ABSTRACT
Dry tetanus toxoid (TTx) patches were formulated without any adjuvant, with excipients to impart antigen stabilization and to enhance skin delivery. The booster effects of the TTx patches were assessed using a guinea pig model. The study revealed significant rises in TTx IgG titers induced by the TTx patches after a low-dose subcutaneous (s.c.) prime with TTx adsorbed to aluminum hydroxide. The TTx patch can therefore be considered an effective alternative to a subcutaneous booster.
Subject(s)
Immunization, Secondary/methods , Tetanus Toxoid/administration & dosage , Tetanus Toxoid/immunology , Tetanus/prevention & control , Administration, Cutaneous , Animals , Antibodies, Bacterial/blood , Antitoxins/blood , Female , Guinea Pigs , Immunoglobulin G/bloodABSTRACT
The preformulation of a trivalent recombinant protein-based vaccine candidate for protection against Streptococcus pneumoniae is described both in the presence and in the absence of aluminum salt adjuvants. The biophysical properties of the three protein-based antigens, fragments of pneumococcal surface adhesion A (PsaA), serine-threonine protein kinase (StkP), and protein required for cell wall separation of group B streptococcus (PcsB), were studied using several spectroscopic and light scattering techniques. An empirical phase diagram was constructed to assess the overall conformational stability of the three antigens as a function of pH and temperatures. A variety of excipients were screened on the basis of their ability to stabilize each antigen using intrinsic fluorescence spectroscopy and circular dichroism spectroscopy. Sorbitol, sucrose, and trehalose stabilized the three proteins in solution. The addition of manganese also showed a drastic increase in the thermal stability of SP1650 in solution. The adsorption and desorption processes of each of the antigens to aluminum salt adjuvants were evaluated, and the stability of the adsorbed proteins was then assessed using intrinsic fluorescence spectroscopy and Fourier transform infrared spectroscopy. All the three proteins showed good adsorption to Alhydrogel. PsaA was destabilized when adsorbed onto Alhydrogel® and adding sodium phosphate showed a stabilizing effect. PcsB was found to be stabilized when adsorbed to Alhydrogel®, and no destabilizing or stabilizing effects were seen in the case of StkP.
Subject(s)
Adjuvants, Immunologic/chemistry , Aluminum Compounds/chemistry , Aluminum Hydroxide/chemistry , Bacterial Proteins/chemistry , Phosphates/chemistry , Pneumococcal Vaccines/chemistry , Streptococcus pneumoniae/immunology , Adhesins, Bacterial/chemistry , Adsorption , Aluminum Compounds/immunology , Aluminum Hydroxide/immunology , Bacterial Proteins/immunology , Chemistry, Pharmaceutical , Circular Dichroism , Excipients/chemistry , Hydrogen-Ion Concentration , Light , Lipoproteins/chemistry , Phosphates/immunology , Pneumococcal Vaccines/immunology , Protein Conformation , Protein Denaturation , Protein Serine-Threonine Kinases/chemistry , Protein Stability , Scattering, Radiation , Sorbitol/chemistry , Spectrometry, Fluorescence , Spectroscopy, Fourier Transform Infrared , Sucrose/chemistry , Technology, Pharmaceutical/methods , Temperature , Trehalose/chemistry , Vaccines, Synthetic/chemistryABSTRACT
Streptococcus pneumoniae is a major human pathogen, causing high morbidity and mortality in children, and also in the elderly, who are particularly susceptible to S. pneumoniae infections due to the dysregulated function of the aged immune system. As the current generation of polysaccharide vaccines do not provide sufficient protection for elderly, new vaccination strategies are urgently needed. To learn whether pneumococcal proteins are able to induce adaptive immune responses in adults in different age groups, we determined serum IgG antibody titers and T cell immunity (IFN-γ, IL-17A and IL-5 production) to three pneumococcal antigens, PcsB, StkP and PsaA, that are components of an investigational protein-based pneumococcal vaccine, IC47. Therefore, sera and PBMCs of 108 healthy adults in three different age groups (young, middle-aged and elderly) were analyzed by ELISA and ELISpot, respectively. We found naturally acquired antibodies to all three proteins in all age groups against all three antigens. However, elderly individuals had significantly lower IgG levels to PcsB and PsaA compared to those of younger donors. There was no significant age-related difference in the overall rate of T cell immunity for the three pneumococcal proteins. We found that the Th17 response was dominant in all age groups and was frequently combined with a Th1 or Th2 response in young and middle-aged subjects. However, in elderly persons there was a lower percentage of PBMC samples producing more than one cytokine upon antigenic stimulation. The narrow cytokine secretion pattern was the most striking difference between elderly and younger adult age groups. Our results demonstrate that in the majority of adults there is a naturally acquired humoral and cellular immune response to the three pneumococcal proteins tested. The dominance of the Th17 response is especially interesting in the light of new insights regarding the role of Th17 cells in mucosal protection against this pathogen.
