乳腺癌组织中Duffy抗原趋化因子受体表达与雌激素受体表达的关系

目的：探讨乳腺癌组织中Duffy抗原趋化因子受体（DARC）与雌激素受体（ER）表达的关系。方法：中国福利会国际和平妇幼保健院乳腺癌临床组织标本85例，患者年龄28～75（52．68±10．51）岁，通过免疫组化技术检测DARC和ER表达，并对其相关性进行分析。结果：对乳腺癌组织标本的免疫组化检测结果显示：ER阳性表达组DARC阳性和DARC阴性表达分别为29例（69．05％）及13例（30．95％），ER阴性表达组DARC阳性和DARC阴性表达分别为15例（34．88％）及28例（65．12％），ER阳性和ER阴性表达组之间的DARc表达差异具有统计学意义（P〈0．01）。DARC表达与ER表达呈正相关（r=0．487，P〈0．05）。结论：人乳腺癌组织中Duffy抗原趋化因子受体（DARC）表达与ER有关。     

ER、PR、PCNA、C-CrbB-2在子宫肌瘤中的表达及其临床意义

目的　探讨雌激素受体 (ER)、孕激素受体 (PR)、增殖细胞核抗原 (PCNA)及癌基因 (C CrbB 2 )在子宫肌瘤中的表达以及与子宫肌瘤生长的关系。方法　应用免疫组化法测定 5 0例子宫肌瘤内ER、PR、PCNA及C CrbB 2表达的阳性率 ,并根据肌瘤的发病病程、临床症状、肌瘤的直径、数量以及不同的月经周期 (卵泡期和黄体期 )进行分组研究。结果　肌瘤内ER、PR表达的阳性率与肌瘤直径之间有统计学差异。卵泡期肌瘤内的ER表达的阳性率高于黄体期 (P <0 .0 5 )。而PCNA及C CrbB 2表达的阳性率与肌瘤无关系。结论　子宫肌瘤的生长、发展与肌瘤内ER、PR含量的水平密切相关 ,而与细胞分裂增殖的关系不大 ,与癌基因的表达无关     

不同基因亚型乳腺癌组织中Duffy抗原趋化因子受体表达的研究

目的：探讨不同基因亚型乳腺癌组织中Duffy抗原趋化因子受体（duffy antigen receptor for chemokines,DARC）的表达差异。方法：中国福利会国际和平妇幼保健院乳腺癌临床组织标本300例,患者年龄28~80（53.74±11.37）岁,通过免疫组织化学技术检测DARC、雌激素受体（estrogen receptor,ER）、孕激素受体（progesterone receptor,PR）、细胞角蛋白5（cytok-eratin5,CK5）、人表皮细胞生长因子受体（epidermal growth factor receptor,EGFR）、人表皮细胞生长因子受体2（epidermalgrowth factor receptor2,HER2）表达,并分析DARC在不同基因亚型乳腺癌组织中的表达差异。结果：38.60%基底细胞样乳腺癌组织中DARC呈高表达,46.15%HER2型乳腺癌组织中DARC呈高表达,76.92%腔内导管上皮A型乳腺癌组织中DARC呈高表达,55.68%腔内导管上皮B型乳腺癌组织中DARC呈高表达,63.16%类正常腺体型乳腺癌组织中DARC呈高表达。各组之间的DARC表达差异有统计学意义（P〈0.01）。结论：不同基因亚型乳腺癌组织中DARC表达有显著差异。     

儿童抽动-秽语综合征HCMV pp65抗原检测的临床意义

目的探讨儿童巨细胞病毒(Hum an cytom egalov irus,HCMV)活动性感染与抽动-秽语综合征(T ourette’s sym-drom e,TS)的关系。方法离心分离64例抽动-秽语综合征患儿和40例正常儿童外周血单个核细胞(PBM C s)和血浆,分别用免疫荧光法和定量PCR检测HCMV pp65抗原和HCMV DNA,并比较2种方法的一致性。结果64例TS患儿HCMV pp65抗原有14例阳性,阳性率21.9%,40例正常儿童HCMV pp65抗原有1例阳性,阳性率2.5%,2组儿童HCMV感染率有显著性差异(2χ=7.49,P<0.01)。免疫荧光法和实时定量PCR有较好的一致性(89.1%)。结论抽动-秽语综合征儿童HCMV感染率显著高于正常儿童,HCMV活动性感染可能诱发抽动-秽语综合征。     

Abnormal chemokine-induced responses of immature and mature hematopoietic cells from motheaten mice implicate the protein tyrosine phosphatase SHP-1 in chemokine responses.

Chemokines regulate a number of biological processes, including trafficking of diverse leukocytes and proliferation of myeloid progenitor cells. SHP-1 (Src homology 2 domain tyrosine phosphatase 1), a phosphotyrosine phosphatase, is considered an important regulator of signaling for a number of cytokine receptors. Since specific tyrosine phosphorylation of proteins is important for biological activities induced by chemokines, we examined the role of SHP-1 in functions of chemokines using viable motheaten (me(v)/me(v)) mice that were deficient in SHP-1. Chemotactic responses to stromal call-derived factor 1 (SDF-1), a CXC chemokine, were enhanced with bone marrow myeloid progenitor cells as well as macrophages, T cells, and B cells from me(v)/me(v) versus wild-type (+/+) mice. SDF-1-dependent actin polymerization and activation of mitogen-activated protein kinases were also greater in me(v)/me(v) versus +/+ cells. In contrast, immature subsets of me(v)/me(v) bone marrow myeloid progenitors were resistant to effects of a number of chemokines that suppressed proliferation of +/+ progenitors. These altered chemokine responses did not appear to be due to enhanced expression of CXCR4 or lack of chemokine receptor expression. However, expression of some chemokine receptors (CCR1, CCR2, CCR3, and CXCR2) was significantly enhanced in me(v)/me(v) T cells. Our results implicate SHP-1 involvement in a number of different chemokine-induced biological activities.     

乙肝核酸疫苗接种小鼠后特异性抗原表达及CTL应答的动态观察

目的:观察乙肝病毒(hepatitis B virus,HBV)核酸疫苗pCMV-S2S免疫小鼠后,接种局部HBV preS2S抗原的表达及特异性细胞毒性T淋巴细胞(CTL)应答的动态变化。方法:将pCMV-S2S接种到BALB/c小鼠胫前肌,分别以免疫组化法检测接种局部肌肉HBV preS2S抗原表达的动态变化,乳酸脱氢酶(LDH)释放法检测特异性CTL活性的动态变化。结果:小鼠接种pCMV-S2S后,局部肌肉细胞3d后已有少量目的抗原HBV preS2S蛋白表达,4周时表达最强,至6月时仍可检出有preS2S蛋白表达,但表达强度明显减弱。小鼠接种pCMV-S2S3天后,特异性CTL活性平均值最高为21·67%,12周时CTL活性平均值最高为35·14%,6月时仍可检出较弱的CTL活性。结论:HBV核酸疫苗pCMV-S2S能在小鼠体内持续表达特异性抗原及诱生特异性CTL应答。     

慢性乙型肝炎患者肝组织胰岛素受体底物2的表达及其酪氨酸磷酸化的研究

目的 观察慢性乙型肝炎(CHB)患者肝组织胰岛素受体底物2(IRS-2)的mRNA和蛋白表达及其酪氨酸磷酸化程度,探讨IRS-2在CHB患者胰岛素抵抗(IR)发生中的作用.方法 选择6例肝功能正常者(对照组)及18例CHB患者,CHB患者再按HOMA模型计算的胰岛素抵抗指数分为CHB非IR(＜2.69)组(10例)和CHB合并IR(＞2.69)组(8例).所有患者于术中及肝穿刺后留取肝组织,采用逆转录-聚合酶链反应检测IRS-2 mRNA表达;采用蛋白质免疫印迹法检测IRS-2蛋白表达;采用免疫沉淀及增强化学发光法检测IRS-2酪氨酸磷酸化程度.结果 CHB非IR组肝组织IRS-2 mRNA(0.38±0.06)、蛋白(0.94±0.18)表达及酪氨酸磷酸化程度(0.78±0.09)较对照组(分别为0.45±0.11、0.99±0.20、1.00±0.23)有所降低,但差异无统计学意义(均P＞0.05);CHB合并IR组IRS-2mRNA(0.26±0.08)、蛋白(0.67±0.11)表达及酪氨酸磷酸化程度(0.63±0.14)均较对照组明显降低,差异有统计学意义(P＜0.05或P＜0.01).结论 CHB合并IR患者肝组织IRS-2的mRNA和蛋白表达及酪氨酸磷酸化程度的降低可能是产生IR的机制之一.     

A Nongenomic Mechanism for Progesterone-mediated Immunosuppression: Inhibition of K+ Channels,Ca2+ Signaling,and Gene Expression in T Lymphocytes

The mechanism by which progesterone causes localized suppression of the immune response during pregnancy has remained elusive. Using human T lymphocytes and T cell lines, we show that progesterone, at concentrations found in the placenta, rapidly and reversibly blocks voltage-gated and calcium-activated K⁺ channels (K_V and K_Ca, respectively), resulting in depolarization of the membrane potential. As a result, Ca²⁺ signaling and nuclear factor of activated T cells (NF-AT)-driven gene expression are inhibited. Progesterone acts distally to the initial steps of T cell receptor (TCR)-mediated signal transduction, since it blocks sustained Ca²⁺ signals after thapsigargin stimulation, as well as oscillatory Ca²⁺ signals, but not the Ca²⁺ transient after TCR stimulation. K⁺ channel blockade by progesterone is specific; other steroid hormones had little or no effect, although the progesterone antagonist RU 486 also blocked K_V and K_Ca channels. Progesterone effectively blocked a broad spectrum of K⁺ channels, reducing both Kv1.3 and charybdotoxin–resistant components of K_V current and K_Ca current in T cells, as well as blocking several cloned K_V channels expressed in cell lines. Progesterone had little or no effect on a cloned voltage-gated Na⁺ channel, an inward rectifier K⁺ channel, or on lymphocyte Ca²⁺ and Cl⁻ channels. We propose that direct inhibition of K⁺ channels in T cells by progesterone contributes to progesterone-induced immunosuppression.     

Expression of Recombination Activating Genes in Germinal Center B Cells: Involvement of Interleukin 7 (IL-7) and the IL-7 Receptor

Mouse germinal center (GC) B cells have been shown to undergo secondary V(D)J (V, variable; D, diversity; J, joining) recombination (receptor editing) mediated by the reexpressed products of recombination activating gene (RAG)-1 and RAG-2. We show here that interleukin (IL)-7 as well as IL-4 was effective in inducing functional RAG products in mouse IgD⁺ B cells activated via CD40 in vitro. Blocking of the IL-7 receptor (IL-7R) by injecting an anti– IL-7R monoclonal antibody resulted in a marked suppression of the reexpression of RAG-2 and subsequent V(D)J recombination in the draining lymph node of immunized mice, whereas RAG-2 expression was not impaired in immunized IL-4–deficient mice. Further, these peripheral B cells activated in vitro or in vivo were found to express IL-7R. These findings indicate a novel role for IL-7 and IL-7R in inducing receptor editing in GC B cells.     

B淋巴细胞刺激因子及其受体在多发性骨髓瘤中的表达及其意义

目的 探讨B淋巴细胞刺激因子(BLyS)及其受体在多发性骨髓瘤(MM)中的表达及其意义.方法 采用流式细胞术(FCM)分析人MM肿瘤细胞系KM3及CZ-1细胞表面BLyS及其受体的表达;RT-PCR及Western blot进一步验证Blys及其受体的表达;采用荧光免疫细胞化学法和激光共聚焦技术鉴定KM3细胞中Blys蛋白的表达与定位;WST细胞增殖实验检测Blys对MM肿瘤细胞生长与存活的影响.ELISA及荧光定量.聚合酶链反应(RTQ-PCR)测定MM患者BLyS蛋白与mRNA的表达水平.同时分析乳酸脱氢酶(LDH)、β2微球蛋白浓度与BLyS蛋白及mRNA表达水平的关系.结果 ①MM细胞能够表达BLyS及其受体;②BLyS定位表达于KM3细胞的浆膜上;③Blys促进MM细胞的生长与存活;④MM患者BLyS蛋白或mRNA表达水平显著高于正常对照组(P值均<0.01);⑤直线相关分析显示LDH、β2微球蛋白浓度与BLyS蛋白及mRNA表达水平呈显著相关性.结论 BLyS及其受体对于MM发生、发展可能起着重要的作用.     

A New Monoclonal Antibody,mAb 4A12, Identifies a Role for the Glycosaminoglycan (GAG) Binding Domain of RANTES in the Antiviral Effect against HIV-1 and Intracellular Ca2+ Signaling

The β-chemokine RANTES (regulated on activation, normal T cell expressed and secreted) suppresses the infection of susceptible host cells by macrophage tropic strains of HIV-1. This effect is attributed to interactions of this chemokine with a 7-transmembrane domain receptor, CCR5, that is required for virus–cell fusion and entry. Here we identify domains of RANTES that contribute to its biological activities through structure–function studies using a new monoclonal antibody, mAb 4A12, isolated from mice immunized with recombinant human RANTES. This monoclonal antibody (mAb) blocked the antiviral activity of RANTES in infectivity assays with HIV-1_Bal, and inhibited the mobilization of intracellular Ca²⁺ elicited by RANTES, yet recognized this chemokine bound to cell surfaces. Epitope mapping using limited proteolysis, reversed phase high-performance liquid chromatography, and mass spectrometry suggest that residues 55–66 of RANTES, which include the COOH-terminal α-helical region implicated as the glycosaminoglycan (GAG) binding domain, overlap the determinant recognized by mAb 4A12. This is supported by affinity chromatography studies, which showed that RANTES could be eluted specifically by heparin from a mAb 4A12 immunoaffinity matrix. Removal of cell surface GAGs by enzymatic digestion greatly reduced the ability of mAb 4A12 to detect RANTES passively bound on cell surfaces and abrogated the ability of RANTES to elicit an intracellular Ca²⁺ signal. Taken together, these studies demonstrate that the COOH-terminal α-helical region of RANTES plays a key role in GAG-binding, antiviral activity, and intracellular Ca²⁺ signaling and support a model in which GAGs play a key role in the biological activities of this chemokine.     

Inflammatory processes in schizophrenia: a promising neuroimmunological target for the treatment of negative/cognitive symptoms and beyond

Emerging evidence indicates that schizophrenia is associated with activated peripheral and central inflammatory responses. Such inflammatory processes seem to be influenced by a number of environmental and genetic predisposition factors, and they may critically depend on and contribute to the progressive nature of schizophrenic disease. There is also appreciable evidence to suggest that activated inflammatory responses can undermine disease-relevant affective, emotional, social, and cognitive functions, so that inflammatory processes may be particularly relevant for the precipitation of negative and cognitive symptoms of schizophrenia. Recent clinical trials of anti-inflammatory pharmacotherapy in this disorder provide promising results by showing superior beneficial treatment effects when standard antipsychotic drugs are co-administered with anti-inflammatory compounds, as compared with treatment outcomes using antipsychotic drugs alone. Given the limited efficacy of currently available antipsychotic drugs to ameliorate negative and cognitive symptoms, the further exploration of inflammatory mechanisms and anti-inflammatory strategies may open fruitful new avenues for improved treatment of symptoms undermining affective, emotional, social and cognitive functions pertinent to schizophrenic disease.