Flow and Image Cytometry

245 Pages·1996·6.851 MB·English

by  Alemseged Truneh, Manjula Reddy, Patricia Ryan, Sally D. Lyn, Ilona Kariv (auth.), Alain Jacquamin-Sablon (eds.)

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Series H: Cell Biology, Vol. 95
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Flow and Image Cytometry
Edited by
Alain Jacquemin-Sablon
Centre National de la Recherche Scientifique
Laboratoire de Cytometrie (UPS 47)
7, rue Guy Moquet
F-94800 Villejuif, France
Springer
Published in cooperation with NATO Scientific Affairs Division
Proceedings of the NATO Advanced Study Institute "Progress in Flow and
Image Cytometry", held at Villejuif, France, May 15-19, 1995
Library of Congress Cataloging-in-Publication Data
NATO Advanced Study Institute "Progress in Flow and Image Cytometry"
(1995 Vi llejuif, France)
Flow and image cytometry I edited by Alain Jacquemin-Sablon.
p. cm. -- (NATO ASI series. Series H, Cell biology; vol.
95)
"Published in cooperation with NATO Scientific Affairs Division."
"Proceedings of the NATO Advanced Study Institute "Progress in
Flow and Image Cytometry" held at Villejuif, France, May 15-19,
1995"--CIP t.p. verso.
Includes bibliographical references and index.
1. Flow cytometry--Congresses. 2. Imaging systems in biology-
-Congresses. I. Jacquemin-Sablon, Alain. II. Title. III. Series.
QH585.5.F5SN38 1995
574.87'028--dc20 95-51230
CIP
ISBN-13: 978-3-642-64701-7 e-ISBN-13: 978-3-642-61115-5
001: 10.1007/978-3-642-61115-5
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© Springer-Verlag Berlin Heidelberg 1996
Softcover reprint of the hardcover 15t edition 1996
Typesetting: Camera ready by authors/editors
Printed on acid-free paper
SPIN 10477102 31/3137 -5 4 3 210
FOREWORD
The second NATO-sponsored International Cytometry course, "Progress in Flow and
Image Cytometry", was specifically structured to provide the most advanced techniques in
flow and image cytometry currently available. The course provided two options for specialized
training in the areas of (a) membrane dynamics and function (Ophon A) and (b) cell
proliferation and sene expression (Option B). Twenty-four candidates were accepted for each
course option which was composed of lecture and practical, "hands-on" laboratory exercices.
The candidates were chosen on the basis of their interest and experience in the respective
subject areas. A core curriculum consisting of topics on data management, control of cell
proliferation, apoptosis, new probes in cytometry, cell sorting, and near field microscopy was
presented to all participants.
Membrane transport and the mechanisms which control molecular traffic across
plasma membranes are of considerable importance in attempts to modulate disease states and
theses topics were addressed in Option A of the Course. Membrane pumping dynamics were
included in the laboratory exercices as were topics on cell-cell interactions and T cell homing.
Techniques using both flow and image cytometry for studying membrane transport and cell
recognition are currently available. Advances in these areas have been made by the
availability of new reagents that facilitate the speed and accuracy of the assays. Membrane
fusion and fission in endocytosis and membrane fluidity and pinocytosis are the focus of many
cytological investigations since the mechanisms involved are important to the understanding
of many normal physiological processes as well as numerous disease conditions.
Recent developments in multiparameter flow cytometry (FCM) that provide for
correlated analysis of a variety of cell kinetic parameters known to regulate and control cell
proliferation were emphasized in Option B of the Course. Bromodeoxyuridine (BrdUrd)
mcorporation studies, for example, have significantly improved analysis of cell cycle
progression, and the use of monoclonal antibodies in FCM immunofluorescence studies are
providing new techniques for examining and quantitating proliferation markers (i.e., PCNA,
Ki-67, pS3 and cyclins) in different phases of the cell cycle. In addition, the role of apoptosis in
regulahon of heterogeneous cell types and in the induction of cell damage by various cycle
perturbing agents is a primary aera of study today. Molecular cytogenetics as assayed both by
flow and Image cytometry have improved our understanding of chromatin organization and
function. Studies on cell differentiation in normal cells are extremely important since
determining the mechanisms involved could, alternatively, lead to a better understanding of
how cells become transformed.
Data handling for analysis of univariate and multivariate distribution continues to be
increasingly more complex, especially as the capabilities for assaying the numbers of variables
increase WIth each new reagent or instrumentation feature that comes on the market. Cluster
analysis, which attempts to analyze and correlate all the variables, is being explored in a
number of laboratories. Data obtained from clinical samples stamed with a battery of
monoclonal antibodies serves as an excellent model for demonstrating the difficulties involved
in such analysis. Coupled with the methods is the requirements for sorting and microscopic
examination for precise identification of each subpopulation. Cell sorting features are
available on many flow instruments but rarely do the users have the opportunity to obtain
the proper instruction for sorting except in courses such as these. The fields of flow and image
cytometry are advancing so qUIckly and -the increase in the number of new techniques is so
rapid only the continuahon of these NATO sponsored courses can serve to provide the training
necessary for the investigators to implement the most up-to-date technology in their research.
Courses such as these, that include both lectures and laboratory experiences, are rare and
expensive, and the support provided by NATO not only provides this unique educational
opportumty but also represents a significant contribution to the international scientific
community.
Co-Directors:
Dr. Alain Jacquemin-sablon
CNRS, Laboratoire de Cytometrie, 94800 Villejuif France
Dr. Harry A Crissman
Los Alamos National Laboratory, Los Alamos, NM 87545 USA
Acknowledgemen t8
Generous support from the following Institutions and Companies is gratefully
acknowledged:
-The Centre National de la Recherche Scientifique (CNRS), the Association pour la
Recherche sur Ie Cancer (ARC, President,jacques Crozemarie)
- BECTON DICKINSON
- BIO-RAD
- CELL ROBOTICS
- COHERENT
- COULTRONICS
- MERIDIAN (D.G.L. Bioscience)
- ORTHO Diagnostic Systems
- PARTEC
We also wish to thank:
- Martine BARDISSA
- Claude BOUCHEIX
- Dominique LAIRD
- Linda PRITCHARD
-Arlette VERVISCH
for their expert assistance during the whole Course.
TABLE OF CONTENTS
PART I : MEMBRANE DYNAMICS AND FUNCTION
Analysis of CDZ8 Interactions with Its Cognate Counter-Receptors 3
CD80 and CD86
A. Truneh, M. Reddy, P. Ryan, S.D. Lyn, I. Kariv, Ch. Eichman, D. Couez, M.R Hurle,
RP. Sekaly, D. Olive and R Sweet
Membrane Transport Dynamics 21
J.V. Watson and C. Dive
Endogenous Lectins in Circulating Cells and Their Glycosylated Ligands : 47
Their Role in T Cell Homing
V. Denis, M. Mitterand and C. Kieda
Use of Different Cytometric Techniques to Study the Cytotoxic Interaction 53
between Human Natural Killer Cells and K56Z Target Cells
K. Radosevic, B.G. de Grooth andJ. Greve
In Vitm Reconstitution of Early Endosome Membrane Dynamics 69
JP. Gorvel and Z. Mishal
Change in Membrane Fluidity After Pinocytosis of a New cytokine IK 81
on Resting and Activated T cells
R Pereno, P. Krief and Z. Mishal
PART II: CELL PROLIFERATION AND GENE EXPRESSION
Cell Cycle and Cell Proliferation Markers 91
H.A. Crissman and A.J. Nastasi
Advances in Flow Cytogenetics: Progress in the Development of a High Speed 103
Optical Chromosome Sorter Based on Photoinduced Adduct Formation Between
psoralens and chromsomal DNA
M.e. Roslaniec, RJ. Reynolds,J.e. Martin, K. Taek Han and L.S. Cram
Analysis of Cell Death by Flow Cytometry 115
Z. Darzynkiewicz and X.Li
Molecular Cytogenetics: Uses of Flow Sorted Chromosomes, Fluorescence 131
in situ Hybridisation (FISH) and Digital Microscopy for the Analysis of Genomes
N.P. Carter
DNA Topoisomerases as Drug Targets and Cell Cycle Checkpoint Effector Molecules 143
P.J. Smith, N. Blunt and S. Soues
VIII
HemopoIetic Cell Differentiation and Death by Retinoids. 155
D. Delia, A. Aiello, M.A. Pierotti
PART III: DATA MANAGEMENT SYSTEMS
CELL SORTING TECHNIQUES
MICROSCOPY
Myc in The Control of Proliferation and Apoptosis 169
1. Brown and G. Evan
flow Cytometry : Analyses for All Sizes 179
J.H.Jett
flow Cytometric Immunophenotyping Using Cluster Analysis and Cluster Editing 191
C.c. Salzman, R.J. Beckman,J.D. Parson, A.M. Nauman, SJ. Stewart and c.c. Stewart
Introduction to High-Speed flow Sorting 213
J.F. Keij
Applications of Near-Field Microscopes to Cell Biology 229
J.Barbet,J. Thimonier andJ. Rocca-Serra
Subject Index 237
P~I
Mem6rane t1Jynamics and l'unction
ANAL YSIS OF CD28 INTERACTIONS WITH ITS COGNATE COUNTER
RECEPTORS CD80 AND CD86.
Alemseged Truneh, Manjula Reddy, Patricia Ryan, Sally D.Lyn, liona Kariv, Christopher
Eichman, Dominique Couez, Mark R.Hurle1, Raffick P. Sekaly\ Daniel Olive3 and
Raymond Sweet.
Department of Molecular Immunology
SmithKline Beecham Phannaceuticals,
King of Prussia, Pennsylvania 19406, USA
Keywords:
CD28, CTLA-4, CD80, CD86, B7.I, B7.2, B70, IgSF, Ig-fold, Site Directed Mutagenesis,
Receptor Recognition, Epitope Mapping.
Summary
CD28 serves as a co-signalling molecule for T cell activation through binding to its cognate
counter-receptors CD80 and CD86, expressed on antigen presenting cells. This report summarizes
studies conducted to determine the regions of CD28 which are involved in its interactions with
CD80 and CD86, using site directed mutagenesis, CD28 mAb epitope mapping, receptor based
adhesion assays and direct binding of Ig-fusion proteins to cell surface receptors. These studies
I Department ofM acromolecular Sciences, SmithKline Beecham Pharmaceuticals, King of
Prussia, Pennsylvania 19406 USA. 2IRCM, Montreal, Canada. 3INSERM Vl19,
Marseille, France
NATO ASI Series. Vol. H 95
Flow and Image Cytometry
Edited by Alain Jacquemin-Sablon
© Springer-Verlag Berlin Heidelberg 1996