During the last decade or so vaccine development has been facilitated by rapid advances in molecular and cell biology. These have laid the foundations of a new generation of vaccines exemplified by subunit vaccines produced through gene cloning and by synthetic peptides mimicking small regions of proteins on the outer coat of viruses. Such peptide~ are capable of eliciting virus-neutralizing antibodies. Unfortunately, subunit and peptide vaccines are only weakly or non immunogenic in the absence of immunological adjuvants that are known to augment specific cell-mediated immune responses to the antigens and to promote the formation of protective antibodies. This book contains the proceedings of the 4th NATO Advanced Studies Institute (ASI) "Vaccines: New Generation Immunological Adjuvants" held at Cape Sounion Beach, Greece, during 24 June -5 . July 1994 and deals in depth with both theoretical and practical aspects of vaccinology. These include the role of antigen presenting cells in the induction of immune responses. immunopotentiation by a variety of new generation immunological adjuvants and vaccine carriers. and recent advances and perspectives in experimental vaccines as well as vaccinatioll with nucleic acids. We express our appreciation to Dr. K. Dalsgaard and Dr. J. L. Virelizier for their cooperatioll in planning the ASI and to Mrs. Concha Pening for her excellent production of the manuscripts. The ASI was held under the sponsorship of NATO Scientific Affairs Division and generously co-sponsored by SmithKline Beecham Pharmaceuticals (Philadelphia).

The NATO Advanced Studies Institute series "Targeting of Drugs" was originated in 1981. It is now a major international forum, held every two years in Cape Sounion, Greece, in which the present and the future of this important area of research in drug delivery is discussed in great depth. Previous ASIs of the series dealt with drug carriers of natural and synthetic origin, their interactions with the biological milieu, ways by which milieu interference is circumvented, approaches to carrier design or modification that contribute to optimal carrier function, and, more recently, with strategies for gene and oligonucleotide delivery in therapy The present book contains the proceedings of the 9th NATO ASI, "Targeting of Drugs: Strategies for Stealth Therapeutic Systems," held in Cape Sounion during 24 June -5 July 1997. As the title implies, the book deals with a variety of approaches to evade the reticuloendothelial system and thus extend the circulation time of a variety of delivery systems including polymers, biopolymers, liposomes, and other nanoparticles or microparticles. We express our appreciation to Mrs. Concha Perring for her assistance with the organization of the AS . The ASI was held under the sponsorship of NATO Scientific Affairs Division and supported by Sequus Pharmaceuticals (Menlo Park, CA, USA), The Liposome Company (Princeton, NJ, USA), NOVO Nordisk (Bagsvaard, Denmark), NeXstar Pharmaceuticals (Boulder, CO, USA), Gene Medicine (Houston, TX, USA), Pfizer Ltd (Sandwich, Kent, UK), Merck (Rahway, NJ, USA), and Biovation Ltd (Aberdeen, UK).

During the last decade or so vaccine development has been facilitated by rapid ad- vances in the molecular and cell biology ofthe immune system. This has laid the foundations of a new generation of vaccines exemplified by subunit vaccines produced through gene cloning and by synthetic peptides mimicking small regions ofproteins on the outer coat ofvi- ruses. However, as subunit and peptide vaccines are only weakly or non-immunogenic, there is areal need for strategies to improve their potency. This book contains the proceedings of the 5th NATO Advanced Studies Institute (ASI), "Vaccine Design: The Role of Cytokine Networks," held at Cape Sounion Beach, Greece, during 24 June-5 July 1996 and deals in depth with the role ofbasic immunology in the regulation of immunity and vaccine design. Special emphasis is given to the use of cyto- kines in conjuction with vaccines with the aim ofimproving their potency or the use ofvac- cines designed to improve cytokine production. We express our appreciation to Dr. J.-L. Virelizier and Dr. G. Kollias for their cooperation in planning the ASI and to Mrs. Concha Perring for her excellent production ofthe manuscripts. The ASI was held under the sponsor- ship of NATO Scientific Affairs Division and generously co-sponsored by SmithKline Beecham Pharmaceuticals (Philadelphia). Financial assistance was also provided by Connaught Laboratories Ltd. (Ontario), Pasteur Merieux (Marcy I 'Etoile), Biochine (Siena), Help SA (Athens), and Avanti Polar Lipids Inc. (Birmingham, USA).

The NATO Advanced Studies Institute series "Targeting of Drugs" was originated in 1981. It is now a major international forum, held every two years in Cape Sounion, Greece, in which the present and the future of this important area of research in drug delivery is discussed in great depth. Previous ASIs of the series dealt with drug carriers of natural and synthetic origin, their interaction with the biological milieu, ways by which the latter influences such interaction, strategies by which milieu interference curtailing the function of drug carriers is circumvented and, more recently, with the application of drug carriers for the delivery of peptides and proteins. The present book contains the of the 7th NATO ASI "Targeting of Drugs: Advances in System Constructs," proceedings held in Cape Sounion during 24 June -5 July 1993. As the title implies, the book deals with a variety of approaches to carrier design or modification that contribute to optimal carrier function. to Mrs Concha Perring for her assistance with the We express our appreciation organization of the ASI. We thank Dr. G. Deliconstantinos who, as chairperson of the Local Committee, contributed to the success of the Institute. The ASI was held under the sponsorship of NATO Scientific Affairs Division and co-sponsored and generously financed by SmithKline Beecham Pharmaceuticals (King of Prussia). Financial assistance was also provided by Liposome Technology Inc. (Menlo Park), Vestar Inc. (San Dimas) and Zeneka (Macclesfield).

The 19 papers explore a variety of systems in terms of their ability to transport nucleic acids to target areas in vitro and in vivo in ways that effectively modify, supplement, correct, or curtail the function of genes in therapy. Among the topics are cystic fibrosis, immune responses with direct g

The NATO Advanced Studies Institute series "Targeting of Drugs" was originated in 1981. It is now a major international forum, held every two years in Cape Sounion, Greece, in which the present and the future of this important area of research in drug delivery is discussed in great depth. Previous ASIs of the series dealt with drug carriers of natural and synthetic origin, their interaction with the biological milieu, ways by which the latter influences such interaction, strategies by which milieu interference curtailing the function of drug carriers is circumvented and, more recently, with the application of drug carriers for the delivery of peptides and proteins. The present book contains the of the 7th NATO ASI "Targeting of Drugs: Advances in System Constructs," proceedings held in Cape Sounion during 24 June -5 July 1993. As the title implies, the book deals with a variety of approaches to carrier design or modification that contribute to optimal carrier function. to Mrs Concha Perring for her assistance with the We express our appreciation organization of the ASI. We thank Dr. G. Deliconstantinos who, as chairperson of the Local Committee, contributed to the success of the Institute. The ASI was held under the sponsorship of NATO Scientific Affairs Division and co-sponsored and generously financed by SmithKline Beecham Pharmaceuticals (King of Prussia). Financial assistance was also provided by Liposome Technology Inc. (Menlo Park), Vestar Inc. (San Dimas) and Zeneka (Macclesfield).

TheNATO AdvancedStudiesInstituteseries"TargetingofDrugs"wasoriginatedin 1981. It is nowamajorinternationalforum,heldeverytwo yearsin CapeSounion,Greece,in whichthepresentandthefutureofthisimportantareaofresearch in drugdeliveryisdiscussed in greatdepth. PreviousASIsoftheseriesdealtwith drugcarriersofnaturalandsynthetic origin,theirinteractionswith thebiologicalmilieu, waysby whichthefunctionofdrugcarriers iscircumvented and,morerecently,with avarietyofapproaches to carrierdesignor modificationthatcontributeto optimalcarrierfunction. Thepresentbookcontainsthe proceedings ofthe8thNATO ASI, "TargetingofDrugs:Strategies for Oligonucleotideand GeneDelivery in Therapy", held in CapeSounionduring24June-5 July 1995. Asthetitle implies,thebookdealswith avarietyofsystemsin termsoftheirability to transportnucleic acidsto targetareasin vitro andin vivo in waysthateffectivelymodify,supplement, correct, orcurtailthefunctionofgenesin therapy. Weexpressourappreciation to Mrs. ConchaPerringfor herassistance with the organizationoftheASI. TheASI washeldunderthesponsorship ofNATO ScientificAffairs Division andco-sponsored andgenerouslyfinancedby SmithKlineBeechamPharmaceuticals (KingofPrussia). Financialassistance wasalsoprovidedby SandozPharma(Baseland Athens),GeneMedicine (Houston,USA), ChironCorporation(Emeryville,USA), BYK GuldenLombergChemische (Konstanz,Gernlany),HelpSA(Athens,Greece),Avanti Polar Lipids Inc (Birmingham,USA), OxfordMolecular(Oxford,UK), Pfizer(Kent,UK), andAlza Corporation(PaloAlto, USA). GregoryGregoriadis BrendaMcCormack v CONTENTS Gene Therapy for Inherited Genetic Disease: Possibilities and Problems c. *Coutelle Gene Delivery and Therapy: The Case for Cystic Fibrosis 15 E. W. F. W. Alton Immune Responses with Direct Gene Transfer: DNA Vaccines and 21 Implications for Gene Therapy H. L. Davis Oligonucleotides: Molecular Versions for Optimal Use in Vivo 31 E. Saison-Behmoaras, A. Van Aerschot, I. Duroux, C. Hendrix, C. Helene, and P. Herdewijn Retrovirus Vectors in Gene Therapy: Targeting to Specific Cells 45 AJ. Kingsman, Y. Bae, J. c. Griffiths, N. Kim, E. E. Ramsdale, G. Romano, Y. Soneoka, P. M. Cannon, and S. M. Kingsman Adenovirus as Vectors for Gene Therapy 53 M. G. Lee Receptor-Mediated Gene Delivery with Synthetic Virus-like Particles 67 E. Wagner, M. Cotten, and K. Zatloukal Controllable Gene Therapy: Recent Advances in Non-Viral Gene Delivery 79 A.

It is widely accepted that vaccination still renains the best answer to ITDst infectious diseases. Recently, vaccine developnent has been greatly facilitated by advances in ITDlecular and cell biology which have laid the foundations of a new generation of vaccines. '!hese are exemplified by submit vaccines produced through gene cloning and synthetic peptides mimicking snall regions of proteins on the outer coat of viruses and capable of eliciting virus neutralizing antibodies. However, submit and peptide vaccines are only weakly or non-inmmogenic in the absence of immunological adjuvants. The latter are a diverse array of agents that augment specific cell-mediated immune responses to the antigens and the formation of protective antibodies. '!his book contains the proceedings of the 3rd NATO Advanced Studies Institute (ASI) "New-Generation vaccines: '!he Role of Basic Irrmmology" held at Cape Sounion Beach, Greece, during 24 June-5 July, 1992. It deals with recent developnents in the understanding of inmmity at the ITDlecular and cellular levels and the application of such knowledge in the search for novel inmmological adjuvants and the fonnulation of new-generation vaccines for experimental and clinical use. We express our appreciation to Professor K. Dalsgaard and H. Snippe for their cooperation in planning the ASI and to Mrs. Concha Perring for her excellent production of the manu- scripts. '!he ASI was held tmder the sponsorship of NATO Scientific Affairs Division and generously co-sponsored by SrnithKline Beecham Pharmaceuticals (Fhiladelp,. ia). Financial assistance was also provided by Pasteur Merieux (Marcy L'Etoile), British Biotechnology Ltd.

The NATO Advanced Studies Institute series "Targeting of Drugs" was originated in 1981. It is now a major international forum, held every two years in Cape Sounion, Greece, in which the present and the future of this important area of research in drug delivery is discussed in great depth. Previous ASIs of the series dealt with drug carriers of natural and synthetic origin, their interactions with the biological milieu, ways by which milieu interference is circumvented, approaches to carrier design or modification that contribute to optimal carrier function, and, more recently, with strategies for gene and oligonucleotide delivery in therapy The present book contains the proceedings of the 9th NATO ASI, "Targeting of Drugs: Strategies for Stealth Therapeutic Systems," held in Cape Sounion during 24 June -5 July 1997. As the title implies, the book deals with a variety of approaches to evade the reticuloendothelial system and thus extend the circulation time of a variety of delivery systems including polymers, biopolymers, liposomes, and other nanoparticles or microparticles. We express our appreciation to Mrs. Concha Perring for her assistance with the organization of the AS . The ASI was held under the sponsorship of NATO Scientific Affairs Division and supported by Sequus Pharmaceuticals (Menlo Park, CA, USA), The Liposome Company (Princeton, NJ, USA), NOVO Nordisk (Bagsvaard, Denmark), NeXstar Pharmaceuticals (Boulder, CO, USA), Gene Medicine (Houston, TX, USA), Pfizer Ltd (Sandwich, Kent, UK), Merck (Rahway, NJ, USA), and Biovation Ltd (Aberdeen, UK).

During the last decade or so vaccine development has been facilitated by rapid advances in molecular and cell biology. These have laid the foundations of a new generation of vaccines exemplified by subunit vaccines produced through gene cloning and by synthetic peptides mimicking small regions of proteins on the outer coat of viruses. Such peptide~ are capable of eliciting virus-neutralizing antibodies. Unfortunately, subunit and peptide vaccines are only weakly or non immunogenic in the absence of immunological adjuvants that are known to augment specific cell-mediated immune responses to the antigens and to promote the formation of protective antibodies. This book contains the proceedings of the 4th NATO Advanced Studies Institute (ASI) "Vaccines: New Generation Immunological Adjuvants" held at Cape Sounion Beach, Greece, during 24 June -5 . July 1994 and deals in depth with both theoretical and practical aspects of vaccinology. These include the role of antigen presenting cells in the induction of immune responses. immunopotentiation by a variety of new generation immunological adjuvants and vaccine carriers. and recent advances and perspectives in experimental vaccines as well as vaccinatioll with nucleic acids. We express our appreciation to Dr. K. Dalsgaard and Dr. J. L. Virelizier for their cooperatioll in planning the ASI and to Mrs. Concha Pening for her excellent production of the manuscripts. The ASI was held under the sponsorship of NATO Scientific Affairs Division and generously co-sponsored by SmithKline Beecham Pharmaceuticals (Philadelphia).

During the last decade or so vaccine development has been facilitated by rapid ad- vances in the molecular and cell biology ofthe immune system. This has laid the foundations of a new generation of vaccines exemplified by subunit vaccines produced through gene cloning and by synthetic peptides mimicking small regions ofproteins on the outer coat ofvi- ruses. However, as subunit and peptide vaccines are only weakly or non-immunogenic, there is areal need for strategies to improve their potency. This book contains the proceedings of the 5th NATO Advanced Studies Institute (ASI), "Vaccine Design: The Role of Cytokine Networks," held at Cape Sounion Beach, Greece, during 24 June-5 July 1996 and deals in depth with the role ofbasic immunology in the regulation of immunity and vaccine design. Special emphasis is given to the use of cyto- kines in conjuction with vaccines with the aim ofimproving their potency or the use ofvac- cines designed to improve cytokine production. We express our appreciation to Dr. J.-L. Virelizier and Dr. G. Kollias for their cooperation in planning the ASI and to Mrs. Concha Perring for her excellent production ofthe manuscripts. The ASI was held under the sponsor- ship of NATO Scientific Affairs Division and generously co-sponsored by SmithKline Beecham Pharmaceuticals (Philadelphia). Financial assistance was also provided by Connaught Laboratories Ltd. (Ontario), Pasteur Merieux (Marcy I 'Etoile), Biochine (Siena), Help SA (Athens), and Avanti Polar Lipids Inc. (Birmingham, USA).