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R C Braylan

Publications and source records attributed to R C Braylan.

At least 19 recordsLinked to original sources
PubMed1995

CD5-expressing B-cell non-Hodgkin's lymphomas with bcl-1 gene rearrangement have a relatively homogeneous immunophenotype and are associated with an overall poor prognosis.

Mantle cell lymphomas (MCLs) are typically CD5-expressing B-cell non-Hodgkin's lymphomas (NHLs) that frequently harbor the chromosomal translocation t(11;14) or bcl-1 gene rearrangements. Insufficient data are available on the biologic features and clinical behavior of rigorously characterized MCL. As these NHLs have been reported to exhibit various histologic and cytologic expressions, and in order to avoid using somewhat arbitrary and subjective morphologic definitions, we chose to study cases of MCL selected on more objective grounds. Specifically, 15 samples (from 14 patients) of CD5-expressing B-cell NHLs with detectable bcl-1 gene rearrangement were included. Overall, these patients had relatively uniform clinical manifestations. Most were older men (mean age, 67 years) who presented with lymphadenopathy, high-stage disease, and bone marrow involvement. All but two patients relapsed, demonstrated residual tumor, or had disease progression after an initial response to various therapies. Nine patients have died; these patients had a median survival of only 19 months. All cases could be classified within the broad morphologic spectrum previously described for MCL, and no predominant histologic subtype was observed. However, cases could be segregated into two major groups according to tissue architecture: one with a purely diffuse pattern and the other with at least a focal nodular component. Patients with purely diffuse tumors had a lower survival rate (0%) than those with tumors having a nodular component (62% survival rate). In contrast to the morphologic variability, these NHL exhibited a rather homogeneous immunophenotypic pattern. All cases demonstrated intense CD20 expression, with typically intense IgM and light chain expression, and relatively weak IgD expression. In no case was CD10 detected on the neoplastic cells. DNA content analysis showed aneuploidy only in three instances, and two groups of cases could be arbitrarily defined on the basis of their S-phase fraction. A relationship between a purely diffuse growth pattern and a high S-phase fraction (greater than 5%) was observed. As expected from this association, patients with tumors having high S-phase fractions fared worse (14% survival rate) than those patients with tumors showing lower S-phase fractions (57% survival rate). Thirteen NHLs from 12 patients had amplifiable bcl-1 gene rearrangements at the major translocation cluster (MTC). The bcl-1 breakpoints aggregated within a 63-bp region of the MTC, and the amplified tumor DNA from each patient had unique N-nucleotide junctional sequences and Ig joining region breakpoint sites.(ABSTRACT TRUNCATED AT 400 WORDS)

Aged↗
PubMed1995

The cell surface antigen and DNA content distribution of lymph nodes with reactive hyperplasia.

Few studies have comprehensively examined the expression and distribution of cell surface antigens and DNA content in non-neoplastic, reactive lymph nodes. We have performed a flow cytometric immunophenotypic and DNA content analysis of 64 lymph nodes from 62 patients (23 male; 39 female; ages 21 months to 80 years) with typical reactive lymphoid hyperplasia as assessed by histology. CD45, a pan-leukocyte marker, was detected on virtually all cells (99 +/- 4%). All pan-T-cell-associated antigens studied were expressed within a narrow range of average values (54 - 64%), and no loss of individual T-cell antigens was observed in any case. Intercase variation of fluorescence intensity for each antigen was minimal. The mean CD4:CD8 ratio was 3.9 +/- 2.6, with only one case (HIV+) showing a marked CD4:CD8 inversion (0.4). The number of cells coexpressing CD4 and CD8 was very low (3 +/- 3%), consistent with the mature phenotypic nature of the majority of T lymphocytes. B-cells, as defined by CD19 and CD20 antibodies, accounted for 36 +/- 16% and 43 +/- 18% of cells analyzed, respectively, and, like the pan-T-cell antigens, displayed minimal intercase variability in antigen expression. Cells coexpressing CD20 and CD5 were noted, although their frequency could not be calculated accurately. IgD- and IgM-bearing cells were both generally well-defined populations: 24 +/- 10% and 31 +/- 14%, respectively. On the other hand, IgG- and IgA-bearing cells displayed broad fluorescence intensity, precluding an exact calculation of their frequencies.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

A knowledge-based system for the interpretation of flow cytometry data in leukemias and lymphomas.

Flow cytometry immunophenotyping and analysis of DNA ploidy and proliferative activity have become extremely helpful techniques for diagnosing and subclassifying hematopoietic cell populations in a modern, full-service hematopathology laboratory. The number of physicians with special training in the interpretation of these studies is limited. A knowledge-based computer system has been designed to aid in the interpretation of immunophenotyping and DNA flow cytometry results in hematopoietic disease. The system, known as "Professor Fidelio," is a heuristic classification system that reasons on the basis of defined diagnostic patterns. In this study, Fidelio was tested as a stand-alone system on 366 specimens from two large tertiary medical centers. Fidelio's interpretation was considered to be appropriate in all cases. In 300 of 366 (82%) specimens, the system's interpretation agreed with the diagnosis of record. Many of the disagreements could be traced to errors in the recording of the original diagnosis and minor differences in diagnostic criteria between Fidelio's knowledge base and the criteria in use at the medical centers. When used in a stand-alone mode, Fidelio's interpretation was less specific than the diagnosis of record in certain lymphoproliferative disorders that require morphologic information for subclassification. Professor Fidelio is one module in a workstation for the diagnostic hematology laboratory. This workstation is designed for interpretive reporting, education, and database functions for clinical research. Clinical and morphologic information are shared between Fidelio and the other modules for peripheral blood analysis, bone marrow morphology, and lymph node interpretation by means of a relational database. The system will be useful in hospitals that lack individuals specially trained in flow cytometry.(ABSTRACT TRUNCATED AT 250 WORDS)

Artificial Intelligence↗
PubMed1994

Antigen expression and polymerase chain reaction amplification of mantle cell lymphomas.

Flow immunophenotyping, DNA content analysis, and polymerase chain reaction (PCR) amplification for t(11;14) and t(14;18) were performed on 11 cases of typical mantle cell lymphoma (MCL), 5 cases of apparent MCL with proliferation centers (MCL-PC), and 5 cases of small lymphocytic lymphoma (SLL). Immunophenotyping showed IgM (P < .001), Ig light (P < .001), and CD20 (P < .001) expression to be more intense in MCL than in SLL. In MCL-PC, the mean intensity of IgM, Ig light chain, and CD20 expression was intermediate to the intensities observed in MCL and SLL. Furthermore, in contrast to SLL, all MCL and 4 of 5 MCL-PC cases exhibited stronger CD20 than CD19 expression. CD10 expression was not observed in any case and CD5 expression was present in all SLL and MCL-PC cases and in 9 of 11 MCL cases. DNA content analysis showed an S-phase fraction of less than 3% in all cases studied and, except for 1 MCL case, all lymphomas were DNA diploid. The t(11;14) breakpoint junctions involving the bcl-1 major translocation cluster were amplified by PCR in 4 of 11 (36%) MCL cases and in none of the MCL-PC or SLL cases. The t(14;18) involving the bcl-2 major breakpoint region was not identified by PCR in any case. We conclude that the level of expression of surface antigens and the rapid detection of t(11;14) by PCR are potentially useful for distinguishing MCL and SLL in the clinical setting. Further investigations as to the biologic relationship between MCL, MCL-PC, and SLL, and the utility of t(11;14) PCR in these lymphomas are warranted.

Aged↗
PubMed1994

Evaluation of sensitivity in DNA aneuploidy detection using a mathematical model.

Detection of DNA aneuploid populations in samples analyzed by flow cytometry generally requires recognition of more than a single G0/G1 peak on the resulting DNA histogram. This assessment is critical in clinical samples if diagnosis or subsequent patient treatment are influenced. In this paper we evaluate three major distribution parameters that contribute to recognition or obscuration of closely spaced peaks on a DNA histogram. These parameters are (1) coefficient of variation (CV), (2) proportion of cells within each peak, and (3) separation of peaks (DNA index, DI). Through the analysis of computer-synthesized DNA histograms, we produced a single chart illustrating the minimum CV required for two peaks (representing DNA diploid and DNA aneuploid cells) in mixtures of various proportions (1:99 to 99:1) and DI values (1.01 to 1.20) to be resolved. This information can be utilized to provide insight into the ability of flow cytometry to resolve a DNA aneuploid population in a histogram.

Algorithms↗
PubMed1994

Pure red cell aplasia associated with angioimmunoblastic lymphadenopathy with dysproteinemia.

Angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) can best be described as a disorder of T-cells resulting in amplification of the B-cell response and clinical symptoms of lymphadenopathy, fever, hepatosplenomegaly, and a variety of blood abnormalities. Pure red cell aplasia (PRCA), an autoimmune disorder resulting in selective aplasia of the erythroid series, has only rarely been associated with AILD. Herein we report three cases of AILD and PRCA. Serum from one patient was available for study and contained a dose-dependent inhibitor of the CFU-E but not CFU-GM cultures from normal bone marrow. This activity was found in the globulin fraction after ammonium sulfate precipitation. Patients with AILD are known to make antibodies to many autologous epitopes, and the most well-characterized mechanism of PRCA involves antibodies to red cell precursors. Our serum data are consistent with the hypothesis that such an antibody existed in our patient. Aggressive treatment of these patients resulted in transient improvement in two; however, all three died without achieving a durable complete remission with two dying of infectious complications.

Aged↗
PubMed1994

Optimal primer selection for clonality assessment by polymerase chain reaction analysis: I. Low grade B-cell lymphoproliferative disorders of nonfollicular center cell type.

Recent polymerase chain reaction (PCR)-based studies focused on the detection of immunoglobulin heavy chain gene (IgH) rearrangements have suggested that clonal populations may be amplified more easily from certain categories of B-cell neoplasia than others and that primer makeup can be a critical factor in successful amplification. However, these particular reports contained relatively few low grade B-cell lymphoproliferative disorders of nonfollicular center cell type (LG-BLPD) and used only a limited panel of available primer sets for PCR amplification of monoclonal B-cell populations. To address this issue more extensively we evaluated 156 samples of LG-BLPD by the PCR to determine optimal primer selection in this setting. All cases were classified according to standard morphological and immunophenotypic criteria, with monoclonality documented by Ig light chain restriction analysis. The LG-BLPD included 33 cases of chronic lymphocytic leukemia (CLL), 57 cases of small lymphocytic lymphoma (SLL), 10 cases of atypical CLL, 32 cases of mantle cell lymphoma (MCL), 17 plasma cell neoplasms (PCNs), and seven cases of hairy cell leukemia (HCL). All primer sets included a 3' IgH joining region consensus primer, whereas the 5' IgH variable region (VH) primer was different in each set. The first-line panel included the following: Set 1, VH-framework III consensus primer, and Set 2, seven separate VH-framework I family-specific primers. A reserve panel of alternate VH consensus primers directed at framework II or III regions was used only when Set 1 showed no evidence of B-cell monoclonality.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed1994

Optimal primer selection for clonality assessment by polymerase chain reaction analysis: II. Follicular lymphomas.

Prior studies have shown variable success in amplification of monoclonal B-cell populations from follicular lymphomas (FLs) of germinal center cell origin using the polymerase chain reaction (PCR). We examined 60 FLs by the PCR to determine optimal primer selection for detection of clonal immunoglobulin heavy chain gene (IgH) rearrangements in this common group of B-cell lymphomas. Each primer set included a 3' IgH joining region consensus primer; the 5' primer was different in each reaction. The first-line panel included the following: Set 1, bcl-2 major breakpoint region (mbr) primer; Set 2, bcl-2 minor cluster region (mcr) primer; Set 3, IgH variable region framework III consensus primer; and Set 4, seven separate IgH variable region framework I family-specific primers. A reserve panel also was used. The efficiency of monoclonal B-cell population amplification differed among primer sets. The bcl-2-targeted primer pairs (Sets 1 and 2) were most efficient and amplified 42 (70%) of 60 cases. Of these, the mbr and mcr primer sets detected monoclonality in 38 and four cases, respectively. Set 3 amplified monoclonal B-cell in 38 and four cases, respectively. Set 3 amplified monoclonal B-cell populations in 31 (52%) cases and Set 4 detected similar populations in only 27 (45%) samples. When results from primer Sets 1, 2, and 3 were combined, 49 of 60 (82%) FLs showed evidence of B-cell monoclonality by the PCR. Three of the 11 negative cases were documented as monoclonal with primer Set 4, and three additional samples were amplified only with our reserve panel.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed1994

The clinical relevance of L26, a B-cell-specific antibody, in Hodgkin's disease.

Although Reed-Sternberg cells and their variants (RS/V) in most cases of Hodgkin's disease (HD) lack lymphocyte-associated antigens, some cases of HD, particularly nodular lymphocyte-predominant HD (NLPHD), contain RS/V cells that appear to be of B-cell origin. This study was designed to test whether differences in clinical presentation and therapeutic response exist between patients with HD of various histologic types whose tumors contain RS/V cells that express antigens recognized by L26, an antibody directed against B lymphocytes, and those that do not. We studied 46 patients with stages I and II HD who were treated with radiotherapy alone between 1969 and 1986. Although the expression of L26 and the other antibodies was not predictive for relapse-free, cause-specific, or absolute survival, the labeling of the RS/V cells with L26, independent of histologic type, was significantly associated with peripheral lymph nodal presentation, male gender, and infradiaphragmatic presentation.

Antigens, CD↗
PubMed1994

Primers frequently used for detecting the t(14;18) major breakpoint also amplify Epstein-Barr viral DNA.

We observed a potentially misinterpretable polymerase chain reaction (PCR) amplification product generated with standard primers used to detect the major breakpoint region (mbr) of chromosomal translocation t(14;18). This unexpected phenomenon was initially detected during attempts to transform follicular lymphomas in vitro with Epstein-Barr virus (EBV). Additional studies were performed using the EBV-producing cell line MCUV5, cell lines from EBV-transformed normal B-lymphocytes, and an excised lymph node from a patient with documented EBV-associated infectious mononucleosis. These samples consistently produced a 167-base pair product, which was indistinguishable from a t(14;18) lymphoma product when viewed on ethidium bromide-stained gels. Through DNA sequencing and gene bank analysis, the product was identified as a portion of the EBV genome. A mbr-specific 20-base oligonucleotide probe was able to discriminate between true translocations and the EBV-related amplifications. These results underscore the importance of employing a specific detection system, and comprehensively screening primers when working with PCR.

Animals↗
PubMed1994

Standard polymerase chain reaction analysis does not detect t(14;18) in reactive lymphoid hyperplasia.

There are conflicting data regarding the detection of t(14;18) in reactive lymphoid hyperplasia (RLH) by the polymerase chain reaction (PCR). Although most studies have not detected t(14;18), several groups have definitively shown that a very low number of cells with this translocation (one in 10(5) to 10(6)) are present in a significant proportion of follicular hyperplasias. Review of the methods from these series reveals that modifications of the PCR assay (ie, enhanced sensitivity steps such as seminesting, lengthy autoradiographic exposure times, multiple aliquot reactions of single samples, and/or high concentrations of template DNA) are probably necessary to detect t(14;18) in RLH. We evaluated a diverse set of 111 RLH (85 lymph nodes, 22 tonsils, and four other sites) from patients of different age groups (age range, 9 months to 80 years) to determine if a standard PCR assay would amplify t(14;18). Of these, 61 (55%) specimens had a prominent follicular hyperplastic component. Fifty-seven follicular lymphomas served as a control group. Polymerase chain reaction was performed as a single-run, two-primer-based assay for major breakpoint region bcl-2 translocations (5' major breakpoint region primer and 3' immunoglobulin heavy-chain gene-joining region consensus primer). Two different types of thermocyclers were employed. A metal block thermocycler was used with 35 cycles of amplification on 500 ng to 1 micrograms of genomic DNA, and a separate air thermocycler was used with 45 cycles of amplification on 50 ng of genomic DNA. Product detection was carried out through ethidium bromide staining and UV gel illumination, along with a digoxigenin-alkaline phosphatase-based, internal major breakpoint region oligonucleotide probe system. We found no amplified t(14;18) products in any RLH. In contrast, 36 (63%) of 57 follicular lymphomas showed t(14;18) (published range for detection of major breakpoint region translocations by PCR, 31% to 74%). Moreover, the assay's sensitivity, estimated through dilution studies, was to one in 10(4) to 10(5) cells. Although theoretically possible, our data suggest that there is practically no risk of amplifying a t(14;18) from RLH when utilizing a standard PCR assay.

Adolescent↗
PubMed1992

Reduced expression of CD20 antigen as a characteristic marker for chronic lymphocytic leukemia.

The surface antigens expressed by the cells of chronic lymphocytic leukemia (CLL) are well known. Most CLL are monoclonal B-cell lymphoproliferative disorders characterized by the coexpression of B-cell antigens and CD5, an antigen present predominantly on T cells. Very little attention, however, has been paid to the quantitative characteristics of the expression of B-cell antigens in CLL. In this study, we used flow cytometry to analyze the expression of CD20, a well-known B-cell-associated antigen, in lymphocytes from 42 cases of CLL and its tissue counterpart, small lymphocytic lymphoma (SLL), and compared the results with results obtained from the analysis of 21 follicular lymphomas, 20 hyperplastic reactive nodes, and 26 samples of normal peripheral blood. The intensity of CD20 expression in the CLL/SLL cells was significantly lower than that of B cells in the other categories. This antigen expression abnormality does not appear to be a universal phenomenon in CLL/SLL, since CD19, another pan-B antigen, was expressed in CLL/SLL at levels higher than those in follicular lymphomas and comparable to those in reactive lymph nodes. These results indicate that the low CD20 expression can be used as a marker for CLL/SLL. The few cases exhibiting intense CD20 expression may represent a biologically different disease. CLL/SLL cells faintly expressing CD20 also show concomitant low CD5 expression in a manner not observed in normal CD5-expressing B cells.

Adult↗
PubMed1992

Alterations of lymphocyte populations during development in the spontaneously hypertensive rat.

OBJECTIVE: Immune system abnormalities have been linked to hypertension in the spontaneously hypertensive rat (SHR). The goal of our study was to examine different lymphocyte subpopulations in the prehypertensive and developmental phases of hypertension in the SHR. DESIGN: Blood samples were obtained from SHR and Wistar-Kyoto (WKY) rats at the following time-points: 2 weeks and 1, 2, 3 and 4 months. Lymphocytes were separated from the whole blood. METHODS: Monoclonal antibodies were used to fluorescently label the following lymphocyte subpopulations; total T cells, T non-helper cells, T helper cells and B cells. Fluorescence-activated cell sorting (FACS) analysis was used to quantify the percentages of the different subpopulations examined. RESULTS: The T non-helper cell population was depressed in SHR from 2 weeks of age. This finding persisted throughout the entire 4-month study period. At the 4-month time-point, the total T cell percentage was also depressed in the SHR. CONCLUSIONS: These results demonstrate that immune system abnormalities are present in the prehypertensive and developmental phases of hypertension in the SHR. This supports the hypothesis that the immune system is involved in the development and maintenance of hypertension in the SHR, preceding not adapting to this state.

Animals↗
PubMed1991

Flow cytometric analysis of terminal deoxynucleotidyl transferase.

Terminal deoxynucleotidyl transferase (TdT) is a useful marker for lymphoid precursor cells. In this study, the authors used flow cytometry (FCM) to analyze TdT expression in human hematopoietic malignancies. Cells were fixed in 0.5% formaldehyde, briefly exposed to nonionic detergent, and subsequently labeled with mouse monoclonal anti-TdT antibodies followed by fluorescein-conjugated antimouse IgG and propidium iodide (PI), which was used for the simultaneous analysis of DNA content. Cells from 20 of 22 acute lymphoblastic leukemias (ALL), 4 of 7 mixed lineage leukemias, 2 of 21 acute myeloid and myelomonocytic leukemias, 1 of 2 chronic myeloid leukemias in blast crisis, 1 lymphoblastic lymphoma, and 1 thymoma were TdT positive. Cells from 13 nonlymphoblastic lymphomas, 3 myelodysplastic bone marrows, and peripheral blood mononuclear cells from 29 normal individuals were negative. An excellent correlation was seen between this assay, the conventional slide immunofluorescence technique, and an enzyme immunoassay method. The FCM assay detected as few as 2% blasts in mixing experiments of TdT-containing leukemic cells with normal peripheral lymphocytes. Furthermore, the combined analysis of TdT and DNA allowed the recognition of aneuploid TdT positive cells in four cases of ALL. The high sensitivity, the quantitative information obtained, and the capability of simultaneously analyzing DNA content make this method of TdT analysis more valuable than conventional techniques.

DNA Nucleotidylexotransferase↗
PubMed1989

High-grade B-cell lymphoma presenting as polyserosal disease. Diagnosis by flow cytometry.

Two patients presenting with anasarca were found to have aggressive B-cell lymphoma. No bulky disease was detected. The diagnosis was rapidly established by the flow cytometric analysis of cell surface immunophenotype and cell cycle fractions of pleural or peritoneal cells. Such presentation of lymphoma is unusual and previously undescribed, and it may have a significant negative prognostic impact. The authors' observations indicate that lymphoma be included in the differential diagnosis of anasarca and that flow cytometry can be useful for a fast confirmation of the diagnosis.

Aged↗
PubMed1989

Flow cytometric analysis of lymphomas.

Flow cytometry is rapidly developing as an important tool in the characterization of lymphomas. Analysis of cell surface antigens and DNA content in these tumors provides useful biologic information that can be applied advantageously to their diagnosis and classification. In this article we examine practical aspects of handling, preparation, and staining of lymphoid samples for flow cytometry and delineate approaches to data analysis and interpretation. We discuss specific applications, advantages, and limitations of the technique in the evaluation of neoplastic lymphoid diseases.

Evaluation Studies as Topic↗
PubMed1987

Correlation of monoclonal antibody phenotyping and cellular DNA content in non-Hodgkin's lymphoma. The Southeastern Cancer Study Group experience.

Flow cytometric measurements of DNA ploidy and synthetic (S) fractions are quantitative parameters that can aid in the diagnosis and classification of non-Hodgkin's lymphomas (NHL). Although the S-fraction correlates with histologic classification, the relationship between specific immunologic phenotypes and DNA ploidy is less well known. We investigated this relationship in 106 cases of NHL. Samples from 17 SEG institutions were sent for flow cytometry and for frozen section immunoperoxidase phenotyping. DNA histograms were analyzed for ploidy changes and cases classified by degree of abnormality. Ninety-eight cases were B-cell and eight were T-cell. B-cell tumors were subdivided by expression of antigens CD24, CD10, CD5, HB31, CD22, CD20, and transferrin receptor. Among B-cell tumors there was no correlation between degree of aneuploidy and phenotype, but B-cell tumors displayed a higher degree of aneuploidy than T-cell tumors (P less than 0.02). There was no difference between the S-fractions of B-cell and T-cell lymphomas. However, the transferrin receptor was more often expressed when the S-fraction was higher than 5%. Cases with S-fractions higher than 5% were more likely to lack any of the Pan-B antigens CD19, CD22 or CD20, and also were more frequently CD24 negative. We conclude that T-cell and B-cell NHL differ in degree of aneuploidy, and that monoclonal antibody phenotyping and DNA ploidy analysis independently define subgroups of B-cell NHL. Within B-cell lymphomas phenotype also correlates with grade of NHL as defined by the S-fraction.

Aneuploidy↗