Marianne Jacobsen

Immunohistochemical investigations of tumors of supposed fibroblastic-histiocytic origin

The aim of this study was to localize alpha 1-antitrypsin, ferritin, and lysozyme by means of the indirect immunoperoxidase technique and to evaluate the significance of these antigens as markers of histiocytic differentiation in tumors of a supposed dual fibroblastic-histiocytic origin. The series comprised 31 malignant fibrous histiocytomas (MFH) of the pleomorphic, spindle cell, and myxoid types, four cutaneous fibrous histiocytomas, and four atypical fibroxanthomas, four dermatofibrosarcoma protuberans, and two osteoclastomas of bone. For comparison, 15 soft tissue sarcomas of various other types were examined. Of the MFHs of the pleomorphic type, 18 of 22 (82 per cent) were positively stained for alpha 1-antitrypsin and 12 of 22 (54 per cent) were positively stained for ferritin. Of the five MFHs of the spindle cell type, none was positively stained for alpha 1-antitrypsin, three were positive for ferritin, and one was positive for lysozyme. None of the myxoid variants (corresponding to grade I-II myxofibrosarcoma) was positively stained for either of the antigens. These results and the observations made on the cutaneous fibrous histiocytomas, atypical fibroxanthomas, dermatofibrosarcoma protuberans, and the various soft tissue sarcomas indicated that 1) alpha 1-antitrypsin is a valuable marker of histiocytic differentiation in both benign and malignant fibrous histiocytomas, 2) ferritin can be visualized in more than half of these fibroblastic-histiocytic tumors, and the presence of ferritin distinguishes the spindle cells of these tumors from fibroblasts of connective tissue and most fibrosarcomas, and 3) lysozyme, although a good marker of histiocytic differentiation in ordinary histiocytes and benign fibrous histiocytomas, is a poor marker of neoplastic histiocytes of malignant tumors. The results further support the concept that MFH is a tumor of a dual fibroblastic-histiocytic origin.

Immunohistochemical evidence for an intracellular localization of plasma proteins in human foetal choroid plexus and brain

The intracellular distribution of plasma proteins in human foetal choroid plexus and brain was investigated by immunohistochemistry. Small groups of cells or single scattered cells in the epithelial layer of the choroid plexus exhibited positive staining for alpha-fetoprotein, albumin and transferrin, whereas prealbumin was found in the majority of the epithelial cells. Numerous nerve cells in the cerebral wall and in various brain stem nuclei were positively stained for alpha-fetoprotein, albumin and prealbumin. All appropriate controls were negative. The presence of plasma proteins within choroid plexus epithelial cells suggest that these proteins are transported from blood to CSF by a transcellular route across the choroid plexus epithelium. The intracellular distribution of plasma proteins in developing neurons may indicate that these proteins play some important role in neuronal differentiation or development.

Immunohistochemical identification of some plasma proteins in human embryonic and fetal forebrain with particular reference to the development of the neocortex.

The histogenesis of the cerebral neocortex has been studied in human embryos and fetuses from the ventricular zone stage at 9-10 mm crown-rump length (CRL) to the well-developed neocortex at 210 mm CRL. The initial proliferation of the neuroepithelial cells in the ventricular zone stage was followed by a stage characterized by a ventricular zone covered by a primordial plexiform layer; the subventricular zone then arose before the cortical plate was formed within the primordial plexiform layer, thus dividing it into an outer marginal zone and an inner subplate zone; finally the intermediate zone appeared between the subventricular and subplate zones. The distribution of cells containing albumin, alpha-fetoprotein, transferrin, prealbumin, IgG and alpha 1-antitrypsin in the cerebral vesicle and developing neocortex was investigated by the indirect immunoperoxidase technique. Alpha-fetoprotein found in the cells of the ventricular zone was the most widespread and prominent of the plasma proteins examined in the early embryos. The cerebral vesicle was negative for all other plasma proteins investigated at this stage. By 15 and 16 mm CRL, a few cells in the ventricular zone were positive for albumin and transferrin whereas AFP exhibited a distribution similar to that of the 9 mm embryo. By 20-25 mm CRL, albumin and AFP had a similar distribution in the telencephalic wall. At 40-150 mm CRL a positive staining reaction for AFP, albumin, prealbumin and transferrin was predominant in the outer half of cortical plate. At 150-170 mm CRL only cells in the inner half exhibited positive staining and at 210 mm CRL the staining reactions were negative. The cells containing plasma proteins did not belong to a single cell line or type; thus plasma proteins were detected primarily in different types of neurons but also in glial cells. Staining with polyvalent antiserum indicated that the same cells may be positive for more than one plasma protein. Positive staining reactions were also observed in or along fiber systems. It is proposed that cells initially take up plasma protein from the CSF and migrate with it towards the cortical plate. After a certain period they lose their plasma protein but when the neuronal cells which represent the majority of the positively stained cells have reached their final position in the cortical plate they commence plasma protein synthesis which continues for a short period during which the neurons establish their pattern of connectivity.

Distribution of tumor-associated antigens in the various histologic components of germ cell tumors of the testis

The various histologic components of 39 germ cell tumors of the testis were studied with indirect labeled immunoperoxidase technique for the presence of the following tumor-associated antigens: alpha-fetoprotein (AFP), alphal-antitrypsin (AlAT), chorionic gonadotropin (HCG), specific pregnancy βl-glycoprotein (SPl), human placental lactogen (HPL), carcinoembryonic antigen (CEA), and ferritin (Fe). Embryonal carcinoma components were positive for AFP in 9/16, for AlAT in 6/16, for CEA in 2/16, and for Fe in 14/16. All yolk-sac tumor components were positive for AFP and Fe, and 8/13 contained AlAT and 2/13 CEA. Epithelium in teratoid components was positive for AFP in 5/14 cases, for AlAT in 8/14, for CEA in 7/14, and for Fe in 8/14. Syncytial trophoblast in choriocarcinoma components and syncytial giant cells were all positive for HCG, SPl, and HPL. In addition, tumor giant cells in two nonseminomatous tumors and in one seminoma contained HCG. Otherwise, all pure seminomas were negative for all antigens, except for Fe, which was positive in 12/16 cases. Demonstration of this functional aspect of germ cell tumors of the testis may clarify problems of classification and elucidate histogenesis. Furthermore, immunohistochemical demonstration of tumor-associated antigens is of value in the management of the patient as an indicator of which tumor markers should be used for monitoring the treatment. In addition, the presence of tumor-associated antigens may be used in prognostic evaluation.

Differential immunocytochemical staining for fetuin and transferrin in the developing cortical plate

SummaryThe distribution of the plasma proteins fetuin and transferrin in fetal sheep neocortex has been studied using the indirect immunoperoxidase method. Most of the cells (presumed to be neurons) in the early cortical plate (35 days gestation) were positive for fetuin. In contrast the fibre systems on either side of the cortical plate were positive for transferrin; some fibres were weakly positive for fetuin. It is proposed that these fibres may originate from brainstem and perhaps from some thalamic nuclei and that the plasma proteins are probably transferred from the neuronal cell bodies to their peripheral processes. It is further suggested that plasma proteins such as fetuin and transferrin may have a role in neuronal differentiation and synaptogenesis.

Intracellular plasma proteins in human fetal choroid plexus during development II. The distribution of prealbumin, albumin, alpha-fetoprotein, transferrin, IgG, IgA, IgM, and alpha1-antitrypsin

Individual choroid plexuses of human fetal brain at different developmental stages were investigated by an indirect immunoperoxidase technique. The number and distribution of prealbumin-, albumin-, alpha-fetoprotein-, transferrin-, IgG-, IgA-, IgM- and alpha 1-antitrypsin-containing choroid plexus epithelial cells were recorded. The distribution of prealbumin in choroid plexus epithelial cells differed from that of other plasma proteins identified in these cells. Prealbumin was present in more than 90% of the cells at all ages examined in contrast to albumin, alpha-fetoprotein, transferrin and IgG, which were found in less than 40% of the cells, and this proportion declined later in gestation to only a few per cent. The telencephalic plexus exhibited a staining pattern for the proteins, except for prealbumin, which was different from that of the diencephalic and myelencephalic plexuses. Double-staining and staining with polyvalent antiserum indicated that the same epithelial cells showed positive reaction for albumin, alpha-fetoprotein, transferrin and IgG. The results are related to the observation of a high concentration of plasma protein in fetal CSF which may result from transcellular transfer from blood to CSF across the choroid plexus epithelial cells.

Interobserver agreement for tumour type, grade of differentiation and stage in endometrial carcinomas

The histopathologic evaluation plays a major role in subdividing endometrial carcinomas into treatment groups. We have evaluated the interobserver agreement regarding tumour type, grade of differentiation, stage and stage I low and high risk cases. A total of 177 cases of endometrial carcinoma in which a hysterectomy and a bilateral salpingo‐oophorectomy were performed, were reviewed by three examiners. A variety of features including tumour type, architectural grade, nuclear grade, FIGO grade, and spread/metastases were recorded, and the FIGO stage was determined. Using two different definitions low and high risk groups in stage I tumours were separated. A kappa value was calculated for each of the various parameters. The current study showed a good strength of agreement for tumour type, myometrial invasion, spread/metastases, and FIGO stage (kappa 0.62–1.00). For two of the examiners good agreement was found as to architectural grade (kappa 0.71) while the kappa value for nuclear grade was lower (0.56). As nuclear grading is included in the revised FIGO recommendation a precise definition of nuclear atypia is needed. In stage I tumours very good agreement was demonstrated as to the defined low and high risk group (Kappa 0.64–0.86).

The choroid plexus in fetal sheep during development with special reference to intracellular plasma proteins

Abstract The developmental stages of the telencephalic, diencephalic and myelencephalic choroid plexuses and the morphology of the choroid plexus epithelial cells in the fetal sheep during development are briefly described. The individual choroid plexuses at different developmental stages were investigated by the indirect immumoperoxidase technique. The number and distribution of albumin-, alpha-fetoprotein-, fetuin-, transferrin-, and alpha1 -antitrypsin-containing cells were recorded. Early in gestation 30–50% of the epithelial cells in the telencephalic choroid plexus were positive for albumin, alpha-fetoprotein and fetuin, and this proportion declined later in gestation to a few percent. In contrast, during the entire gestational period investigated, less than 10% of the epithelial cells in the diencephalic and myelencephalic choroid plexuses showed a positive staining reaction for the various proteins. The cells containing plasma proteins did not belong to a single type. No positive staining reaction for transferrin and alpha1 -antitrypsin was found in the choroidal epithelium in any of the individual plexuses. The results are compared to the staining pattern in human fetal choroid plexus. Furthermore, the results are discussed in relation to the concentration of the proteins in CSF and their transfer from blood to CSF.

Intracellular plasma proteins in human fetal choroid plexus during development I. Developmental stages in relation to the number of epithelial cells which contain albumin in telencephalic, diencephalic and myelencephalic choroid plexus

The developmental stages of telencephalic, diencephalic and myelencephalic choroid plexuses in the human fetus and the morphology of choroid plexus epithelial cells in the various plexuses in different development stages were described on basis of PAS- and toluidine blue-stained material. Six different cell types were identified in various combinations in 4 different stages (I-IV). The number and distribution of albumin-containing epithelial cells in various plexuses in the different stages of development were investigated by indirect immunoperoxidase technique. Albumin-containing cells did not belong to a single cell type. The telencephalic plexus exhibited a staining pattern for albumin which was different from that of the diencephalic and myelencephalic plexuses. In the telencephalic plexus positive epithelial cells were frequent in stage I, whereas only a few positive cells were present in stage II and III. In contrast, 30-40% of the epithelial cells in both diencephalic and myelencephalic plexuses in stage I, II and III showed a positive staining reaction. Later in gestation less than 10% were positive. It is suggested that a main function of the diencephalic and myelencephalic plexuses early in gestation is associated with protein transport rather than with glycogen synthesis and storage which might be a major function of the telencephalic plexus.

Possible liver cell differentiation in testicular germ cell tumours.

Germ cell tumours may imitate various structures of the developing embryo and foetus. Certain structures have, however, very rarely or never been observed in these tumours. Thus the presence of hepatic tissue in testicular germ cell tumours has not been reported. In a series of 37 non‐seminomatous testicular tumours seven tumours contained epithelial structures showing morphological and functional resemblance to liver cell trabeculae. These structures were present in tumours with yolk sac tumour (YST) components and most of the tumours also contained teratoid elements. With the immunoperoxidase technique the epithelial structures were heavily stained for alpha‐foetoprotein (AFP) and ferritin in all cases, while positive staining for albumin, prealbumin and transferrin was occasionally found. Alpha‐i‐antitrypsin and haemoglobin F were demonstrated in few scattered cells. Whether or not these epithelial structures should be included among the various patterns of YST or considered to be teratoid components is uncertain. It is suggested that examinations of the heterogeneity of the concomitant serologic AFP may support one or other assumption.