Interactions between Ricinus Agglutinin and Human Plasma Proteins

Ricinus agglutinin purified to homogeneity precipitates certain serum glyco-proteins containing terminal non-reducing galactose residues. Immunoelec-trophoresis of human sera in agarose gel with ricinus agglutinin in the antibody trough gave 2 fusing precipitin lines due to reaction with IgM and haptoglobin. All of 50 monoclonal IgM proteins precipitated with ricinus agglutinin, and the reactive sites were localized to the Fc μ fragment. By affinity chromatography on Sepharose columns containing insolubilized ricinus agglutinin prealbumin, albumin, α₁-lipoprotein, α₁-antitrypsin, α₁-acid glycoprotein, α₁-antichymotrypsin, α₂HS glycoprotein, Ge-globulin, caerulo-plasmin, β-lipoprotein, β₁ C-globulin, and transferrin were not retained, whereas α₂-macroglobulin, haptoglobin, IgM, IgA, and IgG were retained and could be eluted with lactose. Fused rocket immunoelectrophoresis revealed varying degrees of microheterogeneity among the latter proteins; almost all of IgM but only a small fraction of polyclonal IgG was retained on the ricinus column.     

α-Fetoprotein Synthesis in Yolk Sac Tumor: Sex-dependent Production of α-Fetoprotein by Transplantable Rat Yolk Sac Tumor

Transplantable rat yolk sac tumor cells produce α-fetoprotein (AFP) and their ability to synthesize AFP is maintained for generations in female rats. However, when they are transplanted in male rats, the AFP production decreases markedly.
To examine this sex dependency in AFP production, experiments at cellular and subcellular levels were carried out. In cell incubation studies, yolk sac tumor cells maintained in female rat (YST-F cells) synthesized more AFP than yolk sac tumor cells maintained in male rat (YST-M cells). Using cytosol RNAs prepared from YST-F and YST-M cells, AFP production was studied in cell-free protein synthesizing system derived from wheat germ.
In this system, cytosol RNA from both YST-F and YST-M cells directed AFP synthesis. But the amount of AFP synthesized was smaller in the presence of RNA from YST-M cells. From these results, it was suggested that the reduced AFP synthesis by YST-M cells is, at least partly, due to a quantitative decrease in their cytosol messenger RNA coded for AFP.     

Rat Yolk Sac Tumor: Biological Aspects and Microheterogeneity of AFP

α-Fetoprotein (AFP)-producing yolk sac tumors were established in rats as transplantable tumor lines growing in either solid or ascitic form. AFP levels in the ascites and in the serum of recipients were quite high, but higher in castrated female rats than male rats. The tumor cells were cultured also in vitro and showed synthesis of AFP and albumin. AFP produced by the yolk sac tumors was investigated comparatively with AFP produced by a hepatoma. There was no remarkable difference between two AFPs in behavior in gel filtration; however, AFP of yolk sac tumors migrated more slowly than that of hepatoma in immunoelectrophoresis. However, the mobilities of two AFP preparations became same after desialization by neuraminidase treatment. This fact suggests that AFP of yolk sac tumors contains sialic acids in smaller amount than that of hepatomas.     

Immunostaining for α1-antichymotrypsin and α1-antitrypsin in gliomas

Antisera to α₁-antichymotrypsin, α₁-antitrypsin and lysozyme were reacted with 20 cases of glioblastoma multiforme, seven anaplastic astrocytomas, eight astrocytomas, six oligodendrogliomas, four ependymomas and the cerebral cortex from six normal autopsy brains. In addition, two pleomorphic xantho-astrocytomas and two heavily lipidized malignant gliomas were similarly examined. All astrocytic lesions were confirmed with anti-GFAP antisera. Thirty astrocytic tumours (77%), four oligodendrogliomas (67%) and three ependymomas (75%) reacted positively with anti-α₁-antichymotrypsin; 25 astrocytic tumours (64%), three oligodendrogliomas (50%) and three ependymomas (75%) showed positive staining for α₁-antitrypsin. The pattern of staining with either of these two markers did not correlate with tumour grading. None of the gliomas examined stained positively with anti-lysozyme. Non-neoplastic glial elements did not react with any of the three antisera. The results of this study suggest that staining for α₁-antichymotrypsin and α₁-antitrypsin is of little value in the differential diagnosis of neuroepithelial or mesenchymal lesions in the brain.     

Human Membrane-bound C3 Receptors I. Serological and Immunohistological Demonstration of C3 Receptors

The aim of the present study was to present further evidence of the specific reactivity of an anti-C3 receptor serum (AC3RS), to demonstrate membrane-bound C3 receptors by using this AC3RS in different serological and immunohistological methods, and to investigate the relationship between membrane-bound C3 receptor and α₁-antitrypsin. The AC3RS, or F(ab)₂ fragments of the IgG fraction of this antiserum, stained a percentage on various viable cell populations roughly equivalent to the number of cells that bound EAC3b and or EAC3d; C3 receptor-negative T cells and thymocytes were not stained. On frozen sections of tonsils and kidneys it was found that the AC3RS stained the area to which EAC3b adhered. After absorption with neutrophils or E_hu, the AC3RS Inhibited the agglutination of EAC3d with tonsil cells, not the agglutination of tonsil cells or neutrophils with EAC3b; this absorbed AC3RS still stained tonsil cells but not neutrophils, in frozen tonsil sections it Stained only those areas to which AC3d adhered. The absorbed AC3RS did not stain glomeruli. Antisera to α₁-antitrypsin failed to in-hibit EAC agglutination with C3 receptor-bearing cells or to stain C3 receptor-positive cells either in suspension or in frozen sections. Absorption of the AC3RS with purified α₁-antitrypsin did not affect its specific reactivity.     

Migration of Prebursal Stem Cells from the Early Chicken Embryo to the Yolk Sac

Allogeneic yolk sac—embryo chimaeras were constructed by association of S¹⁵B¹⁵ yolk sac and B²B² embryo on day 2 of incubation. Five days later yolk sac cells from the chimaeras were injected intravenously into 14-day-old irradiated embryos, using recipients of B²B² and B¹⁵B¹⁵ genotypes. One week after hatching, cells in the bursa of Fabricius and peripheral blood erythrocytes were studied for la-like antigens and B alloantigens, respectively, to determine whether they were derived from the embryo or yolk sac part of the chimaera. The results obtained demonstrate that prebursal and erythropoietic stem cells migrate from the early embryo to the yolk sac during the 2nd to the 7th day of incubation. They also exclude the de novo generation of prebursal stem cells in the yolk sac.     

GIANT CELL TUMOUR OF TENDON SHEATH| An Immunohistochemical Study of 28 Cases

An immunohistochemical study was carried out on 28 cases of giant cell tumor of tendon sheath. Although this tumor has been considered to be of histiocytic origin on the basis of light and electron microscopic findings, there remains some debate about the histogenesis of the tumor. To clarify this point, by using the PAP method, each surgical specimen was stained for α₁- antitrypsin, a₁-antichymotrypsin, lysozyme, ferritin, neuron specific enolase, and S-100 protein. Tumor cells in fifteen out of 28 cases were positively stained for α₁-antitrypsin, 19 for α₁-antichymotrypsin, 23 for lysozyme, 22 for ferritin, 22 for neuron specific enolase, but no case for S-100 protein. These results suggest that this tumor is composed of cells with histiocytic character. In addition, from the immunohistochemical point of view, at least two types of giant cells seem to exist in this disease. ACTA PATHOL. JPN. 36:1487-1494, 1986.     

Is the PiF allele of α1-antitrypsin as-sociated with pulmonary disease?

Pulmonary function was studied in thirteen individuals heterozygous for the α₁-antitrypsin allele Pi^F. Respiratory symptoms were present in seven out of twelve individuals with the FM phenotype, of which five had pulmonary function impairment, mostly of the obstructive type. One patient with the phenotype FZ had bronchitic symptoms and a mild obstructive spirometry pattern. The results suggest a relationship between the Pi^F allele and chronic obstructive pulmonary disease, which is independent of the serum α₁-AT level.     

卵黄囊瘤卵黄囊分化的形态学特征及其与血清AFP的关系

本文报道100例卵黄囊瘤免疫组化标记,并与16例患者血清AFP水平相比较。结果表明,脏层卵黄囊分化(88例)AFP阳性,且与血清AFP升高相关;而壁层卵黄囊分化(92例)Laminin基膜样物质阳性。血清AFP测定有助于本瘤术前诊断、术后随访和预后估价。     

BENIGN HISTIOCYTOSIS X OF STOMACH Previously Undescribed Lesion

Histiocytosis X of the stomach of a 47-year-old Japanese woman, who underwent subtotal gastrectomy following a clinical diagnosis of scirrhous carcinoma, was studied by light and electron microscopy as well as by immuno-histochemistry. The histiocytoid cells proliferated monotonously in the lamina propria mucosae of the atrophied mucosa covering the body and fornix. They were arranged in a sheet- or pavement stone-pattern and included some giant cells. The histiocytoid cells had a reniform to irregularly indented nucleus and conspicuous cytoplasm. Ultrastructurally, they were characterized by interdigitating cytoplasmic extensions and abundant tubulovesicular structures including Langerhans granules. S-100 protein, α₁-antitrypsin, and α₁-antichymo-trypsin were immunohistochemically identified in the cytoplasm. Endoscopic biopsies of the extragastric digestive tract, a biopsy of the lymph node, and bone marrow aspiration excluded a systemic disorder. The case is regarded as benign localized histiocytosis X of the stomach, a previously undescribed gastric lesion.     

Expression of Integrin αvβ3 in Gliomas Correlates with Tumor Grade and Is not Restricted to Tumor Vasculature

In malignant gliomas, the integrin adhesion receptors seem to play a key role for invasive growth and angiogenesis. However, there is still a controversy about the expression and the distribution of α_vβ₃ integrin caused by malignancy. The aim of our study was to assess the extent and pattern of α_vβ₃ integrin expression within primary glioblastomas (GBMs) compared with low-grade gliomas (LGGs). Tumor samples were immunostained for the detection of α_vβ₃ integrin and quantified by an imaging software. The expression of α_vβ₃ was found to be significantly higher in GBMs than in LGGs, whereby focal strong reactivity was restricted to GBMs only. Subsequent analysis revealed that not only endothelial cells but also, to a large extent, glial tumor cells contribute to the overall amount of α_vβ₃ integrin in the tumors. To further analyze the integrin subunits, Western blots from histologic sections were performed, which demonstrated a significant difference in the expression of the β₃ integrin subunit between GBMs and LGGs. The presented data lead to new insights in the pattern of α_vβ₃ integrin in gliomas and are of relevance for the inhibition of α_vβ₃ integrin with specific RGD peptides and interfering drugs to reduce angiogenesis and tumor growth.     

Selective Cytotoxicity of Two Rodent T Cell Lymphomas to Rat Yolk Sac Tumours Involves a Retroviral Envelope Protein Expressed by the Lymphoma

A Gross virus induced rat T cell lymphoma G1-Tc1 and a Moloney virus induced mouse T cell lymphoma YAC-1 are shown to exert a strong cytotoxic activity against rat yolk sac tumours but not to various types of rat, mouse or human normal cells or tumour cell lines including carcinomas, sarcomas, lymphomas and gliomas. Both lymphomas are CD3⁺, CD4⁻, CD8⁻ and T-cell receptor (TCR)αβ⁺. The cytotoxicity was not MHC restricted or dependent on the density of MHC class I of the target cells, and the mouse lymphoma killed the rat yolk sac tumour target. The cytotoxic action was fast and up to 80% specific killing was observed in 4-h ⁵¹Cr release assays. A rat B cell hybridoma was established from a Wistar/Furth (WF) rat immunized with the syngeneic lymphoma G1-Tc1 producing an immunoglobulin (Ig)G2c monoclonal antibody (MoAb) 1F2. This binds to the lymphomas G1-Tc1 and YAC-1 and also to a murine non-cytolytic Rauscher lymphoma RMA, but not to any other of several rat, mouse or human cell types tested. The 1F2 completely inhibited the killing of rat yolk sac tumours by the two cytolytic lymphomas, but did not interfere with the killing mediated by natural killer (NK) cells or cytolytic lymphokine-activated killer (LAK) cells. Immunochemical analysis of solubilized cell membranes of the lymphoma G1-Tc1 demonstrates that the 1F2 antibody recognizes an epitope on a retroviral gp 70 envelope protein. This indicates that a retroviral protein is involved in the lytic activity of the two lymphomas.     

Immunohistochemical Characterization of Human Cutaneous Mast Cells in Urticaria Pigmentosa (Cutaneous Mastocytosis)

To clarify the origin and function of human cutaneous mast cells (CMCs), immunohistochemical characterization was done in 19 cases of urticaria pigmentosa (cutaneous mastocytosis) using 9 antibodies (anti leukocyte common antigen, MX-PanB, anti lysozyme, anti α₁, antitrypsin, anti α₁-antichymotrypsin, anti vimentin, anti-neuron specific enolase, anti factor VIII related antigen, and anti-ACTH). CMCs showed positive reactions with anti α₁ anti-chymotrypsin and anti vimentin in almost all of the specimens. In more than half of the specimens, CMCs were stained positively with anti -α₁-antitrypsin, MX-PanB, and anti factor VIII related antigen. Anti-leukocyte common antigen and anti ACTH also showed positive reactions in some specimens. These results confirm the existence of vimentin filaments in CMCs and suggest a functional role of CMCs in hemostasis via factor VIII. Furthermore, immunohistochemical similarity between CMCs and granulocyte/macrophage group cells is also suggested.     

The Influence of Serum on Random Migration and Chemotaxis of Polymorphonuclear Leucocytes: Methodological Evaluation using Sera from Infection-prone Patients and Normals

The effects on normal polymorphonuclear leucocyte (PMN) of chemotactic and chemokinetic factors in sera from normal subjects and Infection-prone patients wan examined by means of the leading-from technique, using a modified Boyden chamber. The analysts schema used made it possible to differentiate between chemotactic and chemokinetic factors and demonstrated that different factors account for the major chemotactic and chemokinetic activities of serum. The major chemotactic activity of unactivated serum was heat-labile, and the chemotactic activity of factor XII-deficient sera was normal, suggesting the major chemotactic activity to be distinct from C5a and factor XII-dependent pathways. The existence of both heat-stable and heat-labile chemokinetic factors was shown. The possibility that the reduced chemokinetic effect of several patient sera was caused in abnormal levels of the serum proteins γ₁-antitrypsin, γ₂macroglobulin and albumin was excluded.     

Association of severe rheumatoid arthritis with heterozygosity for α-antitrypsin deficiency

Genetic types of α₁-antitrypsin (protease inhibitor types, or Pi types) were determined in 108 patients with rheumatoid arthritis. These patients were selected for severely destructive disease and had classical rheumatoid arthritis according to ARA criteria, were seropositive, and had joint erosions shown by X-ray. Heterozygotes for the deficiency Z allele (Pi types MZ, SZ, etc.) were found among 9.2 % of patients and 3.5 % of a control adult population. The increased frequency in patients was statistically significant. Heterozygotes were most frequent among female patients with an early onset of disease. Heterozygosity for α₁-antitrypsin deficiency may be a factor in familial recurrence of rheumatoid arthritis. Among 98 patients with juvenile rheumatoid arthritis not selected for severity, 4.1 % were Z heterozygotes compared with 1.3 % of control children, not a statistically significant difference. Reduced concentrations of α₁-antitrypsin in Z heterozygotes may be inadequate to inhibit the proteolytic enzymes released into the joints of adults with rheumatoid arthritis during phagocytosis of immune complexes. This may be a factor promoting severe joint destruction.     

A Sensitive and Simple Determination of Human α-fetoprotein by Enzyme-electroimmunodiffusion

A simple and sensitive determination method of α-fetoprotein was developed by introducing α-fetoprotein horseradish peroxidase conjugate into electroimmunodiffusion.
The precipitin line formed was directly visualized by staining for peroxidase activities. The sensitivity was 32 ng/ml. Electrophoretic activities of AFP were analyzed by two dimensional electrophoresis applying the present method to the second electrophoresis.
AFP from normal adults and several patients was confirmed to migrate as an α-globulin as AFP of hepatoma, yolk sac tumor and fetuses did.
AFP-like activities were detected in β-γ-globulin region in some samples but these were not due to AFP-anti-AFP interactions.     

PANCREATIC CARCINOMA IN CHILDHOOD

A case of pancreatic carcinoma in a 14-year-old Japanese boy is reported. He complained of general fatigue, anorexia, abdominal distension, and abdominal mass. At autopsy, a whitish tumor was found from the head to the body of the pancreas. Metastasis was found in the liver, lungs, gall bladder, and various lymph nodes such as stomach, hilus, and periaorta. The tumor was histologically determined to be moderately differentiated adenocarcinoma (cribriform type) of duct cell origin. However, the tumors showed PAS-positive diastase-resistant mucus in the cytoplasm. Histocytology showed the positivity for α₁-antitrypsin, secretory component (sc), and CEA, but no S-amylase was detected in the cytoplasm. Electron microscopy revealed zymogen-like granules in the cytoplasm suggesting acinar differentiation.     

Immunohistochemical Investigation of Vimentin, Neuron-specific Enolase, α1-antichymotrypsin and α1-antitrypsin in Adenoid Cystic Carcinoma of the Salivary Gland

Fifty-four adenoid cystic carcinomas (ACC) arising in major and minor salivary glands as well as in normal salivary glands were studied by immunohistochemistry for the presence of vimentin, neuron specific enolase (NSE), α₁-antichymotrypsin (α₁-ACT) and α₁-_- antitrypsin (α₁-_- AT). Five patterns of histological differentiation were found in ACC, and for the cellular components of each, it was possible to establish a special immunohistochemical profile. In ACC, vimentin-positive cells were observed in the outer tubular, cyst-lining and small angular cells. NSE was positive in the myoepithelial cells of normal salivary gland. Neoplastic cells of ACC showed NSE positivity mainly in the small angular cells and partly in the duct luminal cells. α₁-ACT was localized in the intercalated duct cells and serous acinar cells of normal salivary gland, and in the duct luminal cells of ACC. α₁-AT could not be detected in any of the epithelial cells of normal salivary gland. In ACC, eosinophilic hyaline material in the cribriform spaces was positive for α₁-AT, but no positivity was demonstrated in tumor cells. The present study showed that there are at least two populations of tumor cells in ACC: duct luminal cells that express α₁-ACT, thus indicating their ductal character, and small angular cells that express vimentin, characteristic of non-luminal cells. Moreover, our results indicate that α₁-AT is a useful marker of basement membrane-like material.     

Immunoglobulins and Other Serum Proteins in Feces from Infants and Children

IgA in variable amounts was found in extracts of feces from all of 14 infants and children with no apparent diseases involving the gastrointestinal tract or the immune system, whereas IgG and IgM were only found in some. No age variation was evident's even one-month-old infants seemed fully able to secrete intestinal immunoglobulins. Freeze-drying of the feces facilitated the extraction, and the amounts of immunoglobulins were related to the dry weight of fecas. Most of the immunoglobulins were easily dissolved in saline. Of other serum proteins α₁ antitrypsin was constantly present; α₁ antichymotrypsin and α₂ macroglobulin were found in many. Only traces of albumin were sometimes demonstrated.     

An autopsy case of hepatic sarcomatoid tumor: Immunohistochemical comparison with a sarcomatous component of hepatocellular carcinoma

A case of primary hepatic tumor exclusively composed of malignant cells with sarcomatous features is described and compared immunohistochemically with two cases of hepatocellular carcinoma (HCC) with a sarcomatous component. More than 30% of HCC cells were positively stained with anti-cytokeratin (CAM5.2), anti-albumin, anti-fibrinogen and anti-α₁-antitrypsin antibodies, and some with anti-epithelial membrane antigen. The present sarcomatoid tumor and the sarcomatous component with HCC showed similar immuno-histochemistry; many tumor cells were strongly immuno-reactive for vimentin and some positive for cytokeratin, albumin, fibrinogen and u,-antitrypsin. Other immunohistochemical markers, indicating specific differentiations to lineage of macrophages, muscle cells, glial cells, endothelial cells and so forth, were not detected in sarcomatous tumor cells of all cases. These findings suggest that the present sarcomatoid tumor would belong to an anaplastic sarcomatous variant of HCC.