浆膜腔积液中转移性腺癌与间皮细胞的病理学研究

目的探讨浆膜腔积液中转移性腺癌与反应性间皮细胞增生的鉴别.方法选取胸、腹水转移癌48例及反应性间皮细胞增生30例,采用常规涂片HE染色观察两者形态学特点,并应用免疫细胞化学SP法检测HBME-1、calretinin、E-cadherin、MOC-31和BerEP4的表达.结果腺癌和间皮细胞两者形态各有特点,但是分化好的腺癌和明显增生的间皮细胞仅从常规涂片难以鉴别;免疫细胞化学显示反应性间皮细胞HBME-1和calretinin阳性率为86.7%和76.7%;而腺癌细胞只有少量表达,特异性高达95.8%和100%.MOC-31、E-cadherin和BerEP4对转移性腺癌细胞阳性率为70.8%、77.1%和88.4%,间皮细胞表达很少,特异性分别为90%、93.3%、93%.两者对5种抗体的表现差异显著.结论应用常规涂片和免疫细胞化学相结合的方法对鉴别转移性腺癌和间皮细胞增生有很大的帮助,最好应用一组抗体综合分析判断,HBME-1、Calretinin、E-cadherin、MOC-31及BerEP4是目前非常有效的组合.     

Orosomucoid Content of Pleural and Peritoneal Effusions

22 nonneoplastic, noninflammatory effusions (cirrhosis and congestive heart failure), 12 non-neoplastic inflammatory effusions (tuberculosis, lupus erythematosus, rheumatoid arthritis, and idiopathic pleuropericarditis), and 58 neoplastic effusions (cancer of lung, breast, ovary, and pancreas, and lymphoma) were analyzed by radial immunodiffusion for orosomucoid concentration. The average concentration +/-SE was 35+/-4, 65+/-17, and 130+/-13 mg/100 ml in the three types of effusion, respectively. By gel filtration and ion exchange chromatography, orosomucoid was isolated from 12 nonmalignant and 14 malignant fluids. The orosomucoid preparations reacted as single components in acrylamide gel electrophoresis at pH 9.0, and in immunodiffusion and immunoelectrophoresis against antisera to human serum and to human plasma orosomucoid. In radial immunodiffusion, the slope of the line relating concentration to the square of the diameter of the precipitate area was identical for orosomucoid isolated from normal human plasma and from nonneoplastic effusions, but was subnormal for orosomucoid isolated from neoplastic fluids. All orosomucoid preparations had normal amino acid composition. Orosomucoid from the nonmalignant effusions had normal carbohydrate content. 11 of 14 samples of orosomucoid isolated from neoplastic fluids had abnormalities in carbohydrate composition, consisting of subnormal content of sialic acid (11 of 14), hexose (10 of 14), and hexosamine (3 of 14), and abnormally high content of hexosamine (4 of 14).Discriminant analysis showed that concentration of orosomucoid distinguished between neoplastic and nonneoplastic noninflammatory effusions more effectively than concentration of total protein, albumin, alpha(1), alpha(2), beta, or gamma-globulin.     

一组单抗鉴别浆膜转移腺癌细胞与反应性间皮细胞的意义

目的 为提高浆膜转移腺癌细胞的诊断率,探讨一组正反互补单克隆抗体鉴别浆膜转移腺癌细胞与反应性间皮细胞的意义。方法 用低分子量细胞角蛋白（CK^LMW)、癌胚抗原（CEA）及间皮细胞（MC）3种标记物,对50份浆膜腔积液及14份腹腔冲洗液内的细胞进行免疫细胞化学标记。结果 3组细胞与CK^LMW、CEA及MC的表达率分别为：腺癌细胞（95.83%,70.83%,16.67%;可疑癌细胞100%,50.00%,不表达;反应性间皮细胞33.33%,33.33%,66.66%;细胞病理学最初分类的4例可疑癌细胞和2例反应性间皮细胞应归属于腺癌细胞。CK^LMW、CEA及MC3种抗体的敏感度、特异度及可用度分别为96.30%,80.00%,77.10%;70.37%,90.00%,61.60%;90.00%,85.20%,75.30%。结论 免疫细胞化学CK^LMW、CEA及MC联合应用在鉴别浆膜转移腺癌细胞与反应性间皮细胞时,是一项具有重要价值的辅助手段。     

联合检测B72.3、癌胚抗原、神经特异性钙结合蛋白、血栓素在浆膜腔积液细胞学鉴别诊断中的价值

目的探讨B72.3、癌胚抗原、神经特异性钙结合蛋白、血栓素组合检测在浆膜腔积液中恶性间皮瘤和增生间皮细胞与腺癌鉴别诊断的价值。方法取123例浆膜腔积液,经活检或临床资料证实为转移腺癌89例、恶性间皮瘤8例、增生间皮细胞26例,并对细胞学标本做特殊处理后常规涂片,做B72.3、癌胚抗原、神经特异性钙结合蛋白、血栓素免疫细胞化学与苏木素-伊红(HE)染色分析。结果123例积液标本中,B72.3、癌胚抗原诊断腺癌的敏感性分别为80.1%、78.7%,特异性分别为97.1%、97.1%,B72.3、癌胚抗原组合检测诊断转移腺癌的敏感性为95.5%(85/89);神经特异性钙结合蛋白、血栓素诊断间皮源性细胞敏感性分别为88.2%、79.4%,特异性分别为95.5%、91.0%,神经特异性钙结合蛋白、血栓素组合检测诊断间皮源性细胞敏感性为94.1%(32/34),其中恶性间皮瘤为100%(8/8)。结论通过对浆膜腔积液细胞学标本的有效处理和制片技术的改进,选择B72.3、癌胚抗原、神经特异性钙结合蛋白、血栓素组合检测和细致的细胞形态学观察,可进一步提高浆膜腔积液中恶性间皮瘤与转移性腺癌的鉴别诊断。     

附睾原发性恶性间皮瘤2例临床病理特点

目的 探讨附睾原发性恶性间皮瘤的临床病理特征、诊断与鉴别诊断要点。方法 对2例附睾原发性恶性间皮瘤进行临床病理分析、组织形态学、组织化学及免疫组化染色观察，结合文献对其临床表现、病理形态特点及鉴别诊断进行探讨。结果 2例附睾原发性恶性间皮瘤呈上皮样组织学改变(上皮样型间皮瘤)，在纤维性间质背景中，含有片状、灶状或巢状上皮细胞团，局部有腺样结构形成。淀粉酶消化的PAS染色瘤细胞呈(-)，网织纤维染色显示上皮性肿瘤特点，局部呈双向分化；免疫组化染色显示瘤细胞calretinin( )，EMA强( )，vimenfin弱( )，CEA(-)。结论 原发于附睾的恶性间皮瘤是一种罕见的恶性肿瘤，预后差，临床易误诊。其组织形态复杂多样，应注意与附睾的良性增生性病变及腺癌和梭形细胞肿瘤等鉴别。     

Flow cytometric features of angioimmunoblastic T-cell lymphoma

BACKGROUND: The immunophenotypic features of angioimmunoblastic T-cell lymphoma (AILT) have not been well described. METHODS: We retrospectively reviewed our institutional experience with the flow cytometric features of 16 cases of AILT. RESULTS: Multiparameter flow cytometry was able to identify a distinct population of immunophenotypically aberrant T cells in 15 of 16 cases. In 13 lymph node specimens, the neoplastic cells ranged from 1.9 to 87% (median 23%) of cells. The ratio of reactive to neoplastic T cells ranged from 0.01 to 20 (median 1.5); reactive T cells outnumbered neoplastic in 9/13 (69%) cases. The neoplastic populations expressed CD2, CD4, CD5, and CD45RO in all cases, lacked expression of CD8 and CD56 in all cases, and showed negative or dim surface CD3 in most cases. CD10 was expressed by the neoplastic populations in 11 of 14 cases at diagnosis; in 3 of these 11 only a subpopulation of the neoplastic cells was CD10(+). CD10 tended to be absent on neoplastic cells in staging bone marrows. The neoplastic population in all but one of the 15 positive cases possessed multiple immunophenotypic abnormalities and these were generally retained during the follow-up analyses of several cases. CONCLUSIONS: These results indicate the potential utility of flow cytometry in the diagnosis and follow-up of AILT.     

Ber-EP4 and anti-calretinin antibodies: a useful combination for differential diagnosis of various histological types of ovarian cancer cells and mesothelial cells

The differential diagnosis between reactive mesothelial cells and ovarian carcinoma cells is often difficult in cytologic specimens. Immunocytochemical procedures have been utilized in assisting this differential diagnosis, with limitations. Furthermore, previous studies examined only serous type but not other histological types of ovarian carcinoma cases. Therefore, we evaluated the practical value of various epithelial and mesothelial markers in differential diagnosis of these two types of cells. Various types of ovarian carcinoma (serous, n = 22; mucinous, n = 10; endometrioid, n = 7; clear cell, n = 10) and benign mesothelial tissues (n = 15) were studied by immunohistochemistry. We then studied effective panels of antibodies by immunohistochemistry in 43 cytologic specimens of ascites or peritoneal lavage fluid consisting of 20 reactive mesothelium and 23 adenocarcinomas of the ovary. In the tissue specimens, Ber-EP4, a monoclonal antibody of epithelial antigen, and a polyclonal antibody against calretinin, which is expressed in mesothelium, are used in differentiating reactive mesothelial cells from ovarian carcinoma. In cytologic specimens, the sensitivity and specificity of Ber-EP4 were 100% and 90%, respectively. The sensitivity and specificity of the anti-calretinin antibody were 90% and 91%, respectively. Using multiple regression analysis, the correlation coefficient between epithelial antigen and calretinin reactivity was r = 0.938, with a significance level of p < 0.0001. In conclusion, the combined immunostaining of cytologic specimens for Ber-EP4 and the anti-calretinin antibody is helpful for the differential diagnosis between mesothelial cells and not only serous type, but also mucinous, endometrioid and clear cell types of ovarian cancer cells.     

Requirements for the construction of antibody heterodimers for the direction of lysis of tumors by human T cells.

We constructed a series of MAb heterodimers consisting of the J5 (anti-common acute lymphoblastic leukemia antigen [CALLA]) antibody and antibodies to a variety of structures present on the surface of activated human T cells, including CD3 antigen (T cell receptor-associated glycoproteins), CD2 antigen (T11/E-rosette receptor), CD25 antigen (IL-2 receptor), and the transferrin receptor. We tested the ability of these heterodimers to direct a CD2 + CD3 + CD8 + CD4 - CD25 + transferrin receptor + MHC-restricted human cytolytic T lymphocyte (CTL) clone to lyse a CALLA + human tumor in vitro. Only heterodimers containing an anti-CD3 antibody or activating antibodies to CD2 could direct the clone to lyse these human tumor targets, even when the clone was additionally activated with anti-CD3 or anti-CD2 antibodies. Our findings may have implications in the design of strategies for the use of such reagents in the treatment of human neoplasia.     

Effusion cytology

Cytologic evaluation is the best way to detect the presence of malignancy in body cavity fluids. Although a positive diagnosis is highly reliable, a negative result does not rule out a malignant cause. Adenocarcinomas, well-differentiated squamous carcinomas, small-cell carcinomas, malignant melanomas, large-cell lymphomas, and acute leukemias are accurately classified when present in effusions. The definitive diagnosis of malignant mesothelioma, small-cell lymphomas, and chronic leukemias, and subclassification of sarcomas and poorly differentiated neoplasms are difficult and may require additional diagnostic techniques. With a few exceptions, the exact causes of benign effusions cannot be determined by cytologic methods.     

CD40 ligation induces Apo-1/Fas expression on human B lymphocytes and facilitates apoptosis through the Apo-1/Fas pathway

The Apo-1/Fas antigen (CD95) mediates programmed cell death of lymphocytes when bound by Fas ligand or anti-Apo-1/Fas antibody. In contrast, the CD40 antigen provides a potent activation and survival signal to B lymphocytes when it is engaged by its T cell ligand (CD40L, gp39) or cross-linked by anti-CD40 antibody. In this study, we use human tonsillar B cells and the Ramos Burkitt's lymphoma B cell line, which serves as a model for human germinal center B lymphocytes, to study the effectors of Apo-1/Fas expression and apoptosis of human B cells. We found that Apo-1/Fas expression was upregulated on both malignant and normal human B lymphocytes after CD40 ligation induced by (a) cognate T helper-B cell interaction mediated by microbial superantigen (SAg); (b) contact-dependent interaction with CD40L+, but not CD40L- Jurkat mutant T cell clones; and (c) monoclonal anti-CD40, but not any of a panel of control antibodies. Enhanced B cell Fas/Apo-1 expression is functionally significant. Coculture of Ramos Burkitt's lymphoma line cells with irradiated SAg-reactive CD4+ T cells with SAg or CD40L+ Jurkat T cells results in B cell apoptosis, evidenced by reduced cell viability and DNA laddering. This process is augmented by the addition of anti-Apo-1/Fas monoclonal antibody, consistent with an acquired susceptibility to Apo-1/Fas-mediated apoptosis. These data support an immunoregulatory pathway in which seemingly contradictory signals involving the B cell proliferation/survival antigen CD40, as well as the Apo-1/Fas molecule, which mediates programmed cell death of lymphocytes, are linked in the process of human B cell activation.     

Characterization of a chimeric T-cell receptor with specificity for the Hodgkin's lymphoma-associated CD30 antigen

Recombinant receptors with antibody-like specificity for tumor-associated antigens were shown to direct specifically T cells to target tumor cells. Hodgkin and Reed-Sternberg cells, the malignant cell population in Hodgkin's lymphoma, express high amounts of the cell surface antigen CD30. An anti-CD30 T-cell receptor with cellular activation properties is expected to graft T cells with specificity to Hodgkin cells. Here, the authors characterize a chimeric T-cell receptor with an extracellular domain consisting of the single-chain antibody fragment HRS3-scFv with specificity for the CD30 antigen and intracellular domain of the signal transducing part of the Fc-epsilon-I-gamma receptor. The HRS3-scFv was derived from the monoclonal anti-CD30 antibody HRS3 and retained specificity for the CD30 antigen. The recombinant HRS3-scFv-gamma receptor was expressed under control of the RSV-LTR after transfection into MD45 T-cells. The chimeric receptor protein is detected and analyzed by enzyme-linked immunosorbent assay (ELISA) and immunoprecipitation. Expression of the chimeric receptor converts MD45 T cells to specificity for CD30+ lymphoma cells. Specific cross-linking of the chimeric receptor with antigen resulted in cytolytic reactivity against CD30+ tumor cells in vitro. The results demonstrate that the chimeric receptor HRS3-scFv-gamma converts T cells to a specific MHC-unrestricted cytolytic response against CD30+ tumor cells offering an alternative strategy in cellular immunotherapy of Hodgkin's disease.     

Targeting of natural killer-like T immunologic effector cells against leukemia and lymphoma cells by reverse antibody-dependent cellular cytotoxicity

Recently, highly efficient natural killer-like T immunologic effector cells called cytokine-induced killer (CIK) cells have been described. Most interestingly, CIK cells have been shown to eradicate established human lymphoma cells in a severe combined immunodeficient (SCID) mouse xenograft model in vivo. The current study was aimed at increasing the sensitivity of leukemia and lymphoma cells to CIK cells. In particular, the authors wanted to target CIK cells to leukemia and lymphoma cells via reverse antibody-dependent cellular cytotoxicity. Binding of an anti-CD3 monoclonal antibody to CIK cell cultures derived from patients with lymphoma was shown using flow cytometric analysis. For the target side, several B-cell lines were found to express CD19 on the cell surface. There was an impressive increase in sensitivity to CIK-mediated lysis of various lymphoma and leukemia cell lines by preincubation of the targets with a monoclonal antibody against CD3. This increase could be partially blocked by preincubation with anti-CD16 (Fc receptor III) and anti-CD32 (Fc receptor II) antibodies. These data suggest that the increase in cytotoxic activity is caused by Fc receptor-mediated antibody binding. Cytotoxic activity could be further increased by adding an anti-CD28 antibody in addition to anti-CD3. Finally, there was a further increase in sensitivity to CIK-mediated lysis of CD19+ malignant cells using the bispecific OKT3xHD37 antibody with specificity against CD3 and CD19. Interestingly, preincubation of malignant cells with an anti-CD3 monoclonal antibody followed by addition of the bispecific OKT3xHD37 antibody led to a further increase of cytotoxic sensitivity compared with the addition of the bispecific antibody alone. In conclusion, these data suggest that cytotoxic activity of immunologic effector cells can be increased not only by using the bispecific antibody OKT3xHD37 in vitro but also by preincubation of CD19+ leukemia and lymphoma cells with a monoclonal antibody against CD3. In addition, the immunostimulatory effect of the bispecific antibody OKT3xHD37 can be further increased by adding a monoclonal antibody against CD3.     

Better-surviving liver grafts by the injection of anti-CD2 antibody: the important roles of host CD8+ and CD2+CD28+ T cells in chronic graft rejection and beta type platelet-derived growth factor receptor (PDGFR-beta) expression on apoptotic liver grafts.

Syngeneic liver grafts were implanted in the livers of 22 LEW/Sea strain rats. To prolong the graft survival, anti-CD2 monoclonal antibody (MAb) or anti beta type platelet-derived growth factor receptor (PDGFR-beta) antibody (Ab) was injected, or splenectomy was performed in the rats which were then followed until 10 to 11 weeks posttransplantation. The 22 rats with chronic graft rejection showed increased CD8a-like antigen (probably Fas ligand) on the peripheral blood T cells. All the liver grafts had both necrosis and apoptosis. The liver graft apoptosis was indicated by histopathological abnormalities, and by DNA strand breaks and hemosiderin depositions in the cytoplasm. PDGFR-beta expression in the apoptotic liver graft was demonstrated immunohistochemically. Among the 17 rats injected with anti-CD2 MAb, CD2 signaling on host T cells was effectively suppressed by the injection of anti-CD2 MAb in 4 rats with better-surviving liver grafts. In these 4 rats, CD28 antigen on thymic lymphocytes was down-modulated and high numbers (136-233-positive cells per lobe) of the epithelial reticular cells with apoptotic lymphocytes were counted. Anti-PDGFR-beta Ab caused high pulmonary secretions of growth factors and reticular fibrosis in the lungs of 5 rats injected with the Ab. Anti-PDGFR-beta Ab injection reduced the host cell apoptosis in the lung and thymus, but did not prolong the survival of liver grafts. In the 9 rats with both splenectomy and anti-CD2 MAb injection, pulmonary apoptosis was induced with the 6-16% reductions of CD4+ lymphocytes. Prolonged graft survival was observed in only one of the 9 rats. Anti-CD2 MAb was effective for prolonging the liver graft survival with suppressed CD28 antigen, but anti-PDGFR-beta Ab and splenectomy were not.     

Measurement of vitamin D-binding protein in pleural fluids and sera by means of a turbidimetric immunoassay measuring system

BACKGROUND: Vitamin D-binding protein (DBP) has been recognized as a multifunctional plasma protein that can modulate certain immune and inflammatory responses. There may be differences between the DBP concentrations in pleural fluids from various diseases involving a variety of possible responses in the pleural cavity. METHODS: An anti-DBP polyclonal antibody was prepared using commercially available DBP to establish a quantitative measuring system for DBP. With a rabbit antibody, a turbidimetric immunoassay (TIA) was developed for DBP with an automatic analyzer. Using this measuring system, the concentrations of DBP were compared with the protein concentration in pleural fluid and serum specimens from patients with various diseases. RESULTS: The fluid DBP concentrations in transudative (n=11) and exudative (n=41) effusions were 71.9+/-21.2 and 180.7+/-43.7 mg/l, respectively. Among the exudative effusions, the fluid DBP concentrations in the bacterial (n=10), tuberculous (n=13), and malignant (n=18) effusions were 218.8+/-37.3, 186.7+/-26.2, and 155.1+/-41.3 mg/l, respectively. The DBP fluid/serum ratio and the fluid DBP/protein ratio in bacterial effusions were significantly higher than those in tuberculous (p<0.005, p<0.05, respectively) and malignant effusions (p<0.0005, p<0.005, respectively), although no statistically significant differences in the serum DBP/protein ratio between those effusions were found. CONCLUSIONS: Using the TIA assay, the DBP concentrations in bacterial pleural effusions were significantly higher than in tuberculous and malignant effusions.     

Human CD4 “internal antigen” mimicry by anti-idiotypic monoclonal antibodies

Summary Of 1019 hybridomas generated from a BALB/c mouse immunized with the syngeneic anti-CD4 monoclonal antibody HP2/6, 3 were found to secrete anti-idiotypic antibodies. Detailed analysis of anti-idiotypic monoclonal antibodies F16-10F6, F16-14D6 and F16-16D7 showed they recognize idiotope(s) not expressed by any of the anti-CD4 monoclonal antibodies tested, including those which inhibit the binding of HP2/6 to CD4 antigen. The idiotope recognized by the three anti-idiotypic antibodies are within (or closely related to) the antigen combining site of the immunizing antibody and distinct and spatially distant from the idiotope defined by monoclonal antibody F11-2302 which was previously shown to be outside the antigen combining site of HP2/6. Although F16-14D6 and F16-16D7 are indistinguishable in isotype, binding titer to idiotopes, fine specificity on a panel of monoclonal antibodies, relation to the combining site and competitive binding, it is likely that they are structurally different and recognize two distinct combining site-related idiotopes on HP2/6, as they display different spectrotypes and induce antianti-idiotypic (Ab3) immune sera with different specificities. Analysis of the fine specificity of the two Ab3 immune sera suggest they share idiotopes with HP2/6 and contain antibodies reacting with CD4 antigen. Among the latter, those induced with F16-14D6 display a different CD4 epitope specificity than HP2/6. Hence, anti-idiotypic antibodies F16-14D6 and F16-16D7 behave as “network antigen” for human CD4; idiotope-triggered antibody cascade may have a role in changing the specificities of antibody.     

Diagnostic usefulness of tumour marker levels in pleural effusions of malignant and benign origin

Pleural effusion is a common diagnostic problem. The analysis of serum and body fluids for tumor markers has been intensively applied to clinical diagnosis. The aim of the present study was to determine the usefulness of simultaneous quantification of carbohydrate antigen 19.9, carbohydrate antigen 125, neuron specific enolase, mucinous-carcinoma-associated antigen, and ferritin in samples of pleural fluids in the malign pleural effusion and its differentiation from benign effusions. A total of 61 pleural effusions were collected from the patients, who were subjected either to simple needle aspiration or to tube drainage for the diagnosis of pleural effusion. Tumor markers were determined in benign patient groups with nonspecific pleurisy, tuberculous pleurisy, empyema, congestive heart failure and in malignancy groups consisting of adenocarcinoma, small cell lung carcinoma, mesothelioma, epidermoid lung cancer. The tumor markers CA-19.9, CA-125, NSE, and ferritin levels were quantified by the sandwich assay using the streptavidin technology of ELISA in an ES-300 Boehringer-Mannheim analyser. MCA was measured by employing a two-side solid phase EIA method. MCA measurements were done by the Cobas-Core. For all patients, the effusions correctly or incorrectly identified by the different procedures as being malignant or nonmalignant are defined as true positive, false positive, true negative, and false negative, the term ‘positive’ referring to histologically proven malignant pleural effusion while nonmalignant effusions are referred to as ‘negative’. Therefore, sensitivity, specificity, positive predictive value, and negative predictive value were defined as diagnostic parameters. The cut-off values calculated were 352 U/ml for CA-125, 54 U/ml for CA-19.9, 555 for ferritin, 11.1 for MCA and 8.7 for NSE. In our study, the highest sensitivity is found to be MCA with 100%; specificity, CA-19.9 with 97%; PPV, CA-19.9 and MCA with 95% and NPV, MCA with 100%. Our data imply that the co-measurement of MCA+CA-19.9+CA-125 levels may further improve their diagnostic value in malignant pleural effusion compared with that of each tumour marker alone and may be useful in distinguishing malignant from benign pleural effusions.     

脾原发性恶性淋巴瘤11例临床病理及免疫组化分析

目的　对 11例脾原发性恶性淋巴瘤 (PLS)进行临床病理及免疫组化研究。方法　对瘤组织进行常规HE染色及SP法免疫组化染色 ,光镜观察。结果　 11例患者以左上腹疼痛和巨脾为主要症状。 11例瘤细胞CD45均( ) ;10例B细胞性淋巴瘤CD2 0 ( ) ,其中B小淋巴细胞性淋巴瘤 3例 ,CD43、CD79α和bcl 2 ( ) ;脾边缘区B细胞性淋巴瘤 3例 ,CD79α、IgM、ALK( ) ,IgD(± ) ;淋巴浆细胞样淋巴瘤 2例 ,CD43、CD79α、bcl 2 ( ) ,IgD(-) ;弥漫性大B细胞性淋巴瘤 2例 ,ALK( ) ,CD3 0、CD3 (± )。 1例周围T细胞性淋巴瘤 ,无其他特征型CD45RO和CD3 ( ) ,CD3 0 (± )。结论　PLS较罕见 ,应与脾内其他原发性或继发性小细胞性恶性肿瘤鉴别 ,免疫组化染色对诊断、鉴别诊断及分型具有重要意义     

Antitumor efficacy of CD137 ligation is maximized by the use of a CD137 single-chain Fv-expressing whole-cell tumor vaccine compared with CD137-specific monoclonal antibody infusion

Tumor-destructive immune responses can be generated by engaging CD137 (4-1BB) via infusing a monoclonal antibody specific for CD137 or vaccinating with a single-chain Fv (scFv) CD137-expressing whole-cell tumor vaccine. We assessed whether such a vaccine can induce tumor rejection in the neu-transgenic (neu-Tg) mouse breast cancer model and compared the antitumor efficacy of vaccination with the infusion of a CD137-specific antibody. Mammary carcinoma cells (MMC) from a neu-Tg mouse were transfected to stably express surface scFv derived from the anti-CD137 rat hybridoma 1D8 or 3H3. The anti-CD137 scFv-expressing cells were rejected when transplanted into neu-Tg mice by a mechanism that involved both CD4(+) and CD8(+) T cells, and vaccination with such cells delayed the outgrowth of MMC cells transplanted 3 days previously. T cells from neu-Tg mice that had been vaccinated proliferated and produced IFN-gamma when stimulated by MMC but not by antigen-negative variant breast cancer cells that did not express the neu tumor antigen. In addition, antibodies binding to the MMC but not to antigen-negative variant cells were detected in sera from some but not all of the immunized mice. Complete regression of s.c. transplanted MMC tumors was observed in mice repeatedly immunized against MMC-1D8 starting on the day the MMC cells were transplanted. In contrast, repeated administration of either of two different anti-CD137 monoclonal antibodies did not induce complete tumor regression, although tumor growth was delayed.     

Therapeutic effects of tumor reactive CD4+ cells generated from tumor-primed lymph nodes using anti-CD3/anti-CD28 monoclonal antibodies

T-cell activation involves multiple signaling pathways. In this report, we conducted in vitro and in vivo immune function analysis of tumor-draining lymph node (TDLN) cells after anti-CD3/anti-CD28 activation versus anti-CD3 activation alone in a murine tumor model. In cytokine release assays, the doubly activated TDLN cells secreted significantly greater amounts of IFN-gamma and GM-CSF in response to specific tumor antigen compared with anti-CD3 activated cells. In adoptive immunotherapy, the doubly activated TDLN cells were more effective in mediating regression of 3-day pulmonary metastases compared with anti-CD3 activated cells. Although there was predominant proliferation of CD8+ cells after either activation procedure, the mean-fold expansion of CD4+ cells was significantly greater after anti-CD3/anti-CD28 activation than anti-CD3 activation alone. Using magnetic bead-enriched T-cell subsets, we found that either CD4+ or CD8+ doubly activated TDLN cells could independently mediate tumor regression. Furthermore, the doubly activated CD4+ cells were more effective than CD8+ cells in adoptive immunotherapy on a per-cell basis. The antitumor activity mediated by CD4+ or CD8+ cells could be significantly enhanced with the exogenous administration of IL-2. CD28 co-stimulation of tumor-primed lymphoid cells promotes the generation of potent tumor reactive effector cells, particularly CD4+ T cells, with antitumor activity in adoptive immunotherapy.     

CD4+CD25+ suppressor lymphocytes in the circulation of patients immunized against melanoma antigens

Murine studies have suggested that a population of CD4+ T cells expressing the alpha chain of the interleukin (IL)-2 receptor (CD25+) are phenotypically anergic in response to T cell receptor stimulation and can suppress the function of CD4+ and CD8+ T cells. Recent studies of peripheral lymphocytes from healthy human volunteers have identified a similar population, although little is known about the presence and activity of these cells in patients with cancer and their possible impact on anticancer immunization strategies. Thus, the authors have undertaken these studies in patients with metastatic melanoma undergoing immunizations with known melanoma antigens. CD4+ CD25+, CD4+ CD25-, and a 1:1 ratio of these isolated T cells were stimulated with soluble anti-CD3 antibody in the presence of irradiated T cell-depleted PBMCs, and proliferation was assessed by measuring [3H]thymidine incorporation. In 13 patients, isolated CD4+CD25+ T cells proliferated 68% (+/- 5.8%) less than separately cultured CD4+ CD25- T cells. Moreover, CD4+ CD25+ T cells suppressed the proliferation of an equal number of cocultured CD4+ CD25+ T cells in 11 of 13 patients by an average of 60% (+/- 4.9%). Suppression was not seen at day three of culture and became apparent at days five through nine. The degree of suppression was proportional to the numbers of CD4+ CD25+ T cells. Addition of high-dose IL-2 reversed the hypoproliferative phenotype of the CD4+ CD25+ T cells and abrogated their suppressive function. These studies demonstrate that anergic and functionally suppressive CD4+ CD25+ T cells exist in patients with melanoma undergoing tumor antigen immunization and thus may play a role in modifying the magnitude of the T cell response to immunization.