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Biomedical subjects

R D Brunning

Publications and source records attributed to R D Brunning.

At least 19 recordsLinked to original sources
PubMed1992

Methylation status of the major breakpoint cluster region in Philadelphia chromosome negative leukemias.

It has been shown that a 600 bp long cluster of cell lineage specific hypomethylated sites in the major breakpoint cluster region (M-bcr) on chromosome 22 exists in hematopoietic cells. To determine possible relationships between methylation patterns within the M-bcr and the stage of hematopoietic cell development, the M-bcr methylation status of 39 patients with leukemia and lymphoma and two patients with myelodysplastic syndrome with non-rearranged M-bcrs was examined by BgIII-HpaII digestion. In the myeloid malignancies, the presence of a hypermethylated 4.8 kb BgIII-BgIII M-bcr allele was directly proportional to the combined myeloblast and promyelocyte percentage of the specimen, whereas the presence of a 2.5 kb BgIII-HpaII allele was directly proportional to the combined percentage of monocytic cells and neutrophils. All five acute monoblastic leukemias showed a methylation pattern that closely resembled neutrophils. All of thirteen surface immunoglobulin positive B-cell malignancies showed a distinct methylation pattern consisting of three or more BgIII-HpaII restriction fragments of 2.5 kb or less in length. The B-cell precursor leukemias showed heterogeneous M-bcr methylation patterns, with four of seven showing a B-cell pattern and three showing a hypermethylated pattern with 4.8, 3.1/3.0 and/or 2.5 kb BgIII-HpaII M-bcr alleles. It is concluded that the M-bcr methylation status is related to the maturation of the neutrophil series; the surface immunoglobulin positive B-cell malignancies are characterized by a distinct, extreme hypomethylation pattern of the M-bcr; and the B-cell precursor malignancies appear to have a heterogeneous M-bcr methylation pattern.

Acute Disease↗
PubMed1991

Identification of novel B-lineage cells in human fetal bone marrow that coexpress CD7.

In the present study we used multiparameter flow cytometry and cell sorting to evaluate fetal bone marrow, a rich source of cells early in lymphoid development. We found CD7 to be expressed on a subset of CD19+ cells, including some that had matured to cytoplasmic mu+ (pre-B) and surface mu+ (B) cells. In addition, a less mature CD7+19+ population was characterized as mu- and CD34+/-. The CD7+19+ population was clearly distinct from the mature T cells. The CD7+19+ cells were negative for nuclear TdT in contrast to CD7-19+ cells, which frequently contained TdT. CD10, which is coexpressed on the cell surface of more than 90% of CD19+ lymphocytes, was detected in a minority of CD7+19+ lymphocytes. The CD7+19+34+ cell population may be B-lineage committed, or may represent uncommitted lymphoid precursors. The biologic role of the expression of CD7 on immature and mature cells, including those of the B lineage, may indicate (1) the presence of CD7+19+ lymphoid precursor cells and/or (2) an alternate pathway of B-cell development, in which cells coexpress CD7 with other B-lineage markers.

Antigens, CD↗
PubMed1991

The morphologic and immunophenotypic assessment of the lymphocytosis accompanying Bordetella pertussis infection.

Bordetella pertussis (Bp) infection in infants and young children can be associated with a significant increase in small lymphocytes with convoluted and cleaved nuclei (SLCCN) in the peripheral blood (PB). Buffy coat smears were studied that were prepared from the PB of 11 children with documented Bp infection, whose ages ranged from one month to four years. The white blood cell count ranged from 8.4 to 72.9 X 10(9)/L, with a mean of 28.6 X 10(9)/L. In all cases, the percentage of PB lymphocytes was in the normal range; the absolute lymphocyte count ranged from 6.5 to 54.8 X 10(9)/L, with a mean of 20.3 X 10(9)/L. SLCCN represented 12-56% of the lymphocyte population. B and T lymphocytes, identified with monoclonal antibodies with the use of an immunoalkaline phosphatase method, accounted for a mean of 21% and 53%, respectively, of the total nucleated cells (TNCs) on buffy coat smears. The T-helper and T-suppressor subsets represented 38% and 16% of the TNCs, respectively, resulting in a CD4-CD8 ratio of 2.4. Most SLCCN were of the T-helper phenotype; SLCCN of the T-suppressor subset and, rarely, of the B-cell type also were identified. These observations document that the lymphocytosis associated with Bp infection in infants and young children is characterized by the presence of morphologically abnormal cells that are predominantly CD4 positive and appear to represent an expansion of an immunophenotypically normal lymphocyte population.

Antigens, CD↗
PubMed1991

Acute basophilic leukemia. A clinical, morphologic, and cytogenetic study of eight cases.

The authors describe eight cases of acute basophilic leukemia. In six of the eight cases, basophilic involvement was not apparent by light microscopic examination. The cases were identified on the basis of ultrastructural evidence for basophil/mast cell differentiation of the blasts with little or no differentiation into other lineages. Ultrastructural analysis revealed immature basophil granules in blasts in all eight cases and theta granules in blasts in four cases. In three cases, ultrastructural evidence of mast cell differentiation also was present, with rare cells showing evidence for both basophil and mast cell differentiation. No clinical features distinguished this group of patients from others with acute myeloid leukemia. Cytogenetically, the cases were heterogeneous. Three had a Philadelphia chromosome; none had a t(6;9). The authors conclude that ultrastructural analysis usually must be used to diagnose acute basophilic leukemia, that acute basophilic leukemia is associated frequently with the Philadelphia chromosome, and that the ultrastructural findings provide evidence for a common origin of basophils and mast cells.

Adolescent↗
PubMed1991

Transformation of chronic lymphocytic leukemia to small non-cleaved cell lymphoma: a cytogenetic, immunological, and molecular study.

A patient with chronic lymphocytic leukemia (CLL) transforming into a small non-cleaved cell lymphoma (SNCL) with the occurrence of a t(8;22) is described. The SNCL and the CLL were both found to have a germline lambda light chain gene configuration and the same heavy chain and kappa light chain gene rearrangements. The SNCL was CD10 (CALLA) negative and appeared to be CD5 negative. It is concluded that the SNCL is derived from the CLL and that activation of the c-myc oncogene may have played a role in this transformation.

Bone Marrow↗
PubMed1990

Presence of cell lineage-specific hypomethylated sites in the major breakpoint cluster region.

To examine the role of DNA methylation in breakpoint location of chromosomal translocation, HpaII sites in and flanking the M-bcr on chromosome 22 were mapped in DNA from blood granulocytes and lymphocytes, bone marrow cells, thymic tissue, and spermatozoa from normal individuals. Allelic HpaII sites were identified clustered in a 600-base pair genomic area of the M-bcr. Bone marrow cells and blood granulocyte DNA showed identical allelic patterns. Thymic tissue and blood lymphocytes showed identical allelic patterns distinct from bone marrow cells and blood granulocytes. Spermatozoa showed a third methylation pattern. In all individuals, the HpaII sites were present within the BamHI/BglII fragment of the M-bcr, the same area associated with high breakpoint frequency in chronic myelogenous leukemia (CML). Three of 15 patients with chronic phase CML showed fully methylated rearranged BglII/BglII M-bcr restriction fragments not seen in normal bone marrow cells. These methylation patterns of the M-bcr may be important in CML breakpoint location and may be a marker for tissue differentiation.

Blotting, Southern↗
PubMed1990

Oligoclonal T cell receptor gene rearrangements in blood lymphocytes of patients with acute Epstein-Barr virus-induced infectious mononucleosis.

Gene rearrangement studies were performed on blood lymphocytes from eight patients with acute Epstein-Barr virus-induced infectious mononucleosis. The diagnosis in each case was based on characteristic clinical, hematologic, and serologic findings. The blood lymphocytes in each patient consisted predominantly of CD8+ T cells. EBV DNA was detected in seven patients by Southern blot analysis (EBV Bam HI W probe, Bam HI). A germline configuration was found for the immunoglobulin heavy and light chain genes (JH probe, Bam HI and Eco RI; C kappa probe, Bam HI; and C lambda probe, Eco RI). T cell receptor gene rearrangements were detected with J gamma and J beta 1 + 2 probes. Using a J gamma probe with two different restriction enzymes (Bgl II and Eco RI), the blood from each patient showed several bands corresponding to the polyclonal pattern previously described in the blood of normal individuals. Using J beta 1 + 2 probes with two different restriction enzymes (Bgl II and Bam HI), each case showed from 3 to about 12 extragermline bands of varying intensity and in different locations from case to case. In addition, each case showed relative deletion of the J beta 1 germline band. This oligoclonal pattern of T cell receptor gene rearrangements has not been previously reported in benign or malignant T cell populations.

Acute Disease↗
PubMed1989

Refractory anemia with excess of blasts in transformation hematologic and clinical study of 52 patients.

This study consists of 52 patients whose blood and bone marrow findings fulfilled the French-American-British (FAB) Cooperative Group criteria for a diagnosis of refractory anemia with excess of blasts in transformation (RAEB-T). The basis for the diagnosis of RAEB-T included 37 patients (71%) meeting one criterion, 13 patients (25%) meeting two criteria, and two patients (4%) meeting three criteria; 44% met only the criterion of the detection of Auer rods. Clonal chromosome abnormalities were seen in seven of 16 cases studied and included abnormalities of chromosome 5, an extra 8, and complex abnormalities. Thirty-one patients were treated with a chemotherapeutic regimen that included an anthracycline and cytarabine; 13 of 17 patients (77%) 45 years of age or younger and six of 14 patients (43%) over 45 years of age achieved CR. The median survival for the 17 patients 45 years of age or younger is 25+ months; the median survival for the 14 patients over 45 years of age is 12.3 months. The median survival for 13 untreated patients over the age of 45 is 4.5 months. The process in 18 of the 52 patients (35%) evolved to overt acute myeloid leukemia (AML) in 1 to 38 months after diagnosis of RAEB-T, in seven of 20 patients (35%) 45 years of age or younger, and 11 of 32 patients (34%) over 45 years. Fifteen of the 18 patients received antileukemic therapy in the RAEB-T phase. There were no clinical or laboratory features that were reliably predictive of transformation to AML.

Adolescent↗
PubMed1989

Establishment of a leukemic cell model for studying human pre-B to B cell differentiation.

Reproducible models for examining early stages of human B cell differentiation are poorly developed. We now describe the establishment and characterization of a novel human leukemic cell line that recapitulates the pre-B to B cell stage of differentiation. This cell line, designated BLIN-1, was initially established in tissue culture medium containing low m.w. B cell growth factor, and consistently shows a dependency on this cytokine for optimal growth at low density. BLIN-1 cells have a 9p chromosomal abnormality, identical to the abnormality present in the leukemic blasts from the patient's original bone marrow aspirate. The immunologic phenotype of BLIN-1 is consistent with a cell arrested at the pre-B cell stage of development. Analysis of Ig gene rearrangement and Ig expression in a series of BLIN-1 subclones show that the cells spontaneously rearrange kappa light chain genes, leading to the differentiation of surface kappa-negative pre-B cells into surface kappa-positive B cells. The BLIN-1 cell line is, to our knowledge, the first defined human model for examining this critical developmental stage in human B cell ontogeny. As such, it offers a unique resource for examining variables influencing onset of kappa L chain gene rearrangement and expression.

B-Lymphocytes↗
PubMed1989

Immunohistochemical evaluation of neoplasms in bone marrow biopsies using monoclonal antibodies reactive in paraffin-embedded tissue.

A panel of commercially available monoclonal antibodies (MoAbs) including LN1, LN2, MB2, L26, Leu M1, UCHL1, MT1 and L60 was used to evaluate a diverse group of neoplastic processes in 256 Zenker's-fixed, decalcified, paraffin-embedded bone marrow biopsies using the ABC immunoperoxidase technique. LN2 and MB2 were useful in delineating the extent of B-cell lymphoproliferative processes and in identifying interstitial patterns of involvement. The combined application of LN2, MB2 and UCHL1 had utility in differentiating B-cell from T-cell lymphoproliferative processes; in no instance was reactivity with LN2 observed in T-cell processes. The combined application of these three MoAbs was also used in differentiating benign reactive lymphoid aggregates from focal malignant B-cell proliferations. LN2 exhibited positivity with the Reed-Sternberg cells (RSC) of Hodgkin's Disease (HD) and significantly aided in the identification of these cells. Staining of RSC with Leu M1 was inconsistent and was observed in only 50% of cases of HD. Use of the entire panel of MoAbs together with more recently available reagents such as Cathepsin G, MAC 387 and neutrophil elastase was essential in optimally evaluating a particular lesion; none of these MoAbs used singly reliably differentiated myeloid from lymphoid, hematopoietic from metastatic, or reactive from malignant processes.

Antibodies, Monoclonal↗
PubMed1988

Systemic polyclonal immunoblastic proliferations.

This report describes the clinical and pathologic features of four patients with a florid, systemic immunoblastic proliferation. The blood of these patients exhibited a mild to marked leukocytosis with a high percentage of immunoblasts and plasma cells. The bone marrow also was infiltrated extensively by immunoblasts. Lymph node biopsy specimens from two patients showed near total effacement of the nodal architecture by a diffuse infiltration of immunoblasts and plasma cells. The proliferative process was determined to be polyclonal with immunohistochemical techniques. Cytogenetic studies of bone marrow from two patients showed a pseudodiploid abnormal clone, with a translocation involving a break in band 14q32 in each case. The pathogenesis of these proliferative disorders in unclear, although three patients had some evidence of an acute immune disorder. One of these patients was treated with steroids, vincristine, and cyclophosphamide. Another patient was treated with steroids only, and one patient was treated with steroids and cyclophosphamide. All had rapid regression of the disease process. Two patients are alive and apparently free of disease 31 and 48 months after diagnosis. One died of sepsis. The fourth patient had acquired immune deficiency syndrome (AIDS) and died without therapy. The biology of the immunoblastic proliferation of these patients is uncertain. The immunohistochemical results suggest a reactive, polyclonal proliferation, but the cytogenetic abnormalities in two patients indicate the possibility of a cryptic neoplastic clone.

Acquired Immunodeficiency Syndrome↗
PubMed1988

Refined chromosome study helps define prognostic subgroups in most patients with primary myelodysplastic syndrome and acute myelogenous leukaemia.

Based on a 6 1/2-year study of 284 consecutive adult patients with primary myelodysplastic syndrome (MDS) and and acute myelogenous leukaemia (AML), we have found that refined chromosome analysis can be used as an independent prognostic indicator in the great majority of patients with MDS and AML. In MDS, the FAB subtype was also found to have prognostic value and this was enhanced when the chromosomal findings were taken into consideration. In AML, the age of the patient correlated more closely with the chromosomal changes in predicting prognosis in most patients than did the FAB classification. Previously we reported that refined chromosome analysis of bone marrow specimens from 161 adult patients with primary or non-therapy related MDS and AML identified three prognostic chromosomal categories in each disease, representing 40% of all patients (Yunis et al, 1984, 1986). By extending our study to 284 patients, as well as a longer follow-up, it was possible to determine the prognostic implications of two additional chromosomal categories in MDS and five in AML. Since 73% of all patients are now represented in well-defined chromosomal subgroups with prognostic significance, refined chromosome analysis emerges as a tool that could have considerable impact in protocols.

Adolescent↗
PubMed1987

Central nervous system involvement in acute nonlymphocytic leukemia. A prospective study of adults in remission.

To identify adults with acute nonlymphocytic leukemia at risk for the development of central nervous system involvement, we performed periodic cerebrospinal fluid examinations on patients in remission. Among 58 consecutive patients monitored during first remission, central nervous system leukemia developed in nine (16 percent). Four patients, including one who was symptomatic, had central nervous system leukemia detected simultaneously with marrow relapse. Five additional patients were asymptomatic and continue to have bone marrow remission. Following central nervous system and systemic treatment, two of these five patients have never had relapse, and three had relapse in the bone marrow five, 10, and 21 months later. Factors at diagnosis associated with the subsequent development of central nervous system leukemia were elevated leukocyte count, serum lysozyme and lactate dehydrogenase, extramedullary infiltration including splenomegaly, and monocytic (FAB M4 or M5a) morphology. In six of 17 patients (35 percent) with monocytic morphology, central nervous system leukemia developed compared with only three of 41 patients (7 percent) with other subtypes (p = 0.02). Discriminant analysis identified leukocyte count, splenomegaly, and M4 or M5a morphology as the most important risk factors and led to a mathematical formula that correctly identified 90 percent of the patients. Although the risk of central nervous system leukemia in adults with acute nonlymphocytic leukemia is too low to justify routine prophylaxis, those patients recognized to be at a greater risk should receive prophylaxis or be monitored closely with periodic lumbar punctures.

Acute Disease↗
PubMed1987

Hematologic parameters and leukocyte histogram patterns in infectious mononucleosis.

Blood samples from 60 patients with serologically positive infectious mononucleosis were analyzed on the Coulter S-Plus IV. Hemoglobins and platelet counts were essentially normal, and leukocyte counts were elevated in less than half the cases. Reactive lymphocytes were present on blood smears from all patients. The S-Plus IV automated three-part leukocyte differentials showed significantly lower numbers of lymphocytes and more mononuclears and granulocytes than manual microscopic differentials. On the leukocyte histograms, lymphocyte peaks were consistently wider than normal and asymmetrical. The instrument generated R2 flags on 43 of 60 (72%) of the samples. Using a combination of "R" flags, mononuclear backlighting, and "H" flags, the Coulter S-Plus IV identified 59 of 60 infectious mononucleosis samples as requiring further review.

Adolescent↗
PubMed1987

Immunophenotyping of acute myeloid leukemia using monoclonal antibodies and the alkaline phosphatase-antialkaline phosphatase technique.

The leukemic cells from 41 cases of acute myeloid leukemia (AML) and 17 cases of acute lymphocytic leukemia (ALL) were immunophenotyped by the alkaline phosphatase-antialkaline phosphatase (APAAP) immunocytochemical technique utilizing eight monoclonal antibodies (MoAb) reactive with cells of myeloid origin and seven MoAb reactive with lymphoid antigens. Ninety percent of the cases of AML reacted with one or more of the pan-myeloid MoAb, My7, My9, or 20.3. Reactivity of the myeloid panel of MoAb showed some correlation with the French-American-British (FAB) classification of AML. Five of six cases of acute promyelocytic leukemia (APL) were HLA-DR negative; the one HLA-DR-positive APL had a minor population of HLA-DR-negative promyelocytes. OKM5 and/or My4 reacted with 16 of 16 monocytic leukemias. No specific marker of early erythroid development was identified. AP3, a MoAb reactive with platelet glycoprotein (GPIIIa), was specific for acute megakaryoblastic leukemia. Immunocytochemistry was also helpful in classifying seven cases of AML with equivocal or negative routine cytochemistry. Two cases of AML had minor populations of blasts detected by the APAAP technique that were immunologically distinct from the major blast population; these minor populations emerged as the predominant cell type at relapse. Two cases of ALL expressed multiple myeloid and lymphoid antigens. Two other cases that morphologically were ALL reacted with only myeloid MoAb; one consisted entirely of immature basophils on ultrastructural examination. Immunophenotyping results using the APAAP technique were comparable with those obtained with flow cytometry. The APAAP technique is a reliable method for immunophenotyping leukemia that complements other methods of immunologic evaluation. The primary advantages of this method include its use with routinely prepared blood and bone marrow smears and the ability to correlate immunocytochemical reactions with morphology.

Alkaline Phosphatase↗