Obstructive Uropathy Associated with Myelomonocytic Infiltration of The

Total Page:16

File Type:pdf, Size:1020Kb

Obstructive Uropathy Associated with Myelomonocytic Infiltration of The 340 J Clin Pathol 1998;51:340–342 Obstructive uropathy associated with myelomonocytic infiltration of the prostate J Clin Pathol: first published as 10.1136/jcp.51.4.340 on 1 April 1998. Downloaded from B Hope-Gill, J R Goepel, R C Collin Abstract The bone marrow showed myeloid and mono- A 72 year old man was diagnosed with cytic hyperplasia and myeloid precursors were chronic myelomonocytic leukaemia hypogranular. There was no excess of blast (CMML) according to the FAB group cells. Micromegakaryocytes were identified in classification. He presented with symp- marrow smears. toms of anaemia, urinary frequency, hesi- In the context of a persistent peripheral tancy, and nocturia. He was later admitted blood monocytosis of > 1.0×109/litre, these with acute urinary retention and acute findings were consistent with a diagnosis of renal failure, which resolved with treat- chronic myelomonocytic leukaemia as defined ment. A transurethral resection of the by the FAB criteria.1 Elective admission was prostate was performed. Histological arranged for a three unit transfusion of packed examination showed fibromuscular hy- red cells a few days later. perplasia with dense infiltration by myelo- While awaiting admission he became in- monocytes which stained positively with creasingly lethargic and nauseous. Examin- chloroacetate esterase; immunohisto- ation revealed a distended bladder and a chemical staining was positive for lyso- smooth, tender, moderately enlarged prostate; zyme, CD43, CD45, and CD68. Following when catheterised, 2300 ml of urine were treatment with oral etoposide he trans- drained. He had not previously experienced formed to acute myeloid leukaemia and any symptoms of urinary obstruction until two eventually died. Myelomonocytic infiltra- weeks before admission. Investigations at this tion of the prostate has not been reported stage revealed Na 131 mmol/litre, K 5.9 mmol/ before. This case extends the spectrum of litre, HCO3 21 mmol/litre, urea 30 mmol/litre, disease previously recognised in CMML. and creatinine 995 µmol/litre. (J Clin Pathol 1998;51:340–342) Treatment with intravenous fluids was started and a blood transfusion was given. He Keywords: chronic myelomonocytic leukaemia; developed postcatheterisation diuresis and his prostate; urinary retention; myelomonocytes serum biochemistry improved such that the creatinine had fallen to 126 µmol/litre two days http://jcp.bmj.com/ later. He was discharged with a urethral Chronic monomyelocytic leukaemia (CMML) catheter in situ to await an elective transure- as defined by the FAB group1 is classified as a thral resection of the prostate. Subsequently a myelodysplastic disorder characterised by a 30 g prostate was resected and histological peripheral monocytosis and particular bone examination revealed dense infiltration with marrow features. Cytopenia, hepatospleno- myelomonocytes. megaly, and lymphadenopathy are common. Postoperatively, he required multiple packed on September 26, 2021 by guest. Protected copyright. Skin infiltration and serous eVusions are also red cell and platelet transfusions owing to recognised, and gingival infiltration occasion- heavy haematuria. ally occurs, particularly if accompanied by Subsequently, he was entered into the Medi- a high peripheral monocytosis (> 2.6×109/ cal Research Council CMML trial and ran- litre).2–5 However, myelomonocytic infiltration domised to receive oral etoposide (VP-16), 50 of other tissues is not reported. mg/day. However, three months later he was Chesterfield Royal readmitted with rigors, a chest infection, and Hospital, Department Case report of Haematology, general deterioration. Full blood count and Calow, Chesterfield, A 72 year old man was referred to our depart- peripheral film revealed a haemoglobin of 10.8 UK ment in September 1995 with two weeks of g/dl and a white blood cell count of 137×109/ B Hope-Gill increasing exertional shortness of breath. He litre (neutrophils 115.5, lymphocytes 0.5, R C Collin also had a four week history of urinary monocytes 16.7, 14% blast cells with large frequency, hesitancy, and nocturia. numbers of Auer rods) signifying transforma- Royal Hallamshire The general practitioner had performed a Hospital, Department tion to acute myeloid leukaemia. His clinical full blood count—haemoglobin 5.7 g/dl (MCV condition deteriorated progressively and in of Histopathology, 9 Beechill Road, 90.8 fl), white blood cell count 5.6×10 /litre conjunction with the patient and his wife the SheYeld, UK (neutrophils 0.6, lymphocytes 3.1, monocytes decision was made to treat palliatively. He died J R Goepel 1.7), platelet count 69×109/litre—and urea and three weeks later at home. No necropsy electrolyte determinations—sodium 138 examination was performed. Correspondence to: Dr R C Collin, Department mmol/litre, potassium 4.2 mmol/litre, HCO3 of Haematology, Chesterfield 25 mmol/litre, urea 8.9 mmol/litre, creatinine Royal Hospital, Calow, 143 µmol/litre. Prostate specific antigen was Histology report Chesterfield S44 5BL, UK. 12.1 ng/ml. The blood film and bone marrow Numerous prostate fragments were received Accepted for publication smears both showed morphological abnormali- fixed in formalin. These showed bladder neck 12 November 1997 ties with abnormal dysplastic granulopoiesis. and prostate, with hyperplasia of prostate Obstructive uropathy from prostatic disease 341 (table 1). The majority presented with the onset of or worsening of symptoms of urinary J Clin Pathol: first published as 10.1136/jcp.51.4.340 on 1 April 1998. Downloaded from outflow obstruction. The best documented and most recent cases occurred with chronic lymphocytic leukaemia, in which 20 cases of 29 presented with acute urinary retention; subse- quent investigation and treatment of this revealed leukaemic infiltration of the prostate. A necropsy study of 503 leukaemic patients showed that the actual incidence of metastases to the prostate is higher than the number of case reports would suggest.10 Necropsy re- vealed up to 40% prostatic infiltration in acute lymphoblastic leukaemia (ALL), 11–20% in chronic lymphocytic leukaemia (CLL), 11– 20% in acute myeloblastic leukaemia (AML), and between 1% and 10% in chronic myelo- cytic leukaemia (CML).10 These frequencies reflect the tendencies of the diVerent leukae- Figure 1 Prostate infiltrated by chronic myelomonocytic leukaemia cells. (Haematoxylin and eosin, ×149.) mias to metastasise to non-reticuloendothelial tissues. The discrepancy between the fre- quency of case reports and incidence of prostatic involvement found at necropsy re- flects the fact that unless urinary symptoms develop prostatic involvement is unlikely to be detected. The extent to which the leukaemic infiltrate contributes to urinary obstruction is unknown. The best documented cases of CLL infiltration of the prostate associated with urinary retention were combined with histological evidence of benign prostatic hypertrophy or with bladder trabeculation, suggesting chronic obstruction. Certainly others suggest that the presence of benign prostatic hypertrophy is necessary to precipitate retention, and not all cases with leu- kaemic infiltration of the prostate develop 6 obstructive symptoms. Cases of acute retention http://jcp.bmj.com/ in acute leukaemia may be explained by other Figure 2 Chronic myelomonocytic leukaemia in prostate. Many cells are positive for mechanisms, such as leukaemic meningitis. CD68. (Immunoperoxidase PGM 1, ×297.) Patients present with typical symptoms of urinary outflow obstruction.6–9 The prostate is Table 1 Summary of reported cases of leukaemic uniformly enlarged and smooth, as in benign infiltration of the prostate gland prostatic hypertrophy. In CLL, lymphocytic Cases with Cases with infiltration is irregular in distribution, so that on September 26, 2021 by guest. Protected copyright. prostatic urinary preoperative needle biopsy is unreliable. There Leukaemia type histology retention are insuYcient published data to show whether CLL 24 15 this is also true for granulocytic leukaemias. It Acute granulocytic 3 3 Chronic granulocytic 2 2 has been suggested that patients who present Chronic myelomonocytic (present with acute granulocytic leukaemia and urinary case) 1 1 retention tolerate surgical transurethral resec- Cases with no prostatic histology 22 tion of the prostate for alleviation of symptoms less well than patients with CLL, in whom the glands and particularly of the fibromuscular five year survival is good.6 Therefore, where the stroma. Prostatic stromal elements were infil- diagnosis is known in advance, preoperative trated by dense masses and cords of cells (fig 1). prostatic needle biopsy is warranted in these These were of medium size with slight pleomor- patients and other treatment options should be phism. The nucleus was ovoid to rounded and considered. This patient responded to surgical vesicular cytoplasm varied in extent; in some it transurethral resection of the prostate and he was granular. Chloroacetate esterase staining subsequently died after transformation to was positive in many of these cells. Immunohis- AML. tochemical staining was positive in most cells Dajani and Burke reported five previous for lysozyme, in many for CD43 and CD45, cases of granulocytic leukaemia (table 1) caus- and in many for CD68 (fig 2). ing urinary retention.6 However, precise de- fined subtyping of the granulocytic infiltrate is Discussion lacking. We were unable to find a conclusive Case reports concerning leukaemic infiltration report in the English language literature of of the prostate are uncommon. Dajani
Recommended publications
  • Cultured Human Langerhans Cells Resemble Lymphoid Dendritic Cells in Phenotype and Function
    Cultured Human Langerhans Cells Resemble Lymphoid Dendritic Cells in Phenotype and Function Nikolaus Romani, Ph.D., Angela Lenz, Herta Glassel, Ph.D., Hella Stossel, Ursula Stanzl, Otto Majdic, M.D., Peter Fritsch, M.D., and Gerold Schuler, M.D. D~p2rnnent$ of DerlTl2tology and Internal Medicine (HG). University of lnnsbrud:.. Innsbruck. :and Inscirute of Immunology (OM). University of Vienna. Vienn2, Austria Freshly isolated murine epidermal Langerhans cells (LC) are rured human LC resembled human lymphoid dendritic cells weak stimulators of resting T cells. Upon culture their phe­ in morphology, phenotype, and function. Specifically, LC notype changes, their stimulatory activity increases signifi­ became non-adherent upon culture and developed sheet-like cantly. and they come to resemble lymphoid dendritic cells. processes (so-called "veils"), decreased their surface ATP / Resident murine Le, therefore. might represent a reservoi.r ADP'ase acti vity, and lost nonspecific esterase activity. As in of immature dendritic cells. We have now used enzyme cyto­ the mouse, surface expression of MHC class I and 11 an tigens chemistry. a panel of some 80 monoclonal antibodies, and increased significantly. and Fell receptors were significantly immunofluorescence microscopy or two-color flow cytom­ reduced. Markers that are expressed by dendritic cells (like eery, as well as transmission electron microscopy. CO analyse CD40) appeared on LC following culrure. Cultured human the phenotype and morphology of human LC before and LC were potem T-cell stimulators. Our findings support the after 2 - 4 d of bulk epidermal cell culrure. In addition, LC view that resident human Le, like murine Le, represent were enriched from bulk epidermal cell culrures, and their immature precursors of lymphoid dendritic cells in skin­ stimulatory capacity was tested in the allogeneic mixed leu­ draining lymph nodes.] It,vest DermatoI93:600-609, 1989 kocyte reaction and the oxidative mitogenesis assay.
    [Show full text]
  • Induction of Myeloid Colony-Stimulating Activity in Murine Monocyte Tumor Cell Lines by Macrophage Activators and in a T-Cell Line by Concanavalin A1
    [CANCER RESEARCH 38, 1414-1419, May 1978] Induction of Myeloid Colony-stimulating Activity in Murine Monocyte Tumor Cell Lines by Macrophage Activators and in a T-Cell Line by Concanavalin A1 Peter Ralph, Hal E. Broxmeyer,2 Malcolm A. S. Moore, and Ilona Nakoinz Sloan-Kettering Institute for Cancer Research, Rye, New York 10580 ABSTRACT activating agents and in T-lymphomas by T-lymphocyte mitogens. Certain fibrosarcoma lines in culture and the WEHI-3 myelomonocytic leukemia cell line have previously been shown to secrete myeloid colony-stimulating activity MATERIALS AND METHODS (CSA) spontaneously. We describe here other hemato- Murine Tumor Cell Lines. Monocyte and macrophage poietic tumor cell lines in which CSA is either produced tumor cell lines are described in Ref. 32, except for Abelson constitutively or inducible by immunostimulators. CSA leukemia virus-induced line RAW264 (33). T-lymphoma lines production in macrophage and monocyte tumor lines is EL4, RBL-5, BW5147, and S49; myelomas P3 and induced by lipopolysaccharide, zymosan, Mycobacterium MOPC315; mastocytoma P815; lymphoma P388; and Abel- strain Bacillus Calmette-Guerin, tuberculin purified pro son line R8 are described in Ref. 31. Rauscher leukemia tein-derivative preparation from mycobacteria, and dex- virus line RBL-3 and chemically induced leukemia L1210 tran sulfate. Myeloma, mastocytoma, and T-lymphoma (39) were obtained from K. Chang (NIH, Bethesda, Md.); lines do not produce CSA with or without these agents. In fibrosarcoma L929 (5) was obtained from B. Williams contrast, the T-lymphocyte mitogen concanavalin A (but (Sloan-Kettering Institute, Rye, N. Y.); bone marrow fibro- not phytohemagglutinin) induces CSA synthesis in one of blast JLSV9 and Rauscher leukemia virus-infected JLSV9- seven T-lymphomas tested.
    [Show full text]
  • Cddpre6 7 5..5
    Cell Death and Differentiation (1999) 6, 599 ± 608 ã 1999 Stockton Press All rights reserved 13509047/99 $12.00 http://www.stockton-press.co.uk/cdd The role of Ets family transcription factor PU.1 in hematopoietic cell differentiation, proliferation and apoptosis Tsuneyuki Oikawa*,1, Toshiyuki Yamada1, Spi-1: spleen focus forming virus proviral integration-1; TPA: 12-0- Fumiko Kihara-Negishi1, Hitomi Yamamoto1, tetradecanoylphorbol-13-acetate Nobuo Kondoh1,2, Yoshiaki Hitomi1,3 and Yoshiyuki Hashimoto1 Introduction Hematopoietic cell differentiation is thought to be achieved 1 Department of Cell Genetics, Sasaki Institute, 2-2, Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan through promoting proliferation and survival of particular 2 Department of Biochemistry II, National Defense Medical College, 3-2, Namiki, progenitors by hematopoietic growth factors and through Tokorozawa, Saitama 359-0042, Japan differentiation by stochastic and/or hierarchical programmed 3 Investigative Treatment Division, National Cancer Center, Research Institute, cascades of the expression of several tissue-specific East, 6-5-1, Kashiwanoha, Kashiwa 277-0882, Chiba, Japan transcription factors.1,2 During these differentiation pro- * corresponding author: T. Oikawa cesses, however, cells with accidental inappropriate expres- tel: +81-3-3294-3286; fax: +81-3-3294-3290; e-mail: toikawa@gold®sh.sasaki.or.jp sion of the genes that control cell proliferation and/or differentiation might be eliminated by apoptosis, otherwise Received 26.11.98; Revised 7.4.99; Accepted 28.4.99 the cells could be harmful to the host in some cases. This Edited by R Knight hypothesis is supported by the experimental facts that ectopic or inappropriate overexpression of such transcription factors often triggers off development of malignant tumors3,4 and that Abstract most of them are also implicated in the process of apoptotic cell 5 The PU.1 gene encodes an Ets family transcription factor death.
    [Show full text]
  • Japanese Case of Follicular Lymphoma of Ocular Adnexa Diagnosed by Clinicopathologic, Immunohistochemical, and Molecular Genetic Techniques
    Clinical Ophthalmology Dovepress open access to scientific and medical research Open Access Full Text Article CASE REPORT Japanese case of follicular lymphoma of ocular adnexa diagnosed by clinicopathologic, immunohistochemical, and molecular genetic techniques Takaaki Otomo1 Background: Follicular lymphomas of the ocular adnexa are very rare in Japan, with only Nobuo Fuse1 two reported cases. Kenichi Ishizawa2 Case: A 44-year-old woman visited our clinic for treatment of ocular adnexal tumors in both Motohiko Seimiya1 eyes. Masahiko Shimura4 Findings: Histologic examination showed that the neoplastic lesions consisted of atypical Ryo Ichinohasama3 lymphoid cells, and the tentative diagnosis was malignant lymphoma. Immunophenotypic analyses by flow cytometry and immunohistochemistry showed that the atypical lymphoid cells 1 Department of Ophthalmology, expressed CD45, bcl-2, CD10, CD19, CD20, IgM, and kappa light chains. The cells were negative 2Departments of Rheumatology and Hematology, 3Division of for CD5 and other T, natural killer, or myelomonocyte antigens. Southern blot hybridization Hematopathology, Tohoku University demonstrated gene rearrangement bands in the immunoglobulin JH region. Fluorescence in situ Graduate School of Medicine, Sendai, hybridization studies showed a translocation at t(14,18)(q32,q21). Systemic evaluations detected 4Department of Ophthalmology, NTT East Japan Tohoku Hospital, enlargements of both the inguinal lymph nodes and parabronchial lymph nodes. Sendai, Myagi, Japan Conclusion: Our results show that
    [Show full text]
  • Rajendiran Et Al Protocol Biorxiv 120320
    bioRxiv preprint doi: https://doi.org/10.1101/2020.12.03.410894; this version posted December 4, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. A Quantitative Hematopoietic Stem Cell Reconstitution Protocol: Accounting for Recipient Variability, Tissue Distribution and Cell Half-Lives Smrithi Rajendiran1, Scott W. Boyer1, E. Camilla Forsberg1,# Institute for the Biology of Stem Cells, Department of Biomolecular Engineering, University of California, Santa Cruz, Santa Cruz, CA 95064, USA # Corresponding author: Director, Institute for the Biology of Stem Cells, Professor of Biomolecular Engineering, University of California Santa Cruz, Mail stop SOE2, 1156 High Street, Santa Cruz, CA 95064, USA. T: 831-459-2111. E-mail: [email protected] bioRxiv preprint doi: https://doi.org/10.1101/2020.12.03.410894; this version posted December 4, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Abstract Hematopoietic stem and progenitor cell (HSPC) transplantation is the paradigm for stem cell therapies. The protocol described here enables quantitative assessment of the body-wide HSPC reconstitution of different mature hematopoietic cells in mice based on their presence in circulating blood. The method determines donor-derived mature cell populations per mouse, over time, by quantitatively obtaining their absolute numbers in the peripheral blood and utilizing previously assessed tissue-distribution factors. A Markov-based birth/death computational model accounts for the drastic differences in mature cell half-lives. By quantifying the number of cells produced and eliminating host variability, the protocol can be used to directly compare the lineage output of different types of HSPCs on a per cell basis, thereby clarifying the lineage potential and expansion capacity of different cell populations.
    [Show full text]
  • Granzyme M: Behind Enemy Lines
    Cell Death and Differentiation (2014) 21, 359–368 & 2014 Macmillan Publishers Limited All rights reserved 1350-9047/14 www.nature.com/cdd Review Granzyme M: behind enemy lines SAH de Poot1 and N Bovenschen*,1,2 The granule-exocytosis pathway is the major mechanism via which cytotoxic lymphocytes eliminate virus-infected and tumor cells. In this pathway, cytotoxic lymphocytes release granules containing the pore-forming protein perforin and a family of serine proteases known as granzymes into the immunological synapse. Pore-formation by perforin facilitates entry of granzymes into the target cell, where they can activate various (death) pathways. Humans express five different granzymes, of which granzymes A and B have been most extensively characterized. However, much less is known about granzyme M (GrM). Recently, structural analysis and advanced proteomics approaches have determined the primary and extended specificity of GrM. GrM functions have expanded over the past few years: not only can GrM efficiently induce cell death in tumor cells, it can also inhibit cytomegalovirus replication in a noncytotoxic manner. Finally, a role for GrM in lipopolysaccharide-induced inflammatory responses has been proposed. In this review, we recapitulate the current status of GrM expression, substrate specificity, functions, and inhibitors. Cell Death and Differentiation (2014) 21, 359–368; doi:10.1038/cdd.2013.189; published online 10 January 2014 Facts cellular immune response and can exert their protective function by either killing potentially harmful cells or by crippling Granzyme M is a serine protease expressed in multiple viral replication within the host cell. Upon recognition of a cytotoxic lymphocyte subsets. target cell, effector cells engage with the target cell and form Granzyme M prefers to cleave after a Met or a Leu residue an immunological synapse.
    [Show full text]
  • Group 1 CD1 Genes in Rabbit Sandra M
    Group 1 CD1 Genes in Rabbit Sandra M. Hayes and Katherine L. Knight J Immunol 2001; 166:403-410; ; This information is current as doi: 10.4049/jimmunol.166.1.403 of September 23, 2021. http://www.jimmunol.org/content/166/1/403 Downloaded from References This article cites 45 articles, 19 of which you can access for free at: http://www.jimmunol.org/content/166/1/403.full#ref-list-1 Why The JI? Submit online. http://www.jimmunol.org/ • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication *average by guest on September 23, 2021 Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2001 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Group 1 CD1 Genes in Rabbit1 Sandra M. Hayes2 and Katherine L. Knight3 CD1 is an Ag-presenting molecule that can present lipids and glycolipids to T cells. The CD1 genes were first identified in the human, and since then, homologs have been identified in every mammalian species examined to date. Over a decade ago, CD1B and CD1D homologs were identified in the rabbit.
    [Show full text]
  • Inhibition of IRF4 in Dendritic Cells by PRR-Independent
    RESEARCH ARTICLE Inhibition of IRF4 in dendritic cells by PRR-independent and -dependent signals inhibit Th2 and promote Th17 responses Jihyung Lee1, Junyan Zhang1,2,3, Young-Jun Chung1,4, Jun Hwan Kim1, Chae Min Kook1, Jose´ M Gonza´ lez-Navajas3,5,6, David S Herdman1, Bernd Nu¨ rnberg7, Paul A Insel1,8, Maripat Corr1, Ji-Hun Mo4, Ailin Tao2,3, Kei Yasuda9, Ian R Rifkin9,10, David H Broide1, Roger Sciammas11, Nicholas JG Webster1,12*, Eyal Raz1,3* 1Department of Medicine, University of California San Diego, San Diego, United States; 2The Second Affiliated Hospital of Guangzhou Medical University (GMU), The State Key Laboratory of Respiratory Disease, Guangdong Provincial Key Laboratory of Allergy & Clinical Immunology, Guangzhou, China; 3Center for Immunology, Inflammation and Immune-mediated disease, GMU, Guangzhou, China; 4Department of Otorhinolaryngology-Head and Neck Surgery, Dankook University College of Medicine, Chungnam, Republic of Korea; 5Alicante Institute for Health and Biomedical Research (ISABIAL - FISABIO), Alicante, Spain; 6Networked Biomedical Research Center for Hepatic and Digestive Diseases (CIBERehd), Institute of Health Carlos III, Madrid, Spain; 7Department of Pharmacology and Experimental Therapy, University of Tu¨ bingen, Tu¨ bingen, Germany; 8Department of Pharmacology, University of California San Diego, San Diego, United States; 9Boston University School of Medicine, Boston, United States; 10VA Boston Healthcare System, Boston, United States; 11Center for Comparative Medicine, University of California, Davis, Davis, United States; 12VA San Diego Healthcare System, San Diego, United States *For correspondence: [email protected] (NJGW); [email protected] (ER) Abstract Cyclic AMP (cAMP) is involved in many biological processes but little is known Competing interests: The regarding its role in shaping immunity.
    [Show full text]
  • Identification of Red and White Blood Cells from Whole Blood Samples Using the Agilent 2100 Bioanalyzer
    Identification of red and white blood cells from whole blood samples using the Agilent 2100 bioanalyzer Application Note Sylvie Veriac Valérie Perrone Madeleine Avon Abstract This Application Note describes an antibody based method to identify Agilent Equipment: and count blood cells from whole blood samples on the Agilent 2100 2100 bioanalyzer Cell fluorescence assay kit bioanalyzer without a washing step. Application Area: The ability of the Agilent 2100 bioanalyzer to identify peripheral whole Blood cell analysis blood cells is demonstrated and compared to a flow-cytometry-based reference instrument. Introduction Staining of red blood cells WBCs staining protocol The monoclonal antibody anti- 1. Add 2 µL of SYTO 16 (1 mM) to This Application Note describes a human CD235a conjugated with 20 µL of whole blood. method to identify blood cells the fluorochrome Alexa 647, a red 2. Wait 5 minutes and add 200 µL from a whole blood sample using excited fluorescent marker (Abd- of lysing solution. Incubate antibody staining and cell fluores- Serotec), was chosen for RBC 30 minutes at room temperature cence assays on the Agilent 2100 staining. Each sample was diluted in the dark. bioanalyzer without a washing to adjust the cell density with the 3. Dilute the preparation (1:2) step. A broad variety of cell lines Agilent cell buffer, a component of with the Agilent cell buffer. have been tested with the Agilent the cell kit. For prestaining the 4. Load and run the chip in the 2100 bioanalyzer so far. We want RBCs, 50 µL of diluted (1:600) 2100 bioanalyzer according to to extend these tests to whole whole blood were transferred into the reagent kit guide.
    [Show full text]
  • Proapoptotic BID Is Required for Myeloid Homeostasis and Tumor Suppression
    Downloaded from genesdev.cshlp.org on September 25, 2021 - Published by Cold Spring Harbor Laboratory Press Proapoptotic BID is required for myeloid homeostasis and tumor suppression Sandra S. Zinkel,1 Christy C. Ong,1 David O. Ferguson,2 Hiromi Iwasaki,3 Koichi Akashi,3 Roderick T. Bronson,3 Jeffery L. Kutok,3 Frederick W. Alt,2 and Stanley J. Korsmeyer1,4 1Howard Hughes Medical Institute, Departments of Pathology and Medicine, Harvard Medical School, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA; 2Howard Hughes Medical Institute, Division of Immunology, Boston Children’s Hospital, Center for Blood Research, Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA; 3Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115, USA The proper expansion and contraction of hematopoietic cells requires tight regulation of cell death. BID, a “BH3-only” molecule, amplifies death receptor signals connecting the extrinsic to intrinsic pathways by triggering the mitochondrial pathway of apoptosis. Bid-deficient mice, as they age, spontaneously develop a myeloproliferative disorder, which progresses from myeloid hyperplasia to a fatal, clonal malignancy closely resembling chronic myelomonocytic leukemia (CMML). Thus, an apoptotic defect can result in myeloid leukemogenesis. Premalignant Bid−/− myeloid precursor cells are resistant to death receptor-induced apoptosis. Furthermore, a competitive reconstitution assay demonstrates that Bid-deficient long-term repopulating cells give rise to expanded myelomonocytic cells in vivo. Surprisingly, a single BH3-only molecule operating in the extrinsic death receptor pathway proved essential in vivo for physiologic cell death required to maintain myeloid homeostasis. Moreover, progression to CMML indicates that an upstream BH3-only molecule, BID, is required to suppress tumorigenesis.
    [Show full text]
  • Origin and Expansion of the Serine Protease Repertoire in the Myelomonocyte Lineage
    International Journal of Molecular Sciences Review Origin and Expansion of the Serine Protease Repertoire in the Myelomonocyte Lineage Stefanie A. I. Weiss 1 , Salome R. T. Rehm 1, Natascha C. Perera 2, Martin L. Biniossek 3, Oliver Schilling 4,5 and Dieter E. Jenne 1,6,* 1 Comprehensive Pneumology Center (CPC-M), Institute of Lung Biology and Disease (iLBD) Helmholtz Zentrum München and University Hospital of the Ludwig-Maximilians University (LMU), 81377 Munich, Germany; [email protected] (S.A.I.W.); [email protected] (S.R.T.R.) 2 ARTTIC Innovation GmbH, 80333 Munich, Germany; [email protected] 3 Institute of Molecular Medicine and Cell Research, Faculty of Medicine, University of Freiburg, 79104 Freiburg, Germany; [email protected] 4 Institute of Surgical Pathology, University Medical Center Freiburg, Faculty of Medicine, University of Freiburg, 79106 Freiburg, Germany; [email protected] 5 German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany 6 Max Planck Institute of Neurobiology, 82152 Planegg-Martinsried, Germany * Correspondence: [email protected] Abstract: The deepest evolutionary branches of the trypsin/chymotrypsin family of serine proteases are represented by the digestive enzymes of the gastrointestinal tract and the multi-domain proteases of the blood coagulation and complement system. Similar to the very old digestive system, highly diverse cleavage specificities emerged in various cell lineages of the immune defense system during vertebrate evolution. The four neutrophil serine proteases (NSPs) expressed in the myelomonocyte lineage, neutrophil elastase, proteinase 3, cathepsin G, and neutrophil serine protease 4, collectively Citation: Weiss, S.A.I.; Rehm, S.R.T.; display a broad repertoire of (S1) specificities.
    [Show full text]
  • Identification of Dendritic Antigen-Presenting Cells In
    Identification of dendritic antigen-presenting cells in the zebrafish Geanncarlo Lugo-Villarinoa,1, Keir M. Ballaa,1, David L. Stachuraa, Karina Bañuelosa, Miriam B. F. Werneckb, and David Travera,c,2 aSection of Cell and Developmental Biology, Division of Biological Sciences, University of California, La Jolla, CA 92093-0380; bDivision of Cellular Biology, Brazilian National Cancer Institute (INCA), Rio de Janeiro, 22031-050, Brazil; and cDepartment of Cellular and Molecular Medicine, University of California, La Jolla, CA 92093-0380 Edited* by Laurie H. Glimcher, Harvard University, Boston, MA, and approved June 2, 2010 (received for review January 13, 2010) In mammals, dendritic cells (DCs) form the key link between the mammalian LCs, in leukocytes from lymphoid tissues of salmonids innate and adaptive immune systems. DCs act as immune sentries in and from inflammatory lesions in rainbow trout bearing micro- various tissues and, upon encountering pathogen, engulf and traffic sporidial gill disease (10). Here, we have addressed the biology of foreign antigen to secondary lymphoid tissues, stimulating antigen- DCs in the zebrafish, a small cyprinid teleost that features an im- specific T lymphocytes. Although DCs are of fundamental impor- mune system similar to that of mammals (11). tance in orchestrating the mammalian immune response, their presence and function in nonmammalian vertebrates is largely Results and Discussion unknown. Because teleosts possess one of the earliest recognizable Identification of DCs in Zebrafish. We previously determined that adaptive immune systems, we sought to identify antigen-present- the major hematopoietic lineages of the zebrafish could be iden- ing cells (APCs) in the zebrafish to better understand the potential tified by light-scatter characteristics using fluorescence activated origins of DCs and their evolutionary relationship to lymphocytes.
    [Show full text]