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[Hodgkin and Sternberg-Reed cells--cell origin, cell marker, cell function].

The controversy in literature with regard to the origin of Hodgkin's and Reed-Sternberg cells persists, however only two conceptions seem to be plausible at present: the first is based on the relation to histiocytic elements, the second postulates lymphocytic precursors. The significance of surface-markers of these malignant cells in cryostat-sections and in cell-cultures and the relevance of their functional properties are discussed with respect to pathophysiology, clinical appearance, diagnosis and prognosis of Hodgkin's disease. The authors present two tendencies in the classification of malignant lymphomas based on the present knowledge achieved especially by monoclonal antibodies: the first includes aspects of integration between non Hodgkin's lymphomas and Hodgkin's disease, illustrated by the Ki-1-lymphoma, the second is related to separation of entities of the group of Hodgkin's lymphomas (for example the nodular paragranuloma). The aetiopathogenesis of Hodgkin's disease is considered as a causal trinity of virus infection, genetic determination and immunologic predisposition.

Biomarkers, Tumor

Intimal cell mass-derived atherosclerotic lesions in the abdominal aorta of hyperlipidemic swine. Part 1. Cell of origin, cell divisions and cell losses in first 90 days on diet.

Atherosclerotic lesions may originate and develop in a variety of ways. In this study we are focusing our attention on atherosclerotic lesions arising in normally occurring intimal cell masses (ICM) in the abdominal aortas of hyperlipidemic (HL) swine. Times chosen for study were 0, 14, 49 and 90 days on HL diet; mash-fed swine were used as controls. Total numbers of cells in the ICM of HL and mash swine were similar at 14 and 49 days; by 90 days the number of cells had increased dramatically in the HL swine to 8-fold greater than control values. Changes present at 49 days and thus preceding increase in cell numbers included extensive intracellular lipid accumulation with by count nearly half of the ICM cells involved and elevated tritiated thymidine labeling indices (LI) 4-fold greater than control. Differential cell counts by transmission electron microscopy were made on the ICM lesions in the HL swine at 49 and 90 days. More than 95% of all cells were smooth muscle cells (SMC), with relatively few monocytes being present. Calculations from the LI and total cell counts showed that the entire increase in cell numbers could be accounted for by divisions among the resident SMC in the ICM. Further calculations suggested that cell losses (deaths) from the ICM were minimal. Scanning electron microscopy studies reported elsewhere revealed no loss of endothelial integrity. The results suggest: (1) that the lesions arise by stimulation of the resident SMC in the ICM to hyperplastic activity, (2) that the role of monocytes in the early development of these lesions is minimal if any, (3) that in view of the intact endothelium platelets are not likely to play an important role, (4) that ICM cell death is not a major factor, (5) that the most likely candidate for the cell growth stimulatory role (? mitogen) is some component(s) of the excess lipid that accumulates in the ICM.

Animals

Development of the oculomotor nucleus, with special reference to the time of cell origin and cell death.

The developmental pattern of the oculomotor nucleus from day 7 of incubation through two weeks after hatching was studied in white Peking duck embryos. The neuroblasts comprising the nucleus complete their last phase of DNA synthesis on days 4 and 5 and the anlage first appears on day 7. The various subnuclei become identifiable as distinct cell groups on day 8 or 9. There is a cell migration between the ventral-most portions of the two ventromedial nuclei on days 9 through 11, and as a result a well-developed oculomotor commissure is established between these two subnuclei. The maximum number of cells in the nucleus is present on day 11. There is a normally occurring overall loss of approximately 43% of the cells during ontogenesis. Cell death appears to be random, without any gradient, and virtually all of it occurs between days 11 and 15. Although the duration of cell death is essentially similar in all subnuclei, great variations exist in its magnitude. For example, there is a cell loss of approximately 61% in the accessory nucleus, 38% in the dorsolateral nucleus, 40% in the dorsomedial nucleus and 33% in the ventromedial nucleus. Cell loss in the oculomotor nucleus is compared with that observed in the other two eye-muscle nuclei.

Accessory Nerve

Human macromolecular insoluble cold globulin (MICG). I. T-cell origin of T-MICG and null cell origin of N-MICG.

Although surface immunoglobulin characterizes B cells in man, there are few surface markers that distinguish T cells. We have described a new protein synthesized in human T cells, termed T-MICG. This protein is a macromolecule of 225,000 daltons, is insoluble in the cold, and migrates as a beta-globulin on electrophoresis. Separation of human peripheral blood lymphocytes into T and B-cell populations by rosette sedimentation and anti-human-Fab columns clearly demonstrated the T-cell origin of the 225,000 dalton component. Furthermore, null cells were shown to synthesize a protein of 185,000 daltons, termed N-MICG, with physical properties similar to T-MICG, T-MICG and N-MICG were shown to be antigenically dissimilar, employing antiserum to each of these proteins. The present studies demonstrate two novel cell surface markers, T-MICG and N-MICG, which characterize T cells and null cells, respectively.

Antigens, Surface

Adult T cell leukaemia cells are of CD4+ CDw29+ T cell origin and secrete a B cell differentiation factor.

Cells from six cases of adult T cell leukaemia were studied with respect to phenotypical and functional features. All cells were reactive with anti-CD4 and anti-CDw29 monoclonal antibodies (antibody against helper inducer T cells) but were unreactive with anti-CD45R monoclonal antibody (antibody against suppressor inducer T cells). Functionally, these cells secreted a B cell differentiation factor detected by SKW6-CL4 cell differentiation to IgM-producing cells, this secretion being enhanced by culture with recombinant IL-2. Though these results indicate that adult T cell leukaemia cells are of mature helper inducer T cell origin, these cells strongly suppressed PWM-induced B cell differentiation in the absence of CD8+ suppressor effector T cells.

Aged

"B" cell origin of malignant cells in a case of American Burkitt's lymphoma. Characterization of cells from a pleural effusion.

Ninety-eight percent of the cells isolated from a malignant pleural effusion in a case of American Burkitt's lymphoma showed membrane fluorescence to antihuman IgM antisera, while only 2% of the cells formed spontaneous rosettes with sheep red blood cells. It is concluded that the Burkitt cells in the malignant effusion are derived from bursal equivalent lymphocytes. Epstein-Barr (E-B) viral titers to both viral capsule antigen and early antigen were not elevated. The significance of these findings is discussed.

B-Lymphocytes

Use of stimulating capacity of mixed lymphocyte reaction (MLR-S) as a possible marker for the cell-origin of null-cell acute lymphoblastic leukaemia.

The recent introduction of surface marker analysis indicates that the T-cell ALL represents 15--25%, the B-cell ALL represents less than 5% and the null-cell ALL represents more than 70% of cases. The origin of leukaemic null-cells is at present not clear. The present study shows that fresh leukaemic cells from five patients with null-cell ALL exerted a strong stimulating effect while the leukaemic cells from three patients with null-cell ALL failed to stimulate in 'one-way' MLR. Cultured leukaemic cells from three null-cell lines (NALM-16, NALL-1 and MOLT-10) consistently exerted a strong stimulation while leukaemic cells from one null-cell line (REH) exerted little or no stimulation on allogeneic lymphocytes. Leukaemic null-cells from the NALM-,6 line exhibited a lesser but significant stimulation in 'one-way' MLR. These observations lead us to speculate that leukaemic null-cells which possess a stimulating capacity may represent less differentiated leukaemic B lymphoid cells (early B-cell precursors) and leukaemic null-cells which possess no stimulating capacity may represent less differentiated leukaemic T lymphoid cells (early T-cell precursor).

Adult

Single cell origin of bigenotypic and biphenotypic B cell proliferations in human follicular lymphomas.

To investigate the possible relatedness of the subpopulations that make up so-called biclonal lymphomas, we examined five bigenotypic and biphenotypic follicular lymphomas using DNA probes specific for the t(14;18) chromosomal translocation, which is a characteristic feature of these neoplasms. On Southern blot analysis, both subpopulations from four of five lymphomas contained comigrating t(14;18) DNA rearrangements, confirming the single cell origins for these neoplasms. No comigrating t(14;18) DNA rearrangements were observed in the fifth lymphoma, but nucleotide sequence analysis of cloned, breakpoint DNA showed identical t(14;18) crossovers in the two subpopulations. The migration differences of both the Ig and chromosome 18 DNA rearrangements were shown to result from somatically acquired mutations of the Ig genes from the fifth lymphoma. These studies indicate that Ig gene rearrangements and idiotope expression are not consistently stable clonal markers since they are subject to variability as a result of somatic mutation. Although translocated chromosome 18 DNA rearrangements are more reliable, they may also vary among cells of some tumors since somatic mutation can affect, as well, DNA of translocated alleles in follicular lymphomas.

B-Lymphocytes

Cytogenetic and immunohistochemical evidence for the germ cell origin of a subset of acute leukemias associated with mediastinal germ cell tumors.

Most acute leukemias occurring in patients with mediastinal germ cell tumors (MGCTs) appear to be primary rather than therapy-related; however, no data have been obtained to date to confirm the hypothesized germ cell origin of the leukemias in this syndrome. We identified six male patients with this syndrome treated at Memorial Sloan-Kettering Cancer Center: in all six, the leukemia was studied immunohistochemically for various hematologic and germ cell antigens; in four, the karyotype of the leukemia was available; in one, the MGCT had also been karyotyped. In three patients, we found evidence of a germ cell origin of the acute leukemias. A 19-year-old male developed an acute myeloblastic leukemia 11 months after presenting with an MGCT; karyotypes of the two tumors showed them to be clonally related, both showing an i(12p), a cytogenetic marker of germ cell tumors. A 16-year-old male with probable Klinefelter's syndrome presented simultaneously with acute monocytic leukemia and an MGCT; although the MGCT was not karyotyped, the leukemia showed an i(12p). A 23-year-old male developed concurrently an MGCT and acute myelomonocytic leukemia; the leukemia cells coexpressed myelomonocytic antigens (HAM56, My4, My9) and cytokeratin, suggesting dual differentiation, myeloid and germ cell. Evidence for a germ cell origin of the acute leukemias in the three other patients was not obtained, although in all three the MGCT and the leukemia occurred simultaneously, supporting an etiologic relationship. Hence, these leukemias may represent another form of non-germ cell malignancy developing in germ cell tumors. All patients died of the leukemia, with a median survival of less than 2 months. This syndrome may be a useful pathologic model for the study of germ cell differentiation and hematopoietic ontogeny.

Adolescent

[Autoradiography of neurogenesis and morphogenesis of the regio cingularis of the rat. IV. Quantitative studies on the times of cell origins in the cortical lamina].

In 171 embryonic and neonatal rats has been investigated the time of origin of cells in the cortical laminae by means of autoradiographs, after injection of tritiated thymidine. It was made a comparison between the Regio cingularis mesoneocorticalis, Regio cingularis mesoarchicorticalis, Area praecentralis agranularis and Area postcentralis by the laminar differential labeling index. The time of origin indicates for each layer a principal day and three secondary days. In all regions the time of origin of cells is identically for the hylogenetic old layers VI and V, differences exist in the supragranular layers: the cells are generated earlier in the phylogenetic old areas than in the young areas.

Age Factors

Morphologic, immunologic, enzymehistochemical and chromosomal analysis of a cell line derived from Hodgkin's disease. Evidence for a B-cell origin of Sternberg-Reed cells.

Cell lines derived from Hodgkin's disease may provide a clue to the nature of Sternberg-Reed cells. In the current study, the establishment of an Epstein-Barr-virus-negative lymphoblastoid cell line, derived from the pleural fluid of a patient with the nodular sclerosis type of Hodgkin's disease, is described. The morphologic and immunologic cell marker findings indicate that this cell line is derived from Sternberg-Reed cells. The immunologic findings and a chromosomal analysis are in agreement with a B-lymphocyte origin of these cultured cells. Extrapolation of the results to Hodgkin's disease in vivo would indicate that Hodgkin's disease, like most non-Hodgkin's lymphomas, is the result of B-cell proliferation.

Antibodies, Monoclonal

Clinicostatistical study of ovarian tumors of germ cell origin.

Ovarian tumors of germ cell origin consisted of 440 dermoid cysts, 5 struma ovariis, 3 immature teratomas (G1, G2), 2 carcinoids, 8 dysgerminoma, 3 yolk sac tumors, one choriocarcinoma, 4 dermoid cysts with malignant transformation, and one immature teratoma (G3) of a total of 1,387 ovarian tumors. The patients' mean ages ranged between 17.3 years to 31.9 years, but the mean ages of those with carcinoid and dermoid cyst with malignant transformation were 46.0 and 53.0 years, respectively. In dermoid cyst bilaterality was 17.3% and occurrence during pregnancy or puerperium was 19.6%. In benign tumors, torsion of pedicle and association with pregnancy or puerperium were frequent unlike common epithelial carcinoma, but ascites was rare. Abdominal tumor, pain and metromenorrphagia were the main symptoms, but abdominal distension was scarce with dermoid cysts. The rate of malignant transformation of dermoid cysts was 0.9% overall but 4.3% in patients over 40 years. The prognosis for borderline germ cell tumors was good.

Adolescent

Single-cell origin of human mixed hemopoietic colonies expressing various combinations of cell lineages.

We have established single-cell culture for human mixed hemopoietic colonies using a micromanipulator. Mononuclear cells from human umbilical cord blood were cultured at a concentration of 1 X 10(4) cells per milliliter in methylcellulose medium containing medium conditioned by phytohemagglutinin-stimulated leukocytes and erythropoietin. It was possible to identify the single hemopoietic progenitors in situ in methylcellulose culture on the basis of unique morphology and migratory ability after 36 to 60 hours of incubation. Candidate single hemopoietic progenitors from methylcellulose medium were individually micromanipulated to secondary culture dishes and cultured for an additional ten to 14 days. The colonies derived from the single progenitors were individually picked and stained with May-Grünwald-Giemsa for analyses of the cellular composition. A total of 288 single cells were individually transferred to second dishes. Then 186 single cells produced secondary colonies consisting of cells in one to five different lineages. A total of 39 single cells produced mixed hemopoietic colonies consisting of cells in two, three, four, and five different lineages. There were eight types of colonies revealing two different lineages, ie, neutrophil (n)-erythrocyte (E), macrophage (m)-E, m-megakaryocyte (M), eosinophil (e)-basophil (b), eE, bE, bM, and EM lineages. Three types of colonies consisting of cells in three lineages were also seen, ie, nmM, nbE, and ebE. There were six types of colonies consisting of cells in four lineages, ie, nmbM, nmEM, nebE, mebM, and meEM. One type of colony consisted of cells in five different lineages (nmbEM). These results indicate the single-cell origin of human mixed hemopoietic colonies expressing various combinations of cell lineages. It also provides experimental data in support of stochastic mechanisms of stem cell differentiation.

Azure Stains

Ultradian mitotic rhythms in culture of human sarcomatous bone cells originating from the patient before and after chemotherapy.

Bone sarcomatous cells derived from human malignant tumors were cultured. The mitotic index was recorded for 39 hr. When the cultured cells originated from patients with cancer disease before any chemotherapy, ultradian mitotic rhythms of a 6-9-hr period were detected, but in many cases only after a sensitive statistical analysis was performed. When the cultured cells originated from cancer patients undergoing chemotherapy, the mitotic index was decreased, and the amplitudes of the 6-9-hr component oscillations of the mitotic index were highly significantly increased. Damping and fading out of an ultradian mitotic rhythmicity was a bad prognostic portent in bone cancer. With reference to chemotherapy, the restored and amplified ultradian rhythmicity disclosed an appreciable antitumor effect and better survival prospects for the patient.

Bone Neoplasms