Emilio A. Machado

Cyclic neutropenia. A tale of two brothers and their family.

A family history encompassing four generations is presented. Definite cyclic neutropenia has been noted in two generations. In one member, the symptoms abated as she grew older. One 12-year-old member died of clostridial septicemia secondary to gangrene of the terminal ileum. At autopsy he was found to have amyloidosis. A trial of lithium carbonate therapy was carried out in a 7-year-old male but a positive hematologic response was not obtained.

Arrest and extravasation of neoplastic cells - An electron microscopy study of serial sections at sequential stages

The morphological aspects of the arrest and extravasation of malignant cells of human origin (K-562 cell line) in the lungs of athymic (nude) and asplenic-athymic (lasat) mice were studied by electron microscopy examination of serial sections. The specimens were obtained at sequential stages after the sc inoculation into newborn mice of 107 malignant cells. K-562 cells (105) were also injected iv into control groups of nude and lasat mice to assess the influence of the route of inoculation on the in vivo behavior of K-562 cells. Our results demonstrated that K-562 cells were arrested and proliferated within the pulmonary capillaries without the participation of platelets or fibrin. The neoplastic cells extravasated by attrition and penetration of the endothelium (rather than by diapedesis) and continued to proliferate in the interstitial tissue of the lung, developing into neoplastic nodules. Following iv injection, K-562 cells induced the formation of platelet-tumor cell aggregates within the pulmonary capillaries. However, under these conditions, the neoplastic cells did not adhere to the endothelium nor did they proliferate or extravasate. These aggregates were flushed out by the circulation, restoring the permeability of the capillaries.

Psychogenic purpura (autoerythrocyte sensitization).

A 41-year-old woman has had a long history of repeated episodes of recurrent painful ecchymotic lesions. Results of coagulation tests were normal other than a slight decrease in antithrombin III. Skin tests were positive in response to the patients own washed red cells. Light and electron microscopy of both the spontaneous and the induced lesions showed nonspecific changes but failed to reveal immunologic vasculitis. Psychologic evaluation showed hysterical and masochistic traits, depression, anxiety, and inability to deal appropriately with hostile impulses. Placebo was successful on several occasions in controlling or modifying the severity of the ecchymotic lesions.

Immunotherapy of metastases of human neoplastic cells grown in immunodeficient mice

SummaryHereditarily athymic (nude) and asplenic-athymic (lasat) mice were inoculated neonatally with 107 K-562 pluripotential leukemia cells of human origin. Meningeal infiltration and/or multiple metastases were found in the lungs, kidneys, and lymph nodes in nearly 60% of mice. Twice as many lasat mice as nude mice had meningeal and lymph node infiltrations. This result indicates that the spleen of nude mice influences the infiltrations and/or the distribution of metastases. Metastases of K-562 cells were found as early as 10 days and as late as 115 days of age. Goat immune gamma (γ)-globulin, prepared from antiserum to K-562 cells and absorbed with peripheral leukocytes and bone marrow cells from normal individuals, markedly diminished the incidence of metastases of K-562 cells. About 16% of the mice treated with immune γ-globulin had metastases in the lungs only. All mice receiving the immune γ-globulin had peripheral monocytosis and lymphocytosis as well as a hyperplasia of the bone marrow monocytic series. Immune γ-globulin may lyse heterotransplanted leukemia cells by direct binding to leukemia cells in the presence of complement and/or may activate antibody-dependent effector cells, for example macrophages or killer cells, which would destroy the transplanted leukemia cells.

Renomedullary deficiency. A contributory factor in the pathogenesis of experimental renal hypertension

The results indicate that renomedullary deficiency induced by renal artery clipping might contribute to the development of renal hypertension.

Ultrastructural studies on the evolution of amyloidosis in the cyclic hematopoietic (CH) dog.

Electron microscopy studies were made on tissues of cyclic hematopoietic (CH) dogs of various ages presenting a high incidence of spontaneous amyloidosis. The distribution and morphologic characteristics of amyloidosis in this animal model closely correspond to the secondary and familial forms of the disease in humans. Plasma cells and, particularly, macrophages presented marked changes during the evolution of amyloid deposition. Residual bodies in the macrophages contained abundant cell debris, a result of both endocytic and autophagocytic activities. Intracellular amyloid fibrils were not observed by conventional electron microscopy. A few reticular cells contained intracytoplasmic fibrils which were morphologically different from amyloid. There was no correlation between the amount of intracellular fibrils and the size of the extracellular amyloid deposits. On the contrary, a temporal association between the magnitude of the amyloid deposits and cytoplasmic changes in the macrophages at sequential stages of the evolution of the disease was evident. It is suggested that the hematopoietic defect in the CH dog could play an important role in the production of amyloidosis, making this animal an excellent experimental model for studies of that disease.

Ultrastructural changes of bone marrow in canine cyclic hematopoiesis (CH dog). A sequential study.

The pathogenesis of cyclic hematopoiesis (CH) in the grey collie dog is still unknown. It has been proposed that periodic bursts of necrosis of the bone marrow neutrophils would induce cyclic arrests of the stem cell differentiation. In the present study, the sequential changes undergone by the erythroid and neutrophil series of the bone marrow of CH dogs were evaluated by electron microscopy. Erythroid cells presented quantitative periodic oscillations but the morphologic features of both immature and mature cells were normal. On the contrary, nonspecific necrotic changes were observed to occur in the myeloid series. Those abnormalities, which were more marked between days 9 and 11 of the cycle, mainly involved the immature cells and, to a lesser extent, the mature neutrophils. The number of necrotic cells was variable in different cycles, but always represented a small portion of the myeloid cells. In addition, few bone marrow macrophages displayed signs of phagocytic activity containing cell debris. The ultrastructural changes of the myeloid series were accompanied by an abnormal decrease of peroxidase activity and the permanence of large acid phosphatase-positive Golgi complexes in mature neutrophils, as defined by morphologic criteria. Döhle-like arrays of rough endoplasmic reticulum were present in many cells. Our findings suggest that an asynchronic development of myelocytes occurs as a result of regulatory abnormalities related to the congenital defect of the bone marrow which interferes with the differentiation and maturation of the stem cells. Necrosis in some myeloid cells would be a secondary phenomenon rather than a causal factor for the cyclic arrest of cell maturation as has been previously submitted. Furthermore, the small size of the necrotic cell population could not justify the production of “inhibitors” in sufficient amounts as to block the normal evolution of the bone marrow stem cell pool.

Absence of morphological, chromosomal and antigenic changes in the K-562 cell line growing as localized or disseminated tumours in nude mice.

Transplantation of K-562 cells into adult and newborn nude mice led to the development of localized s.c. and disseminated myelosarcomas, respectively. This age-associated, changing pattern of in vivo proliferation of K-562 cells derived from a single aliquot was consistently repeated throughout sequential passages. The only variable in this experimental system was the age of the recipient mice. Not only did the mice have an identical genetic background, but also the transplanted K-562 cells were derived from a single culture passage. As shown by cytological and histological examinations, the characteristic morphology and percentage composition of the subpopulations of the K-562 cell line were preserved in successive in vitro and in vivo passages. The K-562 cells had no prevailing phenotypic traits which could be associated with the growth either in the s.c. tissue or in the viscera. Furthermore, the cells maintained the human karyotype, including their typical chromosomal abnormalities and antigenic determinants, as demonstrated by the binding of a specific antibody, throughout all passages. Our results demonstrate that heterotransplanted K-562 cells may change their behaviour in vivo without undergoing modifications associated with different types of growth. These findings would indicate that the ability of neoplastic cells to proliferate in various environments (metastases) is not the consequence of predetermined cellular characteristics but is functionally conditioned.

Variant of congenital dyserythropoietic anemia.

Nonspherocytic hemolytic anemia was diagnosed in a 34-year-old man with jaundice since childhood. Splenectomy at the age of 8 had no influence on the anemia. Bronze diabetes was diagnosed at age 31, presumably due to hemosiderosis and secondary hemochromatosis. Iron chelation was unsuccessful in controlling iron overload, but phlebotomies proved effective without aggravating the anemia. We believe the anemia represents a variant of congenital dyserythropoietic anemia, type I.

Local and metastatic growth and in vivo differentiation of human myeloid leukemia cells transplanted in nude mice

Cells from the established human myeloid cell lines KG-1, KG-1a, and HL-60, transplanted subcutaneously (sc) into nude mice, developed discrete tumors (myelosarcomas). These myelosarcomas had a hosts age-associated pattern of growth identical to that of experimental tumors produced by sc transplantation of cells derived from malignant solid neoplasias. Thus, leukemia cells yielded either localized myelosarcomas at the site of inoculation or a disseminated neoplastic growth after inoculation inadult (more than 4 weeks old) ornewborn (1–3 days old) nude mice, respectively. Human myeloid leukemia cells proliferating in the nude mice preserved the human karyotype and a surface antigenic determinant and did not influence the hematopoietic tissues of the host. The KG-1 and HL-60 cell lines consistently attained varying degrees of differentiation along the myeloid series in vitro, and these features were maintained during proliferation in the mice. Furthermore, cells of the variant subline KG-1a, which had a blastic morphology, developed signs of differentiation that were not seen in culture. The presence of readily identifiable markers, such as cytoplasmic granules containing myeloperoxidase, in the cell lines tested makes these models particularly useful for studying the influence of a biological environment on cell differentiation and its influence on tumor growth. These experimental systems are also suitable for investigating the mechanism(s) of metastases and for in vivo experimental therapeutic trials.