我国浙南地区正常人血浆可溶性CD44s、CD44v6水平的调查

目的了解正常人体血液中sCD44的表达水平及其变化规律.方法用本研究中心建立的酶联免疫杂交法定量检测112例正常人血浆sCD44s及sCD44v6的浓度.结果成年人sCD44s检测水平为21.6±8.4U/ml,成年人sCD44v6检测水平为14.4±3.3u/ml.结论男女之间sCD44有差异,血浆sCD44的含量随人体生长发育而变化.     

血清sCD44s及sCD44v6变化与乳腺癌的初步研究

目的:测定血清中sCD44s和sCD44v6含量并探讨其在乳腺癌中的临床意义。方法:用酶联免疫吸附试验(ELISA)检测38例乳腺癌患者及15例乳腺良性疾病患者和40例正常人血清中可溶性CD44s(sCD44s)和可溶性CD44v6(sCD44v6)的水平。结果:乳腺癌患者血清sCD44s和sCD44v6水平明显高于正常人和乳腺良性疾病患者(P＜0.01)。Ⅲ、Ⅳ期患者血清sCD44s和sCD44v6明显高于Ⅰ、Ⅱ期(P＜0.05);乳腺癌手术后1周sCD44v6明显低于手术前(P＜0.05),手术后2周sCD44v6更加明显(P＜0.01);手术后2周sCD44s明显低于手术前(P＜0.05)。结论:血清sCD44s和sCD44v6水平可作为诊断、治疗乳腺癌的辅助指标;乳腺癌血清中sCD44s和sCD44v6升高及降低与肿瘤负荷有关。     

大肠癌CD44v6、p53基因突变体的表达

探明CD44v6 与肿瘤转移的关系, 了解p53 基因突变、CD44v6 在大肠癌中的表达。应用银染PCR SSCP检测p53 基因的突变; 应用特异CD44v6 探针进行Southern blot 杂交, 对杂交条带进行X光片暴光, 并对X 光片上的杂交条带进行灰度扫描, 定性、定量分析CD44v6 的表达。结果显示, 正常对照组、腺瘤组、未转移组和转移组的p53 基因突变率分别为:0 % 、50-0 % 、35-3% 和55-6% 。四个实验组的CD44v6 表达阳性率分别为0 % 、25-0% 、64-7 % 和88-9% ; CD44v6 表达量分别为175-87 ±43-16 、2017-31 ±1429-9、2039-88±1568-78 和10004-47±8013-36 , 转移组CD44v6 表达量明显高于其它实验组。CD44v6 与转移有关; 在肿瘤转移组和未转移组, CD44v6 阳性率明显高于p53 基因突变率。与p53 基因相比,CD44v6 不失是一个肿瘤转移的良好标记     

胃癌中CD44s和CD44v6的表达及意义

目的 研究CD44s和CD44v6的表达与胃癌发生及临床病理学相关性.方法 利用各种类型的胃癌56例、非典型增生18例及正常胃黏膜组织22例,通过免疫组化ABC法检测CD44s和CD44v6的表达.结果 在胃癌、非典型增生及正常胃黏膜中CD44s蛋白的阳性表达率分别为80.4%、72.2%和13.6%;CD44v6蛋白的阳性表达率分别为83.9%、77.8%和4.6%.胃癌及非典型增生中CD44s和CD44v6蛋白的表达与正常胃黏膜相比均差异有显著性(P<0.01).CD44s和CD44v6蛋白在进展期胃癌中浸润到肌层时强阳性表达率分别为60.7%和57.1%;浸润到浆膜层的胃癌中强阳性表达率分别为82.4%和88.2%,与早期胃癌的强阳性率18.2%和9.1%相比均具差异有显著性(P<0.05,P<0.01).有淋巴结转移的胃癌中CD44s蛋白的强阳性表达率为90.5%,CD44v6蛋白的强阳性表达率为85.7%,与无淋巴结转移的胃癌中强阳性表达率为40.0%相比均差异有显著性(P<0.01).结论 CD44s和CD44v6蛋白的高表达与胃癌的发生有密切相关,CD44s和CD44v6蛋白与癌组织浸润深度及淋巴结转移有关,可做为胃癌的早期诊断及预后断定的重要指标之一.     

类风湿性关节炎活动期外周血CT4+T细胞的表型变化

目的观察类风湿性关节炎(RA)活动期外周血CT4+T淋巴细胞的表型变化,探讨其免疫病理机制。方法采用ELISA和双标记免疫荧光法检测50例活动期RA患者和50例健康成人外周血CD4+T细胞表面CD154、CD69的表达以及血清、血浆可溶性CD154(sCD154)的含量。结果活动期RA患者的外周血CD4+T细胞CD154(16.8%±7.9%)和CD69(14.1%±8.2%)的表达水平均显著高于健康人,其血清sCD154(18.56±6.32,ng/ml)和血浆sCD154(8.41±3.51,ng/ml)含量亦分别高于健康人血清sCD154(9.56±4.71,ng/ml)和血浆sCD154(2.98±1.13,ng/ml)。结论CD4+T细胞表达的CD154、CD69以及血浆sCD154含量与RA的活动相关,可以作为RA的辅助诊断、疗效评价和预后判断的有价值的实验室参数。     

老年人T细胞膜型CD28及血清中可溶性CD28的表达及其意义

分析老年人(年龄在60岁以上)外周血T细胞膜型CD28(mCD28)及血清中可溶性CD28(sCD28)的表达水平,探讨该分子在老年人免疫功能改变中的意义。分别收集肝素抗凝及不抗凝的老年人外周血,抗凝血经分离获取单个核细胞,间接免疫荧光及流式细胞仪分析,老年人T细胞mCD28的阳性表达率为男性(41.56%±11.23%)及女性(42.97%±12.02%)。与年轻人[男性(61.51%±17.45%)及女性(62.38%±16.22%)]相比,差别具有显著性(P<0.01)。酶联免疫吸附法(ELISA)检测,老年人血清中sCD28的含量为男性(1.08±0.45)及女性(0.98±0.47)ng/ml,与青年人[男性(0.55±0.16)及女性(0.52±0.13)ng/ml]相比,差别具有显著性(P<0.01)。老年人T细胞mCD28的表达水平下降,而血清中sCD28的含量升高。     

CD44s和CD44v6在宫颈鳞癌中表达及其临床意义

目的：探讨宫颈鳞癌中CD44s和CD44v6的表达及其与临床病理资料的关系。方法：应用免疫组化EnVision两步法对31例宫颈鳞标本中CD44s和CD44v6蛋白表达并进行分析。结果：肿瘤原发灶中CD44s阳性表达率为61．3％（19／31）。CD44v6阳性表达率为93．5％（29／31）,CD44v6阳性率高于CD44s,CD44s阳性表达与临床分期,病理分级和分类无关（P＞0．05）,CD44v6阳性表达与肿瘤细胞分化程度无关,但与浸润程度及分期有关（P＜0．05）,结论：CD44v6基因蛋白与宫颈鳞癌的侵袭,转移相关,可作为预测肿瘤进展和预后的一种有用指标。     

可溶性CD40配体酶联检测试剂盒的研制及其运用

目的 :建立灵敏、特异和稳定的人可溶性CD4 0配体 (sCD4 0L)酶联检测试剂盒。方法 :采用鼠抗人CD4 0L单克隆抗体 1B1为包被抗体 ,识别人CD4 0L不同位点的单克隆抗体 4F1经琥珀酰羟基生物素 (BNHS)标记后为检测抗体 ,建立双抗体夹心的人sCD4 0L酶联检测方法。在此基础上 ,测定 2 0 0例健康供血员血清和血浆中sCD4 0L的含量。结果 :成功研制人sCD4 0L酶联检测试剂盒 ,灵敏度为 0 5ng ml。该试剂盒 4℃放置 3个月 ,离散度 (CV) <± 6 9% ,回收率为 94 7%～ 10 4 8% ,提示该检测系统具有良好的灵敏度、稳定性和准确性 ,与国外商品化试剂盒的分析结果相似。应用该试剂盒测得的健康供血员血清和血浆sCD4 0L含量分别为 9 5 6± 4 71、2 98± 1 13ng ml。结论 :研制成灵敏、特异和稳定的人sCD4 0L酶标检测试剂盒 ,能够对血清和血浆中sCD4 0L进行准确定量分析 ,并首次证实血清和血浆中sCD4 0L水平的差异性     

肾综合征出血热患者血浆和血清可溶性IL-2R水平变化的研究

本文应用双McAb ELISA夹心法检测了HFRS患者血清和血浆中SIL-2R水平。结果表明,急性期患者血浆和血清中sIL-2R水平(分别为199±81U/ml和214±122U/ml)显著地高于恢复期患者(分别为104±37U/ml和97±28U/ml)和正常人(分别为88±31U/ml和71±21U/ml)(P<0.01);在急性期中尤以少尿期升高最明显。表明HFRS患者机体免疫细胞处于高度活化的状态,患者血清和血浆中sIL-2R的检测对于判断病期和指导治疗均有一定的意义。     

抑郁伴焦虑症患者血浆中NGF水平的研究

目的:通过测定抑郁伴焦虑症患者血浆中NGF水平,探讨NGF在抑郁焦虑症中的作用。方法:应用NGF放射免疫分析,检测52名患者血浆和32名正常人血浆中的NGF含量。结果:正常组为(314.72±34.14)pg/ml,抑郁伴焦虑症患者血浆中NGF水平,轻度抑郁伴焦虑组为(365.23±47.48)pg/ml、中度抑郁伴焦虑组为(356.17±53.95)pg/ml及重度抑郁伴焦虑组为(362.70±42.52)pg/ml。正常对照组与3组患者血浆NGF水平相比均明显升高(P<0.001);3组患者间相比,相差不显著。结论:NGF参与了抑郁伴焦虑症的发病过程,反映出机体对神经组织的修复和保护的双重效应。这一指标也为该病的诊断和治疗提供了实验依据。为部队掌控训练方式和强度提供实验依据。     

A role for CD44 in T cell development and function during direct competition between CD44+ and CD44- cells

The role of CD44 in T cell biology remains incompletely understood. Although studies using anti-CD44 antibodies have implicated this cell adhesion molecule in a variety of important T cell processes, few T cell defects have been reported in CD44-deficient mice. We have assessed the requirement for CD44 in T cell development and mature T cell function by analyzing mice in which CD44(-/-) and WT cells were produced simultaneously. In mixed (CD44(-/-) + CD44(+/+)) bone marrow chimeras, production of CD44(-/-) T cells was shown to be reduced compared to WT cells due to inefficient intrathymic development. In addition, mature CD44(-/-) CD8(+) T cells generated a substantially lower response than WT T cells after infection of mice with lymphocytic choriomeningitis virus, with the reduction in response apparent in both lymphoid and non-lymphoid tissues. Overall, these results demonstrate a poor capacity of CD44(-/-) T lineage cells to compete with WT cells at multiple levels, implicating CD44 in normal T cell function.     

CD44s and CD44v6 in diagnosis and prognosis of human bladder cancer

The cell adhesion molecules (CAMs) CD44 standard (CD44s) and its variant 6 (CD44v6) are involved in the progression and invasion of human malignancies. However, discrepancies in the prognostic value of CD44s and CD44v6 expression need to be addressed. Aims: To investigate the expression of CD44s and CD44v6 in bladder carcinomas and relate the results to the established prognostic factors. Materials and Methods: 50 bladder carcinoma specimens, 30 cases with transitional cell carcinoma (TCC: 6 bilharzial and 24 nonbilharzial) and 20 cases with squamous cell carcinoma (SCC: 8 bilharzial and 12 nonbilharzial), were included. Immunohistochemical analysis for CD44s and CD44v6 was carried out using avidin-biotin peroxidase method. Results: The level of both CD44s and CD44v6 in TCC was significantly higher in invasive than in preinvasive tumors and normal urothelium (p < .05). A direct association between the percentage of expression of both markers and the grade of TCC (p < .05) was observed. An inverse correlation between CD44s and SCC was seen, where metaplastic urothelium showed higher expression than invasive carcinomas. No association was observed between the expressions of both CD44s and CD44v6 and bilharzial ova, sex and age of the patient, or size of the tumor. Conclusions: The authors report statistically significant correlation between CD44s and CD44v6 expression and increasing grade and stage of TCC. No such correlation with SCC and with bilharzial cystitis, sex and age of the patient, or size of the tumor was documented.     

CD44和CD44v6基因在胃腺癌组织中的表达及其意义

目的： 研究胃腺癌组织中ＣＤ４４ 的表达及其与淋巴结转移和预后的关系。方法： 应用免疫组化方法, 对１０５ 例胃腺癌组织中ＣＤ４４ 的表达进行了观察, 并对其中６２ 例患者做了随访。结果： ＣＤ４４ 和ＣＤ４４ｖ６ 基因的表达率分别为５４－３ ％ 和４８－６ ％ 。ＣＤ４４ｖ６ 在胃腺癌组织中的表达与癌细胞的分化、浸润深度, 以及临床分期和预后有关（Ｐ＜ ０－０５）, 而ＣＤ４４ 的表达则与上述临床病理指标无关。另外, 抗ＣＤ４４ 和抗ＣＤ４４ｖ６ 抗体的阳性反应, 与癌细胞的淋巴结转移有关（ＣＤ４４ｖ６, Ｐ＜ ０－０１ ; ＣＤ４４ , Ｐ＜ ０－０５） 。结论： ＣＤ４４ 的表达可用于胃癌患者的病情监测, 其中ＣＤ４４ｖ６ 有望作为判断预后的一个指标。     

CD44H and CD44V6 expression in different subtypes of Hodgkin lymphoma.

CD44 is a broadly distributed family of cell surface glycoproteins. The expression of CD44H has been documented in both Hodgkin lymphoma and non-Hodgkin lymphoma. CD44V6 has been associated with more aggressive behavior in non-Hodgkin lymphoma, but such a correlation has not been established in Hodgkin lymphoma. In addition, the utility of CD44 and CD44V6 in the subclassification of Hodgkin lymphoma in paraffin-embedded tissues has not previously been evaluated. The current study included formalin- or methacarn-fixed, paraffin-embedded tissue specimens from 42 patients with Hodgkin lymphoma (25 nodular sclerosis, three interfollicular, four lymphocyte-rich classic Hodgkin, six lymphocyte predominant, and four mixed cellularity). The clinical stage of the study population at initial presentation ranged from stage IA to IVB. Evaluation of CD44H and CD44V6 (Novocastra) was performed by ABC immunoperoxidase technique after heat-induced epitope retrieval. In the six cases of lymphocyte predominant Hodgkin, the neoplastic cells lacked reactivity with CD44H reminiscent of their normal germinal center counterparts. On the other hand, classic Hodgkin lymphoma showed variable membranous and Golgi reactivity in the neoplastic cells in all cases irrespective of disease stage at presentation. In all cases, the neoplastic cells lacked reactivity with CD44V6 except for three one lymphocyte-predominant, one interfollicular, and one nodular sclerosis), all of which represented recurrent cases. In conclusion, CD44 evaluation is useful in the distinction between lymphocyte predominant and classic Hodgkin lymphoma. The presence of CD44H expression has no relation to the clinical stage of the disease at presentation or recurrence. CD44V6 is detected in a minority of cases irrespective of the histologic subtype and its presence may be associated with recurrence. There was no correlation between disease stage at presentation and the expression of CD44V6.     

Expression of CD44 molecules and CD44 ligands during human thymic fetal development: expression of CD44 isoforms is developmentally regulated

It has recently been recognized that CD44 comprises a largefamily of alternatively spliced forms.In the thymus, CD44 hasbeen postulated to play an important role in immature T cellmigration and maturation. In this paper, we have studied theexpression of CD44 molecules and two CD44 ligands, hyaluronan(HA) and fibronectin (FN), during human thymic fetal development.We found that mAbs against all CD44 isoforms (A3D8 or A1G3)reacted with both thymic epithelial (TE) cells and thymocytesbeginning at the time of initial colonization of the human thymusby hematopoietic stem cells at 8.2 weeks of fetal gestation.However, mAbs specific for splice variants of CD44 containingmembrane-proximal inserts (11.24, 11.10 and 11.9) reacted onlywith terminally differentiated TE cells in and around Hassall'sbodies beginning at 16–19 weeks of fetal gestation. Studiesof differentiated versus undifferentiated TE cells in vitroconfirmed the selective expression of CD44 variant isoformson terminally differentiated TE cells. Expression of HA andFN was determined by fluorescence microscopy using either biotlnylated-HAbinding protein or an anti-FN mAb. We found that whereas FNwas present throughout the human fetal thymus beginning at 8.2weeks, HA was not present until 16 weeks of gestational age.These data demonstrate the differential expression of standardversus variant CD44 isoforms during thymic ontogeny and implicateCD44 interactions with ligands other than HA as important inthe earlier stages of humanthymus development     

Diagnostic utility of CD44 standard, CD44v6, and CD44v3-10 expression in adenocarcinomas presenting in serous fluids.

CD44 is an 85 to 90 kd integral transmembrane protein encoded by a single 20-exon gene located on the short arm of chromosome 11. In the standard form (CD44s), 10 of the 20 exons are transcribed. Multiple variant isoforms exist (CD44v1-10) which arise from alternate mRNA splicing of the remaining 10 exons. In contrast to the standard form of CD44, which is almost ubiquitously expressed, splice variants are highly restricted in their expression in normal or malignant tissues. The purpose of this study was to evaluate the extent to which metastatic adenocarcinomas in effusions express CD44s, CD44v6, and CD44v3-10 and to assess their diagnostic utility in distinguishing reactive mesothelial cells from adenocarcinomas. Archival paraffin-embedded cell blocks of serous fluids from 23 cases of benign effusions containing reactive mesothelial cells and 45 cases of malignant effusions with metastatic adenocarcinoma (18 ovarian, 11 pulmonary, 9 gastrointestinal, and 7 breast) were retrieved from the surgical pathology files. The cytopathology of all cases was reviewed to confirm the diagnosis. Immunohistochemistry was performed on all cases using antibodies for CD44s, CD44v6, and CD44v3-10 (Bender MedSystems, CA). Positive staining was defined as distinct linear membrane staining. Strong staining in at least 10% of the tumor cells was required to consider the case positive for the particular marker. In benign effusions mesothelial cells expressed CD44s in 22 cases (96%), CD44v6 in 1 cases (4%) and CD44v3-10 in 0 cases (0%). In contrast neoplastic cells in malignant effusions expressed CD44s in 11 cases (24%), CD44v6 in 21 cases (47%), and CD44v3-10 in 39 cases (87%). We concluded that CD44s and CD44v3-10 are useful markers that can be applied to cytologic specimens. CD44s immunostaining can be used as a reliable marker to identify reactive mesothelial cells, meanwhile CD44v3-10 immunostaining can detect majority of adenocarcinomas in malignant effusions.     

Structural Variability of CD44v Molecules and Reliability of Immunodetection of CD44 Isoforms Using mAbs Specific for CD44 Variant Exon Products

CD44 can be considered structurally and functionally one of the most variable surface molecules. Alternative splicing of variant exons as well as posttranslational modifications of the molecule (differences in glycosylation) generate a rich repertoire of CD44 isoforms (CD44v), some of which seem to play a key role in tumor growth and progression. Immunodetection of CD44 isoforms in vivo, using mAbs specific for CD44 variant exon products, is largely used to identify those CD44 molecules involved in tumor growth and progression and to interfere with CD44-mediated processes. In the present work we demonstrate that the immunoreactivity of some mAbs directed to CD44 exon-specific epitopes can be impaired by the structural variability of the molecule. Our findings demonstrate that (1) specific exon assortment and/or posttranslational modifications of CD44v molecules can mask CD44 exon-specific epitopes; (2) glycosaminoglycan side chains, carried by some CD44v isoforms of high molecular weight, may play a critical role in determining the exact conformation of the molecule, which is necessary for the detection of CD44 variant epitopes by specific mAbs; and (3) in a panel of stable transfectants expressing CD44 N-glycosylation site-specific mutants, generated in the constant region of CD44 extracellular domain, asparagine-isoleucine substitution is sufficient per se to impair the immunoreactivity of several mAbs to pan-CD44. Thus, conformational changes due to the alternative splicing of CD44 variant exons and/or posttranslational modifications of the molecule (different degree of glycosylation), which are cell type-specific, are likely to generate CD44 variants that elude immunodetection. These findings strongly suggest that immunohistochemical analysis of CD44 expression in vitro and in vivo, using mAbs specific for CD44 variant exon epitopes, can potentially be impaired by a large number of false negative results.     

New monoclonal antibodies in CD44 and CD58: their use to quantify CD44 and CD58 on normal human erythrocytes and to compare the distribution of CD44 and CD58 in human tissues.

The cell-surface glycoproteins CD44 and CD58 are involved in cell adhesion reactions. In this paper 12 monoclonal antibodies in CD44 and two in CD58 are described. Competitive binding assays using CD44 antibodies identified three distinct epitope groups. Antibodies in Group 1 and, with one exception (BRIC 214), antibodies in group 2, but not antibodies in Group 3, recognized epitopes that are sensitive to reduction and to trypsin or chymotrypsin treatment of intact erythrocytes, and so these epitopes probably reside on the N-terminal disulphide-bonded domain of CD44. Antibodies in CD44 did not inhibit the binding of CD58 antibodies to erythrocytes or vice versa. Quantitative binding studies using radioiodinated IgG measured 1888-5592 copies of CD44 and 1772-3290 copies of CD58 on normal erythrocytes. Similar measurements with radioiodinated Fab fragments gave values of 6508-10,450 (CD44) and 3457-7622 (CD58). Immunocytochemical studies indicated that CD44 is much more widely expressed in non-haemopoietic tissues than CD58. Comparison with previously described CD44 antibodies suggests that antibodies in our Group 1 encompass Hermes 2 and that those in Group 2 encompass Hermes 1. All the CD44 antibodies gave weakened reactions with Lu(a-b-) erythrocytes of the In(Lu) type by one or more methods. BRIC 214 and antibodies in epitope Group 3 were used to demonstrate that CD44 on these variant cells gives membrane-bound trypsin and chymotrypsin cleavage fragments of similar molecular weight to those obtained with normal erythrocytes.     

Expression of CD 44 s, CD 44 v 3 and CD 44 v 6 in benign and malignant breast lesions: correlation and colocalization with hyaluronan

AIMS: To examine the expression of CD 44 s, CD 44 v 3 and CD 44 v 6 in breast lesions, and to correlate it with the expression of hyaluronan (HA). Methods and results: CD 44 expression was studied in 75 breast tissue samples, consisting of benign, premalignant and malignant breast lesions, using immunohistochemistry. CD 44 s, but not CD 44 v 3 or CD 44 v 6, was found in the stromal cells, and it was similar in benign and malignant tumours. In benign lesions CD 4 v 6 was detected in 20-30% of the ductal epithelial cells, while C 44 v 3 and CD 44 s were not expressed. CD 44 s, CD 44 v 3 and CD 44 v 6 were all up-regulated in the in situ carcinomas and invasive carcinomas. The level of CD 44 expression in carcinoma cells did not correlate with the type or differentiation of the tumours. CD 44 and HA expression levels were not closely linked in the benign or malignant breast lesions, because HA was overexpressed later in breast cancer progression than CD 44. However, in breast carcinomas CD 44 and HA positivity was often found in the same areas of the sections, and the dual staining confirmed actual colocalization of CD 44 s and HA in the same cells. Conclusions: CD 44 s, CD 44 v 3 and CD 44 v 6 are up-regulated earlier than HA in breast carcinoma progression, and in later stages they often colocalize with cell surface HA.