement tests showed monoclonal IGH/IGK/IGL and TCRD rearran-gements.Fluorescence in situ hybridization studies did not reveal any BCL2,BCL6 or MYC rearrangements.Furthermore,Epstein-Barr virus was not detected by in situ hybridization in the lesions.Based on the histopathological and imaging examinations,the neoplasm was classified as stage IE ALK-positive LBCL.No further treatments were administered.At the 6,15,and 21 mo postoperative follow-up visits,the patient was in good condition,without obvious discomfort.This case represents the first example of primary extranodal ALK-positive LBCL presenting as a bulbar conjunctival mass,which is extremely rare and shares morphological and immunohistochemical features with a variety of other neo-plasms that can result in misdiagnosis.CONCLUSION Awareness of the condition presented in this case report is necessary for early and accurate diagnosis and appropriate treatment.

TRPV6对结肠癌SW480细胞生物学行为及小鼠模型中结肠肿瘤发生的影响

目的:通过观察沉默瞬时受体阳离子通道亚家族V成员6(transient receptor potential cation channel,subfamily V member 6,TRPV6)对结肠癌SW480细胞生物学行为的影响和细胞内钙的浓度变化,以及1,25(OH)_2D_3、CaCl_2及CuCl_2在SD大鼠结肠肿瘤模型建立中的影响,探索TRPV6在结肠癌发生过程中的相关作用,为结肠癌的防治寻找新的靶点。方法:通过构建慢病毒颗粒感染结肠癌SW480细胞,采用免疫组织化学法、Western bolt、PCR技术检测TRPV6蛋白及m RNA的表达,MTT法、迁移及凋亡实验观察结肠癌SW480细胞增殖、迁移及凋亡变化,高速离子成像系统测定结肠癌SW480细胞内的Ca^(2+)浓度变化。以二甲基肼(dimethyl hydrazine,DMH)建立SD大鼠结肠肿瘤模型,分为实验组(DMH组)和干预组[(DMH+1,25(OH)_2D_3组、DMH+CuCl_2组)]、对照组,干预组分别予1,25(OH)_2D_3(37.5 nmol/kg)、CuCl2(375μmol/kg)对模型进行干预。观察各组大鼠结肠腺瘤及腺癌的发生情况,Western blot检测各组结肠组织中TRPV6蛋白的表达情况。结果:TRPV6-RNAi转染结肠癌SW480细胞后,TRPV6 m RNA及蛋白表达减少,细胞Ca^(2+)浓度水平降低,结肠癌SW480细胞的增殖率、迁移能力下降,细胞凋亡率增加,与空白对照组及阴性对照组比较差异具有统计学意义(P<0.05)。对照组大鼠成瘤率为0,DMH+1,25(OH)_2D_3组为100%,DMH组为84.62%,DMH+CuCl_2组为33.33%。各组大鼠大肠中均有TRPV6蛋白的表达,表达情况为DMH+1,25(OH)_2D_3组>DMH组>DMH+CuCl_2组>对照组,差异具有统计学意义(P<0.05)。结论:降低结肠癌SW480细胞Ca^(2+)浓度水平可抑制结肠癌SW480细胞的增殖、迁移能力,诱导细胞凋亡。1,25(OH)_2D_3可以增加实验鼠结肠组织TRPV6蛋白表达,促进结肠肿瘤的形成。CuCl_2可以使实验鼠结肠组织中的TRPV6蛋白表达降低,并抑制结肠肿瘤的形成。

CML患者外周血TCR Vδ亚家族T细胞的谱系变化研究

目的了解慢性粒细胞白血病(chronic myeloid leukemia,CML)患者慢性期(chronic phase,CP)和缓解期(complete remission,CR)外周血T细胞受体(T cell receptor,TCR)Vδ亚家族T细胞的分布和克隆性情况。方法利用RT-PCR扩增8例CML-CP和8例CML-CR患者外周血T细胞中8个Vδ亚家族基因的互补决定区(CDR3),了解TCR Vδ亚家族T细胞的分布和利用情况;阳性产物进一步经基因扫描分析CDR3长度,从而了解其克隆性。10例健康人外周血作为对照。结果8例CML-CP患者外周血T细胞平均表达(3.25±0.89)个Vδ亚家族,主要以Vδ1、Vδ2和Vδ3的表达为主,而8例CR期患者则平均表达(3.50±0.76)个Vδ亚家族,主要集中在Vδ1、Vδ2、Vδ3和Vδ8中表达;两组与健康对照组(3.50±0.52)比较无统计学意义。此外,Vδ8表达频率在CML-CR患者(5/8)中略高于健康人(3/10)和CML-CP患者(2/8)。基因扫描绝大多数患者和健康人外周血TCR Vδ亚家族T细胞均存在克隆性增殖现象,主要以Vδ1-Vδ3为主,但在CML-CR患者中,表达Vδ7亚家族的两例样本均呈克隆性增殖,这在10例健康人中均未检测到。结论3组样本的外周血T细胞中TCR Vδ谱系和克隆性增殖类似,提示CML患者T细胞免疫抑制可能并不主要累及TCR Vδ谱系变化,并在CML-CR期所出现了一些不同的Vδ表达和克隆模式。

Profiling the repertoire of T-cell receptor beta-chain variable genes in peripheral blood lymphocytes from subjects who have recovered from acute hepatitis B virus infection

The profile of T-cell receptor beta-chain variable （TRBV） genes usually skews in subjects with virus infection or cancer. The gene melting spectral pattern （GMSP） can be used to determine the profile of the TRBV gene family. To explore the portrait of the TRBV family in peripheral blood lymphocytes from subjects who have recovered from acute hepatitis B virus infection （AHI）, peripheral blood mononuclear cells （PBMCs） were separated and further sorted into CD4^＋ and CD8^＋ T-cell subsets. The molecular features of the TRBV complementary determining region 3 （CDR3） motifs were determined using GMSP analysis. When a GMSP profile showed a single peak, the monoclonally expanded TRBV gene was cloned and sequenced. Skewed expansions of multiple TRBV genes were observed among the CD4^＋ and CD8^＋ T-cell subsets and the PBMCs. The frequency of monoclonally expanded TRBV genes in the CD8^＋ T-cell subset was significantly higher than that of the CD4^＋ T-cell subset and the PBMCs. Compared to other members of the TRBV gene family, TRBV11, BV15 and BV20 were predominantly expressed in the repertoire of peripheral blood lymphocytes in recovered AHI subjects. The relatively conserved amino acid motifs of TRBV5.1 and BV20 CDR3 were also detected in the CD4^＋ and CD8^＋ T-cell subsets. These results demonstrate the presence of multiple biased TRBV families in recovered AHI subjects. TRBV11, BV15 and BV20, especially from the CD8＋ T-cell subset, may be relevant to the pathogenesis of subjects with AHh The preferentially selected TRBV5.1 and BV20 with the relatively conserved CDR3 motif may be potential targets for personalized treatments of chronic HBV infection.

甲状腺乳头状癌组织中TRAP1表达、BRAF V600E基因突变情况观察及其相关性分析

目的观察甲状腺乳头状癌(PTC)组织中肿瘤坏死因子受体相关蛋白1(TRAP1)表达、V-raf鼠类肉瘤滤过性病毒致癌基因同源体B1(BRAF)V_(600E)基因突变情况,并分析其相关性。方法 PTC组织100例份(A组)、甲状腺腺瘤组织38例份(B组)、结节性甲状腺肿组织38例份(C组),观察各组TRAP1蛋白表达及BRAFV_(600E)基因突变情况,分析TRAP1表达、BRAFV_(600E)基因突变两者间的相关性及其与PTC临床病理特征的关系。结果A、B、C组TRAP1阳性率分别为71.00%、13.16%、10.53%,BRAFV_(600E)基因突变率分别为52.00%、0、0,A组分别与B、C组比较,P均<0.05。BRAFV_(600E)基因突变与PTC患者年龄及肿瘤直径相关(P均<0.05)。A组中TRAP1表达与BRAFV_(600E)基因突变呈正相关(r=0.445,P<0.05)。结论 PTC组织中TRAP1阳性率及BRAFV_(600E)基因突变率升高,且TRAP1高表达与BRAFV_(600E)基因突变密切相关,两者可能共同参与了PTC的发生发展。

Predominant expression of β-chain variable region genes of T-cell receptor in TILs from human renal cell carcinoma

THE repertoire of T-cell antigen receptor(TcR)can be effectively analysed by an approachwith quantitative polymerase chain reaction(qPCR),which allows to characterize the ex-panding T-cell clones by the patterns of restricted usage of TcR V and J region genes.

T cell receptor Vβ gene bias in rheumatoid arthritis

Objectives To explore the pathogenesis of rheumatoid arthritis (RA) by studying the expression of T cell receptors (TCRs).Methods T cell receptor Vβ (TCR Vβ) gene usage and expression were analyzed from synovial membrane and peripheral blood of 8 RA patients, 2 osteoarthritis patients and 2 accident amputees. The complementary determining region 3 (CDR3) of 25 TCR Vβ subfamily genes in unselected T cell populations were amplified semi-quantitatively by reverse transcription-polymerase chain reaction (RT-PCR). The products were further studied by genescan for frequency of Vβ usage.Results The numbers of Vβ subfamilies expressed by T cells from RA peripheral blood and synovial membrane were not significantly restricted. More importantly, biased Vβ gene expression in RA synovium was observed and Vβ6, Vβ17, and Vβ22 genes were the predominant subfamilies. It was noteworthy that the expression of Vβ17 in RA synovium was significantly increased. Conclusion Our data were consistent with the hypothesis that several antigen or superantigen-driven processes may be involved in the pathogenesis of RA.

Relationship between T-cell receptor α gene polymorphisms and symptomatic differences in patients with narcolepsy type 1

Background:Recent genome-wide association studies have identified an important role of T-cell receptor α(TRA) gene in the development of narcolepsy type 1.However,the role of TRA haplotype polymorphisms in the symptomatic diversity of narcolepsy remains unclear.This study aimed to investigate whether TRA polymorphisms can influence the symptomatic diversity of narcolepsy.Methods:Totally,903 patients with narcolepsy type 1 were included in the study.Patients were divided into different groups according to their symptoms.First,13 genotyped single nucleotide polymorphisms in the TRA were assessed for their association with symptoms of narcolepsy.We used the Chi-square test to determine differences in genotype frequencies in patients with narcolepsy.Further,we identified the haplotypes and variations of the TRA and tested their association with the symptoms of narcolepsy using a logistic regression model.Results:According to the results of the logistic regression,TRA haplotypes TG and CT were significantly associated with auditory hallucination,with odds ratios of 1.235 (95% confidence interval [CI],1.012-1.507) and 1.236 (95% CI,1.012-1.511),respectively (P < 0.05).Contusions:The patterns of haplotype in TRA (haplotypes TG and CT) are associated with hypnagogic auditory hallucination in patients with narcolepsy type 1.However,further studies are needed to confirm our results and explore the underlying mechanisms.

Features of intestinal T-cell lymphomas in Chinese population without evidence of celiac disease and their close association with Epstein-Barr virus infection

Background Intestinal T-cell lymphoma （ITCL） is a heterogeneous lymphoid neoplastic group with variable clinical and pathological features. ITCL in oriental countries is different from enteropathy-type intestinal T-cell lymphoma （ETCL） in relation to celiac disease and Epstein-Barr virus （EBV）. The objective of this study was to investigate the clinicopathological features, immunophenotype, expression of cytotoxic molecule （TIA-1）, T-cell receptor （TCR）-γ gene rearrangement, and Epstein-Barr virus （EBV） latent infection in primary ITCL without celiac disease in Chinese. Methods The clinical data of 42 patients were analyzed, and the patients were followed up. Compared with human reactive lymphoid tissues, in situ hybridization for EBER1/2, polymerase chain reaction for TCR-~/gene rearrangement, and immunohistochemical staining for immunophenotypes, TIA-1 and EBV latent membrane proteins （LMP-1） were investigated. Survival curves of different clinicopathological features, immunophenotypes, expression of LMPI , TCR-γ/gene rearrangement and therapy were analyzed. Results Three fourths of the patients suffered from ITCL in China were men with a peak age incidence in the 4th decade. Common presenting features included fever and hemotochezia. The prognosis was poor with a median survival of 3.0 months. The lesions were mostly localized in the ileocecum and colon. About 38/42 （90. 5% ） patients demonstrated pleomorphic medium-sized on large seen. All 42 patients with ITCL revealed CD45RO cells. Histological features of celiac disease were rarely positive. Neoplastic cells partially expressed T-cell differentiated antigens （CD3ε, CD4, CD8） and NK cell associated antigen （CD56）. The positive frequency of CD3e, CIM, CD8 and CD56 was 28/42 （66.7%） 7/42 （16.7%）, 10/42 （23.8%） and 12/42 （28.6%） respectively. Thirty-nine cells （92. 9% ） expressed TIA-1, but none expressed CD20 and CD68. More than half of the patients （64. 3% , 64.3% and 59.5% ） revealed TCR-γ primers respectively. EBER1/2 was detected in 41 （97.6%） of LMP-1 was 38. 1% （16/42）. gene rearrangement by three different TCR-γ the 42 patients. The expression frequency ofConclusions Primary ITCL without celiac disease in Chinese is a special highly EBV-associated clinicopathological entity. There are few similarities in patients with celiac disease in western countries. A small proportion of primary ITCLs in Chinese and extranodal NK/T-eell lymphoma of nasal type belong to the same speetrum.

Study on Cellular Differentiation and Genes Rearrangement of Malignant Lymphoproliferative Disorders

Malignant lymphoproliferative disorders are common diseases characterized by the clonal proliferation of cells derived from the lymphocytes of different developmental stages. The analyses of routine morphology and cytochemistry fail to determine the origin and differentiation stages. The cell differentiation origin of most of malignant lymphoproliferative disorders can be defined by using monoclonal antibodies(McAbs) against the differentiation antigens of lymphocytes, but it is unable to determine the cell origin of a minority of undifferentiated malignancies and also fails to distinguish the malig-

γδT cells:Major advances in basic and clinical research in tumor immunotherapy

γδT cells are a kind of innate immune T cell.They have not attracted sufficient attention because they account for only a small proportion of all immune cells,and many basic factors related to these cells remain unclear.However,in recent years,with the rapid development of tumor immunotherapy,γδT cells have attracted increasing attention because of their ability to exert cytotoxic effects on most tumor cells without major histocompatibility complex(MHC)restriction.An increasing number of basic studies have focused on the development,antigen recognition,activation,and antitumor immune response ofγδT cells.Additionally,γδT cell-based immunotherapeutic strategies are being developed,and the number of clinical trials investigating such strategies is increasing.This review mainly summarizes the progress of basic research and the clinical application ofγδT cells in tumor immunotherapy to provide a theoretical basis for further the development ofγδT cell-based strategies in the future.

免疫检查点抑制剂治疗驱动基因阳性晚期非小细胞肺癌的研究进展

免疫检查点抑制剂(immune checkpoint inhibitors,ICIs)已成为晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)的重要治疗方法,但其在驱动基因阳性患者中的疗效仍存在争议。现有研究显示:ICIs的疗效可能与不同的驱动基因类型、程序性死亡蛋白-配体1(programmed cell death-ligand 1,PD-L1)表达水平、肿瘤突变负荷(tumor mutational burden,TMB)等相关,同时可能需要参考其他因素,如临床特征、免疫细胞密度等。ICIs单药或联合治疗可能会在一部分驱动基因阳性NSCLC患者中发挥作用,但仍需要进一步研究来筛选出这些患者,这或许是晚期NSCLC治疗未来发展的一个重要方向。

Study on immunological characteristics of T lymphocyte in peripheral blood from patients with sjogren's syndrome

Objective: To study on in immunological characteristics of T lymphocyte in peripheral blood from patients with Sjogren's syndrome (SS), provide the theory basis for the further research of Traditional Chinese medicine (TCM) targeted therapy.Methods:T lymphocyte subsets in peripheral blood from SS, RA and normal control groups were tested with various monocbnal antibodies and multicolor flow cytometry.T lymphocyte subsets were tested in peripheral blood from SS patients of different TCM Syndromes. The relationship between T Lymphocyte subsets and disease activity was measured by erythrocyte sedimentation rate (SR)and serum IgG. Results: The percentage of CD4^+T cells and the ration of CD4^+/CD8^+T cells in peripheral bbod from SS patients decreased significantly, and the percentage of CD8^+T cells increased than that in control groups (P<0.05). Compared with patients in inactive period, the percentage of CD4^+T cells in peripheral bLood from SS patients in the active stage increased (P>0.05), the percentage of CD8^+T cells decreased (P>0.05), and the ration of CD4^+/ CD8^+T cells increased (P<0.05).Compared with control groups, the percentage of CD69 expressed on T cells in peripheral blood from SS patients decreased, and the percentage of CD95's expression increased (P<0.05).The percentage of CD69CD95's expression also increased (P>0.05). There were no differences among CD4^+T cells, CD8^+T ceLs and CD4^+/ CD8^+ in peripheral blood from SS patients of different TCM Syndromes (P>0.05). CD69 and CD95's expression on SS patients of different TCM Syndromes was no significantLy different (P>0.05).T cell receptor Vβ(TCR Vβ) subfamilies gene in peripheral blood from SS patients and control groups were expressed in all. Compared witji control groups, Vβ 5.3,Vβ 13.1,Vβ 12,Vβ 7.2 had significant differences (P<0.05).Comparison of different TCM Syndromes in SS group: Vβ 13.1 and Vβ 14 were significantly different (P<0.05).Conclusion:There was dysimmunity on T lymphocyte subsets in peripheral blood of SS patients, which was related to disease activity;apoptosis after activation of autoreactive T cell may be induced by Chinese medicine;TCR Vβ subfamilies of different TCM syndromes were different, which noted that different autoreactive T cells of abnormal activation tended to result in different expression of TCM Different syndromes.

TRPV1对H9c2心肌细胞缺氧复氧后凋亡的影响及机制

目的探讨瞬时受体电位阳离子通道V亚族成员1(TRPV1)对H9c2心肌细胞缺氧复氧后凋亡的保护作用及可能机制。方法将H9c2心肌细胞随机分为对照组、模型组(缺氧/复氧组)、辣椒素(TRPV1激活剂)组(缺氧/复氧+辣椒素组)、辣椒素+LY组[缺氧/复氧+辣椒素+LY294002(PI3k抑制剂)组]。采用CCK-8法测定心肌细胞存活率,采用流式细胞仪测定各组心肌细胞凋亡率,采用Western blot法和逆转录-聚合酶链反应(RTPCR)法测定各组细胞caspase-3、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、磷酸化-蛋白激酶B(pAkt)、蛋白激酶B(Akt)水平。结果与对照组比较,模型组、辣椒素组、辣椒素+LY组心肌细胞存活率和Bcl-2水平降低(均P<0.05),细胞凋亡率和caspase-3、Bax、p-Akt水平升高(均P<0.05);与模型组比较,辣椒素组心肌细胞存活率和Bcl-2、p-Akt水平升高(均P<0.05),细胞凋亡率和caspase-3、Bax水平降低(均P<0.05);模型组与辣椒素+LY组各指标比较差异均无统计学意义(均P>0.05)。结论 TRPV1可抑制缺氧复氧后H9c2心肌细胞凋亡,其机制可能与TRPV1激活磷脂酰肌醇3-激酶(PI3k)/Akt信号通路有关。

Self-activation of Vγ9Vδ2 T cells by exogenous phosphoantigens involves TCR and butyrophilins

The high cytotoxic activity of Vγ9Vδ2 T lymphocytes against tumor cells makes them useful candidates in anticancer therapies.However,the molecular mechanism of their activation by phosphoantigens(PAgs)is not completely known.Many studies have depicted the mechanism of Vγ9Vδ2 T-cell activation by PAg-sensed accessory cells,such as immune presenting cells or tumor cells.In this study,we demonstrated that pure resting Vγ9Vδ2 T lymphocytes can self-activate through exogenous PAgs,involving their TCR and the butyrophilins BTN3A1 and BTN2A1.This is the first time that these three molecules,concurrently expressed at the plasma membrane of Vγ9Vδ2 T cells,have been shown to be involved together on the same and unique T cell during PAg activation.Moreover,the use of probucol to stimulate the inhibition of this self-activation prompted us to propose that ABCA-1 could be implicated in the transfer of exogenous PAgs inside Vγ9Vδ2 T cells before activating them through membrane clusters formed byγ9TCR,BTN3A1 and BTN2A1.The self-activation of Vγ9Vδ2 T cells,which leads to self-killing,can therefore participate in the failure ofγδT cell-based therapies with exogenous PAgs and should be taken into account.

大麻二酚对载脂蛋白E基因敲除小鼠空间记忆障碍的调节及作用机制研究

目的研究大麻二酚(CBD)对载脂蛋白E(ApoE)基因敲除(ApoE^(-/-))小鼠空间记忆障碍的调节作用及其作用机制。方法将18只ApoE^(-/-)小鼠采用随机数字表法分为ApoE^(-/-)组和ApoE^(-/-)+CBD组,每组9只。ApoE^(-/-)+CBD组小鼠每天予CBD 10 mg/kg腹腔注射,ApoE^(-/-)组小鼠腹腔注射等量生理盐水,连续注射21 d。最后一次注射后,通过旷场实验和新物体识别实验研究CBD对ApoE^(-/-)小鼠学习记忆能力和焦虑情绪的影响;采用免疫印迹法检测小鼠大脑皮层和海马组织中相关蛋白的表达;培养原代神经元,分为野生型(WT)组、WT+CBD组、ApoE^(-/-)组、ApoE^(-/-)+CBD组4组,观察各组神经元的分化情况。结果ApoE^(-/-)+CBD组小鼠的新物体识别指数明显高于ApoE^(-/-)组,差异有统计学意义(P<0.05)。与WT组比较,ApoE^(-/-)组神经元轴突长度和胞体大小明显缩小,差异均有统计学意义(P<0.01);与ApoE^(-/-)组比较,ApoE^(-/-)+CBD组神经元轴突长度和胞体大小均明显增大,差异均有统计学意义(P<0.05)。与ApoE^(-/-)组比较,ApoE^(-/-)+CBD组小鼠大脑皮层中瞬时受体电位阳离子通道亚家族V成员2(TRPV2)蛋白、磷酸化蛋白激酶B(AKT)及磷酸化钙/钙调蛋白依赖性蛋白激酶Ⅱ型亚基α和δ(CaMKIIα/δ)表达水平明显升高,差异均有统计学意义(P<0.05)。结论CBD可抑制ApoE基因敲除小鼠的空间记忆障碍,减弱ApoE基因敲除小鼠大脑神经元分化障碍,这些作用主要是通过TRPV2/AKT/CaMKIIα/δ通路实现的。

抗原受体基因多样性产生机制的起源与演化

动物免疫系统根据其应答方式及存在形式的不同,可以分为天然免疫和适应性免疫.由于适应性免疫的关键器官/分子(比如胸腺//MHC)等只在有颌类脊椎动物中被发现,因此被认为是生命演化到高级阶段的体现,关于适应性免疫系统的起源一直是演化生物学和免疫学研究共同关注的重要问题.本文重点梳理了国内外学者近年来在抗原受体基因多样性产生机制及其演化等方面所取得的进展,尤其是在V(D)J重排机制的转座子起源及RAGL转座子的宿主驯化机制上.本文还总结了无颌类脊椎动物VLR受体及数个进化阶段特有的免疫多样性受体的研究进展.这些研究在完善经典免疫学理论的同时,也为医疗保健领域带来崭新的思路与技术.