23 results on '"Wormsley SB"'
Search Results
2. Loxoribine induces chronic lymphocytic leukemia B cells to traverse the cell cycle
- Author
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Goodman, MG, primary, Wormsley, SB, additional, Spinosa, JC, additional, and Piro, LD, additional
- Published
- 1994
- Full Text
- View/download PDF
3. Characteristics of CD11c+CD5+ chronic B-cell leukemias and the identification of novel peripheral blood B-cell subsets with chronic lymphoid leukemia immunophenotypes [see comments]
- Author
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Wormsley, SB, primary, Baird, SM, additional, Gadol, N, additional, Rai, KR, additional, and Sobol, RE, additional
- Published
- 1990
- Full Text
- View/download PDF
4. Comparative density of the human T-cell antigen T65 on normal peripheral blood T cells and chronic lymphocytic leukemia cells
- Author
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Wormsley, SB, Collins, ML, and Royston, I
- Abstract
A 65,000 dalton T-cell specific antigen previously demonstrated to be present on the surface of normal and malignant T cells, but not normal B cells, has been detected on the surface of leukemic cells from patients wih nonsecretory, surface immunoglobulin-positive chronic lymphocytic leukemia (CLL). By means of immunofluorescence and flow cytometry, the relative surface density of the T65 antigen on CLL cells was compared to that on normal peripheral blood T cells and human thymocytes, as well as cell lines of T-cell lineage. In all cases, the CLL cells had a more homogeneous and a lower median fluorescence intensity than that of normal circulating T cells. Thymocytes were composed of three populations, two with low surface density of T65 resembling the CLL cells and the other with higher density similar to normal T cells. The staining of cell lines varied from bright, heterogeneous staining (8402) to uniform, low-density staining (Molt- 4). The implications of these findings with regard to lymphocyte differentiation are discussed.
- Published
- 1981
- Full Text
- View/download PDF
5. Murine monoclonal antibody therapy in two patients with chronic lymphocytic leukemia
- Author
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Dillman, RO, Shawler, DL, Sobol, RE, Collins, HA, Beauregard, JC, Wormsley, SB, and Royston, I
- Abstract
We infused the murine monoclonal antibody T101 into two patients with advanced refractory chronic lymphocytic leukemia (CLL) after confirming its reactivity with their CLL cells. One patient received doses of 1, 3, and 12 mg; the second patient received 10 mg. Antibody was delivered over 10--15 min. The major observations were: (1) T101 murine monoclonal antibody did bind to cells with T65 surface antigen and saturated these cells in vivo; (2) cells that bound T101 disappeared from the circulation by 2 hr after treatment, as evidenced by a marked drop in lymphocyte counts; (3) T101 serotherapy resulted in some intravascular cell injury associated with sequestration and probably destruction in the liver and lung; (4) free serum T101 was demonstrable, but disappeared by 2--4 hr after infusion; (5) rapid infusion of T101 did not induce significant modulation of T65; (6) rapid infusion of greater than 10 mg of T101 was associated with significant systemic reactions. Monoclonal antibodies may someday have an application in leukemia therapy, but additional experimental trials are clearly indicated.
- Published
- 1982
- Full Text
- View/download PDF
6. CD4(+)CD8(dim) T lymphocytes exhibit enhanced cytokine expression, proliferation and cytotoxic activity in response to HCMV and HIV-1 antigens.
- Author
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Suni MA, Ghanekar SA, Houck DW, Maecker HT, Wormsley SB, Picker LJ, Moss RB, and Maino VC
- Subjects
- Adult, Antigen Presentation, B-Lymphocytes immunology, CD4-Positive T-Lymphocytes cytology, Cell Division, Cells, Cultured, Coculture Techniques, Cytokines immunology, Dendritic Cells immunology, Flow Cytometry, HIV-1 immunology, Histocompatibility Antigens Class I immunology, Histocompatibility Antigens Class II immunology, Humans, Immunologic Memory, Immunophenotyping, Interferon-gamma biosynthesis, Interferon-gamma immunology, Lymphocyte Subsets cytology, Lymphocyte Subsets immunology, Lymphocyte Subsets metabolism, Middle Aged, T-Lymphocytes, Cytotoxic immunology, T-Lymphocytes, Cytotoxic metabolism, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes metabolism, Cytokines biosynthesis, Cytomegalovirus immunology, Cytotoxicity, Immunologic, HIV Antigens immunology, Lymphocyte Activation
- Abstract
CD4(+)CD8(dim) T cells represent a minor subset of the total CD3(+) T cell population in peripheral blood. Although transient and persistent expansions of these cells have been reported in both healthy and diseased individuals, the functional properties of the CD4(+)CD8(dim) population are largely unknown. In this study, we examined antigen-specific cytokine and proliferative responses of the CD4(+)CD8(dim) subset. In whole blood cultures stimulated with the viral antigens HCMV and HIV-1, a significant fraction of the CD4(+)CD8(dim) subset exhibited cytokine expression and proliferation in response to antigen activation. Typically, the CD4(+)CD8(dim) population contained two- to eightfold higher frequencies of antigen-specific cytokine producing cells than the CD4(+)CD8(-) population. Phenotypic analysis of the cytokine expressing CD4(+)CD8(dim) population indicated that these cells are memory T cells, with a high frequency of this population expressing the cytotoxic markers CD56 and perforin. Furthermore, the CD4(+)CD8(dim) cytokine responses to CMV were shown to be MHC class II dependent. Significantly, purified CD4(+)CD8(dim) T cells were found to possess higher CMV-specific cytotoxic activity than purified CD4(+)CD8(-) T cells in a standard (51)Cr-release CTL assay. Thus, CD4(+)CD8(dim) T cells appear to be MHC class II dependent, are capable of cytolytic effector activity, and are highly enriched within the CD4(+) cell populations specific for HCMV and HIV-1.
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- 2001
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7. Enhancement of HIV Type 1 Antigen-Specific CD4+ T Cell Memory in Subjects with Chronic HIV Type 1 Infection Receiving an HIV Type 1 Immunogen.
- Author
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Maino VC, Suni MA, Wormsley SB, Carlo DJ, Wallace MR, and Moss RB
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- 2000
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8. Validation and quality control of immunophenotyping in clinical flow cytometry.
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Owens MA, Vall HG, Hurley AA, and Wormsley SB
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- Cell Survival, Data Interpretation, Statistical, Flow Cytometry methods, Fluorescence, Humans, Immunophenotyping instrumentation, Immunophenotyping methods, Quality Control, Reproducibility of Results, Staining and Labeling, Flow Cytometry standards, Immunophenotyping standards, Pathology, Clinical standards
- Abstract
Clinical flow cytometry has evolved from two-parameter quantitative assessment of peripheral blood lymphocytes to six-parameter qualitative evaluation of bone marrow for hematopathology. Leukemia and lymphoma immunophenotyping represent an extremely important complement to morphology in the diagnosis and monitoring of hematopoietic malignancies. The complexity of five- and six-parameter analyses and the interpretation of the data rely on standardization and validation of the instrument, the reagents and the procedure. In addition, flow cytometry laboratories in the U.S. are required to document proficiency testing, sample preparation, method accuracy, specificity, sensitivity and precision. NCCLS and the U.S.-Canadian Consensus Conference have provided recommendations, but each laboratory is ultimately responsible for validating its own qualitative and quantitative procedures. This paper reviews procedures for validation and quality control of all aspects of the operation of a clinical flow cytometry service.
- Published
- 2000
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9. Multisite comparison of methods for the quantitation of the surface expression of CD38 on CD8(+) T lymphocytes. The ACTG Advanced Flow Cytometry Focus Group.
- Author
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Schmitz JL, Czerniewski MA, Edinger M, Plaeger S, Gelman R, Wilkening CL, Zawadzki JA, and Wormsley SB
- Subjects
- ADP-ribosyl Cyclase, ADP-ribosyl Cyclase 1, Antibodies, Monoclonal analysis, Antigens, CD immunology, HIV Infections blood, HIV Infections immunology, Humans, Laboratories standards, Laboratories statistics & numerical data, Membrane Glycoproteins, Specimen Handling, Statistics, Nonparametric, Time Factors, Antigens, CD analysis, Antigens, Differentiation analysis, CD8-Positive T-Lymphocytes immunology, Flow Cytometry methods, NAD+ Nucleosidase analysis
- Abstract
We evaluated the effect of specimen processing variations and quantitation methods on quantitative determination of CD38 expression on CD8 T lymphocytes. Neither lysing reagent (ammonium chloride versus BD FACSlyse), fixation (paraformaldehyde versus no final fixation step), nor acquisition delay (acquisition within 6 h after fixation versus 24 h after fixation) had a significant effect on CD38 relative fluorescent intensity or CD38 quantitative estimates (RFI or antibodies bound per cell). The only significant difference in fluorescent intensity and CD38 antibodies bound per cell (ABC) was encountered when whole blood was held for 24 h prior to staining and fixation and then acquired after another 24-h hold. However, for all sample processing methods above, the CD4 biologic calibrator and QuantiBRITE bead methods gave significantly different estimates of CD38 intensity. In many cases, however, these differences are relatively small and were more pronounced in certain laboratories. We conclude that there is some flexibility in sample processing methods for quantitative CD38 determination; however, it is preferable for a laboratory to employ one method of fluorescence quantitation calculation consistently because small differences are detected between different methods. Cytometry (Comm. Clin. Cytometry) 42:174-179, 2000., (Copyright 2000 Wiley-Liss, Inc.)
- Published
- 2000
10. Estimation of kinetic cell-cycle-related gene expression in G1 and G2 phases from immunofluorescence flow cytometry data.
- Author
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Jacobberger JW, Sramkoski RM, Wormsley SB, and Bolton WE
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- Computer Simulation, Cyclin B metabolism, Cyclin B1, G1 Phase physiology, G2 Phase physiology, Gene Expression, Humans, Kinetics, Male, Mitosis physiology, Prostatic Neoplasms metabolism, Tumor Cells, Cultured, Cell Cycle physiology, Flow Cytometry methods, Fluorescent Antibody Technique
- Abstract
Background: Flow cytometry of immunofluorescence and DNA content provides measures of cell-cycle-related gene expression (protein and/or epitope levels) for asynchronously growing cells. From these data, time-related expression through S phase can be directly measured. However, for G1, G2, and M phases, this information is unavailable. We present an objective method to model G1 and G2 kinetic expression from an estimate of a minimum biological unit of positive immunofluorescence derived from the distribution of specific immunofluorescence of mitotic cells., Methods: DU 145 cells were stained for DNA, cyclin B1, and a mitotic marker (p105) and analyzed by flow cytometry. The cyclin B1 immunofluorescence (B1) distribution of p105-positive cells was used to model the B1 distribution of G2 and G1 cells. The G1/S and S/G2 interface measurements were used to calculate expression in S phase and test the validity of the approach., Results: B1 at S/G2 closely matched the earliest modeled estimate of B1 in G2. B1 increased linearly through G1 and S but exponentially through G2; mitotic levels were equivalent to the highest G2 levels. G1 modeling of B1 was less certain than that of G2 due to low levels of expression but demonstrated general feasibility., Conclusions: By this method, the upper and lower bounds of cyclin B1 expression could be estimated and kinetic expression through G1, G2, and M modeled. Together with direct measurements in S phase, expression of B1 throughout the entire cell cycle of DU 145 cells could be modeled. The method should be generally applicable given model-specific assumptions.
- Published
- 1999
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11. Age-associated changes in binding of human B lymphocytes to a VH3-restricted unconventional bacterial antigen.
- Author
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Silverman GJ, Sasano M, and Wormsley SB
- Subjects
- Adult, Antigens, CD analysis, CD5 Antigens, Child, Child, Preschool, Gene Rearrangement, Humans, Immunoglobulin G immunology, Immunoglobulin Heavy Chains genetics, Immunoglobulin M immunology, Immunoglobulin Variable Region genetics, Infant, Infant, Newborn, Leukocytes, Mononuclear immunology, Middle Aged, Aging immunology, B-Lymphocytes immunology, Immunoglobulin Fab Fragments immunology, Immunoglobulin Heavy Chains immunology, Immunoglobulin Variable Region immunology, Staphylococcal Protein A immunology
- Abstract
We have recently demonstrated that there is a site on Staphylococcal protein A (SpA) that interacts with B cell Ig receptors in a manner comparable with known T cell superantigens, because this binding specificity is restricted to Fab with VH3 H chains and most VH3 Ig bind SpA. In the present studies, SpA was used as a phenotypic marker for VH3 expression by human lymphoid cells. As expected, this Fab-mediated binding specificity was completely inhibited by certain VH3 antibodies but not by antibodies from other VH families. In multiparameter flow cytometric analyses, this binding activity was demonstrated to be highly prevalent among B cells (14 to 54%), and was more common among IgM-bearing B cells compared with IgG-bearing B cells. In all studies, Fab-mediated binding of SpA was uniformly expressed by a greater proportion of CD5-positive B cells than CD5-negative B cells. The proportion of B lymphocytes with this VH3-restricted binding capacity was found to undergo age-associated changes, because a large proportion of the peripheral B cells of neonates (mean +/- SD, 46.0 +/- 2.9%) bind this site, but two 10-mo-old subjects and older children had significantly lower binding levels (29.0 +/- 3.5%) that were the same as binding levels by adult peripheral B lymphocytes (30.2 +/- 3.3%). In immunohistochemical studies, tonsilar B cells that bind this site on SpA were shown to be common in mantle zones and germinal centers of secondary follicles. We speculate that Fab-mediated SpA binding represents a fundamental and primitive binding capacity that is part of the human preimmune repertoire, and we discuss the implications for the observed age-dependent shift in Fab-mediated binding of SpA by peripheral blood B cells.
- Published
- 1993
12. A chronic illness characterized by fatigue, neurologic and immunologic disorders, and active human herpesvirus type 6 infection.
- Author
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Buchwald D, Cheney PR, Peterson DL, Henry B, Wormsley SB, Geiger A, Ablashi DV, Salahuddin SZ, Saxinger C, Biddle R, Kikinis R, Jolesz FA, Folks T, Balachandran N, Peter JB, Gallo RC, and Komaroff AL
- Subjects
- Adult, Antibodies, Viral blood, Brain pathology, CD4-CD8 Ratio, Cluster Analysis, Female, Herpesviridae Infections cerebrospinal fluid, Herpesviridae Infections immunology, Herpesviridae Infections physiopathology, Human T-lymphotropic virus 1 immunology, Humans, Immunophenotyping, Magnetic Resonance Imaging, Male, Middle Aged, Virus Replication, Fatigue Syndrome, Chronic microbiology, Herpesviridae Infections complications, Herpesvirus 6, Human immunology, Herpesvirus 6, Human physiology
- Abstract
Objective: To conduct neurologic, immunologic, and virologic studies in patients with a chronic debilitating illness of acute onset., Design: Cohort study with comparison to matched, healthy control subjects., Patients: We studied 259 patients who sought care in one medical practice; 29% of the patients were regularly bedridden or shut-in., Main Outcome Measures: Detailed medical history, physical examination, conventional hematologic and chemistry testing, magnetic resonance imaging (MRI) studies, lymphocyte phenotyping studies, and assays for active infection of patients' lymphocytes with human herpesvirus type 6 (HHV-6)., Main Results: Patients had a higher mean (+/- SD) CD4/CD8 T-cell ratio than matched healthy controls (3.16 +/- 1.5 compared with 2.3 +/- 1.0, respectively; P less than 0.003). Magnetic resonance scans of the brain showed punctate, subcortical areas of high signal intensity consistent with edema or demyelination in 78% of patients (95% CI, 72% to 86%) and in 21% of controls (CI, 11% to 36%) (P less than 10(-9)). Primary cell culture of lymphocytes showed active replication of HHV-6 in 79 of 113 patients (70%; CI, 61% to 78%) and in 8 of 40 controls (20%; CI, 9% to 36%) (P less than 10(-8], a finding confirmed by assays using monoclonal antibodies specific for HHV-6 proteins and by polymerase chain reaction assays specific for HHV-6 DNA., Conclusions: Neurologic symptoms, MRI findings, and lymphocyte phenotyping studies suggest that the patients may have been experiencing a chronic, immunologically mediated inflammatory process of the central nervous system. The active replication of HHV-6 most likely represents reactivation of latent infection, perhaps due to immunologic dysfunction. Our study did not directly address whether HHV-6, a lymphotropic and gliotropic virus, plays a role in producing the symptoms or the immunologic and neurologic dysfunction seen in this illness. Whether the findings in our patients, who came from a relatively small geographic area, will be generalizable to other patients with a similar syndrome remains to be seen.
- Published
- 1992
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13. Detection of occult lymphoma cells in bone marrow aspirates by multi-dimensional flow cytometry.
- Author
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Stelzer GT, Shults KE, Wormsley SB, and Loken MR
- Subjects
- Bone Marrow immunology, Flow Cytometry statistics & numerical data, Humans, Leukocyte Common Antigens, Sensitivity and Specificity, Transplantation, Autologous, Bone Marrow pathology, Bone Marrow Transplantation, Lymphoma, B-Cell pathology, Lymphoma, B-Cell surgery
- Published
- 1992
14. Expression of low-, intermediate-, and high-affinity IL-2 receptors on B cell lines derived from patients with undifferentiated lymphoma of Burkitt's and non-Burkitt's types.
- Author
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Benjamin D, Rosolen A, Wormsley SB, DeBault LE, and Colamonici OR
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- Binding, Competitive, Cell Line, Cell Line, Transformed, Cross-Linking Reagents, Flow Cytometry, Humans, Interleukin-2 metabolism, Iodine Radioisotopes, Kinetics, Tumor Cells, Cultured, B-Lymphocytes metabolism, Burkitt Lymphoma immunology, Lymphoma immunology, Receptors, Interleukin-2 biosynthesis
- Abstract
IL-2 receptors on T cells exist in at least three forms which differ in their ligand-binding affinity. The low-affinity IL-2 receptor (IL-2R) consists of the 55-kDa Tac protein (p55 alpha), the intermediate-affinity site corresponds to the 70-kDa molecule (p70 beta), and the high-affinity IL-2R consists of a noncovalent heterodimeric structure involving both p55 alpha and p70 beta. We studied 24 B cell lines (8 EBV-negative and 16 EBV-positive) for IL-2R expression in the presence or absence of the tumor promoter, teleocidin. 125I-IL-2 radioreceptor binding assays and crosslinking studies demonstrated the sole expression of p55 alpha in EBV-negative cell lines only, whereas p55 alpha present in EBV-positive cell lines was always associated with p70 beta to construct high-affinity IL-2R. p70 beta was not detected in any of the EBV-negative cell lines, but was expressed on most of the EBV-positive cell lines (13 of 16). Our data also indicate that the expression of p55 alpha and p70 beta by radiolabeling correlates with their expression in flow cytometry, and that a large excess of p55 alpha is required to construct high-affinity IL-2R. Coexpression of p55 alpha and p70 beta on human B cells contributed to constructing high-affinity IL-2R hybrid complex as shown by (i) rapid association rate contributed by p55 alpha and slow dissociation rate by p70 beta; (ii) teleocidin's ability to induce p55 alpha on cell lines which express p70 beta only, resulting in appearance of high-affinity IL-2R; (iii) blocking p55 alpha by anti-Tac mAb in cell lines which constitutively express high-affinity IL-2R eliminated both high- and low-affinity components. The existence of low, intermediate, and high IL-2R on human B cells bears important future implications for understanding the mechanism of IL-2 signaling and the role of IL-2 in B cell activation, proliferation, and differentiation.
- Published
- 1990
- Full Text
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15. B-cell surface phenotypes of proliferating myeloma cells: target antigens for immunotherapy.
- Author
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Chan CS, Wormsley SB, Pierce LE, Peter JB, and Schechter GP
- Subjects
- B-Lymphocytes pathology, Cell Division, Cell Line, Humans, Interphase, Phenotype, Ploidies, Antigens, Differentiation, B-Lymphocyte analysis, B-Lymphocytes immunology, Multiple Myeloma pathology
- Abstract
Dual-parameter flow cytometric analysis of B-cell antigens and DNA content was used to determine the phenotypes of proliferating tumor cells (S-phase cells) from 30 patients with multiple myeloma. B4 (CD19), J5 (CALLA, CD10), B1 (CD20), and monotypic surface immunoglobulin (Slg) were expressed heterogeneously in 24 patients. J5 and monotypic Slg were found most frequently but were always expressed on a significantly lower percentage of cells than the antigens typically associated with plasma cells, cytoplasmic immunoglobulin (Clg) and T10 (CD38). S-phase cells were found in each antigen(+) subset. B antigen(+) cycling cells were demonstrated in 16 patients whose marrow or blood cells expressed B antigens exclusively in the hyperdiploid fraction and therefore were certainly part of the myeloma clone. Similar to the low level of proliferative activity of the T10(+), Clg(+), and PCA1(+) subsets, the percentages of cycling cells of the preplasma cell B-antigen-bearing myeloma subsets ranged from less than 1% to 12%. The tumor cells of four patients were also studied with dual-color surface antigen analysis and demonstrated independent expression of B antigens, with only rare coexpression of T10 and monotypic Slg, J5, or B4. These findings are consistent with the presence of distinct myeloma subsets bearing differing B phenotypes in the same tumor and provide evidence that the proliferation in myeloma is occurring at various developmental stages in the malignant B lineage. These antigens may be important targets for immunologic therapy aimed at eliminating the entire proliferating compartment of this B-cell tumor.
- Published
- 1990
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16. The use of monoclonal antibodies and flow cytometry to detect peripheral blood and bone marrow involvement of a diffuse, poorly differentiated lymphoma.
- Author
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Shawler DL, Wormsley SB, Dillman RO, Frisman DM, Baird SM, Glassy MC, and Royston I
- Subjects
- Flow Cytometry, Fluorescent Antibody Technique, Humans, Immunoglobulin G analysis, Lymphoma blood, Male, Middle Aged, T-Lymphocytes immunology, Time Factors, Antibodies, Monoclonal, Bone Marrow pathology, Lymphoma pathology
- Abstract
Using monoclonal antibodies and flow cytometry, we were able to characterize the phenotype of a diffuse, poorly differentiated lymphoma and to isolate subpopulations of cells from the blood and bone marrow that expressed the malignant phenotype even though the patient exhibited no absolute lymphocytosis. Because the circulating clone reacted with the anti-T cell monoclonal antibody T101, we initiated serotherapy with T101 as part of a phase I study. A 10 mg infusion of T101 resulted in the rapid clearance of normal T cells from circulation, but the clone showed evidence of modulation and was not cleared. Twenty-four hours following infusion, all cell populations had returned to pre-treatment levels. Our study suggests that, by using monoclonal antibodies and flow cytometry, blood and bone marrow involvement of a lymphoma can be demonstrated in patients without absolute lymphocytosis, a finding which may influence the staging and treatment of the disease.
- Published
- 1985
- Full Text
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17. Cytokines and immune regulation in patients with fibrositis.
- Author
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Wallace DJ, Bowman RL, Wormsley SB, and Peter JB
- Subjects
- Antigen-Antibody Complex analysis, Antigens, Surface analysis, Cytokines, Humans, Immunoglobulin G analysis, Lymphocyte Activation, Lymphocytes immunology, Surveys and Questionnaires, Biological Factors physiology, Fibromyalgia immunology
- Published
- 1989
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18. Dual parameter analysis of myeloma cells by flow cytometry. DNA content of cells containing monotypic cytoplasmic immunoglobulin.
- Author
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Chan CS, Wormsley SB, Peter JB, and Schechter GP
- Subjects
- Flow Cytometry methods, Humans, Multiple Myeloma immunology, Multiple Myeloma pathology, Ploidies, Cytoplasm analysis, DNA, Neoplasm analysis, Immunoglobulins analysis, Multiple Myeloma analysis
- Abstract
Dual-parameter flow cytometric analysis of monotypic cytoplasmic immunoglobulin (CIg) and DNA content on 15 myeloma marrows allowed S-phase determination of the CIg(+) tumor separately from the CIg(-) hematopoietic cell pool. The median percentage of the CIg(+) cells in S-phase was 2% compared with 5% for the CIg(-) cells. The median survival of patients with more than 2%, and those with 2% or less CIg(+) S-phase cells was 2 months and more than 13 months, respectively, from the time of study. Ploidy analysis identified four patterns of plasma cell DNA content: entirely hyperdiploid, entirely diploid, combined diploid and tetraploid stemlines, and tumors containing diploid and aneuploid CIg(+) cells. A monotypic CIg(+) double stemline myeloma was distinguished from two aneuploid tumors containing admixed normal, diploid polyclonal plasma cells. This technique provides an improved and expedient means for determining the proliferating fractions of myeloma cells and enhances recognition of double stemline tumors and clonal evolution in myeloma.
- Published
- 1989
- Full Text
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19. Membrane antigen on Epstein--Barr virus-infected human B cells recognized by a monoclonal antibody.
- Author
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Slovin SF, Frisman DM, Tsoukas CD, Royston I, Baird SM, Wormsley SB, Carson DA, and Vaughan JH
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- Animals, Cell Line, Cell Membrane immunology, Female, Genes, Viral, Herpesvirus 4, Human genetics, Humans, Hybridomas immunology, Mice, Mice, Inbred BALB C, Plasmacytoma, T-Lymphocytes immunology, Antibodies, Monoclonal, Antigens, Surface analysis, B-Lymphocytes immunology, Cell Transformation, Viral, Herpesvirus 4, Human immunology
- Abstract
This paper describes a monoclonal antibody (B532) that detects a membrane antigen present on greater than or equal to 95% of the B cells from lines carrying the Epstein-Barr virus (EBV) genome. Evidence suggesting that B532 is EBV-related was originally obtained by using a cell-binding radioassay with different cell line substrates. Immunofluorescence and cell-sorter analysis confirmed that the antigen was present in high density on all EBV-infected lymphoblastoid B-cell lines, but not on EBV-negative B-, T-, myeloid, or null cell lines. Isolated normal peripheral blood B and T lymphocytes and monocytes failed to bind B532. The monoclonal antibody did not inhibit in vitro EVB infection nor did it block the killing of EBV-infected targets by cytotoxic T lymphocytes. The cell surface antigen recognized by B532 was shown by immunoprecipitation to have a molecular weight of approximately 45,000.
- Published
- 1982
- Full Text
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20. Induction of in vitro and in vivo antigenic modulation by the anti-human T-cell monoclonal antibody T101.
- Author
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Shawler DL, Miceli MC, Wormsley SB, Royston I, and Dillman RO
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- Cells, Cultured, Humans, Immunity, Cellular, Immunotherapy, Kinetics, Leukemia, Lymphoid immunology, Lymphoma immunology, Skin Neoplasms immunology, Temperature, Antibodies, Monoclonal administration & dosage, Antigens, Neoplasm analysis, Antigens, Surface analysis, Leukemia, Lymphoid therapy, Lymphoma therapy, Skin Neoplasms therapy, T-Lymphocytes immunology
- Abstract
Because of its implications for the therapeutic application of monoclonal antibodies, we have studied antigenic modulation in vitro and in vivo in patients receiving T101 monoclonal antibody. Incubation of normal peripheral blood T-cells, chronic lymphocytic leukemia cells, and cutaneous T-cell lymphoma cells with an excess of T101 at 37 degrees induced modulation of the T65 antigen. When assayed by indirect immunofluorescence, a change in cellular reactivity with T101 was seen after 1 hr. After 24 hr, normal T-cells showed a 94 +/- 4% (S.D.) decrease in fluorescence, compared to an 82 +/- 6% decrease for chronic lymphocytic leukemia cells and a 56 +/- 4% decrease for cutaneous T-cell lymphoma cells. When T101 was removed from the culture, the cells reexpressed T65. Modulation was inhibited by cold temperatures, suggesting that it is energy dependent. Patients with chronic lymphocytic leukemia, cutaneous T-cell lymphoma, or T-cell lymphoma have received 24-hr infusions of 3 to 500 mg T101 in therapeutic trials. After infusion, in vivo binding of T101 was observed in 39 of 43 treatments not associated with endogenous host anti-T101 antibodies. T65-target cells were seen in all 39 treatments associated with in vivo bound T101, suggesting that modulation had occurred. When cultured in vitro for 24 hr, these cells reexpressed T65. In vivo, reexpression of T65 occurred following disappearance of the serum T101 titer. The extent and duration of in vivo modulation were related to both the T101 dose and the tumor burden. These data suggest that the rapid rate of antigenic modulation may prevent potential target cell destruction by antibody-mediated cytotoxicity. However, if the process of modulation involves internalization of the antibody:antigen complex, it would be an advantage for the use of cytotoxic immunoconjugates.
- Published
- 1984
21. Analysis of treatment response in chronic lymphocytic leukemia: new approaches.
- Author
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Wormsley SB, Rai KR, and Gale RP
- Subjects
- Biomarkers, Tumor analysis, Clone Cells classification, Clone Cells drug effects, Clone Cells pathology, Gene Rearrangement, B-Lymphocyte, Heavy Chain, Gene Rearrangement, B-Lymphocyte, Light Chain, Humans, Leukemia, Lymphocytic, Chronic, B-Cell classification, Leukemia, Lymphocytic, Chronic, B-Cell genetics, Neoplastic Stem Cells classification, Neoplastic Stem Cells drug effects, Neoplastic Stem Cells pathology, Phenotype, Leukemia, Lymphocytic, Chronic, B-Cell drug therapy
- Abstract
As therapeutic protocols to treat chronic lymphocytic leukemia evolve, new criteria will be required to define remission and identify early relapse. Two techniques that detect low numbers of residual monoclonal B-lymphocytes currently include immunoglobulin (Ig) gene rearrangement and clonal excess analysis. Ig-gene rearrangement studies are time consuming and expensive. Clonal excess analysis by FC may be insensitive because of minimal surface immunoglobulin (SIg) expression. Three antigens, CD5, CD11c and CD14 are expressed on B-lymphocytes of CLL. CD14 is also expressed on most normal B-lymphocytes. In contrast, CD5 and CD11c are expressed on few normal B-lymphocytes. Consequently CD5 and CD11c are useful markers for detecting residual CLL cells. Appropriate selection of the second B-cell marker such as SIg, CD19, CD20 or other, is critical. We evaluated this approach in persons with CLL receiving therapy. Preliminary results will be presented.
- Published
- 1988
22. A comparison of hepatocyte size distribution in untreated and phenobarbital-treated rats as assessed by flow cytometry.
- Author
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Willson RA, Wormsley SB, and Muller-Eberhard U
- Subjects
- Animals, Light, Liver cytology, Male, Rats, Rats, Inbred Strains, Scattering, Radiation, Flow Cytometry, Liver drug effects, Phenobarbital pharmacology
- Abstract
Flow cytometric analysis of narrow forward angle light scatter was used to examine the effect of phenobarbital treatment on hepatocyte size. Light scatter analysis of freshly isolated hepatocytes and of hepatocytes separated by means of centrifugal elutriation into five subpopulations (fractions 1-5) was performed on both untreated and phenobarbital-treated rats. The frequency distribution histogram of forward scatter intensity produced by freshly isolated hepatocytes from untreated rats was used as the baseline. This histogram was arbitrarily divided into four regions; referred to as region 1 (small cells) to region 4 (large cells). Subsequent analysis of the light scatter histograms derived from elutriated fractions of both untreated and phenobarbital-treated animals was performed using these baseline regions. Analysis showed that small cells were enriched in elutriated fraction 1, and large cells in elutriated fraction 5. Phenobarbital treatment was associated with a uniform shift to a higher intensity light scatter (relative increase in cell size) within each of the four selected regions. Our findings suggest that phenobarbital does not have a selective hypertrophic effect on these subpopulations of rat hepatocytes.
- Published
- 1984
- Full Text
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23. Alterations in cell surface phenotype of T- and B-cell chronic lymphocytic leukemia cells following in vitro differentiation by phorbol ester.
- Author
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Shawler DL, Glassy MC, Wormsley SB, and Royston I
- Subjects
- Cell Differentiation drug effects, Histocompatibility Antigens Class II analysis, Humans, Immunoglobulins analysis, Leukemia, Lymphoid pathology, Phenotype, Rosette Formation, Antigens, Surface analysis, B-Lymphocytes immunology, Leukemia, Lymphoid immunology, Phorbols pharmacology, T-Lymphocytes immunology, Tetradecanoylphorbol Acetate pharmacology
- Abstract
The phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) was used for the induction of in vitro differentiation in primary cultures of chronic lymphocytic leukemia cells to study its effects on B-cell antigens [surface IgG, HLA-DR, and the mouse erythrocyte receptor (MR)] and on T-cell antigens [T65 and Lyt-3 (sheep erythrocyte receptor)] found on these cells. Three distinct phenotypes were studied: 1) the common phenotype (slg+, HLA-DR+, MR+, T65+, Lyt-3-); 2) the T-cell phenotype (slg-, HLA-DR-, MR-, T65+, Lyt-3+); and 3) a unique phenotype (slg-, HLA-DR+, MR+, T65+, Lyt-3-). In both the common and unique phenotypes, TPA increased the expression of T65 and HLA-DR, decreased the formation of MR, and induced cytoplasmic immunoglobulin, but it did not induce Lyt-3. In the unique phenotype, TPA also induced surface immunoglobulin. These data suggest that both the common and unique phenotypes are derived from the same lineage, probably B-cell. In the T-cell phenotype, TPA increased the expression of T65 and Lyt-3, but it did not induce any B-cell antigens. These data suggest that the T-cell phenotype is derived from a T-cell lineage distinct from the two B-cell phenotypes.
- Published
- 1984
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