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At least 631 records · Page 35Linked to original sources
PubMed1997

Prediction of therapy-related acute myelogenous leukemia (AML) and myelodysplastic syndrome (MDS) after autologous bone marrow transplant (ABMT) for lymphoma.

Therapy-related acute myelogenous leukemia and myelodysplastic syndrome (t-AML/MDS) are being reported with increasing frequency as a complication of ABMT for Hodgkin's disease and non-Hodgkin's lymphoma. At present there is no method available to predict who is at risk or is destined to develop this nearly universally fatal disorder. We therefore investigated whether clonal growth of cells is predictive of the development of t-AML/MDS. In a patient who developed secondary AML/MDS 18 months after ABMT, X-linked clonality analysis at the human androgen receptor locus was performed on serial banked samples, and documented transition from polyclonal to clonal hematopoiesis. Clonal cells could be identified 6 months after transplant (1 year prior to the diagnosis of t-AML/MDS), at a time when there was no morphologic or clinical evidence of disease. Clonality analysis can be predictive of the development of t-AML/MDS after ABMT and may offer important insights into associated risk factors and strategies to minimize the risk of t-AML/MDS.

Alleles↗
PubMed2003

Growth dynamics underlying petal shape and asymmetry.

Development commonly involves the generation of complex shapes from simpler ones. One way of following this process is to use landmarks to track the fate of particular points in a developing organ, but this is limited by the time over which it can be monitored. Here we use an alternative method, clonal analysis, whereby dividing cells are genetically marked and their descendants identified visually, to observe the development of Antirrhinum (snapdragon) petals. Clonal analysis has previously been used to estimate growth parameters of leaves and Drosophila wings but these results were not integrated within a dynamic growth model. Here we develop such a model and use it to show that a key aspect of shape--petal asymmetry--in the petal lobe of Antirrhinum depends on the direction of growth rather than regional differences in growth rate. The direction of growth is maintained parallel to the proximodistal axis of the flower, irrespective of changes in shape, implying that long-range signals orient growth along the petal as a whole. Such signals may provide a general mechanism for orienting growth in other growing structures.

Antirrhinum↗
PubMed1995

Allocation of epiblast cells to germ layer derivatives during mouse gastrulation as studied with a retroviral vector.

The embryonic ectoderm, or epiblast, is the source of the three primary germ layers that form during gastrulation in the mouse embryo. Previous studies have investigated the fate of epiblast cells in early gastrulation stages using clonal analysis of cell lineage and in late gastrulation stages using transplantation of labeled grafts. In this study, we studied the fate of late gastrulation stage epiblast using a clonal analysis based on a retroviral vector encoding the Escherichia coli lacZ gene. We found that by reducing the volume of viral suspension injected into each embryo, it was possible to achieve single infectious events. Our analysis of 20 embryos singly infected at the late streak stage and 21 at the head fold stage revealed clonal descendants in only a single germ layer in each embryo. These results indicate that allocation of epiblast progenitors to a single germ layer fate has occurred by late gastrulation in mouse embryos.

Animals↗
PubMed1989

Zygotic lethals with specific maternal effect phenotypes in Drosophila melanogaster. I. Loci on the X chromosome.

In order to identify all X-linked zygotic lethal loci that exhibit a specific maternal effect on embryonic development, germline clonal analyses of X-linked zygotic lethal mutations have been performed. Two strategies were employed. In Screen A germline clonal analysis of 441 mutations at 211 previously mapped X-linked loci within defined regions was performed. In Screen B germline clonal analysis of 581 larval and pupal mutations distributed throughout the entire length of the X chromosome was performed. These approaches provide an 86% level of saturation for X-linked late zygotic lethals (larval and pupal) with specific maternal effect embryonic lethal phenotypes. The maternal effect phenotypes of these mutations are described.

Animals↗
PubMed1998

VH gene analysis of clonally related IgM and IgG from human lymphoplasmacytoid B-cell tumors with chronic lymphocytic leukemia features and high serum monoclonal IgG.

An unusual group of human B-cell tumors with cellular features of chronic lymphocytic leukemia or lymphoplasmacytoid leukemia, together with high levels of a monoclonal IgG serum protein, has been investigated. Analysis of tumor-derived VH genes of neoplastic B lymphocytes was used to determine the clonal relationship between the IgM expressed or secreted by the tumor cells and the IgG serum paraprotein. In all five cases, VH gene sequences showed transcripts of IgM and IgG of common clonal origin. Sequences were derived from VH3 (4 of 5) and VH1 (1 of 5) families and were all highly somatically mutated with strong evidence for antigen selection. There was no intraclonal variation detectable in either IgM or IgG sequences. In 3 of 5 cases, in which monoclonal IgM and IgG were found in serum, the VH genes combined to Cmu or Cgamma showed identical mutational patterns. However, in 2 of 5 cases, in which IgM was confined to cell expression with only monoclonal IgG in serum, sequences of the VH transcripts of IgM and IgG showed many shared mutations but also numerous differences. In these cases, the level of mutation was similar in IgM and IgG and both appeared to be antigen selected. In summary, the final neoplastic event in this group of tumors has apparently occurred at the point of isotype switch from IgM to IgG, leading to dual isotype synthesis. In the group that secreted both isotypes, the mutation pattern was identical, indicating either synthesis by a single cell, or silencing of mutational activity before switching. In the group that did not secrete IgM, cells of each isotype were distinct and reflected a divergent mutational history.

Adult↗
PubMed1990

Wing formation in Drosophila melanogaster requires decapentaplegic gene function along the anterior-posterior compartment boundary.

Previous analyses of the decapentaplegic (dpp) gene in Drosophila melanogaster have suggested that its product, a polypeptide of the transforming growth factor-beta family of secreted factors, acts at the level of intercellular communication to control several events in spatial pattern formation. In this report, we use clonal analysis to demonstrate a localized requirement for wild-type dpp expression along the anterior-posterior (A/P) compartment boundary of the developing wing primordium. Clonal analysis reveals that normal wing blade development is solely dependent on dpp+ function in those anterior compartment cells that border the anterior-posterior (A/P) compartment boundary of the wing imaginal disk. Conversely, the wing blade will not develop if these boundary cells lack dpp activity. The localized requirement for dpp coincides with the spatial distribution of dpp transcripts, which accumulate in a stripe of cells at or near the known A/P compartment boundary of the wing imaginal disk. Thus, only a small subset of the cells that normally comprise the wing must express dpp to permit development of the entire structure. We propose that this localized expression of dpp is essential to proximal-distal appendage development. We discuss the possibility that dpp expression serves as a landmark for establishing and/or maintaining positional information in imaginal disks.

Animals↗
PubMed1992

Molecular analysis of clonality in plasma cell dyscrasias.

It has been suggested that multiple myeloma, generally considered a neoplastic disorder of mature plasma cells, may arise from a pluripotent haemopoietic stem cell. The possibility that circulating lymphocytes derive from the same neoplastic progenitor has been tested in a large number of studies in the past few years, as proof of the interest that this subject is raising among scientists, and also of its elusiveness. We studied a group of 29 patients with plasma cell dyscrasias in order to evaluate clonality of haemopoietic cell populations. The X-linked markers hypoxantine phosphoribosyltransferase (HPRT) and phosphoglycerate kinase (PGK) disclosed no monoclonal component in seven heterozygous women. Analysis of immunoglobulin gene rearrangement with four probes showed a germline configuration in samples from 25/29 patients. Only four bone marrow samples from subjects with aggressive disease had rearranged C mu sequence; one had rearrangement of JH and C mu.

Adult↗
PubMed2005

Analysis of clonal immunoglobulin heavy chain rearrangements in ocular lymphoma.

BACKGROUND: The morphologic diagnosis of primary and metastatic intraocular lymphoma (IOL) was made difficult by the paucicellular specimens with fragile populations of lymphocytes retrieved through pars plana vitrectomy (PPV). The analysis of immunoglobulin heavy chain (IgH) gene rearrangements (AIGHR) was used as an adjunct to cytopathology and flow cytometry in systemic lymphoma. In IOL, the sensitivity and specificity of AIGHR are unknown. METHODS: The authors reviewed the clinical records of patients who underwent PPV for suspicion of IOL at the Immunology and Uveitis Service of the Massachusetts Eye and Ear Infirmary (Boston, MA) between 2000 and 2002. AIGHR was performed as a routine diagnostic test on cell lysates isolated from < 0.5 mL of vitreous fluid. The authors used seminested polymerase chain reaction (PCR) with consensus primers for the VDJ-region of the IgH gene. PCR products were analyzed by polyacrylamide gel electrophoresis. RESULTS: Thirty patients (37 specimens) with chronic vitritis and 17 patients (23 specimens) with IOL were included. The specificity of vitreous fluid cytopathology, flow cytometry, and AIGHR was 1.0, and the sensitivity values were 0.24, 0.36, and 0.64, respectively. AIGHR was negative in two patients for whom cytopathology or flow cytometry revealed the diagnosis of lymphoma. Clonal IGHR was found in four specimens classified as negative for lymphoma based on cytopathology and flow cytometry. CONCLUSIONS: AIGHR supplemented cytopathology and flow cytometry to increase the diagnostic yield in IOL.

Adolescent↗
PubMed1976

Commitment to erythroid differentiation by friend erythroleukemia cells: a stochastic analysis.

A method for the clonal analysis of murine erythroleukemia cells has been developed which allows the precise characterization of the number of progeny produced by each cell and the degree of differentiation of each progeny cell. The potential of almost every cell in the culture can be monitored because a plating efficiency close to 100% has been achieved. The effects of treatment with an inducer of differentiation (DMSO) on the proliferative capacity of the treated cells have been studied with this technique. Cells from a mass culture treated with inducer give rise to colonies of differentiated progeny when subsequently cloned in the absence of inducer. Colonies exhibiting this phenotype represent the progeny of cells committed to the differentiation pathway by treatment with inducer. We observe that the commitment decision limits the subsequent proliferative capacity of the cell to four additional cell divisions. A quantitative analysis suggests that the commitment decision for each cell is made in a stochastic manner. Irreversible commitment to the expression of differentiated functions occurs with discrete probability per cell generation for many cell generations. The value for this probability is a function of the concentration of inducer (DMSO). A correlative biochemical study suggests that an irreversible commitment decision by a significant proportion of the population precedes or accompanies increases in cytoplasmic globin mRNA levels, one of the earliest detectable biochemical markers for erythroid differentiation in this system. A specific kinetic model based on these considerations has been developed to predict clonal phenotypes as a function of time and probability of commitment. Quantitative predictions based on this model are in excellent agreement with experimental observations. The effectiveness of a stochastic model in predicting the behavior of this system is discussed in relation to the stochastic behavior of normal hematopoiesis and the biochemical mechanisms which control these differentiation programs.

Cell Division↗
PubMed1991

Effect of transforming growth factor beta-1 on ovine satellite cell proliferation and fusion.

We have evaluated the effect of transforming growth factor beta-1 (TGF beta-1) on proliferation and fusion of cultured ovine satellite cells isolated from 5-month-old wether lambs. The isolation and culture protocols were validated by clonal analysis of the original cell preparation and assessment of proliferation and fusion of control cultures. Approximately 85% of the original cells isolated were myogenic as assessed by clonal analysis. The ovine cells doubled approximately every 18 hours during their exponential growth period and achieved a maximum percent fusion of 39.5% after 144 hours in culture. TGF beta-1 inhibited fusion of these cells in a dose-dependent manner with half-maximal inhibition occurring at .08 ng/ml. Maximal inhibition (95% suppression) occurred between .1 and .5 ng/ml. TGF Beta-1 (.05-3.0 ng/ml) did not inhibit proliferation of cultured ovine satellite cells in serum-containing medium or in serum-free defined medium. In contrast, TGF beta-1 did significantly suppress serum-stimulated proliferation of either porcine or bovine satellite cells that were isolated by using a procedure identical to that used to isolate the ovine satellite cells. Thus, proliferation of ovine satellite cells appears to respond differently to TGF beta-1 than does proliferation of either porcine or bovine satellite cells.

Animals↗
PubMed1997

Evidence for clonal origin of neoplastic neuronal and glial cells in gangliogliomas.

Gangliogliomas are rare tumors of the central nervous system that account for approximately 1% of all brain tumors. Histologically, gangliogliomas are composed of intimately admixed glial and neuronal components, the pathological origins of which remain to be characterized. Clonal analysis through examination of the pattern of the X chromosome inactivation allows one to distinguish monoclonal differentiation of a genetically abnormal progenitor cell from parallel, but independent, clonal expansion of two different cell types during tumorigenesis in biphasic neoplasms, such as gangliogliomas. In the present study, we investigated the clonality of eight gangliogliomas from female patients using both methylation- and transcription-based clonality assays at the androgen receptor locus (HUMARA) on the X chromosome. Among tumors from seven patients who were heterozygous at the HUMARA locus, five were identified as monoclonal with the methylation-based clonality assay, and the results were confirmed by the transcription-based method, whereas two were shown to be polyclonal by the methylation-based clonality assay but monoclonal by transcription-based clonality analysis. We conclude that the predominant cell types in most gangliogliomas are monoclonal in origin and derive from a common precursor cell that subsequently differentiates to form neoplastic glial and neuronal elements.

Adult↗
PubMed1997

Limit theorems and a general framework for risk analysis in clonal forestry.

Use of clonally propagated plantings in reforestation offers management advantages of phenotypic uniformity and high yields. Disadvantages include low genetic diversity and the possibility that the clone or clones chosen are particularly susceptible to attack by an insect or pathogen unforeseen as a problem at the time of clonal selection. In this paper, we continue consideration of the problem of choosing an optimal number of clones to minimize the risk of plantation failure. We present an analysis in which risk of failure for a plantation is represented by the probability that the proportion, S, of ramets that survive until harvest is less than or equal to a prescribed value. Our approach includes most earlier treatments as special cases. We show that the proportion S converges in distribution and, furthermore, that, under general conditions, a moderate number of clones, usually no more than 20 to 40 and often fewer, provides equivalent or better protection against catastrophic loss than does a large number of clones.

Forestry↗
PubMed1991

The autoimmune infiltrate of Basedow's disease: analysis of clonal level and comparison with Hashimoto's thyroiditis.

The availability of high efficiency T-cell cloning techniques recently allowed the identification and characterization of clones derived from the thyroid infiltrate of patients with autoimmune thyroid diseases. Phenotypical and functional analysis of T-cell clones obtained from thyroid infiltrates of patients with Hashimoto's thyroiditis show that most of them are progenies of CD8+ cytolytic T cells with natural killer activity. This phenomenon, of potential importance in tissue damage, is markedly less pronounced in Basedow's disease glands. In both Hashimoto's thyroiditis and Basedow's disease only a minority of clones appear to be specific for autologous thyroid cells and most of them are potent interferon-gamma producers, while increased secretion of tumor necrosis factor-alpha is observed only in Hashimoto's thyroiditis. In contrast with normal lymphoid tissue, only very few T cell clones derived from both BD and HT infiltrates were able to produce detectable amounts of IL-4, suggesting that most of the thyroid-infiltrating T cells represent quite homogeneous populations of Th1-type "inflammatory" T cells. This peculiar potential of lymphokine secretion could play a role in the expression and/or maintenance of thyroid autoimmunity and thyroid functional damage.

CD4-Positive T-Lymphocytes↗
PubMed1997

Routine diagnosis of large granular lymphocytic leukaemia by Southern blot and polymerase chain reaction analysis of clonal T cell receptor gene rearrangement.

AIMS: To compare the polymerase chain reaction (PCR) assay with standard Southern blot (SB) hybridisation for the detection of clonal T cell receptor (TCR) gene rearrangements in large granular lymphocyte (LGL) proliferations; to evaluate the reliability and practicality of the methods for routine diagnostic use; and to determine the sensitivity of the PCR method. METHODS: Blood lymphocytes were isolated from 12 patients with persistent CD3+CD8+ lymphocytosis with LGL morphology. Clonal rearrangements of the TCR gene were demonstrated by SB hybridisation with a TCR beta constant probe, and by PCR amplification of portions of the TCR beta and TCR gamma genes. RESULTS: Monoclonal TCR beta gene rearrangements were detected in eight patients (67%) by PCR analysis and five patients (42%) by SB hybridisation. PCR analysis also showed that seven patients (58%) had monoclonal TCR gamma gene rearrangements. All cases which had TCR beta clonal rearrangements shown by SB hybridisation were similarly identified by PCR. Sensitivity tests suggested that the TCR beta PCR technique was capable of detecting clonality in as little as 50 pg of DNA. The TCR beta primers could detect one clonal cell in approximately 200 or more normal cells (< 0.5%), a sensitivity level that at least doubles that of the SB hybridisation technique. CONCLUSIONS: The use of PCR technology proved to be superior to SB hybridisation for the routine investigation of suspected cases of LGL leukaemia. Nine patients (75%) in this study were found to have TCR beta and/or TCR gamma monoclonal gene rearrangements. This approach is ideal for distinguishing between reactive and clonal LGL proliferation in a routine diagnostic laboratory.

Blotting, Southern↗
PubMed2006

Sequence analysis proves clonal identity in five patients with typical and blastoid mantle cell lymphoma.

Mantle cell lymphoma (MCL) is typically composed of small irregular lymphoid cells. Blastoid variants, composed of lymphoblast-like (classic type) or large (pleomorphic type) cells, arise de novo or in patients with typical MCL. Although it has been assumed that blastoid variant represents histologic transformation of typical MCL, the clonal relationship between the two tumors has rarely been assessed at the molecular level. We identified five patients with typical MCL who subsequently developed the blastoid variant. There were two men and three women with a median age of 65 years (range, 34-70) at diagnosis of typical MCL involving lymph nodes. The median interval between typical and blastoid MCL was 36 months (range, 11-103). Subsequent blastoid variant MCL involved soft tissue (two), lymph node (one), ileum (one), or rectum (one). All typical and blastoid neoplasms were positive for CD20, cyclin D1, and monotypic surface immunoglobulin light chain, and all typical cases were positive for CD5. Two blastoid neoplasms lost CD5 expression, one of which aberrantly expressed CD10. Immunostaining for Ki-67 showed a median proliferative fraction of 20% in typical and 70% in blastoid neoplasms. Sequence analysis of the VDJ regions of the rearranged IgH allele proved clonal identity in each set of paired samples in all five patients. These results support the concept that blastoid MCL arising in patients with typical MCL represents histologic transformation of the original neoplastic clone.

Adult↗
PubMed1988

Expression of a functional CD3-Ti antigen/MHC receptor in the absence of surface CD2. Analysis with clonal Jurkat cell mutants.

To investigate the requirement for CD2 expression in activation of T lymphocytes via the CD3-Ti antigen/MHC receptor complex, we produced and characterized a series of CD2- Jurkat variants. These mutants lack detectable surface CD2 as determined by indirect immunofluorescence, immunoprecipitation analysis, and specific radiolabeled antibody binding assay, but nevertheless, expressed normal numbers of CD3-Ti receptors. As expected, the combination of anti-CD2 antibodies, termed anti-T112 and anti-T113, which are mitogenic for resting T lymphocytes, failed to stimulate activation of these variants. In contrast, triggering of their CD3-Ti components resulted in the normal set of T lymphocyte-associated activation events, including phosphoinositide turnover, elevation in intracellular free calcium, early gene-induction events, and IL-2 production. Assuming that the Jurkat cell line is representative of normal cycling human T lymphocytes, we conclude that the presence of the CD2 molecule on the plasma membrane is not in itself a requirement for an operational CD3-Ti-alpha/beta receptor.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1998

Fine needle aspiration cytology in the investigation on non-Hodgkin's lymphoma.

AIMS: To assess the value of flow cytometry (FCM) in the diagnosis and classification of reactive lymphoid hyperplasia and malignant lymphoma by fine needle aspiration (FNA) cytology. METHODS: Forty six fine needle aspirates of lymphoproliferative disorders were examined by FCM as well as routine cytological assessment. An immunoglobulin light chain ratio (LCR) was calculated for clonality analysis. Additional immunophenotyping was performed in 15 cases. RESULTS: All 25 cases of reactive lymphoid hyperplasia were polyclonal by FCM (LCR < 2/1); 17 of 20 cases of B cell non-Hodgkin's lymphoma were monoclonal (LCR > 3/1). Analysis of cells based on size facilitated detection of small populations of clonal neoplastic cells. Analysis of CD5, CD10, and CD23 expression by FCM facilitated subclassification of mantle cell lymphoma, small lymphocytic lymphoma, and some lymphomas of follicle centre cell origin. One case of T cell non-Hodgkin's lymphoma was correctly classified by FCM. CONCLUSIONS: FNA cytology is a reliable method for investigation of lymphoproliferative disorders. Although excision biopsy and histopathological examination remain the gold standard for primary diagnosis and classification of non-Hodgkin's lymphoma, FNA cytology with clonality analysis and immunophenotyping by FCM is useful for distinguishing reactive from neoplastic lymphoid populations, and can facilitate lymphoma classification.

Adult↗
PubMed1997

Analysis of clonality of lymphocytic leukemia and lymphoma by T-cell receptor gene rearrangement.

OBJECTIVE: To analyse the relationship between the number of T-cell receptor (TCR) gamma gene rearrangement and clonality of malignant cells in lymphocytic leukemia and lymphoma. METHODS: The TCR gamma gene characteristics of 73 cases of lymphocytic leukemia and lymphoma and other diseases that had presented 1 or 2 prominent bands of amplified TCR gamma VI subgroups-J1/2 gene rearrangement (GR) were detected by polymerase chain reaction-restriction enzymes (PCR-RE), heteroduplex formation (HDF), DNA sequencing and single-strand conformational polymorphism (SSCP). RESULTS: Thirty-four percent patients had 2 of TCR gamma GR (biallelic rearrangement); twenty-six percent had 1 of TCR gamma GR; fifty-four percent of acute lymphocytic leukemia's and nineteen percent of non-Hodgkin's lymphomas (NHL) had 2 of TCR gamma GR. HDF could rapidly confirm more than 1 of alleles while more alleles were difficult to be recognized by restriction analysis. Sense and antisense strands of heteroduplex were composed of 2 of TCR gamma GR respectively. By combining HDF method, oligoclonalities in three patients (2 acute lympholytic leukemia and 1 myelodysplastil syndrome and clone evolution in one NHL were found. CONCLUSIONS: The fact that many patients had 2 of TCR GR means that complications of two neoplastic clones can be affirmed only if more than 2 of TCR GR are found. HDF can be used to detect the clonality, oligoclonality and polyclonality of lymphoid cells in lymphocytic leukemia and lymphoma by analysis of the difference of gene segments. HDF is a reliable method for the clone evolution research of lymphoid malignancies in progression.

Base Sequence↗