Abstract
DNA vaccines encoding the idiotype of immunoglobulins of tumour B cells were shown to induce protection in several mouse lymphoma models. The mechanism of rejection of tumour cells has not been fully understood, but there is strong evidence suggesting that engagement of the idiotype by anti-idiotypic antibodies may directly result in inhibition of tumour growth. In this study, we have investigated the structural basis of the idiotypic/anti-idiotypic interaction following immunisation with DNA vaccines. scFvs containing only one of the two tumour-derived V regions recombined to an irrelevant V region partner were generated. These constructs encoding a secretory form of the scFv were used as immunogens to induce anti-Id antibodies. The same scFvs were expressed as membrane-bound molecules on the surface of mammalian cells. Analysis of immune sera on the membrane-displayed idiotypes revealed that DNA immunisation induced a polyclonal antibody response restricted to conformational combined epitopes formed by the parental VL/VH association. Immune sera raised by scFv DNA vaccination did not show any detectable reactivity towards chimeric scFvs containing only one of the two immunising V regions, indicating that the response against combined VL/VHdeterminants is highly dominant. Remarkably, the same immunogen, delivered as scFv protein, induced antibodies also directed against chain-specific determinants. These findings indicate that presentation of properly folded idiotypes results in a highly specific antibody response directed exclusively to private idiotypic determinants of the VL/VH combination of the immunogen.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Campbell M et al. Idiotype vaccination against murine B cell lymphoma: humoral and cellular responses elicited by tumour derived IgM and its molecular subunits J Immunol 1987 139: 2825–2833
Kaminski MS, Kitamura K, Maloney DG, Levy R . Idiotype vaccination against murine B cell lymphoma: inhibition of tumour immunity by free idiotype protein J Immunol 1987 138: 1289–1296
George AJT, Folkard SG, Hamblin TJ, Stevenson FK . Idiotypic vaccination as a treatment for a B cell lymphoma J Immunol 1988 141: 2168–2174
Syrengelas AD, Chen TT, Levy R . DNA immunisation induces protective immunity against B-cell lymphoma Nat Med 1996 2: 1038–1041
Hakim I, Levy S, Levy R . A nine ammino acid peptide from IL-1b aguments anti-tumour immune responses induced by protein and DNA vaccines J Immunol 1996 157: 5503–5511
King CA et al. DNA vaccines with single-chain Fv fused to fragment C of tetanus toxin induce protective immunity against lymphoma and myeloma Nat Med 1998 4: 1281–1286
Benvenuti F, Burrone OR, Efremov DG . Anti-idiotypic DNA vaccines for lymphoma immunotherapy require the presence of both variable region genes for tumour protection Gene Therapy 2000 7: 605–611
Tutt A et al. Monoclonal antibody therapy of B cell lymphoma: signaling activity on tumour cells appears more important than recruitment of effectors J Immunol 1998 161: 3176–3185
Illidge TM, et al. The importance of antibody-specificity in determining successful radioimmunotherapy of B-cell lymphoma Blood 1999 94: 233–243
Ito HO, Ueda Y, Hashimoto T, Koga T . Quaternary structure-dependent idiotype and antigen binding of a monoclonal antibody specific for conformational epitope on type II collagen Cell Mol Life Sci 1997 53: 51–60
Kanda S, et al. Both VH and VL regions contribute to the antigenicity of anti-idiotypic antibody that mimics melanoma associated ganglioside GM3 Cell Biophys 1994 65: 24–25
Hall BL, Zaghouani H, Daian C, Bona A . A single amino acid mutation in CDR3 of the 3–14–9 L chain abolished expression of the IDA 10-defined idiotype and antigen binding J Immunol 1992 149: 1605–1612
Sollazzo M et al. Structural definition by antibody engineering of an idiotypic determinant Prot Eng 1991 3: 531–539
Weissenhorn W et al. VH-related idiotopes detected by site directed mutagenesis J Immunol 1992 149: 1237–1241
Potter KN et al. Molecular characterisation of a cross-reactive idiotope on human immunoglobulin utilizing the VH4–21 gene segment J Exp Med 1993 178: 1419–1428
Haba S, Lascombe MB, Poljak RJ, Nisonoff A . Structure of idiotypes associated with antiphenylarsonate antibodies expressing an intrastrain crossreactive idiotype J Exp Med 1989 170: 1075–1790
Li E et al. Mammalian cell expression of dimeric small immune proteins (SIP) Prot Eng 1997 10: 731–736
Li E, Bestagno M, Burrone O . Molecular cloning and characterization of a transmembrane surface antigen in human cells Eur J Biochem 1996 238: 631–6388
Batista FD et al. The two membrane isoform of human IgE assemble into functionally distinct B cell antigen receptors J Exp Med 1996 184: 2197–2205
Bestagno M et al. Membrane immunoglobulins are stabilised by inter-chain disulphide bonds occurring within the extracellular membrane-proximal domain Biochemistry 2001 (in press)
George AJT et al. Monoclonal antibodies raised against the idiotype of the murine B cell lymphoma BCL1 act primarily with heavy chain determinants Hybridoma 1991 10: 219–227
Syrengelas AD, Levy R . DNA vaccination against the idiotype of a murine B cell lymphoma: mechanism of tumour protection J Immunol 1999 162: 4790–4795
Timmerman JM, Levy R . Linkage of foreign carrier protein to a self-tumour antigen enhances the immunogenicity of a pulsed dendritic cell vaccine J Immunol 2000 164: 4797–4803
Ellgaard L, Molinari M, Heleniuos A . Setting the standards: quality control in the secretory pathway Science 1999 286: 1882–1888
Cragg MS, Zhang L, French RR, Glennie MJ . Analysis of the interaction of monoclonal antibodies with surface IgM on neoplastic B-cells Br J Cancer 1999 79: 850–857
Elliott TJ, Glennie MJ, McBride HM, Stevenson GT . Analysis of the interaction of antibodies with immunoglobulin idiotypes on neoplastic B lymphocytes: implications for immunotherapy J Immunol 1987 138: 981–988
Patricia KA et al. Evidence for an upper affinity threshold for anti-IgM-induced apoptosis in a human B-cell lymphoma Blood 1998 92: 3756–3771
Slavin S, Strobin S . Spontaneous murine B-cell leukaemia Nature 1978 272: 624–626
Batista FD, Efremov D, Burrone OR . Characterization of a second secreted IgE isoform and identification of an asymmetric pathway of IgE assembly Proc Natl Acad Sci USA 1996 93: 3399–3404
Acknowledgements
F Benvenuti was supported by a SISSA predoctoral fellowship. We are grateful to Mauro Sturnega and Dario Pippan for excellent technical assistance. We thank Professor Freda Stevenson for providing the BCL1 specific mAb 10/4A12 and the 123bcl1 hybridoma cell line. We also thank Facundo Batista and Luca Vangelista for helpful comments on the manuscript.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Benvenuti, F., Burrone, O. Anti-idiotypic antibodies induced by genetic immunisation are directed exclusively against combined VL/VH determinants. Gene Ther 8, 1555–1561 (2001). https://doi.org/10.1038/sj.gt.3301546
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3301546
Keywords
This article is cited by
-
Construction of miniantibodies for the in vivo study of human autoimmune diseases in animal models
BMC Biotechnology (2007)
-
BCL1 lymphoma protection induced by idiotype DNA vaccination is entirely dependent on anti-idiotypic antibodies
Cancer Immunology, Immunotherapy (2005)