抗CD3单抗和rIL-2共刺激诱生扩增人CD3AK的实验研究Ⅴ CD3AK介导的MHC非限制性溶瘤作用的机制分析

MHC(Major histocompatibility complex，MHC)Ⅰ类抗原缺失是一些肿瘤细胞逃逸抗原特异性CTL攻击的重要机制。CTL能否通过MHC非限制性，MHC Ⅰ类非依赖性机制杀伤肿瘤细胞是值得深入探讨的。我们用抗CD3单抗和rIL-2共刺激法诱导的CD3AK作为效应细胞，研究了CD3AK对两株MHC Ⅰ类阴性肿瘤细胞株K562和Daudi杀伤作用，

抗CD3单抗和rIL-2共刺激诱生扩增人CD3AK的实验研究Ⅱ CD3AK的免疫生物学特征

本实验采用倒置显微镜、光学显微镜、电子显微镜和荧光激活的细胞分类器(FACS)分析了抗CD3单抗和rIL-2共刺激诱生扩增的人CD3AK的免疫生物学特征。经抗CD3单抗和rIL-2共刺激后细胞形态学发生明显的变化，细胞异型性明显，有些体积增大，呈圆形或不规则形，往往形成集落：有些细胞体积变小。

抗CD3单抗和rIL-2共刺激诱生扩增人CD3AK的实验研究Ⅲ CD3AK与常规LAK细胞的比较

抗CD3单抗激活的杀伤细胞(Anti-CD3 monoclonal antibody activated killer cells．CD3AK)是用抗CD3单抗(Anti-monoclonal antibody，Anti-CD3 mAb)激活的一类杀伤细胞，具有较强的增殖能力和细胞毒活性。在CD3AK诱生扩增过程中除抗CD3单抗外也需要rIL-2作为生长因子，这就提出一个问题。

The human leucocyte differentiation antigens (HLDA) workshops: the evolv-ing role of antibodies in research, diagnosis and therapy

The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8^th Workshop. We consider what remains to be achieved （including an estimate of the number of leucocyte surface molecules still to be discovered）, and how the field can best move forward.

Expression of Caspase－3 in Cord Blood CD34^＋ Cells during Culture in vitro

Objective: To investigate the expression and significance of caspase-3 protein in CD34^+ cells from cord blood (CB) during culture in vitro with different growth factors. Methods: RT-PCR, Western blot and flow cytometry techniques were used to detect the expression of caspase-3 in CD34^+ CB cells during culture in vitro. Results: Caspase-3 mRNA was constitutively expressed at a low level in freshly isolated CD34^+ cells. The expression of caspase-3 mRNA and protein was upregulated when these cellswere first expanded in suspension culture with growth factors for 3 days. However, only the 32 kDa inactive caspase-3 proenzyme was detected in the freshly isolated CD34^+ cells as well as during the first 3 days expansion with cytokines. With longer culture time in vitro, especially in the presence of the combination of IL-3, IL-6 and GM-CSF, caspase-3 was activated and a cleavage product of 20 kDa became detectable.Conclusion: Caspase-3 is involved in apoptosis of primitive CB CD34^+ cells during expansion in vitro.

Sequencing and expression analysis of CD3γ/δ and CD3ε chains in mandarin fish, Siniperca chuatsi

The genomic and cDNA sequences of the CD3γ/δ and CD3ε homologues in the mandarin fish, Siniperca chuatsi, were determined. As in other vertebrate CD3 molecules, the deduced amino acid sequences of mandarin fish CD3γ/δ and CD3ε contained conserved residues and motifs, such as cysteine residues and CXXC and immunoreceptor tyrosine-based activation motifs. However, mandarin fish CD3γ/δ and CD3ε showed some differences to their mammalian counterparts, specifically the absence of a negatively charged residue in the transmembrane region of CD3γ/δ. Additionally, while an N-glycosylation site was present in CD3c, the site was not observed in CD3γ/δ. The CD3γ/δ and CD3ε subunit sequences contain six and five exons, respectively, consistent with homologues from Atlantic salmon, Salmo salar. Phylogenetic analysis also revealed that CD3γ/δ and CD3ε in mandarin fish are closely related to their counterparts in Acanthopterygian fish. Real-time PCR showed CD3γ/δ and CD3ε were expressed mainly in the thymus and spleen in normal healthy fish and, to a lesser extent, in mucosal-associated lymphoid tissues, such as the intestine and gills. When lymphocytes isolated from head kidney were treated with the mitogens phytohemagglutinin, concanavalin, and polyriboinosinic polyribocytidylic acid, mRNA expression levels of CD3γ/δ and CD3ε were significantly elevated within 12 h of treatment. This indicated the presence of T lymphocytes in the head kidney of teleost fish, and also the recognition of mitogens by the lymphocytes. Mandarin fish infected with the bacterial pathogen Flavobacterium columnare also showed an increase in the expression of CD3γ/δ and CD3ε mR_NA, indicating that CD3γ/δ and CD3ε lymphocytes are involved in the immune response of this species.

Mn doping effects on the gate-tunable transport properties of Cd3As2 films epitaxied on GaAs

The Mn doping effects on the gate-tunable transport properties of topological Dirac semimetal Cd3As2 films have been investigated.Mn-doped Cd3As2 films are directly grown on GaAs(111)B substrates by molecular-beam epitaxy,during which the single crystal phase can be obtained with Mn concentration less than 2%.Shubnikov-de Haas oscillation and quantum Hall effect are observed at low temperatures,and electrons are found to be the dominant carrier in the whole temperature range.Higher Mn content results in smaller lattice constant,lower electron mobility and larger effective band gap,while the carrier density seems to be unaffected by Mn-doping.Gating experiments show that Shubnikov-de Haas oscillation and quantum Hall effect are slightly modulated by electric field,which can be explained by the variation of electron density.Our results provide useful information for understanding the magnetic element doping effects on the transport properties of Cd3As2 films.

Preparation and Bioactivity Assay of CD3AK Cell from Lmbilical Cord Blood：—Clinic Report of 10 Tumor Cases

Objective To study the anti-tumor effect of CD3AK cell prepared from umbilical blood,to explore the short-term curative effect on tumor cases and seek better immune index for biotherapy.Methods IL-2 and IL-2+CD3Ab were used to induce LAK cells and CD3AK cells isolated from umbilical blood mononuclear cells(UBMC).The expanding number and bioactivity of LAK cells and CD3AK cells were examined at different time points after culture,the NK activity of peripheral blood mononuclear cells(PBMC)of 10 cases of malignant tumor were determined before and after CD3AK cell adopting immune therapy as well.Results The number and bioactivity(NK killing K562 cell)of LAK and CD3AK cells reached their peaks on 11 th day.The number of LAK and CD3AK cells were 18 folds and 24 folds of that before culture;The NK activities of LAK and CD3AK against K562 were 2.6 folds and 3.2 folds of those before culture respectively.The nK activity for killing K562 cells of malignant tumor patient's PBMC was increased from 63%-81% by CD3AK cell transfusing,rising mean 28%.Conclusion (1)The UBMC is a potential and better source of predecessor for LAK and CD3AK;(2)The NK activity of LAK and CD3AK cells from UBMC reached their peaks at 11 th day after culture,and the NK aftivity of CD3AK cells in much greater than that of LAK cells;(3)The NK activity of malignant tumor patient's PBMC can be obviously elevated by transfusing CD3AK cell(4)The test of NK activity of PBMC of malignant tumor patient may become an objective immune index for tumor biotherapy.

The study of special killing effect of CD3AK on anti-CEA-positive tumor enhanced by DC loaded with CEA-rV

Objective： To survey the special killing activity of CD3AK on anti-CEA-positive tumor enhanced by umbilical cord blood dendritic cell （DC） loaded with CEA recombinant vaccinia virus （CEA-rV）. Methods： Freshly isolated umbilical blood mononuclear calls （UBMC） were cultivated for 3 h. Suspension cells and attached calls were used to induce CD3AK calls and DC separately. DC was loaded with CEA-rV on the 3rd day to prepare CEA-rV＋DC. CD3AK cells were co-cultured with CEA- rV＋DC on the 8th day, to prepare CEA-rV＋DC＋CD3AK. The killing activity of each effector＇s cell, which included UBMC, CD3AK, DC＋CD3AK and CEA-rV＋DC＋CD3AK, was measured respectively by MTT reduction assay. Results： （1） 4 target cells were con- firmed by CEA monoclonal antibody of rabbit anti-human. Lovo and A549 were really CEA positive cell lines, while Bel-7402 and K562 were CEA negative cell lines. （2） It was showed by flow-cytometry that the mature DC cultured at 10th day expressed MHC I, II molecules such as CD86, CDS0, CD83 and CD40 highly, but CD123 lowly. The expression rates of CD86, CDS0, CD83 and CD40 was 82.7%, 51.1%, 57.5% and 69.4%, respectively. The appearances and intra-cellular structures of DC were observed through light and electron microscope. The diameter of mature DC was 15-20 μm presented the irregular morphologic appearanca, much prominences and pseudopodium. There were abundant mitochondria and endoplasmic reticulum in DC endochylema. （3） The rates of CD3, CD4, CD8 and CD28 in CD3AK cells group were 2 folds higher than that in UBMC group by FACS. It was said that the numbers of the mature T lymphocyte in CD3AK cells group were much greater than that in UBMC group. （4） The killing activities to 4 target cells of 3 effector＇s cells, which included CEA-rV＋DC＋CD3AK, DC＋CD3AK and CD3AK, were much greater than that of UBMC （P〈0.01）. Moreover, comparing with the killing activities of 4 effector＇s： CEA-rV＋DC＋CD3AK group 〉 DC＋CD3AK group 〉 CD3AK group 〉 UBMC group. It showed that, cytokine, DC and CEA-rV could efficiently elevate the killing activity of UBMC on broad-spectrum tumor cells. （5） Comparing with the killing activities of CEA-rV＋DC＋CD3AK and CD3AK cells to CEA positive and negative cells, the killing activities of CEA-rV＋DC＋CD3AK to CEA positive tumor calls, Lovo and A549 calls （P〈0.01） were remarkably better than that to CEA negative tumor cells BEL-7402 and K562 cells （P〈0.05）. It was said that the CEA-rV＋DC could obviously enhance the killing activity of CD3AK on CEA positive tumor cells. Comparing with the killing activities of CEA-rV＋DC＋CD3AK and DC＋CD3AK cells, the killing activity of CD3AK on CEA negative tumor cells was no statistical difference （P〉0.05）. However, the killing activity to CEA positive cells of CEA-rV＋DC＋CD3AK group was notably higher than that of DC＋CD3AK group. Namely, CEA-rV could distinctly promote the special killing activity to CEA positive tumor cells of CD3AK, but could not do it to CEA negative tumor cells. Conclusion： CEA-rV＋DC could obviously enhance the special killing activity of CD3AK on CEA positive tumor cell lines, while the DC only couldn＇t. The results indicated that the CEA-rV played an important role during the special killing activity of CD3AK cells to CEA positive tumor cells.

GLTSCR1, ATM, PPPIR13L and CD3EAP Genetic Variants and Lung Cancer Risk in a Chinese Population

Genetic variants in glioma tumor suppressor candidate region gene 1 （GLTSCR1） and ATM serine/threonine kinase （ATM） have been associated with various cancer risks. Epidemiological studies also revealed the association of variants of GLTSCR1 and ATM genes with different brain tumors. However, little is known about the relationship between both gene polymorphisms and lung cancer risk. We conducted a Chinese hospital-based casecontrol study involving 384 lung cancer cases and 387 cancer-free controls. No significant differences in the single polymorphism （GLTSCR1 rs1035938 and ATM rs11212592） association were found in five genetic models （co-dominant, dominant, recessive, overdominant and log-additive models） （adjusted by smoking duration）. Join effect of three SNPs （PPPIR13L rs1970764, CD3EAP rs967591, GLTSCR1 rs1035938） on chromosome 19q 13.3 showed that the designated haplotype2 （rs 1970764 G-rs967591 A-rs 1035938 C） [OR （95% CI）=1.60 （1.11-2.32）, P=0.012] and haplotype8 （rs 1970764 G-rs967591 G-rs 1035938 T） [OR （95% CI）=2.45 （1.17-5.12）, P=0.018] were associated with increased risk of lung cancer （adjusted by smoking duration）. The analysis ofmultifactor dimensionality reduction revealed that two 3-way models were the best fit models in analyses of 2 loci （P〈0.001） or 4 loci （P=0.015-0.016）. The entropy-based analysis indicated the strongest synergistic effect between PPPIR13L rs1970764 and ATM rs11212592 in analysis of four genes. In conclusion, our study suggests that haplotypes consisting of PPPIR13L rs1970764- CD3EAP rs967591-GLTSCR1 rs1035938 on Chr19q13.3, interaction of smoking and GLTSCR1 rs1035938-ATM rs11212592, and synergistic action of PPPIR13L rs1970764 and ATM rs 11212592 may associate with lung cancer risk in the Chinese population.

Immunohistochemical CD3 staining detects additional patients with celiac disease

AIM: To investigate whether performing immuno-histochemical CD3 staining, in order to improve the detection of intra-epithelial lymphocytosis, has an additional value in the histological diagnosis of celiac disease.METHODS: Biopsies obtained from 159 children were stained by hematoxylin and eosin(HE) and evaluated using the Marsh classification. CD3 staining was subsequently evaluated separately and independently.RESULTS: Differences in evaluation between the routine HE sections and CD3 staining were present in 20(12.6%) cases. In 10(6.3%) patients the diagnosis of celiac disease(Marsh Ⅱ and Ⅲ) changed on examination of CD3 staining: in 9 cases, celiac disease had initially been missed on the HE sections, while 1 patient had been over-diagnosed on the routine sections. In all patients, the final diagnosis based on CD3 staining, was concordant with serological results, which was not found previously. In the other 10(12.3%) patients, the detection of sole intra-epithelial lymphocytosis(Marsh Ⅰ) improved. Nine patients were found to have Marsh Ⅰ on CD3 sections, which had been missed on routine sections. Interestingly, the only patient with negative serology had Giardiasis. Finally, in 1 patient with negative serology, in whom Marsh Ⅰ was suspected on HE sections, this diagnosis was withdrawn after evaluation of the CD3 sections.CONCLUSION: Staining for CD3 has an additional value in the histological detection of celiac disease lesions, and CD3 staining should be performed when there is a discrepancy between serology and the diagnosis made on HE sections.

Community Normal Reference Values for CD3+, CD4+, CD8+T Lymphocytes and Leucocytes among Immunocompetent Adults in Coastal Kenya

Background: Studies on the reference values of CD4 and CD3 T cells in healthy individuals have continued to gain significance because of the importance of these immunological markers in the initiation of antiretroviral therapy and prophylactic drugs for opportunistic infections. These ranges tend to vary across populations. The CD4:CD8 ratio is used to measure of how balanced immune function is. Therefore, this study aimed at determining normal reference values for CD4+ and CD3+T-lymphocytes and leucocytes in healthy adults in Coastal Kenya. Methods: A cross-sectional study was carried between May 2015 and February 2016 in Coast General Referral hospital, Tudor, Port-Reitz, Mlaleo, Likoni and Sub-County hospitals. Participants were recruited from voluntary HIV counselling and testing clinics. Patients were counselled for HIV test and those who consented were tested for HIV. They were screened for diseases that potentially cause lymphocyte homeostasis perturbation. CD4+, CD3+ CD8+cells/μl were analyzed using a BD FACSCount flow cytometer (Becton-Dickson, NJ). Results: We enrolled 500 participants, two hundred and forty (48.0%) were males and two hundred and sixty (52.0) females. The mean CD4 cell count was 1054.9 ± 95% CI 1041.2 - 1068.6 cells/mm3, absolute CD8 was 688.4 ± 95% CI 679.1 - 697.7 cells/mm3, absolute CD3 cell count was 1945.1 ± 95% CI 1907.4 - 1982.2 cells/mm3 absolute leukocyte count 5.19 ± 95% CI 5.12 - 5.19, absolute lymphocyte count 1.85 ± 95% CI1.83 - 1.88 and haemoglobin level 12.76 ± 95% CI 12.65 - 12.87. Females had significantly higher mean CD4 and CD8 T cell counts than males (p < 0.05). The mean values of white blood cells 4.7 (3.0 - 7.9) × 109/l, platelets 239 (77 - 353) × 109/l and erythrocytes 4.65 (3.51 - 5.40) × 109 were significantly higher in males than females (p Conclusion: Immunohaematological markers found in this study were different from the standard values for the western countries. Females had significantly higher mean CD4+T and CD3+T cell counts but lower mean haemoglobin level, erythrocytes, white blood cells and platelets than males. Our findings provide new insight in the CD4 and CD3 T cell reference values of Kenyans.

The effect of anti-CD28 on the CD3-AK proliferation and tumoricidal activity

Objective The aim of this study was to costimulate the CD3-AK cells with anti-CD28 monoclonal antibody (mAb), observe the effect of cell proliferation and cytotoxicity and explore the regulatory role of CD28 mAb on the CD3-AK tumoricidal activity.

Research on the effects of CD137 signaling on the function of CD3^-CD56^+NK cells

Objective： To investigate the effects of CD137 signaling on the regulation of CD3-CD56＋NK cells function. Methods： CD3 CD56＋NK cells were treated with CD137 mAb or mouse IgG 1 isotype control to study the effects of CD 137 signaling on the function of CD3-CD56＋NK cells. Cytotoxicity was measured by LDH activity in the supernatants of cell cultures; NKG2D and LFA-I expression on CD3-CD56＋NK cells were analyzed by flow cytometry. Results： CD137 was expressed on activated CD3-CD56＋NK cells. The CD137 mAb enhanced the ability of CDB-CD56＋NK cells to kill lung cancer cells（A549）; Further studies revealed that the expression of NKG2D and LFA-1 was significantly increased in activated cells, and blockade of NKG2D and LFA-1 dramatically attenuated CD3CD56＋NK cytolysis of A549 cancer cells. Conclusion： CD 137 signaling increases the ability of CD3-CD56＋NK cells to kill cancer cells via up- regulating the expression of NKG2D and LFA-1.

不再让顶级货爱家们苦苦空等 Gamut CD3 CD播放机

来自丹麦的Hi-End品牌Gamut推出新品的速度一向非常慢,让很多爱家们等得难受。不过,自从Lars Goller在两年前接手这个品牌后,这种情况有所改变了,平均每年都有二到三样产品问世。现在,Gamut的前任顶级CD播放机CD1已经被新皇者CD3所替代。从外形上讲,CD3和本厂的DI 150合并式放大器风格一致,不过所有人关注的是：CD3的音质能够达到并超过CD1的水平吗？

永远的怀想 Micromega CD3与Marantz CD-63SE CD机赏析

经常有发烧友请我推荐超值靓声的CD 机,在过去我会毫不犹豫地推荐 Marantz CD-63SE。今天,我则又会 加上一款Micromega(美格)的MiniumCD3。推荐使用它们的理由是:价格平实,性价比高,能在一定程度上代表该品牌的声音风格,有较为广泛的使用和认可的发烧群体。

Audio Research CD3唱机

国外以真空管设计制作放大器的厂家很多，若要请资深音响迷随便举例，Audio Research绝对会是前几个被提到的品牌。Audio Research以制造真空管放大器闻名于全世界，不过在进入数字化时代之后，他们除了坚持真空管前后级放大器的设计生产之外，也开始尝试制造数字信号源。从最早的CD1到后来的CD2，都获得不错的评价，温润厚实的声音走向，更是让许多音响迷深爱不已。

CH50 CD3^(+)及CD4^(+)在新生儿急性呼吸窘迫综合征患儿中的表达及意义

目的探讨新生儿急性呼吸窘迫综合征患儿血清总补体活性(CH50)、CD3^(+)及CD4^(+)的表达特点,分析CH50、CD3^(+)、CD4^(+)与新生儿急性呼吸窘迫综合征的关系。方法选取2019年4月—2022年5月丽水市中心医院收治的96例新生儿急性呼吸窘迫综合征患儿为呼吸窘迫组,依据病情严重程度分为轻度组、中度组、重度组。选取同期在丽水市中心医院进行体检的54例健康儿童为健康对照组。采用酶联免疫吸附试验检测血清CH50水平,使用流式细胞仪检测CD3^(+)、CD4^(+)数量。结果呼吸窘迫组CH50、CD3^(+)及CD4^(+)水平均低于健康对照组,差异均有统计学意义(均P<0.05)。轻度、中度、重度急性呼吸窘迫综合征患儿CH50、CD3^(+)及CD4^(+)水平比较,差异均有统计学意义(均P<0.05)。Spearman相关性分析结果显示:CH50与CD3^(+)表达呈正相关(r=0.457,P<0.05),CH50与CD4^(+)表达呈正相关(r=0.549,P<0.05),CD3^(+)与CD4^(+)表达呈正相关(r=0.438,P<0.05)。CH50、CD3^(+)及CD4^(+)预测新生儿急性呼吸窘迫综合征的受试者工作特征曲线下面积分别为0.797(95%CI:0.726~0.868,P<0.05)、0.725(95%CI:0.627~0.822,P<0.05)、0.680(95%CI:0.592~0.769,P<0.05)。三项联合检测对新生儿急性呼吸窘迫综合征的预测价值最高,曲线下面积为0.815(95%CI:0.763~0.895,P<0.05)。结论新生儿急性呼吸窘迫综合征患儿CH50、CD3^(+)、CD4^(+)水平均显著下降,以截断值为判定标准,低水平的CH50、CD3^(+)、CD4^(+)是新生儿急性呼吸窘迫综合征的风险预测因子,可能与病情预测和发展有一定关联。