Skip to main content
SpringerLink
Log in

Development of a Transgenic Mouse Model Immune Tolerant for Human Interferon Beta

  • Research Paper
  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

Purpose

Therapeutic proteins may induce antibodies that inhibit their efficacy or have other serious biological effects. There is a great need for strategies to predict whether a certain formulation will induce an immune response. In principle, conventional animals develop an immune response against all human proteins no matter how they are formulated, which restricts their use. The aim of this study was to develop a mouse model immune tolerant for human interferon beta (hIFNβ).

Methods

A transgenic mouse model immune tolerant for hIFNβ was developed by making C57Bl/6 mice transgenic for the hIFNβ gene. To evaluate the model, both wild-type and transgenic mice were immunized with recombinant human interferon beta 1a (rhIFNβ-1a) and recombinant human interferon beta 1b (rhIFNβ-1b). Serum antibodies against rhIFNβ were detected by ELISA.

Results

The genetically modified mice were shown to be immune tolerant for mammalian cell–derived rhIFNβ-1a, which has a relative low immunogenicity in patients. However, Escherichia coli–derived rhIFNβ-1b, known to have a relatively high immunogenicity in patients, was shown not only to be immunogenic in the wild-type mice but could also break the immune tolerance of the genetically modified mice.

Conclusions

This animal model offers the possibility to study the many factors influencing the immunogenicity of hIFNβ and test new formulations before going into clinical trials. The model also provides the first evidence that the rhIFNβs differ in the immunological mechanisms responsible for the development of antibodies.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

ABTS:

2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)

BAb:

binding antibody

CHO cells:

Chinese hamster ovarian carcinoma cells

E. coli :

Escherichia coli

ELISA:

enzyme-linked immunosorbent assay

HSA:

human serum albumin

IFNβ:

interferon beta

MS:

multiple sclerosis

NAb:

neutralizing antibody

PBS:

phosphate-buffered saline

PCR:

polymerase chain reaction

hIFNβ:

human interferon beta

i.m.:

intramuscular

i.p.:

intra-peritoneally

i.v.:

intravenously

polyICLC:

polyinosinic:polycytidylic acid complexed with poly-L-lysine and carboxymethylcellulose

rhIFNβ:

recombinant human interferon beta

s.c.:

subcutaneous

References

  1. N. Casadevall J. Nataf B. Viron A. Kolta J. J. Kiladjian P. Martin-Dupont P. Michaud T. Papo V. Ugo I. Teyssandier B. Varet P. Mayeux (2002) ArticleTitlePure red-cell aplasia and antierythropoietin antibodies in patients treated with recombinant erythropoietin N. Engl. J. Med. 346 469–475 Occurrence Handle1:CAS:528:DC%2BD38XhtFCktr8%3D Occurrence Handle11844847

    CAS  PubMed  Google Scholar 

  2. S. Hermeling D. J. A. Crommelin H. Schellekens W. Jiskoot (2004) ArticleTitleStructure-immunogenicity relationships of therapeutic proteins Pharm. Res. 21 897–903 Occurrence Handle1:CAS:528:DC%2BD2cXktlykurY%3D Occurrence Handle15212151

    CAS  PubMed  Google Scholar 

  3. P. Perini A. Facchinetti P. Bulian A. R. Massaro D. D. Pascalis A. Bertolotto G. Biasi P. Gallo (2001) ArticleTitleInterferon-beta (INF-beta) antibodies in interferon-beta1a- and interferon-beta1b-treated multiple sclerosis patients. Prevalence, kinetics, cross-reactivity, and factors enhancing interferon-beta immunogenicity in vivo Eur. Cytokine Netw. 12 56–61 Occurrence Handle1:CAS:528:DC%2BD3MXivFGltLs%3D Occurrence Handle11282547

    CAS  PubMed  Google Scholar 

  4. G. Antonelli F. Dianzani (1999) ArticleTitleDevelopment of antibodies to interferon beta in patients: technical and biological aspects Eur. Cytokine Netw. 10 413–422 Occurrence Handle1:CAS:528:DyaK1MXms1ajt7c%3D Occurrence Handle10477398

    CAS  PubMed  Google Scholar 

  5. C. Mayorga G. Luque F. Romero R. Guerrero M. Blanca O. Fernandez (1999) ArticleTitleAntibodies to commercially available interferon-beta molecules in multiple sclerosis patients treated with natural interferon-beta Int. Arch. Allergy Immunol. 118 368–371 Occurrence Handle1:CAS:528:DyaK1MXjsV2qtL8%3D Occurrence Handle10224450

    CAS  PubMed  Google Scholar 

  6. G. Giovannoni F. E. Munschauer F. Deisenhammer (2002) ArticleTitleNeutralising antibodies to interferon beta during the treatment of multiple sclerosis J. Neurol. Neurosurg. Psychiatry 73 465– 469 Occurrence Handle1:STN:280:DC%2BD38nns1Wnug%3D%3D Occurrence Handle12397132

    CAS  PubMed  Google Scholar 

  7. G. Antonelli E. Simeoni F. Bagnato C. Pozzilli O. Turriziani R. Tesoro P. Di Marco ParticleDi C. Gasperini C. Fieschi F. Dianzani (1999) ArticleTitleFurther study on the specificity and incidence of neutralizing antibodies to interferon (IFN) in relapsing remitting multiple sclerosis patients treated with IFN beta-1a or IFN beta-1b J. Neurol. Sci. 168 131–136 Occurrence Handle1:CAS:528:DyaK1MXms1KgtL0%3D Occurrence Handle10526196

    CAS  PubMed  Google Scholar 

  8. A. Bertolotto S. Malucchi E. Milano A. Castello M. Capobianco R. Mutani (2000) ArticleTitleInterferon beta neutralizing antibodies in multiple sclerosis: neutralizing activity and cross-reactivity with three different preparations Immunopharmacology 48 95–100 Occurrence Handle1:CAS:528:DC%2BD3cXlsFequ7s%3D Occurrence Handle10936507

    CAS  PubMed  Google Scholar 

  9. L. Runkel W. Meier R. B. Pepinsky M. Karpusas A. Whitty K. Kimball M. Brickelmaier C. Muldowney W. Jones S. E. Goelz (1998) ArticleTitleStructural and functional differences between glycosylated and non-glycosylated forms of human interferon-beta (IFN-beta) Pharm. Res. 15 641–649 Occurrence Handle1:CAS:528:DyaK1cXjt1ars7s%3D Occurrence Handle9587963

    CAS  PubMed  Google Scholar 

  10. D. F. Mark S. D. Lu A. A. Creasey R. Yamamoto L. S. Lin (1984) ArticleTitleSite-specific mutagenesis of the human fibroblast interferon gene Proc. Natl. Acad. Sci. USA 81 5662–5666 Occurrence Handle1:CAS:528:DyaL2MXhvVKrsg%3D%3D Occurrence Handle6091102

    CAS  PubMed  Google Scholar 

  11. S. D. Cook J. R. Quinless A. Jotkowitz P. Beaton (2001) ArticleTitleSerum IFN neutralizing antibodies and neopterin levels in a cross-section of MS patients Neurology 57 1080–1084 Occurrence Handle1:CAS:528:DC%2BD3MXnsVCnsbk%3D Occurrence Handle11571337

    CAS  PubMed  Google Scholar 

  12. S. Malucchi A. Sala F. Gilli R. Bottero A. Di Sapio ParticleDi M. Capobianco A. Bertolotto (2004) ArticleTitleNeutralizing antibodies reduce the efficacy of betaIFN during treatment of multiple sclerosis Neurology 62 2031–2037 Occurrence Handle1:CAS:528:DC%2BD2cXktVKgt7o%3D Occurrence Handle15184610

    CAS  PubMed  Google Scholar 

  13. A. Bertolotto S. Malucchi A. Sala G. Orefice P. B. Carrieri M. Capobianco E. Milano F. Melis M. T. Giordana (2002) ArticleTitleDifferential effects of three interferon betas on neutralising antibodies in patients with multiple sclerosis: a follow up study in an independent laboratory J. Neurol. Neurosurg. Psychiatry 73 148–153 Occurrence Handle1:STN:280:DC%2BD38zns1Gkuw%3D%3D Occurrence Handle12122172

    CAS  PubMed  Google Scholar 

  14. C. Ross K. M. Clemmesen M. Svenson P. S. Sorensen N. Koch-Henriksen G. L. Skovgaard K. Bendtzen (2000) ArticleTitleImmunogenicity of interferon-beta in multiple sclerosis patients: influence of preparation, dosage, dose frequency, and route of administration. Danish Multiple Sclerosis study group Ann. Neurol. 48 706–712 Occurrence Handle1:CAS:528:DC%2BD3cXosFSgt74%3D Occurrence Handle11079533

    CAS  PubMed  Google Scholar 

  15. D. Hanahan (1989) ArticleTitleTransgenic mice as probes into complex-systems Science 246 1265–1275 Occurrence Handle1:CAS:528:DyaK3cXntV2msw%3D%3D Occurrence Handle2686032

    CAS  PubMed  Google Scholar 

  16. J. L. Ottesen P. Nilsson J. Jami D. Weilguny M. Duhrkop D. Bucchini S. Havelund J. M. Fogh (1994) ArticleTitleThe potential immunogenicity of human insulin and insulin analogues evaluated in a transgenic mouse model Diabetologia 37 1178–1185 Occurrence Handle1:CAS:528:DyaK2MXjvVKnt7o%3D Occurrence Handle7895946

    CAS  PubMed  Google Scholar 

  17. A. Braun L. Kwee M. A. Labow J. Alsenz (1997) ArticleTitleProtein aggregates seem to play a key role among the parameters influencing the antigenicity of interferon alpha (IFN-alpha) in normal and transgenic mice Pharm. Res. 14 1472–1478 Occurrence Handle1:CAS:528:DyaK2sXntFehtL0%3D Occurrence Handle9358564

    CAS  PubMed  Google Scholar 

  18. T. A. Stewart P. G. Hollingshead S. L. Pitts R. Chang L. E. Martin H. Oakley (1989) ArticleTitleTransgenic mice as a model to test the immunogenicity of proteins altered by site-specific mutagenesis Mol. Biol. Med. 6 275–281 Occurrence Handle1:CAS:528:DyaK3cXhtlemu70%3D Occurrence Handle2482920

    CAS  PubMed  Google Scholar 

  19. K. G. Loewen J. B. Derbyshire (1988) ArticleTitleInterferon induction in piglets with polyinosinic:polycytidylic acid complexed with poly-L-lysine and carboxymethylcellulose Res. Vet. Sci. 44 132–133 Occurrence Handle1:CAS:528:DyaL1cXitFKjsr4%3D Occurrence Handle2453906

    CAS  PubMed  Google Scholar 

  20. K. F. Tang P. O. Ts’o S. A. Lesko (1989) ArticleTitleProduction of interferon-beta upon induction with polyinosinic acid:polycytidylic acid during the cell cycle of human fibroblasts Exp. Cell Res. 181 432–441 Occurrence Handle1:CAS:528:DyaL1MXhvVGmur8%3D Occurrence Handle2647497

    CAS  PubMed  Google Scholar 

  21. A. Bertolotto F. Deisenhammer P. Gallo P. S. Sorensen (2004) ArticleTitleImmunogenicity of interferon beta: differences among products J. Neurol. 251 15–24

    Google Scholar 

Download references

Acknowledgments

Biogen-Idec Inc. is acknowledged for kindly providing the test materials. Susan Goelz is greatly acknowledged for her valuable discussions and suggestions.

Author information

Authors and Affiliations

  1. Central Laboratory Animal Institute, Utrecht University, Utrecht, The Netherlands

    Suzanne Hermeling & Huub Schellekens

  2. Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht, The Netherlands

    Suzanne Hermeling, Wim Jiskoot & Daan Crommelin

  3. Department of Biology and Pharmacology, Maxygen ApS, Hørsholm, Denmark

    Claus Bornæs

Authors
  1. Suzanne Hermeling
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wim Jiskoot
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Daan Crommelin
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Claus Bornæs
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Huub Schellekens
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Suzanne Hermeling.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hermeling, S., Jiskoot, W., Crommelin, D. et al. Development of a Transgenic Mouse Model Immune Tolerant for Human Interferon Beta. Pharm Res 22, 847–851 (2005). https://doi.org/10.1007/s11095-005-4578-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11095-005-4578-z

Key words

Search

Navigation