Preclinical and Phase I Clinical Trial Design Issues 101 US Food and Drug Administration Regulations
No detailed or specific guidelines exist in the United States for assessing the safety of adjuvant preparations for use in humans. Only two guidelines refer to adjuvants. The first guideline formally issued by the FDA, which includes adjuvanted vaccines (86), refer to tests of the final container lot of all biological products. These FDA standards are paraphrased in Table 7 for ease of understanding. It is unclear if adjuvants, such as QS-21, which are added to the vaccine immediately before...
Materials
Protein antigen The handling of the antigen before incorporation is important. The antigens most suited for ISCOM formulation are hydrophobic or amphipathic. Such antigens must be purified in the presence of a detergent to prevent micelle or aggregate formation because pure protein micelles or aggregates are very difficult to dissociate to make their hydrophobic part accessible for hydrophobic interactions. There are many examples where antigens that theoretically should incorporate well, in...
Duncan E S Stewart Tull 1 Introduction
There are no officially recognized regulations for the design and toxicity testing of adjuvants or adjuvant formulations the former are also referred to as immunomodulators and immunopotentiators. At the Immunological Adjuvants and Vaccines meeting held in Greece in 1988, however, immuno-adjuvant researchers discussed experimental toxicological tests that might be used to monitor new immunomodulators (1). The usefulness of these tests for the range of immunomodulators and adjuvant formulations...
The Use of MF59 as a Vaccine Adjuvant
The MF59 emulsion adjuvant was developed with the objective of generating a broad spectrum of recombinant vaccines for human use. The specific aim was to elicit neutralizing titers in humans significantly greater than those obtainable with the alum adjuvants in common usage. An extensive body of preclinical efficacy data has been obtained based on ELISA titers obtained with a variety of subunit antigens in a spectrum of animal model systems. A summary of this data is shown in Table 2. Data are...
References
G. (1951) Nouveau vaccin antiaphteux a complexe glucoviral. Gac. Vet. 13, 268. 2. Dalsgaard, K. (1974) Saponin adjuvants. III. Isolation of a substance from Quillaja saponaria Molina with adjuvant activity in foot-and-mouth disease vaccines. Arch. Gesamte Virusforsch. 44, 243-254. 3. Kensil, C. R., Patel, U., Lennick, M., and Marciani, D. (1991) Separation and characterization of saponins with adjuvant activity from Quillaja saponaria Molina cortex. J. Immunol. 146, 431-437. 4. Ma,...
Notes
At this time, there is no availability of threonyl-MDP. This material had been manufactured by or for Syntex or its licensees. This compound is no longer being synthesized. A description of threonyl-MDP and related analogs can be found in U.S. Patent 4,082,735, authored by Jones, G. H., Moffett, J. H., and Nestor, J. J. The literature also suggests other MDP analogs of potential interest (14). Any inquiries regarding threonyl-MDP should be directed to Dr. Gary Ott of Chiron (Emeryville, CA,...
Measurement of the Toxicity of Adjuvants
Cytotoxicity Assay in Cultured Monolayers of Human or Animal Cell Lines This type of assay has the great merit of reducing the number of animals which must be used to comply with standardized toxicity tests. The MTT assay (13) was adapted for determining cell survival and proliferation by a number of workers (14-16) with different cell types and toxins.The assay compares favorably with other similar systems (17), is less time-consuming and objective than microscopic examination of cells,...
Immunopotentiating Reconstituted Influenza Virosomes
Twenty years after the discovery of the immunological adjuvant properties of liposomes (25) and the ensuing multitude of related animal immunization studies (26), liposomes as adjuvants have come of age (27,28) with the first liposome-based vaccine against hepatitis A being licensed for use in humans. Vaccines based on novasomes (nonphospholipid biodegradable, pausilamellar vesicles formed from single-chain amphiphiles, with or without other lipids) have also been licensed for the immunization...
Summary and Conclusion
IRIVs have been shown to act as an efficient and highly effective means of enhancing the immune response to a variety of vaccine antigens, thus illustrating their use as vaccine delivery systems. Their use is not limited to a certain type of antigen. IRIV-based vaccines comprising viral glycoproteins, bacterial Kinetics of the Immune Response Following Immunization with Various Hepatitis A Vaccine Formulations Kinetics of the Immune Response Following Immunization with Various Hepatitis A...
Composition of Mineral Oil Adjuvants
As mentioned, the classical Freund's incomplete adjuvant consists of a mixture of mineral oil and emulsifier in a ratio of 85 v v oil and 15 v v emulsi-fier. This mixture will then be mixed with an equal volume of aqueous solution of antigen and subsequently emulsified prior to use. Ratios of 90 oil and 10 emulsifier have also been described. Both components will be discussed below. The oil component of mineral oil adjuvants is normally light mineral oil of a highly purified quality. Some...
Protein Cochleates
Cochleates as carriers for protein and peptide antigens effectively induce antibody- and cell-mediated immune responses. Protection from lethal and Summary of Cochleate Vaccine Studies Summary of Cochleate Vaccine Studies Intestinal intraepithelial lymphocytes infectious dose challenge with viruses administered parenterally and mucosally has also been achieved (see Table 1 for summary) (14-19, and manuscripts in preparation). For example, cochleate vaccines containing the glycoproteins and...
Hemolysis Test
At very low concentrations, adjuvants should not be hemolytic. This is particularly relevant for the crude, triterpenoid plant saponins, which reportedly destroy erythrocytes if injected intravenously, although this effect may be owing to contaminatory substances (7). The immune stimulating complexes (ISCOMs see Chapter 14) contain a saponin, which is also used to produce the positive 100 lysis of erythrocytes in the method below. It is often stated that such complexes cannot be used in...
Rajesh K Gupta and Bradford E Rost 1 Introduction
Aluminum compounds, including aluminum phosphate (AlPO4), aluminum hydroxide (Al(OH)3), and alum precipitated vaccines, historically referred to as protein aluminate, are currently the most commonly used adjuvants with human and veterinary vaccines (1-6). These adjuvants are often referred to as alum in the literature, which is misleading because (1) two most widely used adjuvants from this group, aluminum hydroxide and aluminum phosphate, have very different physical characteristics (7) and...
Sarah C Gilbert 1 Introduction
Adjuvants are available to promote the generation of antibodies to an antigen following immunization. However, many of these adjuvants do not enhance priming of cytotoxic T lymphocytes (CTL). The reason for this lies in the existence of two alternative antigen processing pathways, leading to stimulation of CD4+ T cells, and, in turn, to the generation of antibodies, or stimulation of CD8+ CTL. In general, exogenous proteins enter the antigen presenting cell (APC) by endocytosis. Peptides...
Final Vaccine Formulation
A variety of antigens including HSV-2 gB gD, HBV, HIV gp120 and or p24, CMV gB, and influenza hemagglutinin (HA) have been formulated with Fig. 4. Long-term particle size stability of the three representative MF59C.1 bulk lots at 2-8 C. Fig. 4. Long-term particle size stability of the three representative MF59C.1 bulk lots at 2-8 C. the adjuvant emulsion MF59C.1 in one of two formats either in single containers or in separate vials (admixed prior to administration and therefore referred to as...
Gregory M Glenn Tanya Scharton Kersten and Russell Vassell 1 Introduction
Transcutaneous immunization (TCI), the introduction of antigens using a topical application to intact skin, is a new technology that has both practical and immunological merits. Practically speaking, a needle-free method of vaccine delivery will decrease the risk of needle-borne diseases, reduce the complications related to physical skin penetration, improve access to vaccination by eliminating the need for trained personnel and sterile equipment, and provide a simple means for multivalent or...
Historical Background
The first evidence of the use of oil emulsions as adjuvants in vaccine formulations dates back to 1916. At that time, Le Moignic and Pinoy immunized mice with heat-inactivated Salmonella typhimurium in an emulsion of water and vaselin oil, using lanolin as an emulsifier (1). The oil emulsions as adjuvants, however, did not receive much attention before Jules Freund and coworkers (2,3) decades later combined a paraffin (mineral) oil emulsion and heat-killed mycobacteria to produce an extremely...
Examples of Modern Vaccine Adjuvants Used in Animals and
Agents listed in Table 1 are examples of the many varieties of immuno-potentiators used during the past 30 years. The majority of adjuvants are being developed and tested by industry. The list of adjuvants is incomplete, because I have not conducted an exhaustive literature search, because the results have appeared in abstracts in nonindexed publications, and because many studies are proprietary. The adjuvants marked by an asterisk in Table 1 have completed trial in man, or they are now...
Erik B Lindblad 1 Introduction
Freund's adjuvants are water-in-mineral oil emulsions (W O emulsions) without heat-killed mycobacteria added (Freund's incomplete adjuvant) or with heat-killed mycobacteria added (Freund's complete adjuvant). Freund's adjuvants are, in a way, historic adjuvants and for many researchers the first and most obvious association to the word adjuvant at all. The adjuvants have been used extensively in experimental immunology owing to their high efficacy, and Freund's incomplete adjuvant has been used...
Preclinical Evaluation of Vaccines Adsorbed onto Aluminum Compounds
To achieve consistency in manufacture of aluminum adjuvants, it is important to characterize these adjuvants for physicochemical and adjuvant properties. Though aluminum adjuvants have been used widely with human vaccines, there are still a number of unanswered questions about their mechanism of action (see Note 5), such as optimal size of the gel particles, degree of adsorp tion of antigens onto adjuvant, amount of gel used as a dose for humans and animals (see Note 3) and even type of...
Terry Ulrich 1 Introduction
Interest in new methods of potentiating the immune response against vaccine antigens has increased considerably over the past decade. In part, this interest is in response to vaccine initiatives that have established aggressive goals for improving existing vaccines and for developing much-needed new vaccines. Many of the candidate vaccine antigens being developed as part of this effort are synthetic or recombinant subunit structures that are often poorly immunogenic and as such, are unable to...
General Adjuvant Properties
QS-21 stimulates strong antibody- and cell-mediated responses in animals. QS-21 has been shown to stimulate strong antibody responses to T-dependent protein antigens in mice 3-5 , guinea pigs 6 , rhesus monkeys 7 , and baboons 8 . Antibody responses induced by QS-21-adjuvanted vaccines were shown to be higher than antibody responses with aluminum hydroxide-adjuvanted vaccines and were similar to those obtained with Freund's com-plete-adjuvanted FCA vaccines 3,6 . QS-21 is also a strong adjuvant...
General Structure and Physicochemical Properties
QS-21 was shown by 1H and 13C-NMR and fast-atom bombardment-mass spectrometry to be an acylated 3,28-o-bisdesmodic triterpene saponin of molecular weight 1990 16 . The structure of QS-21 is shown in Fig. 4. QS-21 is anionic at neutral pH, because of the carboxyl group on glucuronic acid. It is insoluble in aqueous solution in the protonated form, but soluble in the sodium salt form where QS-21 forms micelles in solution. The critical micellar concentration of QS-21 was estimated to be 26 M in...
Aluminum Phosphate
Like aluminum hydroxide, aluminum phosphate is the most widely used adjuvant with routine human vaccines. Most of the vaccine manufacturers throughout the world prepare this adjuvant in-house. Usually, antigens are adsorbed onto a preformed gel, which can be made by several methods two methods are described in this chapter . Adsorption of antigens is also carried out on freshly prepared aluminum phosphate gel 23 . In situ adsorption of antigens onto aluminum phosphate is preferable to aluminum...
Center for Biologics Evaluation and Research Cber Fda
The CBER, FDA, Rockville, MD is responsible for regulating vaccines and other biologics in the United States. In addition to meeting the general standards before public release Table 7 , each vaccine and adjuvant are tested for safety on a case by case basis, preferably with the help and guidance of the CBER as noted before. Such guidance, informal in nature but quite helpful, was published in 1993 in response to the needs of HIV-1 vaccine development 52 . The principles laid down by that...
Vaccine Formulation
Mix the sterile stock solution of QS-21 with a sterile stock of soluble antigen with the dose of each adjusted to the optimum for the animal to be immunized. A typical effective QS-21 dose is 10-20 g for mice, 25-50 g for guinea pigs or rats, and 50-100 g for rabbits, rhesus monkeys, and baboons. The volume of the vaccine is then adjusted to the final formulation volume 0.2 mL for mice, 0.5-1.0 mL for larger animals with sterile saline or sterile-buffered saline. Although QS-21 is a...
