Monoclonal antibodies against human granulocytes and myeloid differentiation antigens

Monoclonal antibodies (MCA) were obtained by immunizing BALB/c mice with 99% pure granulocytes from normal donors or with a whole leukocyte suspension obtained from a chronic myelogenous leukemia (CML) patient, and then fusing the mouse spleen cells with a 315–43 myeloma cell clone. Four MCA were selected and studied using ELISA, immunofluorescence, cytotoxicity assays, and FACS analysis. Antibodies 80H.1. 80H.3. and 80H.5 (from normals) and 81H.1 (from CML) detected antigens expressed on neutrophils. Antibodies 80H.1 and 80H.3 (lgG) also reacted with monocytes but not with other blood cell subsets. Antibodies 80H.5 and 81H.1 (lgM) were cytotoxic and reacted strongly with most of the cells of the neutrophil maturation sequence. i.e., myeloblasts, promyelocytes, myelocytes, and mature granulocytes. Antibodies 80H.5 and 81H.1 also inhibited BFU-GM and CFU-E. Antigens recognized by 80H.3. 80H.5, and 81H.1 were expressed both on a proportion of cells from HL.60, KG.1, ML.1, and K562 myeloid cell lines, and on a proportion of blast cells isolated from patients with acute myelogenous leukemia. They were not found on lymphoid cell lines or lymphoid leukemia cells. These MCA recognize either late differentiation antigens expressed on mature neutrophils and monocytes (80H.1 and 80H.3) or early differentiation antigens (80H.5 and 81H.1) specific to the granulocytic lineage. They may be useful for a better definition of those antigens specific to hematopoietic stem cells and their relationship with normal or neoplastic hematopoiesis.     

Aussagekraft der präoperativen Thrombozytenmarkierung bei Patienten mit idiopathischer thrombozytopenischer Purpura

Summary ⁵¹Cr-platelet kinetic studies were performed in 77 patients with idiopathic thrombocytopenic purpura. The sequestration site was splenic for 63, splenic/hepatic for 7 or hepatic for remaining 7 patients. In 20 patients platelet survival was extremely shortened to 0–3 h, whereas only 26 patients had a survival time of more than 24 h. Those patients with low platelet counts also had a very short platelet survival time, whereas patients with higher platelet counts (>50×10⁹/l) had longer platelet survival times. 51 patients (66%) were splenectomized following the kinetic studies. 25 patients who had a splenic sequestration site had normalized platelet counts and 6 patients had platelet counts between 80–149×10⁹/l 12 months after splenectomy (i.e. in 92% of cases with splenic sequestration site a full or partial remission). Of the 11 patients with a hepatic or splenic/hepatic sequestration site, 2 patients had full remission, 1 partial remission, 3 patients had minimal improvement and 5 other patients were treatment failures in respect to the splenectomy.

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Abkürzungen ACD-A Acid-Citrat-Dextrose Lösung A - HBsAge Hepatitis-B Oberflächen-Antigen - ITP idiopathische thrombozytopenische Purpura - PAIgG plättchenassoziierte IgG-Antikörper - TRP thrombozytenreiches Plasma     

Postmortem Diagnosis of Acute Megakaryocytic Leukemia Usefulness of lmmunohistochemistry and Tissue Hemogram

An autopsy case of acute megakaryocytic leukemia (AMKL) is presented. The bone marrow was hypercellular with proliferation of three lineages, especially megakaryocytes. Immunohistochemical examination revealed many platelet glycoprotein IIb/IIIa (GP IIb/IIIa)- positive blast cells in bone marrow. The proportion of the blasts was 26.4% by tissue hemogram. GP IIb/IIIa-positive blasts and megakaryoblasts were deposited massively in lymph nodes. lmmunohistochemistry against GP IIb/IIIa and tissue hemograms by paraffin section are needed to diagnose AMKL by postmortem examination, since the identification of ultra-structural platelet peroxidase in autopsy materials is difficult.     

Tumor cells induced by the v-src oncogene are heterogeneous for expression of markers of mesenchyme differentiation

The observation that v-src-induced tumors contain tumor cells of differing morphology, notably fibroblastoid or polygonal, raised the question as to whether the tumor cells are also heterogeneous with respect to expression of markers of cellular differentiation. Of the markers tested here, consistent reactivity for tumor tissue was noted only for antibody probes reactive to muscle actin (HHF35, sm-1) or to procollagen type I (SP1. D8); for any given tumor, whether induced by v-src DNA or by Rous sarcoma virus, each of these markers was found only in a subpopulation of tumor cells. The observation of marker heterogeneity in the one v-src DNA-induced tumor examined here that typed as monoclonal suggests that v-src-induced transformation is consonant with a degree of plasticity in the phenotypes of the clonal progeny of a single transformant.     

Comparison of radioimmunoassay and ELISA methods for detection of antibodies to chromatin components

A solid phase radioimmunoassay has been compared with an enzyme-linked immunosorbent assay (ELISA) for efficacy in measuring anti-chromatin antibodies. The low backgrounds achieved with the radioimmunoassay method produced a high signal-to-noise ratio and enabled detection of the human test antiserum at a dilution of 1:102,400. By contrast, the ELISA could detect the same antiserum only at a dilution of 1:3200 and above. The radioimmunoassay was consistently more sensitive than the ELISA for detection of anti-chromatin antibodies in a number of human and mouse sera and ascites fluid containing a monoclonal antibody. Factors affecting sensitivity in both assays are discussed.     

脑啡肽与生长抑素在鸡视网膜无长突细胞共存的免疫组织化学研究

本文介绍用免疫组织化学的单标和双标技术研究脑啡肽(ENK)和生长抑素(SOM)在鸡视网膜无长突细胞的定位和共存。单标的实验结果表明,一些SOM免疫反应阳性无长突细胞的形态、胞体在内核层的位置及其突起在内网层的分支式样与某些ENK免疫反应阳性无长突细胞相似,虽然其突起在内网层的第3、4亚层形成的丛网不象ENK免疫反应阳性突起那样丛密,在内网层的第5亚层也未见SOM免疫阳性突起。双标的实验结果表明,一些无长突细胞显示ENK和SOM两种免疫阳性反应,而另一些无长突细胞分别只显示ENK或SOM阳性免疫反应。文中还对视网膜神经多肽间或与经典神经递质的共存进行了讨论。     

维生素A阻抑大鼠实验性肝纤维化的病理研究

利用光镜、电镜、免疫组化和形态学定量技术动态研究维生素A对大鼠四氯化碳肝纤维化的抑制作用。结果表明,维生素A可减少四氯化碳中毒大鼠肝内纤维连接蛋白和Ⅰ、Ⅲ型胶原沉积,抑制贮脂细胞向成纤维细胞转化,并可明显地减轻肝纤维化程度。本文还对维生素A抑制肝纤维化的机理及意义作了初步探讨。     

大鼠下丘脑及邻近区域催产素免疫阳性神经元与垂体后叶的关系

本文运用免疫细胞化学PAP及ABC法,显示大白鼠下丘脑内OXT免疫阳性神经元,并于垂体后叶注射WGA-HRP,显示下丘脑中逆行标记细胞,结合免疫细胞化学方法,观察下丘脑及其邻近区域内HRP与OXT双标记细胞,证实下丘脑视上核、室旁核、穹窿前核和后核、血管周细胞群、下丘脑视前区、下丘脑前区及外侧区、背侧副细胞群内、室周部、第三脑室侧壁室管膜细胞下及室间孔部室管膜细胞下,均有OXT免疫阳性神经元,其中至少部分神经元可发出向垂体后叶的投射纤维。位于第三脑室侧壁室管膜下及室间孔部室管膜下的神经元,可能监测脑脊液中各种因素的变化,调节垂体后叶OXT的分泌,也可能直接通过共树突向脑脊液内释放OXT。