Simultaneous genotypic and immunophenotypic analysis of interphase cells using dual-color fluorescence: a demonstration of lineage involvement in polycythemia vera

Abstract Fluorescent in situ hybridization has become a useful technique by which chromosomal abnormalities may be shown in interphase cells. We present a dual-fluorescence method whereby a chromosomal and immunophenotypic marker can be visualized simultaneously in the same interphase cell. Two patients with the myeloproliferative disorder polycythemia vera and trisomy for chromosome 8 have been studied using this technique and selective involvement of the myeloid and erythrocyte lineages has been shown by the detection of the trisomy in immunophenotyped cells. Simultaneous analysis of genotype and immunophenotype in individual cells from patients with myeloproliferative disorders or leukemia may help identify the developmental and lineage status of cells in which molecular alterations have resulted in clonal advantage.

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Simultaneous genotypic and immunophenotypic analysis of interphase cells using dual-color fluorescence: a demonstration of lineage involvement in polycythemia vera

Blood ◽

10.1182/blood.v80.4.1033.bloodjournal8041033 ◽

1992 ◽

Vol 80 (4) ◽

pp. 1033-1038 ◽

Cited By ~ 1

Author(s):

CM Price ◽

EJ Kanfer ◽

SM Colman ◽

N Westwood ◽

AJ Barrett ◽

...

Keyword(s):

Polycythemia Vera ◽

Fluorescent In Situ Hybridization ◽

Chromosomal Abnormalities ◽

Myeloproliferative Disorders ◽

Chromosome 8 ◽

Dual Fluorescence ◽

Interphase Cell ◽

Molecular Alterations ◽

Interphase Cells

Fluorescent in situ hybridization has become a useful technique by which chromosomal abnormalities may be shown in interphase cells. We present a dual-fluorescence method whereby a chromosomal and immunophenotypic marker can be visualized simultaneously in the same interphase cell. Two patients with the myeloproliferative disorder polycythemia vera and trisomy for chromosome 8 have been studied using this technique and selective involvement of the myeloid and erythrocyte lineages has been shown by the detection of the trisomy in immunophenotyped cells. Simultaneous analysis of genotype and immunophenotype in individual cells from patients with myeloproliferative disorders or leukemia may help identify the developmental and lineage status of cells in which molecular alterations have resulted in clonal advantage.

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Detection of 11q13 rearrangements in hematologic neoplasias by double- color fluorescence in situ hybridization

Blood ◽

10.1182/blood.v87.4.1512.bloodjournal8741512 ◽

1996 ◽

Vol 87 (4) ◽

pp. 1512-1519 ◽

Cited By ~ 50

Author(s):

LJ Coignet ◽

E Schuuring ◽

RE Kibbelaar ◽

TK Raap ◽

KK Kleiverda ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Hematologic Malignancies ◽

Current Data ◽

Lymphocytic Leukemia ◽

Cell Nuclei ◽

Interphase Cell ◽

Interphase Cells ◽

Chromosomal Breaks

Rearrangements within the chromosome 11q13 region are frequent in hematologic malignancies. 50% of 75% of mantle cell lymphomas (MCLs) carry a translocation t(11;14) (q13;q32). Using Southern blot analysis, a BCL1 breakpoint can be detected in approximately 50% of MCLs. It is not known whether other MCLs harbor also breakpoints at 11q13. Breakpoints in this region not involved in t(11;14), are detected in chronic lymphocytic leukemia and acute myeloid leukemia. To detect and localize breakpoints at 11q13 more accurately, we have developed fluorescence in situ hybridization using two probe sets of differently labeled cosmids, symmetrically localized at either side of the major translocation cluster of BCL1. These probes span a region of 450 to 750 kb. We applied this assay to a series of hematologic malignancies with 11q13 abnormalities identified by classical cytogenetics. All four samples with a t(11;14) (q13;q32) showed dissociation of the differently colored signals in metaphase and interphase cells, thereby indicating a chromosomal break in the region defined by the probe sets. The frequency of abnormal metaphase and interphase cells was comparable with that observed in any of the 13 malignancies with other chromosomal 11q13 abnormalities, indicating that these chromosomal breaks occurred outside the 450- to 750-kb region covered by the probes. One patient showed triplication and one patient showed monoallelic loss of this region. The current data show that double-color fluorescence in situ hybridization is a simple and reliable method for detection of the t(11;14)(q13;q32) in interphase cell nuclei and that is can be used to distinguish this translocation from other 11q13 rearrangements in hematologic malignancies.

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The Prognostic Impact of Fluorescent-in Situ Hybridization (FISH) and Conventional Karyotying in Korean Multiple Myeloma Patients: A Retrospective Multicenter Study.

Blood ◽

10.1182/blood.v114.22.4902.4902 ◽

2009 ◽

Vol 114 (22) ◽

pp. 4902-4902 ◽

Cited By ~ 1

Author(s):

Min Jae Kim ◽

Suk Joong Oh ◽

Chang Ki Min ◽

Chong Won Park ◽

Hwi-Joong Yoon ◽

...

Keyword(s):

Bone Marrow ◽

Overall Survival ◽

Fluorescent In Situ Hybridization ◽

Plasma Cells ◽

Chromosomal Abnormalities ◽

Albumin Level ◽

Complex Karyotype ◽

Independent Risk Factors ◽

Univariate Analyses

Abstract Abstract 4902 Introduction Cytogenetics and fluorescent-in situ hybridization (FISH) are important outcome predictors in multiple myeloma (MM). There were only few small studies that investigated prognostic implication of FISH and/or conventional karyotyping in Korean MM patients. We investigated the incidences and prognostic significances of chromosomal abnormalities detected by FISH and/or conventional karyotyping among Korean MM patients. Patients and Methods We collected data of patients from Korean Myeloma Registry and performed retrospective analysis. We compared the survival of patients with chromosomal abnormalities and other clinical findings. Results From 2000 to 2009, total of 801 newly diagnosed myeloma patients were enrolled in this study. Median age of patients was 62 years. Median overall survival was 82 months, and median follow up of time was 92 months. Among the patients who had conventional karyotype analysis, 17.1% were complex karyotype, followed by del13q (7.4%), hyperdiploidy (7.6%), hypodiploidy (3.0%), and t(11;14) (3.9%). Among the patients who had FISH analysis, 22.8% were del 13q, followed by t(11;14) (18.2%), t(4;14) (13.7%), del17p (11.8%) and t(14;16) (5.9%). Univariate analyses revealed that complex karyotype (p<0.01), hypodiploidy (p=0.01), del13q (p<0.01) by conventional karyotyping, and t(4;14) (p=0.04) by FISH negatively impacted the overall survival. Other genomic aberrations did not affect the overall survival. Clinical parameters that impact on overall survival were percentage of plasma cells in bone marrow, serum beta2-microglobulin, creatinine, low hemoglobin, and low albumin levels. On multivariate analysis, percentage of plasma cells in bone marrow (p<0.01) and low serum albumin level (p<0.01) were independent risk factors for overall survival. Conclusions Our results showed that complex karyotype, hypodiploidy, t(4;14), and del13q by FISH and/or conventional karyotyping were negative prognostic factors for overall survival in univariate analyses. On multivariate analysis, low serum albumin level and percentage of plasma cells in bone marrow were independent risk factors for overall survival. In future, prospective trial with laboratory standardization is warranted for more reliable results from FISH and/or conventional karyotyping in MM patients. Disclosures Suh: Janssen Korea: Research Funding.

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Clonal analysis of myelodysplastic syndrome: monosomy 7 is expressed in the myeloid lineage, but not in the lymphoid lineage as detected by fluorescent in situ hybridization

Blood ◽

10.1182/blood.v80.1.217.217 ◽

1992 ◽

Vol 80 (1) ◽

pp. 217-224 ◽

Cited By ~ 76

Author(s):

WR Gerritsen ◽

J Donohue ◽

J Bauman ◽

SC Jhanwar ◽

NA Kernan ◽

...

Keyword(s):

T Cells ◽

In Situ Hybridization ◽

B Cell ◽

Cell Lines ◽

Fluorescent In Situ Hybridization ◽

Normal Cells ◽

Monosomy 7 ◽

Cell Lineages ◽

Interphase Cells

Abstract Conflicting results have been published on whether or not myelodysplastic syndromes (MDS) affect all cell lineages. Involvement of myeloid and erythroid cell lineages has been regularly observed, but it remains controversial whether the different lymphoid cell lineages are involved. In this study of eight patients with MDS associated with monosomy 7, fluorescent in situ hybridization (FISH) was used to enumerate the chromosomes 7 in interphase cells. With the probe D7Z1, the rate of false-positive detection of monosomy 7 was 3% +/- 2% in normal cells. T- and B-cell lines were established from eight patients with MDS and monosomy 7. As determined by FISH in interphase cells, 1.9% (0% to 3%) of the cells in the B-cell lines showed one fluorescent spot and 1.1% (0% to 2.9%) of the cells in the T-cell lines. These values do not differ from normal values. However, the possibility that normal cells were selected when the T- and B-cell lines were established could not be excluded. Therefore, peripheral blood cells were obtained, separated according to surface markers specific for lymphoid and myeloid cell lineage with a cell sorter, and analyzed for the expression of monosomy 7 by FISH. Antibodies recognizing T cells (CD3), B cells (CD20), natural killer (NK) cells (CD57), monocytes and granulocytes (low and high expression of CD11b antigen), and myeloid progenitors (CD33) were used to separate cells. The expression of monosomy 7 in the T cells, NK cells, and B cells did not differ from control values. These results in the lymphoid subpopulations are in stark contrast with the observations in the myeloid populations; the percentage of cells with monosomy 7 ranged from 9% to 78% (controls: 6% +/- 2%) in cells with low CD11b expression, 20% to 89% in cells with a high expression of the CD11b antigen (controls: 7% +/- 3%), and 23% to 91% in the CD33 positive cells (controls: 5% +/- 3%). The results of this study suggest that monosomy 7 does not usually affect lymphoid subpopulations but is restricted to committed progenitor cells with the capacity to differentiate into mature myeloid cells.

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Assessment of Chromosome 8 Copy Number in Cervical Cancer by Fluorescent in situ Hybridization

Experimental and Molecular Pathology ◽

10.1006/exmp.1999.2256 ◽

1999 ◽

Vol 66 (2) ◽

pp. 157-162 ◽

Cited By ~ 9

Author(s):

H.F.L. Mark ◽

D. Feldman ◽

M. Samy ◽

C.-L. Sun ◽

Sam Das ◽

...

Keyword(s):

Cervical Cancer ◽

In Situ Hybridization ◽

Copy Number ◽

Fluorescent In Situ Hybridization ◽

Chromosome 8

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The sugary-type isoamylase gene from rice and Aegilops tauschii: characterization and comparison with maize and Arabidopsis

Genome ◽

10.1139/g02-130 ◽

2003 ◽

Vol 46 (3) ◽

pp. 496-506 ◽

Cited By ~ 16

Author(s):

S Rahman ◽

Y Nakamura ◽

Z Li ◽

B Clarke ◽

N Fujita ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescent In Situ Hybridization ◽

Aegilops Tauschii ◽

Rice Chromosome ◽

Proximal Region ◽

Chromosome 8 ◽

D Genome ◽

Debranching Enzyme ◽

Cdna Sequences

Genes for an isoamylase-like debranching enzyme have been isolated from rice and Aegilops tauschii, the donor of the D genome to wheat. The structures of the genes are very similar to each other and to the maize SU1 isoamylase gene and consist of 18 exons spread over approximately 7.5 kb. Southern analysis and fluorescent in situ hybridization showed the Ae. tauschii gene to be located in the proximal region of the short arm of chromosome 7D, thus showing synteny with the localization of the rice isoamylase gene on rice chromosome 8. Analysis of the expression pattern of wheat sugary isoamylase genes indicates that they are strongly expressed in the developing endosperm 6 days after flowering. Three distinct Sugary-type cDNA sequences were isolated from the wheat endosperm that are likely to correspond to the products of the three genomes. The deduced amino acid sequence of rice and wheat Sugary-type isoamylase is compared with other sequences available in the database and the results demonstrate that there are three types of isoamylase sequences in plants: those containing 18 exons (the Sugary-type isoamylase gene), those containing 21 exons, and those containing only 1 exon. It is possible that different combinations of isoamylase genes are expressed in different tissues.Key words: isoamylase, rice, wheat, sugary, FISH.

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