Cu rrent Topics in Microbiology 183 and Immunology Editors A. Capron, Lille . R. w. Compans, Atlanta/Georgia M. Cooper, Birmingham/Alabama· H. Koprowski, Philadelphia . I. McConnell, Edinburgh· F. Melchers, Basel M. Oldstone, La Jolla/California . S. Olsnes, Oslo M. Potter, Bethesda/Maryland . H. Saedler, Cologne P. K. Vogt, Los Angeles· H. Wagner, Munich· I. Wilson, La Jolla/California N.J. Dimmock Neutral ization of Animal Viruses With 10 Figures Springer-Verlag Berlin Heidelberg New York London Paris Tokyo Hong Kong Barcelona Budapest NIGEL J. DIMMOCK Department of Biological Sciences University of Warwick Coventry CV4 7 AL U.K. Cover illustration: Influenza virus, freeze-dry negative stain ing. (Picture by courtesy of Dr. M. V. Nermut, National Institute for Biological Standards and Control, Potters Bar, UK.) Cover design: Harald Lopka, IIvesheim ISBN-13: 978-3-642-77851-3 e-ISBN-13: 978-3-642-77849-0 001: 10.1007/978-3-642-77849-0 This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broad-casting, reproduction on microfilms or in other ways, and storage in data banks. Duplication of this publication or parts thereof is only permitted under the provisions of the German Copyright Law of September 9. 1965, in its current version, and a copyright fee must always be paid. Violations fall under the prosecution act of the German Copyright Law. © Springer-Verlag Berlin Heidelberg 1993 Softcover reprint of the hardcover 1s t edition 1993 Library of Congress Catalog Card Number 15-12910 The use of registered names, trademarks etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. Product liability: The publisher can give no guarantee for information about drug dosage and application thereof contained on this book. In every individual case the respective user must check its accuracy by consulting other pharmaceutical literature. Typesetting: Thomson Press (India) Ltd, New Delhi; Offsetprinting: Saladruck, Berlin; 23/3020-54 3 2 1 0 - Printed on acid-free paper. Contents 1 Introduction ..................... . 1 2 Immunoglobulin G Neutralization by Inhibition of Attachment of Virus to the Cell .......... . 3 3 Immunoglobulin G Neutralization Which Does Not Inhibit Attachment of Virus to the Cell ................ 10 4 Immunoglobulin G Neutralization by Aggregation of Virions 14 5 Immunoglobulin G Neutralization Mechanisms Which Operate After Attachment of the Virus-Antibody Complex to a Cell Receptor Unit ................. '. 17 5.1 Inhibition of Fusion atthe Plasma Membrane. . . . . 17 5.2 Inhibition of Endocytosis ................ 17 5.3 Inhibition of Fusion of Viral and Cellular Membranes 17 5.4 Inhibition of Non-fusion Uncoating .......... 21 5.5 Inhibition of Events Which Occur After Primary Uncoating 21 6 Neutralization Which Occurs by Virus Binding Antibody After It Has Attached to a Cell . . . . . 24 7 Role of the Cell in Neutralization ............. 26 8 Antibody-Dependent Enhancement of Infectivity by Neutralizing Antibody: Fc and Complement Receptors 30 9 Neutralization by Polymeric Immunoglobulin A 32 10 Neutralization by Immunoglobulin M 34 11 The Relevance of Immunoglobulin Isotype to Neutralization 37 12 Viral Carbohydrates, Proteins and Neutralization 39 12.1 Carbohydrates and Neutralization ............ . 39 12.2 Proteins and Neutralization ............... . 40 13 Properties of Protein and Peptide Antigens Which Elicit Neutralizing Antibody 44 14 Neutralization In Vivo 49 15 Complement and Neutralization 55 VI Contents 16 Neutralization by Inhibition of Release of Progeny Virus from the Infected Cell ................... . 58 17 Changes in Virus Proteins and Virion Structure on Binding Antibody, Including Synergistic Neutralization 59 18 Reversibility of Neutralization ...... . 64 19 Neutralization by Fragments of Antibody 67 20 Quantitative Aspects of Neutralization .. 71 21 Unconventional Neutralization ..... . 74 21.1 Genetic Engineering of Antibodies and Viruses 74 21.2 Anti-idiotype Antibodies and Neutralization .. 76 22 The Evolutionary Significance of Neutralization Sites 79 22.1 Why Do Viruses Have Neutralization Sites? ..... . 79 22.2 Strategies Which Avoid or Minimize Expression of, or Response, to Neutralization Sites 81 22.2.1 Relating to the Virus Particle ............. . 81 22.2.2 Relating to the Immune System ............ . 84 23 Neutralization of Poliovirus and Rhinovirus: A Summary 88 23.1 Introduction . 88 23.2 Attachment 90 23.3 Internalization ... 91 23.4 Post-internalization 92 23.5 Aggregation .. . . 92 23.6 Conformational Changes on Binding Antibody 95 24 Neutralization of Type A Influenza Virus by Immunoglobulins M, A and G: A Summary 98 24.1 Introduction .... 98 24.2 IgM Neutralization 100 24.3 IgA Neutralization 100 24.4 IgG Neutralization 101 24.5 Discussion ..... 102 25 Neutralization of HIV-1: A Summary 107 26 Conclusions ............. . 111 References . 112 Subject Index 147 Abbreviations FMDV, foot-and-mouth disease virus HA, Haemagglutinin HCMV, human cytomegalovirus H IV, H IV -1, human immunodeficiency virus type 1 HSV -1, herpes simplex virus type 1 influenza virus, influenza virus type A LCMV, lymphocytic choriomeningitis virus LDV, lactate dehydrogenase-elevating virus mab, monoclonal antibody M HV -4, mouse hepatitis virus type 4 NA, neuraminidase NANA, N-acetyl neuraminic acid NDV, Newcastle disease virus PIV-3, parainfluenza virus type 3 poliovirus, poliovirus type 1 RSV, respiratory syncytial virus SLEV, Saint Louis encephalitis virus TBEV, tick-borne encephalitis virus TG EV, transmissible gastroenteritis virus TM EV, Theiler's murine encephalomyelitis virus VEEV, Venezuelan equine encephalomyelitis virus VSV, vesicular stomatitis virus 1 Introduction Understanding neutralization is particularly relevant to an appreciation of the interaction between a virus and its antibody-synthesizing host since it is likely that viruses and the antibody system have evolved in response to reciprocally imposed selective pressures. Neutralization of viruses which only infect non-antibody-synthesizing hosts, while of considerable interest from a number of points of view is de facto without any such evolutionary signifi cance. In this second category are viruses of plants, invertebrates, vertebrates below fish in the evolutionary scale which do not synthesize antibody and most bacteria. Viruses of organisms parasitic on or commensal with antibody synthesizing vertebrates, such as enteric bacteria, protozoa or metazoan parasites, will be in contac, with antibody at some stage of their existence, and arthropod-borne viruses which have a higher vertebrate as second host are obviously bona fide members of the first category. There is an urgent need to understand the principles by which antibodies inactivate virus infectivity since, at present, we are unable to rationally construct effective vaccines against new agents like the human immuno deficiency viruses or to improve existing vaccines. The intention of this volume is to comprehensively review neutralization and where possible to construct a unifying theory which can be tested by experimentation. The major conclusion is that it is not possible to say that a virus is neutralized by anyone mechanism, but rather that there is a mechanism peculiar to the particular permutation of conditions prevailing; these include properties of the virus itself other than the relevant neutralization epitope, the neutralization epitope, the isotype of immunoglobin, the cell receptor and the virus: immuno globulin ratio. Thus a virus may be classified as being neutralized according to several different mechanisms, and the appearance of a virus under only one category of neutralization mechanism probably means that other mech anisms operating under different circumstances have not yet been uncovered. Some early work suffered from the (false) assumption that there was only one mechanism of neutralization, and interpretation of much of the early work was ambiguous because of the uncertainty of the composition of polyclonal antisera even when monospecific reagents were available. Nonetheless there is a wealth of important data in early reviews (FAZEKAS DE ST GROTH 1962; SVEHAG 1968; DANIELS 1975; DELLA-PORTA and WESTAWAY 1978; MANDEL 1979) and in some more recent articles (DIMMOCK 1984, 1987; MANDEL 1984, 1985; MCCULLOUGH 1986; COOPER 1987; IORIO 1988). Other reviews concentrate on specific groups of viruses: Arenaviridae (HOWARD 1986), 2 Introduction rotaviruses (MATSUI et al. 1989), foot-and-mouth disease virus (FMDV; MCCULLOUGH et al. 1992), human immunodeficiency virus (H IV; NARA et al. 1991; McKEATING 1992; PUTNEY 1992), polioviruses and rhinoviruses (MOSSER et al. 1989), Paramyxoviridae (NORRBY 1990), type A influenza viruses (OUTLAW and DIMMOCK 1991) and lentiviruses (PEDERSEN 1989) More general treatments of antibody-antigen interactions are GETZOFF et al. (1987, 1989), and of antibody structure BURTON (1990). 2 Immunoglobulin G Neutralization by Inhibition of Attachment of Virus to the Cell There are surprizingly few documented examples of immunoglobulin G (lgG) inhibiting attachment of virus to its cell receptor (Table 1). For this to occur it would seem that the IgG has to bind directly to or close by the viral attach ment site or to change its conformation so that it no longer functions. Table 1 includes only data which show some indication of direct proportionality between neutralization and inhibition of attachment. For this reason some data where authors have ascribed neutralization to inhibition of attachment have been excluded, for example Venezuelan equine encephalomyelitis virus (VEEV) where there was a 90% reduction in attachment for 99.998% neutrali zation (ROEHRIG et al. 1988), poliovirus where monoclonal antibody (mab) ICJ27 caused an 80% reduction in attachment but 99% neutralization (EMINI et al. 1983a) and transmissible gastroenteritis virus (TG EV) where mabs to site A of the major surface protein inhibited attachment by 78%-96% but neutralized by up to 99.999999% (SUNE et al. 1990). Another problem is that experimenters sometimes use only one concentration of IgG (e.g. LEE et al. 1981), which renders the conclusion that the observed inhibition of attach ment is the primary cause of neutralization open to doubt. This is aptly il lustrated by the interaction of TGEV with anti-site A mabs, where there is inhibition of attachment of already neutralized virus by a vast excess of antibody (SUNE et al. 1990). Thus much of the work cited in Table 1 should be appraised with caution. However, the study by IORIO et al. (1989) on Newcastle disease virus (N DV) pays very careful regard to the correspondance between inhibition of attachment and neutralization and shows clearly that some neutralizing mabs directed against the haemagglutinin-neuraminidase (HN) protein block attachment while others do not; the latter inhibit neither haemagglutination nor neuraminidase (NA) activity. Another group of mabs gives a degree of inhibition of attachment which is less than the observed neutralization. It is also noteworthy that attachment of rhinovirus 14 was diminished even by Fab fragments of IgG (COLONNO et al. 1989) and, further more, this occurred with Fabs reactive with any of the four major antigenic sites. Neutralizing IgG to reovirus is directed against the 0'1 protein, which is the attachment protein (WEINER and FIELDS 1977; LEE et al. 1981), but the relationship between the neutralization site and the attachment site(s) on 0'1 is not yet clear. Again there is the unresolved problem of quantitation mentioned above where a mab (G-5) with a neutralization titre of 1/12500 (BURSTIN et al. 1982) inhibited attachment at a dilution of 1 /1 0 by only 60% (MARATOS- ~ 3' 3 c: ::J 0 cc 0' 0" c: 5' C) z CD S. iil N' '" ..... 0' ::J 0" -< :;-:::T C' ;:j: 0' -::J 0 ~ '" 0 :::T 3 CD a 7 8 9 1 References SUNE et al. 1990 KINGSFORD et al. 1991 NEMEROW et al. 1987 MILLER and HUTT-FLETCHER 1988 a; FULLER and SPEAR 1985HIGHLANDER et al. KUHN et al. 1990 OUBUISSON et al. 1990c,d EISENLOHR et al. 1987; OUTLAW et al. 1990e; OUTLAW and OIMMOCK 1993" aIORIO et al. 1989, 1991 a KELLER 1966; see Fig. 7 BAXT et al. 1984"; Fox et al. 1989 COLONNO et al. 1989 LEE et al. 1981 SABARA et al. 1985; KUKUHARA et al. 1988 RUGGERI and GREENBERG 1991 KENNEDy-STOSKOPF and NARAYAN 1986 MASSEY and SCHOCHETMAN 1981 aC aFIELDS et al. 1988; Ho et al. 1988; SUN et al. 1989POSNER et al. 1991 1, HANSEN et al. 1991 c Table 1. IgG neutralization by inhibition of attachment of virus to the cell Comment Family Virus Anti-B; anti-C, anti-O Corona TGEV Bunya La Crosse virus Site A a Anti-gp350 mabsHerpes Epstein-Barr virus Anti-gp350, anti-gp220 aAnti-gC, anti-gOa HSV-1 Anti-gBa; anti-gOa Bovine herpes virus 4 Myxo Influenza virus Paramyxo NOV Anti-HN Picorna Poliovirus types 2 and 3b FMOV Anti-VP1,141-160 Rhinovirus Reo Reovirus Rotavirus Anti-VP7 (38K)a, anti-VP4 (VP8*)" Retro Maedi-visna virus Fibroblasts only Mouse mammary tumour virus HIV-1 Anti-gp120a Anti-gp120 carbohydrate Only some antibodies; others do not inhibit attachment. " No mabs to type 1 inhibit attachment. b Inferred as mabs fail to neutralize when added after attachment. C Neutralization with pairs of mabs; maximum 25%-30% neutralization. d Contribute to but do not fully account for neutralization. e Some mabs neutralize poorly. 1