scholarly journals Combined method of PAP immunocytochemistry and autoradiography: application to cell kinetic study in plasma cell dyscrasias.

1987 ◽  
Vol 35 (10) ◽  
pp. 1157-1160
Author(s):  
T Kanoh ◽  
T Ohnaka ◽  
H Uchino
Keyword(s):  
Plasma Cell ◽  
Plasma Cells ◽  
Cell Labeling ◽  
Combined Method ◽  
Cell Kinetic ◽  
Diagnosis And Prognosis ◽  
Plasma Cell Dyscrasias ◽  
Pap Method ◽  
Combined Technique

The plasma cell labeling index (LI), in spite of being a reliable indicator for diagnosis and prognosis of multiple myeloma, has been measured in a limited number of laboratories because of technical difficulties. We have developed a new combined technique, using the peroxidase-antiperoxidase (PAP) method and autoradiography, which has several advantages over previously described methods. The primary advantages of our method in the determination of lymphoid-plasma cell LI% are: (a) no damage to slides during storage of more than 1 year; (b) an exact LI measurement in each morphological variety of pleomorphic immunoglobulin-containing cells; (c) no problem in differentiation of lymphoid plasma cells from early red cell precursors; and (d) a separate LI measurement for those lymphoid-plasma cells composed chiefly, if not exclusively, of monoclonal or neoplastic cells. Because of these advantages, this accurate and less difficult technique will facilitate performance of lymphoid plasma cell LI in a number of laboratories.

Improved method for measurement of inorganic phosphate in serum with a centrifugal analyzer.

1976 ◽  
Vol 22 (2) ◽  
pp. 257-260 ◽  
Author(s):  
P W Wentz ◽  
J Savory ◽  
R E Cross
Keyword(s):  
Plasma Cell ◽  
Data Reduction ◽  
Continuous Flow ◽  
Inorganic Phosphate ◽  
High Speed ◽  
Improved Method ◽  
Plasma Cell Dyscrasias

Abstract A direct mehtod [Clin. Chim. Acta 46, 113 (1973)] for determination of inorganic phosphate in serum was adapted for use with a centrifugal analyzer. Contamination is minimized and analysis rate maximized by doing the reaction in the reagent wells of the transfer disc and by utilizing the high-speed spectrophotometric and data-reduction capabilities of the centrifugal analyzer. Hemolysis, icterus, and moderate lipemia cause no interference. Grossly lipemic sera and sera from patients with plasma cell dyscrasias can be analyzed by incorporating appropriate blanking and dilution techniques. The method exhibits excellent sensitivity and precision and results correlate well with those from a continuous-flow procedure.

Myeloma with t(11;14) and CD20+ Plasma Cells: Response to Rituximab.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5178-5178
Author(s):  
Patricia T. Greipp ◽  
Rafael Fonseca ◽  
Dewald Dewald ◽  
S. Vincent Rajkumar ◽  
William G. Morice ◽  
...  
Keyword(s):  
Cyclin D1 ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Early Stage ◽  
Strong Association ◽  
Cell Labeling ◽  
Response To Treatment ◽  
History Of ◽  
M Spike ◽  
Anti Cd20

Abstract Background: Multiple myeloma (MM) is a plasma cell malignancy involving complex cytogenetic dysregulation of genes and heterogeneous expression of multiple B cell and plasma cell (PC) surface antigens. 60% of MM patients have cytogenetic translocations of the IGH locus. In 16% of myeloma patients, the partner oncogene is cyclin D1 on 11q13 causing t(11;14)(q13;q32) (Fonseca et al. Blood. 2002). This translocation is more common in patients with a lymphoplasmacytic morphology and confers a more favorable prognosis (Fonseca et al. Blood. 2003). A subset of MM patients (20%) harbor a clonotypic population of CD20+ bone marrow plasma cells (BMPC). These patients have a shortened survival time (SanMiguel et al. Br J Haematol. 1991). There is a strong association with CD20+ surface expression and t(11;14)(Robillard et al. Blood. 2003) and CD20+ PC has been suggested to be clonogenic and important in the pathogenesis of MM. A small Phase II study showed only a modest response to the humanized anti-CD20 monoclonal antibody rituximab in MM patients with CD20+PC (1 PR and 5SD) (Treon et al. J. Immunother. 2002). We report a response to treatment with rituximab in a MM patient with CD20+ BMPC and t(11;14)(q13;q32) by FISH. Case: The patient is a 68 year old male with a 1.5 year history of MM. Initial diagnosis of ISS stage I disease was made after a three year history of IgA kappa monoclonal gammopathy of undetermined significance (MGUS). At diagnosis of MM, he had symptomatic anemia Hgb10.5g/dL, rising serum IgA of 3950 mg/dL, serum m-spike of 4.1 g/dL (IgA kappa), PCLI (plasma cell labeling index) of 1%, and a small monoclonal IgA kappa with kappa fragment in the urine. BM showed 44% monoclonal kappa plasma cells with 90% CD20+,clonal PC on flow cytometry. Karyotype was normal; 46, XY, but FISH showed evidence of an IGH translocation t(11;14)(q13;q32) in 60% of PC. Immunohistochemical staining was positive for cyclin D1. Given the marked elevation of CD20+ PC and early stage of disease, initial treatment was employed with four weekly cycles of rituximab (375mg/m2). IgA fell from 4750 mg/dL to a nadir of 2990 mg/dL, (m-spike; 4.5g/dL to 3.2 g/dL) five weeks after completion of treatment. The monoclonal protein (MP) in the urine disappeared. Hgb improved to 12.5g, with erythropoietin. Three months later, IgA rose to 3260mg/dL (m-spike 3.2 g/dL) and urinary MP returned. The patient remained asymptomatic. A second course of weekly rituximab was started. IgA level and serum m-spike remained fairly stable over the next 7 months. All treatment was tolerated well except for initial transient rigors, fever, and nausea. Despite bi-monthly maintenance rituximab, the patient devloped a rising IgA of 4940 mg/dL, m-spike of 4.7, falling Hgb and a rising PCLI of 1.5%, 15 months after his original diagnosis of MM. Of particular concern is the development of new, unfavorable, cytogenetic aberrations by FISH with BMPC now showing deletion 13 in 93% and mutation of p53 (17p13) in 98%.100% show fusion of CCND1 and IGH and PC now only partially express CD20. Conclusion: This report suggests that MM patients with CD20+ BMPC and t(11;14)(q13;q32) may represent a target population for anti-CD20 therapy. Our report supports the concept proposed that targeting this clonotypic subset of B cells possibly interrupts a critical oncogenic pathway that is important in the pathogenesis of the clonogenic development of MM.

Cyclooxygenase-2 (COX-2) Is Frequently Expressed in Multiple Myeloma (MM) and Is Associated with Poor Outcome.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2450-2450
Author(s):  
Marco Ladetto ◽  
Sonia Vallet ◽  
Andreas Trojan ◽  
Maria Dell’Aquila ◽  
Luigia Monitillo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Plasma Cell ◽  
Cell Separation ◽  
Plasma Cells ◽  
Mrna Level ◽  
Prognostic Indicator ◽  
Disease Stage ◽  
Plasma Cell Dyscrasias ◽  
Cox 2 ◽  
Ht 29

Abstract Introduction. A perturbed microenvironment with secretion of inflammatory cytokines is typical of MM. Prostaglandins (pg) are implicated in inflammation and angiogenesis and play a role in the pathogenesis of several solid malignancies. Expression of COX-2, the key enzyme of pg synthesis in inflamed tissues, is common in many of these cancers and plays a major role in their development. Moreover, it often acts as a poor prognostic indicator. Despite a large amount of data concerning COX-2 expression in solid tumors, few data are currently available in hematological malignancies. In MM there are several biological, epidemiological and clinical considerations suggesting a potential involvement of the pg pathway. Aim of this study is to verify the involvement of COX-2 in MM and to assess its prognostic role. Patients and methods. COX-2 expression has been assessed by western blotting (WB) as previously described (Du Bois RN, et al, Gastroenterology, 1996). Our positive control was the COX-2 positive cell line HT-29, while bone marrows (BM) from 15 healthy donors were our negative controls. We assessed a panel of 124 samples obtained by 113 patients with plasma cell dyscrasias. Sixteen samples belonged to subjects with MGUS, 80 to patients with MM at diagnosis, and 28 to patients with relapsed/refractory MM. In 11 patients, samples taken at different treatment phases were available. To confirm WB findings and to demonstrate that COX-2 expression occurs in malignant plasma cells immunohistochemistry (IC), and flow cytometry for COX-2 were also performed in 31 and four patients, respectively. Finally, COX-2 expression has been assessed in BM cells from four COX-2 positive patients following selection for the CD138 antigen using the Miltenyi cell separation system. COX-2 expression at the mRNA level has also been assessed by real time quantitative PCR. Results. A dilution test showed that our technique is sensitive enough to detect 2% HT-29 cells in a background of COX-2 negative cells. The 15 normal BM were COX-2 negative. In contrast, COX-2 expression was noticed in 12.5% of MGUS, 34.6% of MM at diagnosis and 56% of MM at relapse. COX-2 positivity at diagnosis and relapse was unrelated to disease stage, BM plasmacytosis, creatinine, Hb levels and ß2 microglobulin. COX-2 expression appeared to be of prognostic relevance: at diagnosis the median time to progression was 14 months in COX-2 positive and 40 months in COX-2 negative subjects (p<0.001). At relapse, of 14 patients showing COX-2 expression, 10 have already died of MM, and four are still alive. In contrast, of 11 COX-2 negative patients only one patient died while 10 are still alive (p<0.001). IC, cell separation and flow cytometry studies indicate that COX-2 expression is related to the malignant plasma cell population. COX-2 mRNA was overexpressed in patients showing increased COX-2 protein expression. Conclusions: a) COX-2 is frequently expressed in plasma cell dyscrasias; b) COX-2 expression is more frequent in advanced disease phases; c) COX-2 expression correlates to a worse outcome. Future studies are required to verify whether COX-2 might be clinically useful as a prognostic marker and/or therapeutic target.

Inhibition of Interleukin-6 Signaling with CNTO 328 Sensitizes Plasma Cell Dyscrasias to the Effects of Melphalan in Association with Suppression of Signaling through Protein Kinase B/Akt.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1836-1836
Author(s):  
Sally A. Hunsucker ◽  
Valeria Magarotto ◽  
Jairo A. Matthews ◽  
Michael Wang ◽  
Veerabhadran Baladandayuthapani ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Protein Kinase ◽  
Cell Lines ◽  
Plasma Cell ◽  
Stromal Cells ◽  
Plasma Cells ◽  
Protein Kinase B ◽  
Myeloma Cell ◽  
Ortho Biotech ◽  
Plasma Cell Dyscrasias

Abstract Abstract 1836 Poster Board I-862 Background: The neutralizing anti-interleukin (IL)-6 monoclonal antibody (MAb) CNTO 328 acts in an additive to synergistic manner to enhance the activity of bortezomib and dexamethasone against models of multiple myeloma by suppressing several IL-6-induced anti-apoptotic signaling pathways. We therefore sought to evaluate the possibility that blockade of IL-6 signaling could also augment the activity of melphalan, and to determine the potential mechanisms underlying this interaction. Methods: A panel of myeloma cell lines was studied both in suspension and with bone marrow stromal cells to evaluate the activity of CNTO 328 with and without melphalan. The CNTO 328 + melphalan combination was also tested in primary cells from patients with a variety of plasma cell dyscrasias. Results: Treatment of IL-6-dependent KAS-6/1, INA-6, and ANBL-6 myeloma cell lines with CNTO 328 + melphalan reduced plasma cell viability in an additive-to-synergistic manner compared to melphalan with a control MAb. Isobologram analysis demonstrated that the combination was synergistic in KAS-6/1 cells regardless of the sequence of drug treatment (combination indices (CIs) from 0.275-0.607), although the strongest synergy was seen with CNTO 328 pretreatment (CIs from 0.275-0.493). These anti-proliferative effects were accompanied by an enhanced activation of drug-specific apoptosis, and this increased cell death was not rescued by the trophic effects of co-culture of plasma cells with the human-derived stromal cell line HS-5. CNTO 328 increased melphalan-mediated induction of both extrinsic, caspase-8-mediated apoptosis, as well as intrinsic, caspase-9-mediated death, which converged to produce increased levels of caspase-3 activity. Apoptosis was enhanced in part by CNTO 328-stimulated cleavage of Bid to tBid, and alterations in the phosphorylation status of BimEL, as well as increased conversion of Bak and, to a lesser extent, of Bax, to their active forms. Neutralization of IL-6 by CNTO 328 also suppressed signaling through the protein kinase B/Akt pathway, as evidenced by decreased levels of phospho-Akt, and decreased activation of several downstream Akt targets, including p70 S6 kinase and 4E-BP1. Importantly, CNTO 328 + melphalan showed enhanced anti-proliferative effects compared to melphalan and a control MAb against primary CD138+ plasma cells derived from patients with multiple myeloma, monoclonal gammopathy of undetermined significance, and amyloidosis, while demonstrating less toxicity to stromal cells. The enhanced effect of the CNTO 328 + melphalan combination was statistically significant compared to either drug alone (p<0.05) in CD138+ cells isolated from patients who had not received prior melphalan therapy. Conclusions: These studies provide a rationale for translation of CNTO 328 into the clinic in combination with melphalan-based therapies, including either high dose therapy in transplant-eligible patients, or standard dose melphalan-containing induction regimens in transplant-ineligible patients, such as with the combination of bortezomib, melphalan, and prednisone. Disclosures: Voorhees: Millennium Pharmaceuticals: Speakers Bureau; Celgene: Speakers Bureau. Xie:Centocor Ortho Biotech Inc.: Employment. Cornfeld:Centocor Ortho Biotech Inc.: Employment. Nemeth:Centocor Ortho Biotech Inc.: Employment.

A high bone marrow plasma cell labeling index in stable plateau–phase multiple myeloma is a marker for early disease progression and death

Blood ◽  
2001 ◽  
Vol 97 (8) ◽  
pp. 2522-2523 ◽  
Author(s):  
David P. Steensma ◽  
Morie A. Gertz ◽  
Philip R. Greipp ◽  
Robert A. Kyle ◽  
Martha Q. Lacy ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Disease Progression ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Group Versus ◽  
Labeling Index ◽  
Cell Labeling ◽  
Plateau Phase ◽  
Bone Marrow Plasma ◽  
High Bone

Abstract The plasma cell labeling index (PCLI) is a measure of plasma cell proliferative activity and is an important prognostic factor in newly diagnosed multiple myeloma (MM). Occasionally patients have been observed with stable, plateau phase MM with minimal numbers of residual light-chain–restricted monoclonal plasma cells, but a high PCLI. No data are available on the outcomes for such patients. Data from 57 patients with plateau phase MM and a marrow PCLI of more than 1.0% were compared with 105 matched control patients with MM with a marrow PCLI of less than 1.0%. All patients had less than 10% total plasma cells on marrow aspirate and biopsy. The median time to progression and overall survival were 8 months and 20 months, respectively, in the high PCLI group versus 39 months and 56 months, respectively, in the low PCLI group (P &lt; .0001). These findings suggest that a high PCLI in patients with apparently stable, plateau phase MM is an adverse parameter that may predict a short time to disease progression and death.

scholarly journals Neutrophil Extracellular Traps (NETs) and Hypercoagulability in Plasma Cell Dyscrasias—Is This Phenomenon Worthy of Exploration?

2021 ◽  
Vol 10 (22) ◽  
pp. 5243
Author(s):  
Olga Ciepiela ◽  
Milena Małecka-Giełdowska ◽  
Emilia Czyżewska
Keyword(s):  
Plasma Cell ◽  
Deoxyribonucleic Acid ◽  
Plasma Cells ◽  
Neutrophil Extracellular Traps ◽  
Related Factors ◽  
Neoplastic Diseases ◽  
Extracellular Traps ◽  
Cancer Metastases ◽  
Induce Activity ◽  
Plasma Cell Dyscrasias

Plasma cell dyscrasias (PCDs) are neoplastic diseases derived from plasma cells. Patients suffering from PCDs are at high risk of hypercoagulability and thrombosis. These conditions are associated with disease-related factors, patient-related factors, or the use of immunomodulatory drugs. As PCDs belong to neoplastic diseases, some other factors related to the cancer-associated hypercoagulability state in the course of PCDs are also considered. One of the weakest issues studied in PCDs is the procoagulant activity of neutrophil extracellular traps (NETs). NETs are web-like structures released from neutrophils in response to different stimuli. These structures are made of deoxyribonucleic acid (DNA) and bactericidal proteins, such as histones, myeloperoxidase, neutrophil elastase, and over 300 other proteins, which are primarily stored in neutrophil granules. NETs immobilize, inactivate the pathogens, and expose them to specialized cells of immune response. Despite their pivotal role in innate immunity, they contribute to the development and exacerbation of autoimmune diseases, trigger inflammatory response, or even facilitate the formation of cancer metastases. NETs were also found to induce activity of coagulation and are considered one of the most important factors inducing thrombosis. Here, we summarize how PCDs influence the release of NETs, and hypothesize whether NETs contribute to hypercoagulability in PCDs patients.

Plasma Cell Dyscrasias

2019 ◽  
pp. 33-38
Author(s):  
Kevin B. Hoover
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Clinical Trials ◽  
African Americans ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Organ Damage ◽  
Disease Staging ◽  
Plasma Cell Dyscrasias ◽  
Urine Testing

Chapter 76 discusses plasma cell dyscrasias, which are currently incurable diseases resulting from the proliferation of plasma cells and the secretion of immunoglobulins with associated anemia and end-organ damage. These diseases are more common in men than women and more common in African Americans than whites. Multiple myeloma is the most common of the plasma cell dyscrasias. Blood and urine testing, bone marrow biopsy, and radiography are the primary tests used for diagnosis. Radiographs are the standard tools in disease staging and monitoring with advanced imaging used primarily for evaluating symptomatic patients with negative radiographs and patients in clinical trials.

Gene Expression Profiling of Plasma Cell Dyscrasias: The Role of IGH Translocations in the Heterogeneity of Multiple Myeloma.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4845-4845
Author(s):  
Antonino Neri ◽  
Michela Mattioli ◽  
Luca Agnelli ◽  
Sonia Fabris ◽  
Luca Baldini ◽  
...  
Keyword(s):  
Gene Expression ◽  
Multiple Myeloma ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Germ Line ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Plasma Cell Dyscrasia ◽  
Transcriptional Level ◽  
Plasma Cell Dyscrasias

Abstract Multiple Myeloma (MM) is the most common form of plasma cell dyscrasia, characterized by a marked heterogeneity of genetic lesions and clinical course. It may develop from a premalignant condition (monoclonal gammopathy ofundetermined significance, MGUS) or progress from intra-medullary to extra-medullaryforms (plasma cell leukemia, PCL). To provide insights into the molecular characterization of plasma cell dyscrasias and to investigate the contribution of specific genetic lesions to the biological and clinical heterogeneity of MM, we analyzed the gene expression profiles of plasma cells isolated from 7 MGUS, 39 MM and 6 PCL patients by means of DNA microarrays. MMs resulted highly heterogeneous at transcriptional level, whereas the differential expression of genes mainly involved in DNA metabolism and proliferation distinguished MGUS from PCLs and the majority of MM cases. The clustering of MM patients was mainly driven by the presence of the most recurrent translocations involving the immunoglobulin heavy-chain locus. Distinct signatures have been found to be associated with different lesions: the overexpression of CCND2 and genes involved in cell adhesion pathways was observed in cases with deregulated MAF and MAFB, whereas genes upregulated in cases with the t(4;14) showed apoptosis related functions. In addition, we identified a set of cancer germ-line antigens specifically expressed in a sub-group of MM patients characterized by an aggressive clinical evolution, a finding that could have implications for patient classification and immunotherapy.

Multi-Parameter Flow Cytometry(FC) Can Detect Proliferating Myeloma Cells After Treatment: Implications for Targeting the Myeloma Proliferative Component

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 4995-4995
Author(s):  
John Lust ◽  
Shaji Kumar ◽  
Michael Timm ◽  
Kathleen Donovan ◽  
Philip R. Greipp ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Plasma Cell ◽  
Research Funding ◽  
Plasma Cells ◽  
Bone Marrow Cells ◽  
Autologous Transplantation ◽  
Cell Labeling ◽  
Growth Fraction ◽  
Proliferating Cells ◽  
Myeloma Cells

Abstract Abstract 4995 Background: Multiple myeloma results from an accumulation of monoclonal nonproliferating plasma cells arising from a small subpopulation of proliferating myeloma cells. In an effort to optimize detection of the human myeloma cell growth fraction and obviate the need for slide review, a novel FC strategy was developed combining elements of light chain restriction, surface antigen expression, and ploidy analysis. Methods: Bone marrow cells from 41 patients with plasma cell proliferative disorders were lysed with ACK and resuspended in 3% BSA. Cells were stained using a 6-color assay with anti-CD45, anti-CD38, anti-CD138, and anti-CD19. Cells were washed and 100 ul Caltag solution A was added for 15 min. Cells were washed and 100 ul of Caltag solution B, anti-kappa and anti-lambda were added for 10 min. Cells were washed and 100 ul PBS and 3 ul RNAse are added for 15 min. Cells were washed and 400 ul of a 1:1000 solution of 3uM DAPI in Tris 0.1% NP-40 was added. Cells were incubated at 4°C for 45 minutes before running on a FACSCanto instrument for ploidy determination. All patients were analyzed both by flow cytometry and the slide based plasma cell labeling index. Results: Forty-one patients were studied; 34 demonstrated a proliferative fraction and 7 had too few plasma cells for analysis after therapy. Of the 34 patients, 6 had MGUS/SMM, 5 newly diagnosed MM, 7 amyloid, and 16 were treated MM. The mean percent proliferating cells were 1.1% (range 0 – 8.6%) with PCLI and 1.4% (range 0.1 – 12.7%) by flow. The correlation between PCLI and flow gave a RSquare value of 0.54. Twelve patients with a PCLI of 0% had a flow proliferation between 0.1 – 1.4% (mean 0.49%). Treated patients received lenalidomide, dexamethasone, bortezomib, and/or autologous transplantation. All 16 treated myeloma patients with adequate plasma cells had a flow proliferation between 0.2 – 12.7% (mean 2.2%). Conclusion: Flow cytometry offers a useful way to detect the proliferative myeloma component at diagnosis and after treatment. The continued presence of proliferating myeloma cells after treatment may explain why most patients relapse and offers another important marker to monitor and cell population to target in patients with active disease. Disclosures: Kumar: Celgene: Consultancy, Research Funding; Millennium: Research Funding; Merck: Consultancy, Research Funding; Novartis: Research Funding; Genzyme: Consultancy, Research Funding; Cephalon: Research Funding.

Sign in / Sign up

Export Citation Format

Share Document