Dual Cell Population Research Articles

Malignant eccrine spiradenoma (spiradenocarcinoma): a clinicopathologic study of 12 cases.

Malignant eccrine spiradenoma is a very rare tumor. The clinicopathologic features of 12 cases are reported herein. Six patients were men, six were women, and the average patient age was 62 years. Seven tumors were located on the trunk, three on the extremities, and two in the head and neck region. All tumors were large (average size-7.5 cm). Lesions had been present from 7 months to 30 years before surgical removal. In all cases, continuity between benign eccrine spiradenoma and areas with malignant change was observed. Malignancy was evidenced by increased mitotic rate, necrosis, nuclear atypia, pleomorphism, and hyperchromasia, loss of nested and trabecular growth patterns, and absence of a dual cell population. In most cases (8 of 12), the malignant component comprised the bulk of the lesion. Two distinctive histologic patterns were observed. Five of 12 tumors exhibited abrupt transition between a benign eccrine spiradenoma and a high-grade carcinoma component. The others lacked a clear-cut transition between benign and malignant components and were diagnostically challenging. Diagnosis in such cases was established based on the loss of two cell populations, increased nuclear to cytoplasmic ratio, hyperchromasia, and marked mitotic activity. Two tumors showed focal squamous differentiation. Five of seven patients on whom there was follow-up information were free of disease (average duration of follow-up = 3.4 years). One patient developed metastases to local lymph nodes 5 years after the primary tumor was resected. This patient had no evidence of disease 16 months after resection of her lymph node metastases.

HyperImmunoglobulin E Syndrome: Clinical and Immunologic Evaluation within One Family 41

Hyper IgE syndrome (HIE) is characterized by recurrent staphylococcal deep-seated infections, mucocutaneous candidiasis, chronic eczematous dermatitis, bone fractures, and coarse facies. A father and two daughters (now 1 and 5 yrs old) with classical manifestations of HIE provided a unique opportunity for early and sequential evaluation of clinical and immunologic parameters. Extremely high serum IgE may be caused by abnormalities in T lymphocyte function and cytokine production. Measurements of T cell populations and cytokines that control IgE production, IL-4 and IFN-γ, have found variable results in HIE. In this family, IgE levels were 0 - 100 IU/ml in the first year of life, rising to >36000 at 4 yrs in one child. T cell responses to mitogens and antigens were already decreased. In the first year, the absolute CD 4 counts were increased (mean 5670) with normal percentages due to elevated total lymphocyte counts. CD8 counts near the lower limit of normal (mean 600) resulted in elevated CD4:CD8 ratios (range 5 to 11). Naive CD4CD45RA and committed CD4CD45RO showed a normal pattern: CD45RA decreased with age while CD45RO increased. Neutrophil dysfunction, especially in chemotaxis, also is reported in HIE. Assessment of neutrophil receptors showed a significantly decreased expression of the CD11b/CD18 heterodimer at 38%/35% for the younger child and 33%/45% for the older girl. In contrast, neutrophils from their father expressed CD11b/CD18 at normal levels. FcRIII(CD16) expression on neutrophils showed a strikingly different pattern: both children had a dual population of cells. 25-45% of neutrophils had decreased expression at 50% control, but 75-55% of cells had greatly upregulated FcRIII from 3.4 to 67 times normal. The father had a single population of FcRIII at 180% of control. Changes in IgE levels, T lymphocytes, and the expression of neutrophil receptors within this family suggest an age-related pattern. These findings may help in understanding the functional abnormalities, and potentially to identify the genetic defect in this rare primary immunodeficiency syndrome.

Maintenance of polarity and a dual cell population in adenoid cystic carcinoma of the breast: an immunohistochemical study.

Adenoid cystic carcinoma (ACC) of the breast, although quite uncommon, is histologically distinct and has an excellent prognosis. Our purpose was to characterize molecular markers of cellular differentiation and polarity to define better the role that these combined features might play in the excellent prognosis of ACC in this site. We performed immunohistochemical stains for expression of vimentin (VM), cytokeratin (CK), smooth muscle actin (SMA), laminin (LAM), E-cadherin (E-cad), beta-catenin (beta-cat) and fodrin in 14 examples of ACC. Two types of intercellular lumens and defining lining cells were detected: true epithelial lumens were lined by CK positive cells that maintained the normal glandular luminal cells with an intact polarity and basolateral membrane compartment indicators: fodrin, E-cad and beta-cat. The second type of intercellular space was a pseudolumen, surrounded by myoepithelial cells that were strongly positive for VM and SMA, and lined by LAM. Double staining for VM and CK, VM and SMA, and CK and SMA detected occasional cytoplasmic co-expression of these markers in the myoepithelial cell compartment, but VM and SMA did not stain epithelial lumenal cells. ACC of the breast is characterized by the presence of two types of intercellular lumens, one conserving basolateral markers of normal polarity (epithelial) with the other containing basement membrane material (abutted by myoepithelial cells). Dominance of VM staining may be a hallmark of ACC, along with maintained epithelial and myoeithelial compartments. This status of advanced, normal differentiation despite local invasion may be responsible for lack of distant metastasis.

Occurrence of subtypes of gustatory cells in cat circumvallate taste buds.

Transmission-electron microscopy of cat taste buds confirmed that, as in other mammals, each taste bud comprised four distinct types of cells: Type I, Type II, Type III (gustatory), and Type IV (basal) cells. Gustatory cells made synaptic contacts with nerves to which synaptic vesicles were gathered. The following are the main findings on the cat gustatory cells: 1) The synaptic vesicles of gustatory cells were essentially all dense-cored in type; small clear vesicles, which usually are intermingled in other mammals, could not be found. 2) The vesicles were accumulated not only in the synaptic area but also in the basal cytoplasm. This implies endocrine and paracrine functions. 3) On the basis of the fine structure of the vesicles, two subtypes of gustatory cells were discriminated. A large part of the cells contained vesicles measuring about 180 nm in diameter, while a small part had smaller ones of about 100 nm in diameter. This is the first demonstration of a dual population of gustatory cells in mammals, suggesting different messenger substances utilized.

Flow cytometry analysis of dual red blood cell populations after bone marrow transplantation.

Flow cytometry represents an alternative method to agglutination assays for the accurate quantification of mixed field populations of erythrocytes observed after bone marrow transplantation. Murine monoclonal antibodies directed against the blood group ABH antigens were selected and processed in order to prepare ready-to-use fluorescent reagents. Anti-A (NaM87-1F6; IgG3), anti-B (NaM9-2E11; IgG3) and anti-H (NaM19-7E11; IgM) were purified, labelled with fluorescein isothiocyanate, and used in a direct flow cytometry assay. Anti-A1 (NaM1-1C9; IgG3) was no longer active after FITC-labelling and then was used in an indirect assay. The agglutination was prevented by formaldehyde pretreatment of erythrocytes. Using artificially-made double populations of erythrocytes, measured values with mixtures of 1-100% of cells were very closely related to expected values, showing both the sensitivity and the accuracy of the method. From careful investigation of a series of bone-marrow transplanted patients, we conclude that engraftments could be demonstrated earlier by flow cytometry than by agglutination, because minor populations (1-10%) of cells could be determined accurately only with labelled reagents. In addition, the disappearance of the donor cells on a long-term follow-up of patients enabled an earlier detection of graft failure in one case. The proposed method provides appreciable help to follow engraftment in patients and may have more general applications for the study of other haemopoietic chimaeras.

Basal cell adenocarcinoma of the submandibular gland

Basal cell adenoma is a well recognized histopathologic variant of monomorphic adenoma of salivary gland, but in recent years there have been occasional reports of malignant basal cell tumors of major salivary glands. We present a case report of one such basal cell adenocarcinoma arising in the submandibular gland and discuss the differential diagnosis and distinction from a basal cell adenoma. We also review the published literature on basal cell adenocarcinoma. This is the first case of basal cell adenocarcinoma to be studied in detail by both immunohistochemistry and electron microscopy. There has been debate in the past as to the cell of origin of salivary gland basal cell tumors and specifically as to whether myoepithelial differentiation occurs in these tumors. The immunohistochemical and ultrastructural features of this case provide evidence for a dual population of ductal and myoepithelial cells. A flow cytometric analysis of nuclear deoxyribonucleic acid content showed the presence of DNA aneuploidy within one tissue block.

Bovine Pulmonary Blastomas: Histomorphologic Description and Immunohistochemistry

Three adult female dairy cattle with pulmonary blastomas were evaluated. The gross lesions at postmortem were described as multiple white circumscribed masses throughout the lungs, with pleural involvement in one cow and lymph node metastasis in the other two cows. Histologically, the tumors contained a dual population of mesenchymal and epithelial neoplastic cells. Epithelial cells formed nests, tubules, and formations resembling bronchioles of normal lung. Mesenchymal cells were spindle shaped with oval nuclei and fibrillar eosinophilic cytoplasm, were large rounded cells with multiple round nuclei and granular eosinophilic cytoplasm, or appeared blast-like, with large hyperchromatic nuclei and amphophilic cytoplasm. The tumors varied greatly in appearance from one field to another within the same tumor. Epithelial cells stained positively with anticytokeratin antibodies. Some spindle-shaped mesenchymal cells exhibited smooth muscle cell differentiation with positive staining with anti-vimentin, anti-muscle-specific actin, and anti-smooth muscle actin antibodies, whereas other rounded mesenchymal cells expressed striated muscle cell differentiation with multiple nuclei and positivity to anti-neuron-specific enolase and anti-muscle-specific actin antibodies. The variable expression of the intermediate filaments and cytoplasmic enzymes indicates multiple pathways of differentiation in the pulmonary blastomas.

Critical observations on neuroblastoma treatment with 131‐I‐metaiodobenzylguanidine at diagnosis

Results of treatment with 131-I-Metaiodobenzylguanidine (131-I-MIBG) in patients with resistant neuroblastoma appear encouraging if one considers that most of the patients had far advanced, intensively pre-treated disease. To further explore the potential role of this new drug in untreated patients, we treated 3 children with stage III neuroblastoma. All three of our cases received 131-I-MIBG at relatively low dose with the complete disappearance of the tumor mass in case 1, whereas in cases 2 and 3 CT scan showed a significant reduction of the tumor mass and, interestingly enough, no evidence of 131-I-MIBG uptake of a tracer dose in the remaining tumor. Particularly, in case 2, the persistence and subsequent progression of part of the tumor mass without 131-I-MIBG uptake after a therapeutic dose of 131-I-MIBG, which apparently destroyed the 131-I-MIBG-positive cell population, clearly suggest heterogeneity at diagnosis, with a dual neuroblastoma cell population, one with 131-I-MIBG uptake and the other without. Besides the biological implications of the 131-I-MIBG uptake heterogeneity in neuroblastoma at diagnosis, our findings suggest that in stage III neuroblastoma patients even a relatively small dose of 131-I-MIBG administered at diagnosis is sufficient to destroy either the primary tumor completely (case 1) or the part of the tumor (case 2 and 3) which shows 131-I-MIBG uptake, without any significant hematologic toxicity. Furthermore, a single course of 131-I-MIBG at the dosage employed does not appear to jeopardize the subsequent use of chemotherapy. In conclusion, if our data are confirmed by further investigation, 131-I-MIBG may be included as a front line drug shortly followed by chemotherapy in future treatment strategies of advanced neuroblastoma without or with minimal bone marrow infiltration.

Epithelial-myoepithelial carcinoma of salivary glands.

Epithelial-myoepithelial carcinomas comprise approximately 1% of all salivary gland neoplasms. They are preponderantly tumors of the parotid glands with a relatively low mortality but a decided locoregional aggressiveness. Histopathologically, the carcinomas are characterized by a dual cell population of epithelial (ductal) cells and myoepithelial cells. These cells vary in their dominance and phenotypic expression.

Histochemical differences in expression of X-linked glucose-6-phosphate dehydrogenase between ectoderm- and endoderm-derived embryonic and extra-embryonic tissues.

We examined the activity of X-linked glucose-6-phosphate dehydrogenase (G6PD) in concepti of the enzyme-deficient mutant and wild-type C3H mice. By using different crosses between the G6PD-deficient homozygous, heterozygous, or wild-type females and hemizygous or wild-type males, we confirmed the inactivation of one of the two X chromosomes in female concepti by a histochemical method. With this technique, a dual (G6PD + or -) cell population could be observed in the tissue sections. We demonstrate that the paternal X chromosome is inactivated in the endoderm of parietal and visceral yolk sac and in the trophoblast, whereas in the embryo and in the yolk sac mesoderm this inactivation is random. Our results confirm biochemical observations showing that only the maternal X chromosome is expressed in all derivatives of trophectoderm and primitive endoderm, whereas derivatives of the primitive ectoderm show random X chromosome expression.

Endodermal origin of yolk-sac-derived teratomas

Mutant mice deficient in glucose-6-phosphate dehydrogenase were used to induce teratomas. This enzyme is linked to the X chromosome, which can be inactivated in female embryo. The differences in the enzyme activity between yolk sac mesoderm and embryo versus yolk sac endoderm can be detected in female concepti by using appropriate crosses of wild-type and G6PD-deficient mice. Histochemical study showed that the dual cell population was observed in heterozygous embryos and in the embryomas derived from them. The teratomas derived from the corresponding yolk sac, however, were G6PD-positive from wild-type and G6PD-negative from homozygous enzyme-deficient mothers. We conclude that yolk-sac-derived teratomas are of endodermal origin because of the fact that the paternal X chromosome is inactivated in the yolk sac endoderm, whereas in the yolk sac mesoderm, as in the embryo, the inactivation is at random.

Characterization of the erythropoietic progenitor cells (BFU‐E) in paroxysmal nocturnal haemoglobinuria (PNH)

In order to investigate erythropoiesis in paroxysmal nocturnal haemoglobinuria (PNH), the relative proportions of complement-mediated lysed erythroblasts in individual bursts of two cases of PNH were analysed by a modified complement lysis sensitivity test using the trypan blue dye exclusion method. All bursts contained both complement-sensitive and complement-insensitive erythroblasts and the ratio between them varied from burst to burst. The means +/- SD of the haemolytic percentages of individual bursts in cases 1 and 2 were 30.9 +/- 10.8% (n = 20, ranging from 13.2% to 52.8%) and 54.8 +/- 16.4% (n = 31, ranging from 26.4% to 82.5%), respectively. In addition, the cumulative frequency distributions of the proportions of complement-mediated lysed erythroblasts in individual bursts approached normal frequency distributions. These results suggest that two red cell populations with different susceptibilities to complement in PNH are derived from a single population of erythropoietic progenitor cells rather than a dual population of normal and PNH erythropoietic progenitor cells.

The demonstration of tissue clonality by X-linked enzyme histochemistry.

We report here the first demonstration in normal, as opposed to chimaeric mice, of the clonal architecture of the small intestine, oesophagus, breast, and thyroid, using polymorphism of the X-linked enzyme glucose-6-phosphate dehydrogenase (G6PD, EC 1.1.1.49) as a clonal marker. Reproducible results using X-linked histochemistry were found to depend upon the simultaneous study of tissue from normal mice, mice homozygous for abnormal enzyme levels, and heterozygous mice. In tissues from homozygous animals where levels of enzyme activity normally vary, the use of a physiological stimulus to reduce this metabolic heterogeneity allowed interpretation of clonality in the heterozygotes. The details of the histochemical technique used require modification for each of the different tissues studied. A dual population of cells was seen in the heterozygous animals in all tissues studied. Oesophageal epithelium showed sharply separated alternating patches of positive and negative cells. Small intestinal crypts were always monophenotypic, while villi were polyphenotypic, indicating that crypts are monoclonal and villi derive from more than one crypt, confirming work in allophenic animals. In contrast, thyroid follicles and breast acini showed a mixture of mono- and polyphenotypia, consistent with a polyclonal origin. These results, which have important implications for the study of the clonal origins of normal and neoplastic growth, require the direct in situ demonstration of the cellular phenotype.

Mosaicism of red cell ABO type without recognizable cause

A 19 year old woman has been found whose red cells are a mixture of groups A 1B (60%) and A 1 (40%). Tests for many other genetic markers show no other evidence of mosaicism. She is not a twin and her parents and a sib are of normal ABO type. The cause of her dual population of red cells cannot be established, but may be an effect of somatic crossing over or mutation.

Haematological changes associated with the McLeod phenotype of the Kell blood group system.

The McLeod phenotype is inherited as an X-linked characteristic. The red cells have weak antigenicity in the Kell blood group and lack Kx, a precursor-like substance that appears to be necessary for proper biosynthesis of Kell antigens. Kx antigen is also required for establishment of normal cell morphology. Absence of Kx antigen causes a membrane abnormality, in which the most prominent feature is acanthocytosis, and a compensated haemolytic state. The X-linked gene that determines normal Kx production is called X1k. Inheritance of a variant allele at the Xk locus is responsible for lack of Kx synthesis and the McLeod phenotype. The Xk locus is inactivated by the Lyon effect, and female carriers of the variant gene exhibit blood group mosaicism in the Kell system and have a dual red cell population of acanthocytes and discocytes.

An evaluation of the mixed lymphocyte culture reaction in marmosets.

Marmosets are unique in that all members may be considered to be natural blood chimeras because of the high frequency of fraternal twinning and placental vascular anastomoses between the fetuses. The mixed lymphocyte culture (MLC) reaction utilizing blood lymphocytes was evaluated to determine whether this in vitro test could detect histocompatibility differences among related and unrelated marmosets. It was found that the responses could be correlated with the probable immunogenetic relationships of these animals. Thus, an allogeneic MLC reaction in which the responding (R) and stimulating (S) cells were obtained from unrelated animals within one subspecies of marmosets (Saquinus fuscicollis illigeri) yielded a lower response than a semi-xenogeneic reaction involving R and S cells from two different subspecies of marmosets (S.f. illigeri (R) versus S.f. lagonotus (S). In contrast, MLC reactions between cells from chimeric marmoset co-twins were essentially negative, indicating specific immune tolerance. Under certain experimental conditions, however, it was suggested that the dual lymphocyte cell population from a chimeric marmoset could undergo "autostimulation" as measured by label incorporation studies. The necessity of further experiments was recognized before the data could be truly accepted as reflecting an in vitro manifestation of incompatibility between the two different genetic populations of blood lymphocytes from a chimera.

Acute myeloid leukemia with two Philadelphia chromosomes in forty-six stemline. Remarks on the karyotypic analysis and chemotherapy.

Serial cytogenetic studies of bone marrow in a 59-year-old male with a 14-month history of acute myeloid leukemia revealed a Ph1 disomic diploid clone (46, XY, 2Ph1). This clone responded to a cytostatic drug regimen of prednisone, vincristine and daunomycine. A dual population of normal and leukemic cells was observed during the course the disease.

Some observations on the ultrastructure of a malignant islet cell tumor associated with duodenal ulceration and severe diarrhea

Ultrastructural study of a hepatic metastasis from a malignant islet tumor—associated initially with duodenal ulceration and subsequently with severe diarrhea and hypokalemia—has revealed a dual cell population. The first cellular category comprised cells believed to be alpha cells. The nature of the second group is not clear; they may have represented modified alpha cells. In view of recent evidence incriminating gastrin as the factor responsible for gastric hypersecretion, and in view of the postulated existence of a separate hormone responsible for the severe diarrhea encountered in some patients with islet cell tumors, it is suggested that these two cellular variants may, respectively, have been responsible for the elaboration of factors mediating the physiologic disturbances present in this patient, viz., secretion of gastrin by the alpha cells, and of an intestinal stimulant by cells of the second category, whatever their derivation.