HUMAN LEUKEMIAS

Developments tn Oncology Series

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HUMAN LEUKEMIAS

Cytochemical and Ultrastructural Techniques in Diagnosis and Research

Edited by

Aaron Polliack, M.D.

Martinus Nijhoff Publishing A member 0/ the Kluwer Academic Publishers Group

BOSTON THE HAGUE DORDRECHT LANCASTER

DISTRIBtrrORS

For {he Uni{ea States and Canada

Kluwer Bosmn, In(. 190 Old Derby Streel Hingham, MA 020H

For all other countries

Kluwa Academic Publishers Group DiwibUlion Centre

P,O. Bo~ 322 3300 AH Dordrecht

The Netherlands

Libn.ry of Congress Cataloging in Publication Dala Main entry under title:

Human leukemias,

(Developments in oncology) Includes index. \. Leukemia-Addresses, es"'ys, leuures

2, Diagnosis, Electron micros<opic-Addres,es, e"3\'S, leclures. 3. Cytochemistry-Technique-Addresses, essays, 1e<:lures, I. Po!liack, Aaron, 1939-II. Series. lD NLM: \. Leukemia-UltrastruClurc. 2. Leukemia-Diasnosis. l HistocYlochemistr\,. WI DE998N v. 14 I QZ 3~O H918) RC643.H85 1983 616.99'419 83-8240

ISBN-13: 978- 1-46 12-9785-7 c- ISBN- 13: 978- 1-46 13-2823-0 001 : 10.1 0071978-1-46 13-2823-0

Copyright 1984 0 by Martinu! Nijhoff Publisbing, Boston. Solkover reprint uflhe hardcover (s\ ~-ditiun (984

AU rights reserved , No part of this publicat ion may be reproduced, stored in a ret rieval system, or tnonsmit ted in any form or by any means, mechan­

ical, photocopying. recording. or olherwisc, withoul wrillen permission of the publish"r, Marlinus Ni jhoff Publishers, P.O. Box '66, nOI eN The

Hague, The Netherlands.

CONTENTS

Contributing Authors IX

Preface XUI

1. CYTOCHEMISTRY, LIGHT MICROSCOPY 1

1. Contribution of Cytochemistry in Leukemia 3 G.A. Pangalis

2. Cytochemical Distribution of 5-Nucleotidase Activity in Hu­man Leukemic cells 15 G.A. Pangalis, P.P. Roussou, and N.B. Tsavaris

3. Significance of Improved Cyto­chemical Methods for Hemo­protein Enzymes in Diagnosis and Classification of Leuke-mIa 23 ].S. Hanker, P.E. Yates, W.W. Am­brose, ].0. Moore,]. Laszlo, W.H Starkweather and B.L. Giammara

II. TRANSMISSION ELECTRON MI­CROSCOPY, ULTRASTRUCTURAL CYTOCHEMISTRY 47

4. Pathology of Neutrophil Gran­ules: Selective Abnormalities of Azurophil and Specific Granules of Human Neutro-philic Leukocytes 49 D.F. Bainton

5. Review of the Ultrastructure and Cytochemistry of Mega­karyoblastic Leukemia 63 ]. Breton-Gorius

6. The Ultrastructure of Mono-cytic Leukemia 93 G. Lambertenghi-Deliliers, N. Polli, and M.T. Nava

7. Blastic Transformation of Chronic Granulocytic Leuke­mia and other Myeloprolifera­tive Disorders: A study of 26 cases Emphasizing the Impor­tance of Cytochemistry and Ultrastructure in Defining the Cell Phenotype 105 A. Polliack, R. Leizerowitz, A. Korn­berg, D. Gur/el, and H Gamliel

8. Ultrastructural Observations in Therapy-Related Panmy-elosis 123 R.W. McKenna,].L. Parkin, and R.D. Brunning

9. A comparison of the Ultra­structure of Secondary Leuke­mias with de Novo Acute non-Lymphoblastic Leukemia and Blast Crisis of Chronic Granulo-cytic Leuke-mia 159 ]. Vardiman, M.A. Biller, and HM. Golomb

10. Acute Lymphoblastic Leukemia 173 R.D. Brunning, J.L. Parkin, and R. W. McKenna

11. Acute Lymphoblastic Leuke­mia: Cytochemistry and Ultra­structure with Emphasis on the Hand Mirror Variant 219 HR. Schumacher, ].D. Cotelingam, and W.K. Hummer

12. Prolymphocytic Leukemia of B- and T-cell Types: Morpho­logical Differences by Light and Electron Micros-copy 251 D. Catovsky, E. Matules, A.D. Crock­ard, M. O'Brien, and C. Costello

vii

V1I1 CONTENTS

13. Ultrastructural Analysis of B­and T-lymphocyte Subsets with Immunogold Technique: Its relevance to the Study of Chronic Lymphocytic Leuke-mias 261 E. Matutes, D. Robimon, and D. Catot'sky

14. Cytochemistry and Ultrastruc­ture of T-cell Chronic Lymphocytic Leukemia 271 D. Huhn

15. Fine structure of leukemic T­and B-cells, with Special Ref­erence to Adult T-cell Leuke-mia in Japan 281 K. Kataoka, Y. Sakano, N. Imamura, S. Kamihira, and I Minowada

16. Ultrastructure of Adult T-cell Leukemia in Japan 297 M. Shamoto

17. Plasma Cell Leukemia: Ultra­structural Aspects in Diagno­sis 309 M. Djaldetti

18. Sezary Syndrome: Cytochemis-try and Ultrastructure 327 G. Flandrin and M.T. Daniel

19. Hairy Cell Leukemia: Cyto­chemistry and Ultrastructure (TEM) in Diagnosis 331 G. Flandrin and M.T. Daniel

20. Ultrastructure of the Leukemic Phase in Non-Hodgkin's Lymphoma 339 E. Kaiser/ing and H.K. Miiller-Hermelink

III. TRANSMISSION IMMUNO ELEC-TRON MICROSCOPY 381

21. Immuno Electron Microscopic Observations in Human B-cell Leukemias 383 F. Reyes, M.F. Gourdin, and IP. Farcel

IV. FREEZE-FRACTURE, ULTRASTRUCTURE 391

22. Principles of Freeze-Fracture Technique and Implications in Studies of Normal and CLL Lymphocytes 393 C.S. Cody and S.D, Douglas

V. SCANNING ELECTRON MICROSCOPY 403

23. Surface Morphology of Leukemic Cells as Seen by Scanning Electron Microscopy: Interpretation and Attempted Correlation with Cell Type in 212 Cases of Leuke-mia 405 A, Polliack, D. Gur/el, R, Leizerowitz. and H Gamliel

24. Scanning Electron Microscopy and Leukemia-Lymphoma Characterization 419 D, Dantchev and G, Mathe

25. Scanning Electron Microscopy of Hairy Cells from 15 Pa­tients with Hairy Cell Leuke­mia: Morphologic Subtypes 435 R,M, Hamilton, S. De Meester, and HM, Golomb

CONTENTS IX

VI. SCANNING IMMUNO-ELECTRON MICROSCOPY 451

26. Immuno Scanning Electron Mi­croscopy of Human Leukemic Cells: A Study of 50 Cases of Leukemia and 14 Established Leukemia-lymphoma Cell Lines 453 H. Gamliel. D. Gurlel. and A. Polliack

VII. SCANNING ELECTRON MICROS-COPY, CYTOCHEMISTRY 467

27. Scanning Electron Microscopic Cytochemistry of Leukemic Cells 467 D. Soligo. E. Pozzoli. and G. Lambertenghi-Deliliers

VIII. ULTRASTRUCTURE, IN-VITRO STUDIES OF LEUKEMIA 481

28. Ultrastructure of Cultured Leukemia Cell Lines. 483 c. Sundstrom

29. Viruses and Human Leukemia in Vitro. 495 A. Karpas

CONTRIBUTING AUTHORS

W. W. Ambrose. Laboratory of Cytostruc­ture and Cytochemistry, Dental Re­search Center, University of North Carolina, Chapel Hill, N.C. 27 514 .

D.F. Bainton. Department of Pathology, University of California School of Medi­cine, San Francisco, California 94143, U.S.A.

M.A. Bitter. Department of Medicine, Sec­tion of Hematology-Oncology, 950 East 59th Street, Chicago, Illinois 60637, U .S.A.

I Breton-Gorius. Unite INSERM U-91, Hopital Henri Mondor, 940101, Creteil, France.

R.D. Brunning. Department of Laboratory Medicine and Pathology, Box 198, Mayo Memorial Building, University of Min­nesota Hospitals, Minneapolis, Min­nesota 55455, U.S.A.

H.W. Carter. Department of Pathology, Duke University Medical Center, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27514, U .S.A.

D. Catovsky. MRC Leukaemia Unit, Royal Postgraduate Medical School, University of London, Ducane Road, London W12 OHS, United Kingdom.

C.S. Cody. The Children's Hospital of Philadelphia, Division of Allergy­Immunology-Pulmonology, University of Pennsylvania, Department of Pedi­atrics, School of Medicine, Philadelphia, Pennsylvania 19104, U.S.A.

C. Costello. MRC Leukaemia Unit, Royal Postgraduate Medical School, University of London, Ducane Road, London W12 OHS, United Kingdom.

x

ID. Cotelingham. Hematopathology, De­partment of Laboratory Medicine, Na­tional Naval Medical Center, Bethesda, Maryland 20814, U .S.A.

A.D. C rocka rd. Royal Victoria Hospital, Belfast, Ireland, United Kingdom.

M. T. Daniel. Laboraroire Central d'Hematologie, Hopital Saint-Louis, 2, Place Dr. A Fournier, 754 75 - Paris Cedex 10, France.

D. Dantchev. Institut de Cancerologie et d'Immunogenetique,Groupe Hospi­talier Paul-Brousse, 14 & 16 Avenue Paul Vaillant-Couturier, 94800 Villejuif, France.

S. De Meester. Department of Medicine, Section of Hematology-Oncology, 950 East 59th Street, Chicago, Illinois 60637 , U.S.A.

M. Vjaldefti. Department of Medicine B, Hematology Clinic and Electron Micros­copy Unit, Hasharon Hospital, Petah Tiqva and Tel-Aviv University Sackler School of Medicine, Tel-Aviv, Israel.

S.D. Douglas. The Children's Hospital of Philadelphia, Division of Allergy­Immunology-Pulmonology, University of Pennsylvania, Department of Pediat­rics, School of Medicine, Philadelphia, Pennsylvania 19104, U.S.A.

IP. Farcet. INSERM 0.91 and Laboratoire de Medicine Experimentale, Hopital Henri Mondor, 94010 Creteil, France.

G. Flandrin. Laboratoire Central d'Hematologie, Hopital Saint-Louis, 2, Place Dr. A. Fournier 75475 - Paris Cedex 10, France.

H. Gamliel. Deparcment of Hematology,

CONTRIBUTING AUTHORS Xl

Hadassah University Hospital, Jerusalem, Israel.

B.L. Giammara. Microelectronics Center of North Carolina, Research Triangle Park, N .C. 27514, U.S.A.

H.M. Golomb. Department of Medicine, Section of Hematology-Oncology, 950 East 59th Street, Chicago, Illinois 60637 , U.S.A.

M.F. Gourdin. INSERM U 91 , Hopital Henri Mondor, 94010 Creteil, France.

D. Gur/el. Lymphoma-Leukemia Unit De­partment of Hematology, Hadassah U ni­versity Hospital, Jerusalem, Israel.

R.M. Hamilton. Department of Medicine, Section of Hematology-Oncology, 950 East 59th Street, Chicago, Illinois 60637, U.S.A.

}.S. Hanker. Laboratory of Cytostructure and Cytochemistry, Dental Research Center, University of North Carolina, Chapel Hill, N.C. 27514, U.S.A.

D . Huhn. Medizinische Klinik III, Klinikum, Grophadern der Universitiit Munchen, Marchionistrasse 15, 8000 Munich 70, West Germany.

W.K. Hummer. Hematopathology, Depart­ment of Laboratory Medicine, National Naval Medical Center, Bethesda, Mary­land 20814, U.S.A.

N. Imamura. Department of Anatomy, Hiroshima University School of Medi­cine, Kasumi 1-2-3, Hiroshima, Japan.

E. Kaiser/ing. Department of Pathology, Christian Albrecht University, 2300 Kiel, West Germany.

S. Kamihara. Department of Anatomy, Hiroshima University School of Medi­cine, Kasumi 1-2-3, Hiroshima, Japan.

A . Karpas . Department of Haematological Medicine, Cambridge University Clin­ical School, Hills Road, Cambridge CB2 2QL, England.

K. Kataoka. Department of Anatomy, Hiroshima University School of Medi­cine, Kasumi 1-2-3, Hiroshima, Japan.

A. Kornberg. Department of Hematology, Hadassah University Hospital, Je­rusalem, Israel.

G. Lambertenghi-Defifien. Institute of Clinica Medica I, University of Milan, Via Francesco Sforza, 35, 20122 - Mi­lan, Italy.

}. Laszlo. Laboratory of Cytostructure and Cytochemistry, Dental Research Center, University of North Carolina, Chapel Hill, N.C. 2751 4, U.S.A.

R. Leizerowitz. Department of Hematol­ogy, Hadassah University Hospital, Jerusalem, Israel.

G. Mathe. Institut de Canccrologie et d'Im­munogenetique, Groupe Hospitalier Paul-Brousse, 14 & 16 Avenue Paul Vaillant-Couturier, 94800 Ville;uif, France.

E. Matutes. Hospital Clinico Provincial, Barcelona, Spain.

R.W . McKenna . Department of Laboratory Medicine and Pathology, Box 198 Mayo Memorial Building, University of Min­nesota Hospitals, Minneapolis, Min­nesota 55455, U.S.A.

}. Minowada . Departments of Pathology and Surgery, Loyola University, Stritch School of Medicine, Maywood, Illinois 60153, U.S.A.

}.D. Moore. Duke University Comprehen­sive Cancer Center, Duke University Medical Center, Durham, N .C. 277 10, U.S.A.

H.K. Muller-Hermelink. Department of Pathology, Christian Albrecht Univer­sity, 2300 Kiel, West Germany.

M. T. Nava . Institute of Clinica Medica I, University of Milan, Via Francesco Sforza, 35, 20122 - Milan, Italy.

Xli CONTRIBUTING AUTHORS

M. O'Brien. MRC Leukaemia Unit, Royal Postgraduate Medical School, University of London, Ducane Road, London W12 OHS, United Kingdom.

G.A. Pangalis. Leukemia and Lymphoma Clinic, Hematology Unit, First Depart­ment of Medicine, University of Athens School of Medicine, "Laikon" Hospital of Athens, Athens 609, Greece.

].L. Parkin. Department of Laboratory Medicine and Pathology, Box 198 Mayo Memorial Building, University of Min­nesota Hospitals, Minneapolis, Min­nesota 55455, U.S.A.

N. Pollio Institut of Clinica Medica I, Uni­versity of Milan, Via Francesco Sforza, 35, 20122 - Milan, Italy.

A. Polliack. Lymphoma-Leukemia Unit, Hematology-Oncology Department, Hadassah University Hospital and Hadassah-Hebrew University Medical School, Jerusalem, Israel.

E. Pozzoli. Institut of Clinica Medica I, University of Milan, Via Francesco Sforza, 35, 20122 - Milan, Italy.

E.S. Rappaport. Department of Pathology, Duke University Medical Center, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27514, U.S.A.

F. Reyes. INSERM U.91 and Laboratoire de Medicine Experimentale, Hopital Henri Mondor, 94010 Creteil, France.

D. Robinson. MRC Leukaemia Unit, Royal Postgrad uate Medical School, University of London, Ducane Road, London W12 OHS, United Kingdom.

P.P. Roussou. Leukemia and Lymphoma Clinic, Hematology Unit, First Depart­ment of Medicine, University of Athens School of Medicine, "Laikon" Hospital of Athens, Athens 609, Greece.

Y. Sakano. Department of Anatomy, Hiroshima University School of Medi­cine, Kasumi 1-2-3, Hiroshima, Japan.

M. Shamoto. Department of Pathology, Fujita-Gakuen University School of Medicine, Katsukake-cho, Toyoake­City, Aichi-prefecture 470-11, Japan.

H.R. Schumacher. Hematopathology, De­partment of Laboratory Medicine, Na­tional Naval Medical Center, Bethesda, Maryland 20814, U.S.A.

D. Soligo. Institute of Clinica Medica I, University of Milan, Via Francesco Sforza, 35, 20122 - Milan, Italy.

C. Sundstrom. Department of Pathology, University Hospital, S-751 85, Upp­sala, Sweden.

N.B. Tsavaris. Leukemia and Lymphoma Clinic, Hematology Unit, First Depart­ment of Medicine, University of Athens School of Medicine, "Laikon" Hospital of Athens, Athens 609, Greece.

]. Vardiman. Department of Pathology Section of Hematology-Oncology, 950 East 59th Street, Chicago, Illinois 60637, U.S.A.

P.E. Yates. Laboratory of Cytostructure and Cytochemistry, Dental Research Center, University of North Carolina, Chapel Hill, N.C. 27514, U.S.A.

PREFACE

In recent years excellent progress has been made in the development of improved cytochemical and ultrastructural tech­niques for electron microscopy in hematol­ogy and in particular in leukemia and lymphoma. Subtypes of these disorders, which are essentially classified according to cell phenotype and the degree of matura­tion, can be well defined today using a combination of immunological methods, light microscopy, cytochemistry, and elec­tron microscopy. Ultrastructural cyto­chemistry is now utilized routinely in leukemia in most major centers today and can also be performed using scanning elec­tron microscopy (SEM), as described in this book. Immuno electron microscopy used with or without simultaneous ultra­structural cytochemistry can also be em­ployed in the accurate classification of undifferentiated or unclassified leukemias, and studies of this nature are illustrated in this book. Undoubtedly some cases of so­called unclassified leukemia will be diag­nosed as early myeloblast leukemia, on the basis of positive ultrastructural myeloper­oxidase staining or as megakaryoblastic in nature if positive platelet peroxidase activ­ity is present on transmission electron microscopy (TEM). A few cases of so­called acute lymphoblastic leukemia (ALL) may also be reclassified after myelo- and platelet peroxidase staining is performed using TEM.

In some cases surface architecture of the leukemic cells as seen by scanning electron microscopy may contribute to the diag­nosis. The techniques for performing im­muno SEM and SEM cytochemistry are now available and may be utilized more in

the study of these disorders in the future. Recently freeze fracture techniques have also been employed in the study of normal and leukemic lymphocytes and much new information concerning leukemic cell membranes may be obtained from the use of this technique in the future.

The purpose of this book is to demon­strate the utility of these techniques in defining leukemic cell types and to illus­trate the potential of some of the newer ultrastructural methods, including trans­mission and scanning immuno electron microscopy, SEM cytochemistry, and freeze fracture techniques, in the study of leukemia. Advances in cytochemical tech­niques and their use in the definition of nonlymphoid cells and in the classification of lymphoid subtypes are clearly demon­strated in this book. The importance of the detection of platelet and myeloperoxidase activity and nonspecific esterases on the ultrastructural level, in acute unclassified leukemia and blastic crisis of chronic granulocytic leukemia (CGL) is clearly out­lined. Ultrastructural characteristics of the various types of ALL and in particular T­type leukemias and prolymphocytic leuke­mias are vividly illustrated in the chapters on ALL, and the contribution of ultrastruc­ture in the accurate definition of therapy­related panmyelosis, secondary leukemia, and blastic crisis of CGL is clearly stated.

Some of the chapters dealing with ALL contain overlapping information, which is unavoidable when different authors contribute on related subjects in an edited book. However, on the whole, these chap­ters are complementary, adding different dimensions and personal perspectives to

Xlll

XIV PREFACE

the morphology of these disorders. Of particular interest is the use of combined ultrastructural cytochemistry and immuno TEM as described by Breton-Gorius and Catovsky and co-workers. Scanning EM vividly illustrates in three dimensional views, the spectrum of surface features seen in the different leukemic cells while immuno SEM and SEM cytochemistry show again how combined ultrastructural approaches can be effectively used to de­fine different cell types. Ultrastructural aspects of cultured leukemic cells and some observations on virus production and

differentiation in vitro are also reviewed in this book. This also demonstrates how TEM can contribute to the understanding of cell differentiation and etiology in these disorders.

In general it is hoped that this volume will focus more attention on the impor­tance of ultrastructure in the definition of the leukemic cell phenotype and highlight the fact that much practical information can be obtained from these methods, which will influence decision making for patients with these disorders.