几种癌基因、层粘连蛋白及其受体和纤维粘连蛋白与正常人早孕绒毛相关性的免疫组化研究(摘要)

为了探讨层粘连蛋白（ＬＮ）及其受体与纤维粘连蛋白（ＦＮ）和几种癌基因在正常人早孕绒毛中的分布状况，本文采用免疫组化方法，观察了ＬＮ、ＬＮＲ、ＦＮ、ｂｃｌ－２、Ｐ１６、Ｐ５３和Ｐ２１在正常人早孕绒毛中的表达。结果显示：ＬＮ主要阳染于细胞滋养细胞（ＣＴ）...     

层粘连蛋白片段对体外培养成纤维细胞及肝细胞增殖的影响

层粘连蛋白片段对体外培养成纤维细胞及肝细胞增殖的影响上海长征医院（上海２００００３）王杰军，孔宪涛，李石，连兆瑞，张玲珍层粘连蛋白（ｌａｍｉｎｉｎ，ＬＮ）作为细胞外间质中（ｅｘｔｒａｃｅｌｌｕｌａｒｍａｔｒｉｘ，ＥＣＭ）一种重要的非胶原性糖蛋白成分，...     

小鼠肺腺癌体内外生长、运动和侵袭与细胞外基质相关性的研究

利用小鼠肺腺癌（ＬＡ＿（７９５））肾包膜下移植模型，采用间接免疫酶标和间接免疫荧光方法，观察了肿瘤发展过程的不同时期纤维粘连蛋白（ＦＮ），层粘连蛋白（ＬＮ）和ＩＶ胶原的分布情况。结果表明，随着肿瘤移植后时间的延长，间质ＦＮ量不变，ＬＮ和ＩＶ型胶原量增多，且三种成分各自的分布图象在不同时期也存在差异，瘤细胞侵袭基底膜处ＬＮ和ＩＶ型胶原缺失。通过体外实验，观察到ＬＡ＿（７９５）细胞母系有内源性ＦＮ和ＬＮ的产生，无ＩＶ型胶原ｍＲＮＡ的表达，ＦＮ、ＬＮ或ＩＶ型胶原尤其是ＬＮ能促进瘤细胞的运动，ＦＮ或ＬＮ能加快瘤细胞集落的生长，表明ＦＮ、ＬＮ和ＩＶ型胶原与恶性肿瘤的侵袭和转移有关。     

银屑病患者血浆透明质酸和层粘连蛋白含量的变化

已认识到细胞外间质（ＥＣＭ）不仅作为一个支架，而且具有许多生物活性。透明质酸（ＨＡ）和层粘连蛋白（ＬＮ）是ＥＣＭ的重要成份，与银屑病表皮细胞过度增生以及细胞的生长、游走和粘附有关。为此我们检测银屑病患者血浆ＨＡ和ＬＮ含量，旨在探讨其在银屑病发病中的作...     

层粘连蛋白受体单克隆抗体对人肺癌细胞形态、粘着及铺展的影响

应用体外细胞培养，观察了层粘连蛋白受体单克隆抗体（ＭｃＢ＿１）加入培养液或预处理人肺巨细胞癌（ＰＧ）和肺腺癌（ＰＡａ）后某些生物学行为的改变。在体外ＭｃＢ＿１预孵育可抑制ＰＧ和ＰＡａ在层粘连蛋白基质上的粘着和铺展。培养液中加入ＭｃＢ＿１经扫描电镜观察，ＭｃＢ＿１明显改变该两种细胞的表面形态。说明层粘连蛋白受体在肿瘤浸润和转移中可能起重要作用。层粘连蛋白受体单克隆抗体在阻断肿瘤转移的发生上可能具有应用价值。     

上皮细胞膜抗原和层粘连蛋白在大肠癌中的表达

上皮细胞膜抗原和层粘连蛋白在大肠癌中的表达冯树宋今丹上皮细胞膜抗原（ＥＭＡ）已作为一种肿瘤标志物用于肿瘤的研究中。层粘连蛋白（ＬＮ）是基底膜的主要成分，对细胞的增殖、生长、分化及癌细胞的浸润和转移等方面均起着重要的调节作用〔１〕。用免疫组化法对６３例...     

不同类型胶质瘤移动性的比较及其与细胞外基质成分关系的研究

用３株不同类型的恶性胶质瘤进行细胞体外移动性及其与细胞外基质成分关系的比较研究。结果表明，髓母细胞瘤移动性最强，多形性胶质母细胞瘤次之，ＳＷＯ－３８胶质瘤最弱。３株胶质瘤体外移动性受Ⅰ型胶原，Ⅳ型胶原，层粘连蛋白（ｌａｍｉｎｉｎ，ＬＮ）和纤维结合蛋白（ｆｉｂｒｏｎｅｃｔｉｎ，ＦＮ）等细胞外基质成分的影响基本上是一致的。故不能以此解释其移动性差异的原因。Ⅳ型胶原蛋白和ＬＮ能促进瘤细胞在体外的移动，但Ⅳ型胶原和ＬＮ两者之间不表现出叠加作用，也无拮抗作用。Ⅰ型胶原网能明显地阻遏肿瘤细胞的体外移动性。ＦＮ对这３株胶质瘤的移动性影响不大，但对非神经系统肿瘤（ＧＬＣ－８２肺腺癌）却有较明显的趋触作用。     

层粘连蛋白受体和nm23蛋白在乳腺癌组织中的表达及其与间 …

探讨乳腺癌组织层粘连蛋白（ＬＮ）及其受体（ＬＮ－Ｒ）和ｎｍ２３蛋白的表达与间质微血管密度及肿瘤转移的关系。方法应用免疫组织化学ＬＳＡＢ方法检测７３例乳腺癌力ｎｍ２３蛋白的表达,在第Ⅷ因子相关抗原染色切片上检测其微血管密度（ＭＶＤ）。结果病理分级越高,乳腺癌ＬＮ表达程度越强,即病理Ⅰ级而ＬＮ分布Ⅲ级者为８．３％,病理和ＬＮ分布均Ⅲ级者为５８．８％,两者间差异有极显著意义。ＬＮ－Ｒ表达程度则与转移有关     

层粘连蛋白与葡萄胎及侵蚀性葡萄胎相关性的免疫组化研究

为了探讨层粘连蛋白（ＬＮ）在非侵蚀性（ＨＭ）和侵蚀性葡萄胎（ＩＨＭ）的发生和转化过程中的作用，本研究采用免疫组化法，观察了ＬＮ在ＨＭ和ＩＨＭ的分布。结果显示：ＬＮ在葡萄胎中的游离绒毛合体滋养细胞（ＬＮＲ）呈阳性着色、ＬＮ呈阴性；游离绒毛细胞滋养细胞ＬＮ呈阴性；游离绒毛细胞滋养层基底膜ＬＮ呈阴性；游离绒毛基质细胞ＬＮ呈阴性；绒毛外滋养细胞ＬＮ呈阴性；绒毛外滋养细胞基底膜ＬＮ呈弱阳性；固定绒毛柱滋养细胞ＬＮ呈阴性；近绒毛滋养层柱基底膜ＬＮ呈弱阳性；远绒毛滋养层柱基底膜ＬＮ呈弱阳性。在侵蚀性葡萄胎中游离绒毛合体滋养细胞、游离绒毛细胞滋养细胞、游离绒毛细胞滋养层基底膜和游离绒毛基质细胞ＬＮ呈阴性；绒毛外滋养细胞和绒毛外滋养细胞基底膜ＬＮ呈强阳性；固定绒毛柱滋养细胞ＬＮ呈阴性     

层粘连蛋白受体抗体及蛋白激酶C抑制剂体内阻断肿瘤转移的实验研究

目的探索阻断肿瘤转移新的途径。方法利用内源性层粘连蛋白（ＬＮ）及其受体（ＬＮＲ）强阳性的小鼠纤维样肉瘤细胞,分别在体外经外源性ＬＮ、蛋白激酶Ｃ（ＰＫＣ）抑制剂（Ｓｔａｕｒｏｓｐｏｒｉｎｅ,ＳＰ）、抗ＬＮＲ、抗ＬＮ处理后,一次性注入Ｃ５７小鼠尾静脉（７７只）,使之在肺内成瘤。各组瘤细胞在未注入前,均经流式细胞仪（ＦＣＭ）定量测定其表面ＬＮＲ水平。结果空白对照组、ＬＮ组、ＳＰ组、抗ＬＮＲ组、抗ＬＮ组,瘤细胞表面ＬＮＲ阳性率顺次为５６５％、７６３％、５０２％、３１９％、２０６％,小鼠肺内瘤灶（含原发及转移灶）体积均值顺次为７６ｍｍ３／只,１１０ｍｍ３／只,１４ｍｍ３／只,５ｍｍ３／只,２０ｍｍ３／只,小鼠肺内淋巴管血管腔内瘤栓形成率顺次为８１％、９４％、３６％、４３％、６８％。结论外源性ＬＮ组的病变及转移程度重于空白对照组,而其余各组均轻于空白对照组;统计学分析表明ＬＮＲ水平与瘤灶大小及瘤栓形成率（病变程度及转移程度）均呈正相关;体内瘤细胞内磷脂肌醇传导系统的功能状态亦与之密切相关。     

Carcinoma-associated perisinusoidal laminin may signal tumour cell metastasis to the liver

Summary The perisinusoidal space of the liver shows extensive modulation of the extracellular matrix in response to various pathological conditions. We studied perisinusoidal laminin expression immunohistochemically using polyclonal and monoclonal antibodies in 110 human liver specimens obtained at autopsy. In normal adult liver the perisinusoidal spaces contained only minimal amounts of immunoreactive laminin. In 86% of patients dying from cancer with liver metastasis, however, a distinct increase in the amount of perisinusoidal laminin could be demonstrated. The perisinusoidal space also contained laminin in cancer patients without liver metastasis. In 3 cases of leukaemia sinusoids were laminin negative. In cirrhosis and chronic passive congestion there was, as expected, laminin immunoreactivity in the perisinusoidal space. The results obtained using polyclonal antibodies against laminin were confirmed using chain-specific monoclonal antibodies against B2 laminin. In an ex vivo assay, viable tumour cells (Panc-1 and clone A) were found to bind with remarkable specificity to frozen sections of liver tissue containing perisinusoidal laminin as opposed to liver tissues without laminin. We suggest that this perisinusoidal laminin may directly on indirectly mediate tumour cell metastasis to the liver.     

Monoclonal antibodies evoked by the free oligopeptide (Gly)5 reacting specifically with peptidoglycan from staphylococci

Murine monoclonal antibodies reactive with the Staphylococcus aureus peptidoglycan (PG) epitope (Gly)5 were obtained using the synthetic oligopeptide (Gly)5 in its free form as immunogen. The selected monoclonal antibodies were of the IgM kappa isotype and reacted specifically with PG from S. aureus and Staphylococcus epidermidis, but gave no reaction with PG from Streptococcus pyogenes, Bacillus subtilis and Micrococcus lysodeikticus. Affinity chromatography showed that the antibodies were reactive with the N-terminus of the (Gly)5 peptide. These monoclonal antibodies can be used for the detection of staphylococcal PG in solution.     

Monoclonal antibodies to Staphylococcus aureus laminin-binding proteins cross-react with mammalian cells.

We and others have previously shown that some microorganisms, including bacteria, express on their surfaces receptors that specifically recognize extracellular matrix proteins, such as laminin, fibronectin, or both. The ability of microorganisms to adhere and to invade might depend on the existence of receptors which could, thus, be correlated with pathogenicity. In the present paper, we report the isolation of five stable cell lines that were producers of monoclonal antibodies to Staphylococcus aureus laminin receptors. One of these antibodies, which was of the immunoglobulin M isotype, blocked the binding of laminin to bacteria before and after fixation and recognized the putative 52-kilodalton laminin-binding protein in whole bacterial extracts. Also, purified receptor was isolated by immunoaffinity chromatography and shown to bind laminin. Furthermore, the same antibodies bound the 67-kilodalton putative receptor from mouse melanoma cells and gave positive immunofluorescence reactions against mammalian tumor cells. These data strongly suggest either the evolutionary conservation of at least some sequences in both procaryotic and eucaryotic laminin-binding proteins or convergent evolution and positive selection of epitopes cross-reacting with laminin. Some of these antibodies to the procaryotic protein could therefore become useful markers for the expression of laminin receptors by cancer cells.     

Monoclonal antibodies against the GP-2 subunit of laminin

Two stable rat X mouse hybridoma lines have been isolated. These hybridoma lines produce IgG antibodies directed against the polypeptide portion of the GP-2 subunit of laminin. Antibodies produced by the hybridomas have been shown to be IgG 2b (lambda) and IgG 2a (kappa), respectively. In competition ELISA assays the monoclonal antibodies exhibited different binding affinities for laminin. Furthermore, the two antibodies were partially additive in their reactivity to laminin. Preliminary results also indicate that the antibodies recognized different antigenic determinants in laminin as determined by their reactivity to basement membranes in human and mouse tissues. The monoclonal antibody designated LAM-I stained a broad spectrum of human and mouse tissues; the other monoclonal antibody LAM-II reacted with mouse, but not human tissues. The results indicate that these monoclonal antibodies could be utilized to explore the organization of laminin in basement membranes of different tissues and species.     

A role for the laminin receptor in leukocyte chemotaxis

Previous studies on the mechanism of leukocyte traversal of basement membranes showed that rabbit peritoneal exudate polymorphonuclear cells (PMN) preferentially used laminin, a major constituent of basement membrane, to attach to another component, type IV collagen. PMN also responded chemotactically to nanomolar levels of laminin. We have now determined that PMN possess a receptor for laminin. Scatchard analysis using 125I laminin indicates a single class of saturable high affinity binding sites (kd = 6.15 nM/L) on PMN and 3.6 X 10(4) sites per cell. A chymotryptically derived fragment of laminin, C1, which lacks both the long arm and the globular end regions of the short arm (i.e., matrix binding sites) but retains the laminin receptor binding region, gave similar results. Immunoperoxidase studies using monoclonal antibodies to the laminin receptor (mAbLR) indicated the presence of the receptor on the surface of PMN. These cells responded chemotactically to nanomolar levels of C1 and laminin, a result consonant with binding data. PMN chemotaxis to a formyl peptide was markedly inhibited by mAbLR, suggesting that the laminin receptor may be required for PMN chemotaxis in general. Our results suggest that PMN extravasation across basement membranes is aided both by reversible attachment of the cells to laminin in the matrix and by chemotaxis to a gradient of soluble intact and possibly degraded laminin. These characteristics have much in common with those of highly metastatic tumor cells.     

PERSISTENT and ENHANCED EXPRESSION OF c-fos GENE PRODUCTS IN VARIOUS KINDS OF HUMAN HEMATOPOIETIC CELL LINES. Immunofluorescence Study Using Prepared Monoclonal Antibodies

Two kinds of monoclonal antibodies recognizing fos proto-oncogene (c-fos) products were prepared using a synthetic oligopeptide corresponding to amino acids 127-152 of the fos oncogene products. These monoclonal antibodies (FO-120 & FO-145) detected fos gene products induced in a human monocyte cell line (U-937) by phorbol acetate (TPA) and induced in both human and mouse fibroblast cell lines (284, BALB/c 3T3) by serum-stimulation. One of the monoclonal antibodies (FO-120) reacted with 50-kDa and 42-kDa proteins and the other antibody (FO-145) reacted with a 30-33-kDa protein. The expression of the fos gene in various human hematopoietic cell lines was investigated using these prepared monoclonal antibodies. While almost all hematopoietic cell lines tested reacted with these monoclonal antibodies to various degrees, the majority of normal peripheral blood lymphocytes cultured with lectin (PHA) and interleukin 2 (IL-2) did not, suggesting that cells of some permanent hematopoietic cell lines, irrespective of their lineage specificity and growth factor dependency, continuously express the fos oncogene. These monoclonal antibodies may be useful for detecting early neoplastic changes in hematopoietic cells. ACTA PATHOL JPN 38: 1523-1536, 1988.     

Deduction from Wilms' tumour that glomerular podocytes produce the basement membrane material bearing Goodpasture determinants

Using an indirect immunoperoxidase technique, we tested frozen specimens from one Wilms' tumour composed of numerous glomeruloid bodies devoid of blood vessels, with monoclonal antibodies directed against vimentin, cytokeratin, CALLA/CD10, CD24, CR1/CD35, endothelium factor VIII, class I and II MHC molecules, laminin, fibronectin, and non-collagenic domain NC1 of type IV collagen. Two reagents against Goodpasture determinants were used: P1 monoclonal antibody and serum IgG (GP antibodies) from a biopsy-proven Goodpasture patient. Glomeruloid bodies comprised two cell types: a peripheral layer of parietal epithelial cells (cytokeratin and CD24-positive) and central cell clumps of podocytes (vimentin and CALLA-positive). The basal lamina surrounding the glomeruloid bodies contained laminin and NC1 domain of type IV collagen, while that present between the podocytes reacted strongly with laminin, and P1 and GP antibodies. Endothelium factor VIII was not detected within the glomeruloid bodies and CR1 molecules bound to the basement membrane material within them. These data favour the hypothesis that podocytes produce the basement membrane material which bears Goodpasture determinants recently identified as a novel chain, named the alpha 3 chain, of type IV collagen.