Solitary mastocytoma of the eyelid

Summary Solitary mastocytoma (mast cell naevus) of the skin represents a relatively rare dermal tumour. Its occurrence on the lower eyelid is exceptional. We report the case of a 4 month old male infant who exhibited a firm, yellowish nodule (1 cm in maximum diameter) on the lower lid of the right eye from birth. Histologically, the tumour consisted of strongly metachromatic tissue mast cells (TMC) infiltrating the whole dermis, the adjacent subcutaneous tissue and the lid muscle. Since comparable skin lesions in other sites were not observed, a diagnosis of solitary mastocytoma was made. Immunocytological investigations revealed strong reactivity of the TMC to antisera against vimentin, common leucocyte antigen (CLA),1-antitrypsin (1-AT) and 1-antichymotrypsin (1-ACT). A minor proportion of the TMC reacted to antisera against lysozyme and KiB3. Surprisingly, the TMC also reacted to antisera against certain regulatory peptides (RP), namely adrenocorticotropic hormone (ACTH), peptide histidine isoleucine (PHI), leu-enkephalin and met-enkephalin. However, absorption controls revealed that the immunostaining for ACTH and the two enkephalins was non-specific. The immunocytological phenotype of TMC suggests a close relationship to the myeloid-monocytic lineage, but a possible relationship between TMC and the diffuse neuroendocrine system needs further investigation.     

Immunohistochemical localization of lysozyme, lactoferrin, alpha 1-antichymotrypsin, and alpha 1-antitrypsin in salivary gland of human fetuses.

26 human fetuses were examined to elucidate the immunohistochemical distributions of lysozyme, lactoferrin, alpha 1-antichymotrypsin, and alpha 1-antitrypsin in prenatal salivary glands. Development of fetal salivary glands was divided into 4 stages: The early developmental stage (EDS), the early intermediate developmental stage (EIDS), the late intermediate developmental stage (LIDS), and the late developmental stage (LDS) and were used to compare antigen localization during salivary gland development. Lysozyme (LY) staining was prominent in serous or demilune cells of the mucous acinar compartment. Lactoferrin (LF) was rarely seen in the fetal glands; only trace amounts were seen in serous cells, alpha 1-antichymotrypsin (alpha 1-ACT) was diffusely positive particularly in glandular ducts, alpha 1-antitrypsin (alpha 1-AT) was also diffusely distributed in all salivary gland elements and was more abundant in ductal cells than acinar cells. During the EDS, immunohistochemical staining of LY, LF, alpha 1-ACT, and alpha 1-AT could be observed with glandular intensity increases corresponding to the advance of cytodifferentiation of granular epithelium occurring in the subsequent EIDS and LIDS. Staining intensities were continuous during the LDS even though the amount of those materials in the fetal salivary glands was not of the extent seen in the adult salivary gland. These results suggest that production of LY, LF, alpha 1-ACT, and alpha 1-AT was positive during prenatal development of human salivary glands. The present study discusses the protective roles and defense mechanisms of LY, LF, alpha 1-ACT, and alpha 1-AT in developing human salivary glands.     

Immunohistochemical investigation of vimentin, neuron-specific enolase, alpha 1-antichymotrypsin and alpha 1-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), alpha 1-antichymotrypsin (alpha 1-ACT) and alpha 1-antitrypsin (alpha 1-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. alpha 1-ACT was localized in the intercalated duct cells and serous acinar cells of normal salivary gland, and in the duct luminal cells of ACC. alpha 1-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 alpha 1-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 alpha 1-ACT, thus indicating their ductal character, and small angular cells that express vimentin, characteristic of non-luminal cells. Moreover, our results indicate that alpha 1-AT is a useful marker of basement membrane-like material.     

Immunohistochemical study of malignant fibrous histiocytoma

Malignant fibrous histiocytomas (MFH) and other soft-tissue tumors were examined immunochemically (by the peroxidase-antiperoxidase technique) for the presence and location of alpha 1-antitrypsin (alpha 1-AT) and alpha 1-antichymotrypsin (alpha 1-ACT). alpha 1-AT and alpha 1-ACT were found in substantial amounts in 19 and 20 of the 20 MFH tumors examined, respectively. In the other soft-tissue tumors, they occurred inconsistently and in less than 50% of cells, mostly those with signs of damage. It is concluded that assays for alpha 1-AT and alpha 1-ACT may aid in the diagnosis and characterization of malignant fibrous histiocytoma.     

Distribution of tissue markers in acinic cell carcinomas of salivary gland.

Eight cases of acinic cell carcinoma of the salivary glands were histologically reclassified and their immunohistochemical expression and distribution for various tissue antigens were examined. The epithelial elements were divided into tubuloglandular components, microcystic patterns and solid nests. The authors' results indicated the following: 1) The duct luminal cells of tubuloglandular components have distinct epithelial features with cytokeratin (KL 1), alpha 1-antichymotrypsin (alpha 1-ACT), transferrin, lactoferrin, IgA, and carcinoembryonic antigen (CEA) positivity. 2) The cyst-lining cells of microcystic pattern expressed immunophenotypes similar to those of the duct luminal cells. 3) The acinic cells in solid nests had positive results for KL 1, alpha 1-antitrypsin (alpha 1-AT), transferrin, lactoferrin and vasoactive intestinal polypeptide (VIP). 4) The clear cells in solid areas had positive results for KL 1, alpha 1-AT, transferrin and VIP. Both the clear cells and the neoplastic acinic cells showed a rather similar pattern of immunoreactivity. Therefore, the clear cells may transform from the neoplastic acinic cells. 5) Secretory products in tubuloglandular and microcystic patterns had positive results for alpha 1-ACT, lactoferrin, IgA and CEA. 6) The basement membrane-like material between the neoplastic islands has distinct positivity for alpha 1-AT. The result suggests that alpha 1-AT is a useful marker of basement membrane-like material.     

An immunohistochemical study of the distribution of lysozyme, lactoferrin, alpha 1-antitrypsin and alpha 1-antichymotrypsin in salivary adenoid cystic carcinoma.

The immunohistochemical expression of lysozyme (Ly), lactoferrin (La), alpha 1-antitrypsin (alpha 1-AT), and alpha 1-antichymotrypsin (alpha 1-Ach) was described, and their distributions were compared to each other in 28 cases of adenoid cystic carcinoma (ACC) of the salivary glands. ACC materials were obtained from the parotid gland (7), the submandibular gland (4), the sublingual gland (8), and minor oral salivary glands (9). Histopathologically, ACC was classified into cribriform (14), tubular (3), and basaloid or solid patterns (11). Positive staining for Ly was found in 1 case of solid ACC in the sublingual gland; La was found in 4 cases (2 cribriform, 1 tubular, 1 basaloid) in the sublinguals (3) and parotid glands (1); alpha 1-AT was found in 6 cases and alpha 1-Ach in 17 cases. The immunohistochemical localization of Ly and La was usually confined to luminal tumor cells of tubulo-ductal structures, irrespective of the pathologic types. Positive staining for alpha 1-AT and alpha 1-Ach appeared in tumor cells of cribriform, tubular and solid ACC. Tumor cells with positive La staining coincided with a positive reaction to alpha 1-AT and alpha 1-Ach, and tumor cells with alpha 1-AT positive deposition were also positive for alpha 1-Ach. The contents of pseudocysts in the cribriform pattern showed a positive reaction to La, alpha 1-AT, and alpha 1-Ach. Of the 28 cases of ACC, positive expressions for Ly, La, alpha 1-AT and alpha 1-Ach were found with a high frequency of alpha 1-Ach staining (17 in 28 cases were positive). In sublingual ACC (8), 7 cases were positive for immunohistochemical reactions. Co-expression or simultaneous expression for Ly, La, alpha 1-AT, and alpha 1-Ach in ACC suggest that tumor cells are protected from proteolysis or degradation.     

Acute leukemias with both myeloid and lymphoid surface markers. Cytoplasmic alpha-1-anti-chymotrypsin and alpha-1-anti-trypsin as possible indicators of early granulocytic differentiation

Blast cells from ten patients (seven adults, three children) with acute lymphoblastic leukemias (ALL) contained immunoreactive cytoplasmic alpha-1-anti-trypsin (alpha-1-AT) and alpha-1-antichymotrypsin (alpha-1-ACT). Cytochemically positive reactions for block-like periodic acid-Schiff and a localized acid phosphatase suggested that the cells were of lymphoid origin rather than myeloid origin: negative for sudan black-B, nonspecific esterase, chloroacetate esterase, and myeloperoxidase. By surface phenotype, the leukemia showed positive reactions for both lymphoid (common acute lymphoblastic leukemia antigen, Ia, OKT-10) and myeloid (OKM-1, Leu M-1) antigens. Three of three patients tested portrayed the Philadelphia chromosome. Nine patients were Mexican-American and one was Japanese: all were of Asian ethnic derivation. Both myeloid and lymphoid treatment regimens were employed, with survival less than expected. Early granulocytic differentiation detectable by cytoplasmic alpha-1-AT and alpha-1-ACT in lymphoid blasts is discussed.     

恶性组织细胞增生症与免疫缺陷

Phenotypes of the tumor cells of malignant histiocytosis (MH) were studied by using monoclonal and polyclonal antibodies in 18 autopsy cases. The tumor cells expressed different antigens in various degrees. Almost all tumor cells showed positive reaction for alpha 1-ACT; partially for alpha 1-AT, LCA and a few for lysozyme as well as LeuM1. It was most likely that the tumor cells of MH originated from the mononuclear phagocytic system (MPS). In order to reveal the relationship between MH and immunodeficiency, morphological changes of the lympho-reticular system in 18 cases of MH were studied. It was found that the lymphoid tissues, including lymph nodes, thymus, tonsil, spleen, bone marrow, lymphoid tissues of GI tract and lung etc showed severe depletion. These findings indicate that MH usually combine with immunodeficiency which is also closely related to the pathogenesis and pathological changes of MH.     

Monoclonal antibody to macrophages (EMB/11) labels macrophages and microglial cells in human brain.

Normal and diseased human central nervous system (CNS) tissues were studied immunohistochemically by a monoclonal antibody to human macrophages (EBM/11), antisera to glial fibrillary acidic protein (anti-GFAP), and alpha-1-antichymotrypsin (alpha 1-ACT). EBM/11 reacted with brain macrophages located mainly around blood vessels in normal brain; it also reacted with resting microglia in normal brain and with numerous reactive microglia and macrophages in brain tumours and inflammatory lesions. Microglia did not react with anti-GFAP or alpha 1-ACT. An EBM/11 positive phenotype, therefore, is shared by microglia and macrophages and suggests that microglial cells form a specialised part of the mononuclear phagocyte system.     

Immunohistochemical characterization of basal cell adenomas of the salivary gland

Seven cases of basal cell adenomas of the salivary gland were analyzed by immunohistochemical methods with a broad panel of routinely used antibodies. Histologically the epithelial elements were classified as tubuloglandular, trabecular and solid patterns. The authors' results indicated the following: 1) The duct lining cells of tubuloglandular and trabecular patterns have distinct epithelial features with cytokeratins (KL 1, PKK 1, *PKK 2 and PKK 3), alpha-one-antichymotrypsin (alpha 1-ACT), carcinoembryonic antigen (CEA) and S-100 alpha subunit positivity. 2) The basaloid cells in the trabecular and solid patterns expressed two immunophenotypes: one had actin, neuron-specific enolase (NSE), S-100 protein and S-100 beta subunit patterns typical of myoepithelial cells in normal glands. The other basaloid cells had vimentin and S-100 protein patterns. The former cell type could be found in 4 of 7 cases and the latter was found in 7 cases. This represents a minor participation of the myoepithelial cells in the basal cell adenoma. 3) The basement membrane and stromal connective tissue around the neoplastic cells were positive for alpha-one-antitrypsin (alpha 1-AT). This antibody is a good marker in identifying the basement membrane-like material.     

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.     

Secretory component, α1-antitrypsin and lysozyme in IgA deficient children. An immunohistochemical evaluation of intestinal mucosa

Nine children with IgA deficiency were studied in order to evaluate by the immunoperoxidase technique the behaviour of secretory component (SC), alpha 1-antitrypsin (alpha 1-AT), lysozyme and esterase in biopsies of intestinal mucosa. In none of the studied patients was SC found to be lacking, suggesting that the epithelial transport mechanism of IgA across enterocytes was relatively normal. The distribution of SC activity in immunodeficient children differed however from that seen in control intestinal mucosa in its non-uniform distribution on the villus, abnormal retention in the Golgi region of enterocytes or exclusive activity confined to the proliferating compartment of the villus. The staining of alpha 1-AT in enterocytes was clearly obvious in all studied cases with no alteration in zonal distribution when compared with normal human mucosa. The lysozyme staining pattern was seen exclusively in Paneth cells. The non-specific esterase positive enterocytes observed in control mucosa failed to stain in biopsies from IgA deficient children. The results of this study of SC, alpha 1-AT, lysozyme and esterase may indicate that IgA deficiency is not related to a defect in enterocyte transport of immunoglobulins and confirms previously reported findings indicating the lymphoid B-cell compartment to be altered.     

Osteoclast-type giant cell tumour of the pancreas

Summary Two cases of osteoclast-type giant cell tumour of the pancreas (OGTP) are presented and compared with similar tumours of other locations and pancreatic carcinomas. One of the tumours was analyzed by immunohistochemical methods. The mononuclear stromal cells and osteoclast-like giant cells, which characterize this very rare neoplasm, reacted with an antibody against vimentin, but were not decorated by antibodies against lysozyme, alpha-1-ACHT, alpha-1-AT. Pleomorphic mononuclear cells in osteoid additionally contained osteonectin and could thus be identified as osteoblasts. Only the tumour glands stained positively with panepithelial keratin antibodies and antibodies against the keratin polypeptides 7, 18, 19. These results demonstrate for the first time the mesenchymal differentiation of the OGTP, which in some cases is also able to form epithelial structures. The immunohistochemical reactions and the characteristic morphology of the tumour show the OGTP to be an entity which must be differentiated from pancreatic carcinoma, especially from its giant cellular subtype.     

Immunohistochemical demonstration of alpha1-antichymotrypsin and alpha1 -antitrypsin in the normal pituitary gland and its tumors

The immunohistochemical distribution of the protease inhibitors alpha₁-antichymotrypsin (alpha₁-ACT) and alpha₁-antitrypsin (alpha₁-AT) has been documented in the normal human pituitary gland and in a series of pituitary tumors. In normal gland, alpha₁-ACT was localized mainly in the dendritic folliculostellate cells, identified by immunopositivity for S 100 protein. A minority of endocrine cells also stained in 3 of 10 autopsy glands. Folliculostellate cells were identified in 11 of 28 tumors, and again, the distribution of alpha₁-ACT positivity corresponded to these cells. In 4 cases, there was staining of a small minority of tumor cells. Alpha₁-AT was localized to colloid in the microfollicles of the anterior lobe. In I normal gland, there was granular staining of endocrine cells. Alpha₁-AT was present in 5 tumors, in microfollicles and in scattered endocrine cells in 2 adenomas. These data would support a physiological role for alpha₁-ACT and alpha₁-AT in the pituitary gland. Their differing distribution might reflect different functions.     

Stromal cells in cerebellar haemangioblastomas: an immunocytochemical study

The nature of the stromal cells in formalin-fixed paraffin-embedded material from 23 cerebellar haemangioblastomas was investigated using antisera to intermediate filaments (glial fibrillary acidic protein, vimentin and desmin), histiocytic markers (alpha 1-antitrypsin, alpha 1-antichymotrypsin and lysozyme), glycolytic enzymes (alpha and gamma enolase and aldolase C4) and the endothelial markers, factor VIII related antigen and Ulex europaeus I lectin. Most stromal cells stained positively for vimentin and the glycolytic enzymes. Occasional process-bearing cells within the stroma stained strongly for glial fibrillary acidic protein, alpha 1-antitrypsin and alpha 1-antichymotrypsin. No stromal cell staining for desmin, lysozyme or the endothelial markers was observed, although the latter stained the vascular endothelium within all neoplasms. The findings do not support previous suggestions of an endothelial or histiocytic origin for the stromal cells. They appear to be a heterogeneous population including entrapped reactive astrocytes and locally-derived non-angiogenic cells of neuroectodermal (pial) origin.     

Polymerized alpha-antitrypsin is present on lung vascular endothelium. New insights into the biological significance of alpha-antitrypsin polymerization

AIMS: The damage to lung tissue in chronic obstructive pulmonary disease (COPD) may involve the progressive loss of pulmonary vascular endothelial cells. Endothelial binding of alpha1-antitrypsin (alpha1-AT) derived from plasma has been identified, and alpha1-AT deficiency is a known genetic risk factor associated with alpha1-AT polymerization and COPD development. Therefore, in the present study we aimed to investigate if alpha1-AT is present on the lung vascular endothelium, and if it is in a polymeric form. METHODS AND RESULTS: Postmortem paraffin-embedded tissue specimens from 15 COPD (chronic bronchitis and emphysema) cases with and without Z alpha1-AT (Glu342Lys) deficiency and from 10 cases without signs of COPD were studied. Immunohistochemistry was performed using the streptavidin-biotin method with a monoclonal ATZ11 antibody specific for polymeric alpha1-AT, and polyclonal antibodies against human alpha1-AT and neutrophil elastase. Vascular endothelium showed intense staining for alpha1-AT with the ATZ11 antibody in all cases; however, intensity of staining in patients with alpha1-AT deficiency was greater. No endothelial staining was observed with the anti-elastase antibody. CONCLUSIONS: This is the first demonstration that alpha1-AT bound to the vascular endothelium of lungs is in a polymeric form, which also suggests a possible previously unknown role for polymeric alpha1-AT in vivo.     

胰腺囊实性肿瘤八例临床病理学观察

目的 观察胰腺囊实性肿瘤的临床病理特点,分析其分化表型,探讨其组织发生。方法 组织学、免疫组化和电镜技术。结果 ８例患者均为女性,年龄１４￣３３岁,平均２５．３岁,手术后均无复发。肿瘤较大,有包膜,由实性肿瘤与囊性坏死区混合组成。组织学上,肿瘤细胞较一致,均由实性片块、假乳头样及两者的过渡型生长方式组成。瘤细胞退变、出血、泡沫细胞和胆固醇裂隙常见。免疫组化：８例均表达α－１－抗胰蛋白酶和溶菌酶,６     

Reactivity of ACTH and synthetic ACTH peptides with antisera to human calcitonin.

We studied reactivity of highly purified pituitary hormones in our human calcitonin (hCT) radioimmunoassay (RIA) which can detect 1 pg of hCT. ACTH at doses of greater than 1 microgram of peptide per RIA tube reacted in the hCT assay, as did beta-endorphin (beta EPH) at a dose of 10 micrograms per tube. No reactivity was observed with comparable concentrations of all other known pituitary hormones. ACTH also reacted at doses greater than 1 microgram per tube with 7 other hCT antisera which recognized differing antigenic determinants in the calcitonin molecule but it was not reactive with 2 antisera against porcine calcitonin or 2 antisera against salmon calcitonin. This slight degree of cross-reactivity of hACTH and beta EPH in the hCT RIA cannot account for the presence of immunoreactive CT in pituitary glands. Nevertheless, antisera used for the localization of peptides must be rigorously tested for the existence of cross-reactivities with other possible substances, especially if such antisera detect the peptide in unexpected tissues.     

Substance P and opioid peptidergic innervation of the anterior eye segment of the rat: an immunohistochemical study

Recently discovered endogenous opioid peptides such as nociceptin are known to modulate neurotransmitter release of primary afferent neurons (especially substance P, SP) and they have also been demonstrated in peripheral nerve fibres. The aim of this study was to investigate the opioid peptidergic innervation of the anterior eye segment and to compare it with the innervation pattern of SP in order to shed light on the functional relationship between these peptides. Anterior eye segments of 20 rat eyes were cut in a tangential plane and the sections stained with antibodies against SP, nociceptin, nocistatin, endomorphin 1 and 2, leu-enkephalin and met-enkephalin. Sections of the spinal cord or brain were used as positive controls. Numerous SP-immunoreactive nerve fibres were found in the conjunctiva, cornea, episclera, trabecular meshwork, iris and ciliary body. A weak staining for met-enkephalin and leu-enkephalin could only be found in the iris and anteriormost ciliary body. Nerve fibres immunoreactive for nociceptin, nocistatin, and endomorphin 1 or 2 could not be detected in any part of the anterior eye segment. It is tempting to speculate that the opioid peptidergic innervation of the anterior ciliary body may play a role in the modulation of intraocular inflammation.     

Tumor cell line characterization of a malignant histiocytosis transplanted into nude mice

Summary The successful transplantation of a human malignant histiocytosis into nude mice allowed the examination of its atypical histiocytic cell proliferation. Histiocytic type cells were identified by positive reactions with acid phosphatase and non-specific esterase and with anti human DR or OKI1 antisera. Presence of OKT9 antigen and negative results obtained with most OKT antisera, rosettes, erythrophagocytosis and lysozyme corroborate the histiocytic immature state of the cells and preclude another type of tumor. All positive tests to prove a mature mononuclear phagocytic origin were attributable to the murine host cell reaction.