Oil‐Based Emulsion Vaccine Adjuvants

Virgil E.J.C. Schijns1, Marius Strioga2, Stephane Ascarateil3

1 Epitopoietic Research Corporation (ERC), Isnes, Belgium, and ERC, Schaijk, 2 Department of Immunology and Immunotherapy, Institute of Oncology, Vilnius University, Vilnius, 3 Biologicals and Injectable Division, SEPPIC, Paris
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 2.18
DOI:  10.1002/0471142735.im0218s106
Online Posting Date:  August, 2014
GO TO THE FULL TEXT: PDF or HTML at Wiley Online Library

Abstract

Vaccine adjuvants are critical components in experimental and licensed vaccines used in human and veterinary medicine. When aiming to evoke an immune response to a purified antigen, the administration of antigen alone is often insufficient, unless the antigen contains microbial structures or has a natural particulate structure. In most cases, the rationale to use an adjuvant is obvious to the experimental immunologist or the professional vaccinologist, who is familiar with the nature of the antigen, and the aim of the vaccine to elicit a specific antibody response and/or a specific type of T cell response. In this unit, we describe protocols to formulate antigens with oil‐based emulsions. Such emulsions represent a major prototype adjuvant category that is frequently used in experimental preclinical vaccines, as well as veterinary and human vaccines. Curr. Protoc. Immunol. 106:2.18.1‐2.18.7. © 2014 by John Wiley & Sons, Inc.

Keywords: emulsions; vaccines; adjuvants

     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Table of Contents

  • Introduction
  • Basic Protocol 1: The Syringe‐and‐Needle Procedure to Produce W/O Emulsions
  • Alternate Protocol 1: The Two‐Syringes‐and‐Connector Procedure to Produce W/O Emulsions
  • Alternate Protocol 2: Homogenizer Procedure to Produce W/O Emulsions
  • Basic Protocol 2: Syringe Procedure to Create Oil‐in‐Water (O/W) Emulsions
  • Alternate Protocol 3: Homogenizer Procedure to Produce O/W Emulsions
  • Commentary
  • Literature Cited
     
 
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Materials

Basic Protocol 1: The Syringe‐and‐Needle Procedure to Produce W/O Emulsions

  Materials
  • Antigen to be emulsified dissolved in aqueous diluent (also referred to as the aqueous phase)
  • Adjuvant to be used (blend of oil and surfactant also referred to as the oil phase; see introduction to this protocol)
  • Syringe, latex‐, and silicon oil–free (two pieces, luer‐lock), e.g.:
  • 2‐ml Injekt (Braun, ref. 46060701V)
  • 5‐ml Injekt F (Braun, ref. 4606710VV)
  • 2‐ml Norm‐Ject (Henke Sass Wolf GmbH, ref. 4020.000V0)
  • 5‐ml Norm‐Ject (Henke Sass Wolf GmbH, ref. 4050.000V0)
  • 3‐ml crimp vial (e.g., Fisher Scientific)
  • 20‐G needle

Alternate Protocol 1: The Two‐Syringes‐and‐Connector Procedure to Produce W/O Emulsions

  Additional Materials ( protocol 1)
  • Two 1‐ml syringes (see protocol 1 materials list)
  • Syringe connector, e.g.:
    • Green Peptide connector (Japan; http://www.green‐peptide.com/english/news/?action_enews_view=1&nid=22
    • Didanorm : (France‐Europe): http://www.didanorm.fr/achat/index.php?catid=1044
    • Smiths Medical (U.S.): http://www.smiths‐medical.com/catalog/iv‐disposables/iv‐disposables‐components‐accessories/caps‐connectors‐adapters‐luer/connectors‐components.html
    • Air‐Tite Products (U.S.): http://www.air‐tite‐shop.com/p‐215‐double‐female‐luer‐lock‐adapter.aspx
    • Promepla (Worldwide): reference no. ODG0015ST
  • 13‐ or 20‐mm vial adapter (West Pharmaceutical Services, http://www.westpharma.com/ or Promepla, http://www.promepla.com/catalog) or 20‐G needle
  • Sterile needle (usually 26‐G) suitable for vaccination procedure

Alternate Protocol 2: Homogenizer Procedure to Produce W/O Emulsions

  Materials
  • Antigen to be emulsified dissolved in aqueous diluent (also referred to as the aqueous phase)
  • Adjuvant to be used (blend of oil and surfactant also referred to as the oil phase; see introduction to this protocol)
  • 1‐ml sterile syringe (see protocol 1 materials list)
  • 3‐ml crimp vial
  • 20‐G needle
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library

Figures

Videos

Literature Cited

Literature Cited
   Ascarateil, S. and Dupuis, S. 2006. Surfactants in vaccine adjuvants: Descriptions and perspectives. Vaccine 24:S283‐S285
   Dang, Y. , Wagner, W.M. , Gad, E. , Rastetter, L. , Berger, C.M. , Holt, G.E. , and Disis, M.L. 2012. Dendritic cell‐activating vaccine adjuvants differ in the ability to elicit antitumor immunity due to an adjuvant‐specific induction of immunosuppressive cells. Clin. Cancer Res. 18:3122‐3131.
   Flemming, A. 2013. Tumour vaccination: Devil in the details of adjuvant choice. Nat. Rev. Drug Discov. 12:343.
   Hailemichael, Y. , Dai, Z. , Jaffarzad, N. , Ye, Y. , Medina, M.A. , Huang, X.F. , Dorta‐Estremera, S.M. , Greeley, N.R. , Nitti, G. , Peng, W. , Liu, C. , Lou, Y. , Wang, Z. , Ma, W. , Rabinovich, B. , Schluns, K.S. , Davis, R.E. , Hwu, P. , and Overwijk, W.W. 2013. Persistent antigen at vaccination sites induces tumor‐specific CD8+ T cell sequestration, dysfunction and deletion. Nat. Med. 19:465‐472.
   Jansen, T. , Hofmans, M.P.M. , Theelen, M.J.G. , and Schijns, V.E.J.C. 2005. Structure–activity relations of water‐in‐oil vaccine formulations and induced antigen‐specific antibody responses. Vaccine 23:1053‐1060.
   Jansen, T. , Hofmans, M.P.M. , Theelen, M.J.G. , Manders, F. , and Schijns, V.E.J.C. 2006. Structure‐and oil type‐based efficacy of emulsion adjuvants. Vaccine 24:5400‐5405.
   Koh, Y. , Higgins, S.A. , Weber, J.S. , and Kast, W.M. 2006. Immunological consequences of using three different clinical/laboratory techniques of emulsifying peptide‐based vaccines in incomplete Freund's adjuvant. J. Transl. Med. 4:42.
   Moreira, L.O. , Smith, A.M. , DeFreitas, A.A. , Qualls, J.E. , El Kasmi, K.C. , and Murray, P.J. 2008. Modulation of adaptive immunity by different adjuvant‐antigen combinations in mice lacking Nod2. Vaccine 26:5808‐5813.
   Ott, G. , Barchfeld, G.L. , Chernoff, D. , Radhakrishnan, R. , van Hoogevest, P. , and Van Nest, G. 1995. MF59. Design and evaluation of a safe and potent adjuvant for human vaccines. Pharm. Biotechnol. 6:277‐296.
   Ramon, G. 1925. Sur l'aumentation anormale de l'antitoxine chez les chevaux producteurs de serum antidiphterique. Bull. Soc. Centr. Med. Vet. 101:227‐234.
   Schijns, V.E.J.C. and Lavelle, E.C. 2011. Trends in vaccine adjuvants. Expert Rev. Vaccines 10:539‐550.
   Schijns, V.E.J.C. and O'Hagan, D. (Editors). 2006. Immunopotentiators in Modern Vaccines. Elsevier/Academic Press, London.
   Schijns, V. , Tartour, E. , Michalek, J. , Stathopoulos, A. , Dobrovolskiene, N.T. , and Strioga, M. 2014. Immune adjuvants as critical guides directing immunity triggered by therapeutic cancer vaccines. Cytotherapy 16:427‐439.
GO TO THE FULL PROTOCOL:
PDF or HTML at Wiley Online Library