Table Of ContentHUMAN INTERFERON
Production and Clinical U se
ADV ANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY
Editorial Board:
Nathan Back S''''e University 01 New York", Bul/alo
N. R. Di Luzio Tulane University School 01 Medicine
Bemard Halpern Colle«e de FrtJllCe and Ins,itu'e ollmmuno-Biology
Ephraim Katchalski The W' eizmGlln/ns'itute 01 Scienee
David Kritchevlky W'isear Insiitu'e
Abel Lajtha New York Sea'e Research Ins'i'u'e lor Neurochemis,ry and DruB A.ddic'iol
Rodolfo Paoletti Universi,y 01 Milan
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HUMAN INTERFERON: Ptoduction and C1inical Use
Edited by Warren R. Stinebring and Paul J. Chappie
HUMAN INTERFERON
Production and Clinical Use
Edited hy
Warren R. Stinebring
The University of Vermont
Burlington, Vermont
and
Paul J. Chapple
W. Alton Jones Cell Science Center
Lake Placid, New Yo rk
PLENUM PRESS • NEW YORK AND LONDON
Library of Congress Cataloging in Publication Data
Interferon Workshop on Production of Human Interferon and Investigations of its Clinical
Use, W. Alton J ones Cell Science Center, 1977.
Human interferon: Production and clinical use.
(Advances in experimental medicine and biology; v. 1l0)
"Proceedings of the Interferon Workshop on Production of Human Interferon and In
vestigations of its Clinical Use, held at the W. Alton Jones Cell Seien ce Center, Lake Placid,
NY., May 19-22, 1977."
Includes index.
1. Interferons-Congresses. 2. Interferons-Therapeutic use-Congresses. I. Stinebring,
Warren R. II. Chapple, Paul J. III. Tide. IV. Series. [DNLM: 1. Interferon-Congresses. Wl
AD559 v. 1l0/QW800 161p 1977)
QR187.5.I572 1977 616.01'94 78-13393
ISBN-13: 978-1-4615-9082-8 e-ISBN-13: 978-1-4615-9080-4
DOI: 10.1007/978-1-4615-9080-4
Proceedings of the Interferon Workshop on Production of Human Interferon
and Investigations of its Clinical Use held at the W. Alton Jones Cell
Science Center, Lake Placid, New York, May 19-22, 1977
© 1978 Plenum Press, New Yo rk
Softcover reprint ofthe hardcover 1st edition 1978
A Division of Plenum Publishing Corporation
227 West 17th Street, New York, N. Y. lOOll
All righ ts reserved
No part of this book may be reproduced, stored in a retrieval system, or transmitted,
in any form or by any means, electronic, mechanicaI, photocopying, microfilming,
recording, or otherwise, eithout written permission from the Publisher
PREFACE
This vo1ume sterns from a workshop held at the W. A1ton Jones Ce11
Science Center, Lake P1acid, New York, May 19-20, 1977. The Cen
ter is an operationa1 unit of the Tissue Cu1ture Association and
offers, in co11aboration with the Association's Education Commit
tee, a wide range of activities. One of the ear1iest symposia
held at the Center was one concerned with Interferon, (waymouth,
C. (Ed.) 1973. The Production and Use of Interferon for the Treat
ment and Prevention of Human Virus Infections. In Vitro, Mono
graph No. 3. Tissue Cu1ture Association, Rockvi11e, Mary1and.) In
the intervening years since that symposium, many deve10pments
have taken p1ace. Much has been 1earned of the mo1ecu1ar bio1ogy
of interferon. However, the focus of this particu1ar workshop,
as the tit1e indicates, was the production of human interferon and
investigation of its c1inica1 use.
Evidence has accumu1ated that interferon is an effective antiviral
and, possib1y, an anti tumor agent. The production of interferon
requires human ce11s and proper inducing agents. Present produc
tion faci1ities are 1imited. The quantities of interferon needed
for c1inica1 trials and eventual routine demand the deve10pment of
new techniques in the growth and handling of ce11s and inducers.
Safety is an important consideration in the production of bio1ogica1
products, and interferon is no exception. All components of the
system and the final product must satisfy stringent safety standards.
The participants of the workshop dealt with all of the above pro
blems on a practica1 level. The next few years will see advances
in the techno1ogy of production of interferon. The future will
also see more widespread investigation of the antiviral and anti
tumor ro1es of interferon. In addition, the 1ymphokine-1ike pro
perties of interferon, the ability to affect ce11 growth and the
ability of interferon to affect lymphoid ce11s will all be in
vestigated with the under1ying purpose of uti1izing interferon
in the treatment of human disease.
v
PREFACE
Thus, there will be an ever increasing need for larger and larger
quantities of purified interferon. The editors and all those who
took part in the workshop sincerely hope that this volume will act
as a catalyst in bringing about the availability in sufficient
quantity that it can be used for therapeutic purposes in a routine
manner.
The editors wish to acknowledge the support and assistance of the
Education Committee of the Tissue Culture Association and the
financial support of the Bureau of Biologics of the Food and Drug
Administration, National Institute of Allergy and Infectious
Diseases, National Cancer Institute, and the Fogarty International
Center.
This volume would not have been possible without the hard work
and perseverance of Mrs. Ellen Anastos-Dorchak, who kept the editors
on track and who supervised the typing of all the manuscripts in a
form ready for publication.
CONTENTS
Large Scale Production and Properties of Human Leukocyte
Interferon Used in Clinical Trials . . . . 1
H.-L. Kauppinen, G. Myllylä, and K. CanteIl
New Microcarriers for the Large Scale Production of
Anchorage-Dependent Mammalian Cells . 15
D.W. Levine, W.G. Thilly, and D.I.C. Wang
Effect of Low Levels of Cyclic Ribonucleotides on Mitogen
and Virus Induced Production of Interferon 25
H.M. Johnson and S. Baron
Production of Interferon in Human Cell Cultures by a New,
Potent Viral Inducer . . . . . . . . . . . 37
P. Jameson and S.E. Grossberg
Human Interferon: Large Scale Production in Embryo
Fibroblast Cultures •..• .•.• 55
V.G. Edy, J. VanDamme, A. Billiau, and P. DeSomer
Factors Influencing Production of Interferon by Human
Lymphoblastoid Cells . • . • 61
M.D. Johnston, K.H. Fantes, N.B. Finter, and B. Chir
Antigenie Properties and Heterospecific Antiviral
Activities of Human Leukocyte Interferon Species.. 75
K. Paucker, B.J. Dalton, and E.T. Torma
Membrane Alterations Following Interferon Treatment • . . .. 85
E.H. Chang, E.F. Grollman, F.T. Jay, G. Lee, L.D. Kohn,
and R.M. Friedman
Selection of New Human Foreskin Fibroblast Cell Strains
for Interferon Production . . . . . . . . . . 101
J. Vilcek, E.A. Havell, M.L. Gradoville, M. Mika-Johnson,
and W.H.J. Douglas
vii
viii CONTENTS
Tissue Culture Models of -In -Viv-o Interferon Production
and Action . . • . . . . . . . . • . . . 119
F. Dianzani, I. Viano, M. Santiano, M. Zucca,
P. Gullino, and S. Baron
Thermal and Vortical Stability of Purified Human Fibroblast
Interferon . . •. ....... . 133
J.J. Sedmak, P. Jameson, and S.E. Grossberg
Interferon Assay Anomaly Variation of Interferon Response
with Cell Type and Sialic Acid Content . . . . 153
A.A. Schwartz and D. Villani-Price
Interferon Therapy for Neoplastic Diseases in Man In Vitro
and In Vivo Studies. ...... ..... 159
---
S. Einhorn and H. Strander
Results of a Five-Year Study of the Curative Effect of Double
Stranded Ribonucleic Acid in Viral Dermatoses and
Eye Diseases .. . . . . . . . • . . . . . . 175
L. Borecky, J. Buchvald, E. Adlerova, I. Stodola,
E. Obrucnikova, Z. Gruntova, V. Lackovic, and
J. Doskocil
The Possibility of Interferon Production in Tumors 193
W.R. Stinebring and S. Jenkins
The Future of Interferon as an Antiviral Drug . . . . . . .. 201
J.K. Dunnick and G.J. Galasso
Considerations in Our Search for Interferons for Clinical
Use 213
R.Z. Lockart, Jr. and E. Knight
Index 217
LARGE SCALE PRODUCTION AND PROPERTIES OF HUMAN LEUKOCYTE
INTERFERON USED IN CLINICAL TRIALS
H.-L. Kauppinen, M.Sc., G. Myllylä, M.D.,
and K. CanteIl, M.D.
Finnish Red Cross Blood Transfusion Centre and
Central Public Health Laboratory, Helsinki, Finland
INTRODUCTION
Human leukocyte interferon has been produced for clinical use
in our laboratories for several years (I). A routine procedure
for the production and purification has been established (2,3),
but studies to improve the recovery and quality of the clinical
interferon continue. Since the availability of fresh leukocytes
is the factor limiting production, maximization of the interferon
yield per blood unit is of prime importance.
In clinical trials with interferons some side-effects including
pyrogenicity have appeared. These have prompted us to analyse the
impurities of the interferon preparations to see whether the side
effects could be eliminated by further purification. This paper
describes our re cent studies on the production of the human
leukocyte interferon for clinical use and on the properties of the
interferon preparations employed in the current clinical trials.
PRODUCTION
Storage of Blood
Routinely, only buffy coats from fresh blood are collected.
In practice the collection takes place 2-5 hours after donation.
The buffy coats are stored overnight at 4°C and used the next day
for interferon production. If production is started immediately
after harvesting the cells, the interferon titers are only slightly
higher. If the buffy coats are harvested from one-day-old blood and
in addition stored overnight so that the total storage time is two
2 H.·L. KAUPPINEN, G. MYLL YLÄ, AND K. CANTELL
days, the interferon titers are clearly reduced. With additional
storage the titers are reduced further (Fig. 1).
8
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z 4
0
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Figure 1
Production of interferon by leukocytes stored for different per iods
of time at 4°C.
When the blood is stored before harvesting the buffy coat, the
leukocytes separate in centrifugation as a more distinct layer
and more leukocytes can be recovered from the same volume. The
maximum yield of leukocytes counted after purification of the cells
is gained when the blood is stored for one day before harvesting
