Development of novel fusogenic vesosomes for transcutaneous immunization.

Transcutaneous immunization (TCI) is a novel vaccination strategy based on the application of antigen together with an adjuvant onto hydrated bare skin. This simple and non-invasive immunization procedure elicits systemic and cell mediated immune responses and therefore, it provides a viable and cost-effective strategy for disease prevention. In the present study, novel fusogenic vesicular carrier constructs, i.e. vesosomes were developed and evaluated for topical delivery of vaccines using tetanus toxoid (TTx) as a model antigen. Prepared vesosomes were characterized for size, shape, entrapment efficiency and zeta potential. The prepared novel systems were examined for in process antigen stability and long-term storage stability studies. In vitro skin permeation and fluorescence microscopy study were also preformed for prepared novel vesicular systems for the evaluation of skin penetration efficiency. The immune stimulating activity of these vesicles was studied by measuring the serum anti-tetanus toxoid IgG titer and isotype ratio IgG2a/IgG1 following topical immunization in three different protocols and results were compared with the alum adsorbed tetanus toxoid given intramuscularly and topically administered plain tetanus toxoid solution, plain liposomes and cationic fusogenic liposomes. Serum IgG titers after three consecutive topical administrations were significantly better (*P < 0.05) than single administration of TTx antigen with vesosomal systems, suggesting an effective stimulation of serum immune response. Furthermore, notable serum anti-TTx antibody titers also occurred in animals primed with alum adsorbed TTx and subsequently boosted with topical administration of novel vesosomal systems. In each immunization studies, the vesosomal systems could elicit combined Th1 and Th2 immune responses following topical administration. These results suggest that the investigated vesosomal systems can be effective as topical delivery of vaccines.

[1]  A. Rawat,et al.  Tetanus toxoid‐loaded transfersomes for topical immunization , 2005, The Journal of pharmacy and pharmacology.

[2]  P. Ahl,et al.  Interdigitation-fusion: a new method for producing lipid vesicles of high internal volume. , 1994, Biochimica et biophysica acta.

[3]  F. Szoka,et al.  Destabilization of phosphatidylethanolamine liposomes at the hexagonal phase transition temperature. , 1986, Biochemistry.

[4]  Sanyog Jain,et al.  Topical immunization: Mechanistic insight and novel delivery systems , 2004 .

[5]  M. Iwaki,et al.  Mechanism for enhancement effect of lipid disperse system on percutaneous absorption : Part II , 1997 .

[6]  Leaf Huang,et al.  Folate-targeted, Anionic Liposome-entrapped Polylysine-condensed DNA for Tumor Cell-specific Gene Transfer (*) , 1996, The Journal of Biological Chemistry.

[7]  C Caux,et al.  Immunobiology of dendritic cells. , 2000, Annual review of immunology.

[8]  T. Kupper.  Immune and inflammatory processes in cutaneous tissues. Mechanisms and speculations. , 1990, The Journal of clinical investigation.

[9]  N. Weiner,et al.  Liposomes as carriers for topical and transdermal delivery. , 1994, Journal of pharmaceutical sciences.

[10]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[11]  C. Alving,et al.  Transcutaneous immunization: A human vaccine delivery strategy using a patch , 2000, Nature Medicine.

[12]  Gregory Gregoriadis,et al.  Preparation of liposomes , 1984 .

[13]  G. Cevc,et al.  Transdermal immunisation with an integral membrane component, gap junction protein, by means of ultradeformable drug carriers, transfersomes. , 1998, Vaccine.

[14]  S. Constant,et al.  Induction of Th1 and Th2 CD4+ T cell responses: the alternative approaches. , 1997, Annual review of immunology.

[15]  Simon C Watkins,et al.  DNA–based immunization by in vivo transfection of dendritic cells , 1996, Nature Medicine.

[16]  Y. Y. Wang,et al.  Effect of liposomes and niosomes on skin permeation of enoxacin. , 2001, International journal of pharmaceutics.

[17]  K. Kono,et al.  Design of fusogenic liposomes using a poly(ethylene glycol) derivative having amino groups. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[18]  N. Weiner,et al.  Topical delivery of liposomally encapsulated interferon evaluated in a cutaneous herpes guinea pig model , 1989, Antimicrobial Agents and Chemotherapy.

[19]  G. Cevc,et al.  Transdermal immunization with large proteins by means of ultradeformable drug carriers , 1995, European journal of immunology.

[20]  J. White,et al.  Rapid separation of low molecular weight solutes from liposomes without dilution. , 1978, Analytical biochemistry.

[21]  R. New,et al.  Liposomes : a practical approach , 1990 .

[22]  T. Morokata,et al.  Antigen dose defines T helper 1 and T helper 2 responses in the lungs of C57BL/6 and BALB/c mice independently of splenic responses. , 2000, Immunology letters.

[23]  Sanyog Jain,et al.  Non-invasive vaccine delivery in transfersomes, niosomes and liposomes: a comparative study. , 2005, International journal of pharmaceutics.

[24]  S. O. Bryant,et al.  Fusogenic properties of Sendai virosome envelopes in rat brain preparations , 1993, Neurochemical Research.

[25]  J. Kraehenbuhl,et al.  Antigen sampling across epithelial barriers and induction of mucosal immune responses. , 1996, Annual review of immunology.

[26]  J. Philippot,et al.  Design of a short membrane-destabilizing peptide covalently bound to liposomes. , 1994, Biochimica et biophysica acta.

[27]  B. Shroot,et al.  Site-Specific Drug Delivery to Pilosebaceous Structures Using Polymeric Microspheres , 1993, Pharmaceutical Research.

[28]  G. Gregoriadis,et al.  Liposome-entrapped plasmid DNA: characterisation studies. , 2000, Biochimica et biophysica acta.

[29]  C. Valenta,et al.  Evaluation of novel soya-lecithin formulations for dermal use containing ketoprofen as a model drug. , 2000, Journal of controlled release : official journal of the Controlled Release Society.

[30]  J. Grivel,et al.  Immunization through dermal delivery of protein‐encoding DNA: a role for migratory dendritic cells , 1999, European journal of immunology.

[31]  J. Philippot,et al.  Non-phospholipid fusogenic liposomes. , 1996, Biochimica et biophysica acta.

[32]  Kazuo Maruyama,et al.  Transferrin-modified liposomes equipped with a pH-sensitive fusogenic peptide: an artificial viral-like delivery system. , 2004, Biochemistry.

[33]  M. Klein,et al.  The interaction of murine IgG subclass proteins with human monocyte Fc receptors. , 1985, Journal of immunology.

[34]  A. Schätzlein,et al.  TRANSDERMAL DRUG CARRIERS - BASIC PROPERTIES, OPTIMIZATION AND TRANSFER EFFICIENCY IN THE CASE OF EPICUTANEOUSLY APPLIED PEPTIDES , 1995 .