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Comparison of immune responses to a native viral antigen and a synthetic peptide derived from it: implications for vaccine development.

Murine immune responses to the hepatitis B surface antigen (HBsAg) and a synthetic peptide derived from it were compared at the humoral level. Six of nine strains used responded to either peptide or HBsAg, though restriction profiles were not superimposable. Two of three strains non-responsive to HBsAg produced an antibody response on immunization with peptide which was cross-reactive with both peptide and HBsAg. In in vitro lymphocyte stimulation assays, lymphocyte from all six peptide-immunized mouse strains could be induced to proliferate on challenge with HBsAg. However, of the HBsAg-immunized groups, lymphocytes from only three of six responder strains proliferated on in vitro HBsAg challenge. Cumulatively, these results suggest that a vaccine formulation that includes both protein antigens and synthetic peptides derived from these proteins may be more effective at eliciting an immune response in a broader cross-section of target population.

Animals↗

A short history of the use of animals in vaccine development and quality control.

Man has been using animals since early times to gain an insight into health, illness and death. The oldest known medical standard work, the Corpus Hippocraticum (circa 350 BC), contains descriptions of experiments on pigs. Although the first attempt at immunoprophylaxis dates as far back as the 6th century (variolation was practised in China to protect people against smallpox), it was not until the middle of the 19th century that animal experimentation acquired full scientific status in the development and quality control of immunobiological products. It was Louis Pasteur and Robert Koch who, through studies on animals, succeeded in underpinning the causal relationship between infectious diseases and micro-organisms, thus opening the way to the discovery of effective therapeutic and prophylactic agents for a number of these diseases. In several respects, the experimental animal work carried out in the last decade of the 19th century to find an effective and reliable way of treating and preventing diphtheria determined the use of animals. Many common routine animal tests in the quality control of immunobiologicals arose from diphtheria research. Conversely, diphtheria was one of the first diseases where experimental animal research laid the foundation for effectively reducing child mortality. This had a very profound impact on the attitude of society towards animal experiments in those days and almost completely eliminated the growing influence of the antivivisection movement. The interest in the possibilities of replacement, reduction and refinement (the Three-Rs concept) of the use of laboratory animals is increasing for several reasons, including concern about animal welfare. The root of animal welfare can be traced back to the 18th century with the formulation of utilitarian ethics. One characteristic feature of these ethics was that the interests of any creature which is submitted to any procedure should be taken into consideration. This presentation sets out some major historical contributions of animal experiments to the development and quality control of immunobiologicals. Attention is also paid to the changing attitude of society towards animal experiments and its impact on the development of alternative methods. It is concluded that, although animal experiments have played an important part, a new area is now beginning in which increasing emphasis will be placed on in vitro methods.

Animal Testing Alternatives↗

Immunomodulation of atherosclerosis: implications for vaccine development.

A number of studies have shown activation of the immune system throughout various stages of atherosclerosis. Recent observations have suggested that activation of immune responses may promote atherosclerosis on one hand by inducing and perpetuating arterial inflammation, whereas on the other hand, selective activation of certain immune functions may inhibit atherosclerosis and arterial inflammation. These observations suggest the possibility that selective suppression of proatherogenic immune responses or selective activation of antiatherogenic immune responses may provide new approaches for atherosclerosis prevention and treatment. Several antigens activating immune responses affecting development of atherosclerosis have been identified. These immune responses may be modulated by presenting the antigens together with different types of adjuvants as well as through the route of administration. In this review, we summarize recent experimental studies using immunomodulatory approaches for treatment of atherosclerosis.

Animals↗

Protective efficacy of monoclonal antibodies to class 1 and class 3 outer membrane proteins of Neisseria meningitidis B:15:P1.16 in infant rat infection model: new prospects for vaccine development.

The protective efficacy of monoclonal antibodies to class 1 and class 3 outer membrane proteins of Neisseria meningitidis B:15:P1.16 was tested in an infant rat infection model. Four monoclonal antibodies to class 1 protein had bactericidal titres exceeding 20,000 and they protected infant rats completely against bacterial challenge with meningococci carrying the same class 1 protein, P1.16. One monoclonal antibody to class 3 protein was highly bactericidal (titer greater than 20,000), whereas two others had no bactericidal activity. All these antibodies gave some protection from infection, resulting in mortalities varying from 66 to 83% as compared to 100% in control rats who had received either unrelated monoclonal antibody or saline. These results strongly speak for class 1 outer membrane protein as a vaccine candidate for meningococcus group B.

Animals↗

Transfer of neutralizing IgG to macaques 6 h but not 24 h after SHIV infection confers sterilizing protection: implications for HIV-1 vaccine development.

Passive transfer of high-titered antiviral neutralizing IgG, known to confer sterilizing immunity in pig-tailed monkeys, has been used to determine how soon after virus exposure neutralizing antibodies (NAbs) must be present to block a simian immunodeficiency virus (SIV)/HIV chimeric virus infection. Sterilizing protection was achieved in three of four macaques receiving neutralizing IgG 6 h after intravenous SIV/HIV chimeric virus inoculation as monitored by PCR analyses of and attempted virus isolations from plasma, peripheral blood mononuclear cell, and lymph node specimens. In the fourth animal, the production of progeny virus was suppressed for >4 weeks. A delay in transferring NAbs until 24 h after virus challenge resulted in infection in two of two monkeys. These results suggest that even if a vaccine capable of eliciting broadly reactive NAbs against primary HIV-1 were at hand, the Abs generated must remain at, or rapidly achieve, high levels within a relatively short period after exposure to virus to prevent the establishment of a primate lentivirus infection.

AIDS Vaccines↗

Vaccine development for Alzheimer's disease.

Alzheimer's disease (AD) is the most common cause of age-related cognitive decline. Both active and passive immunization paradigms have illustrated the potential to prevent and reverse established AD pathology in transgenic and non-transgenic animal models of AD. Follow-up studies have shown that changes in amyloid burden observed with immunization could rescue cognitive deficits in both young and aged mice. Despite the success of immunotherapy in animal models, clinical trials were halted early. It has become clear that more preclinical work was needed before initiating trials, as most of the adverse events observed in patients could have been predicted using animal models. Despite these setbacks, clinical trials have demonstrated the utility of amyloid-beta (Abeta) vaccination in reducing amyloid pathology and potentially reducing cognitive decline. Several novel approaches to immunotherapy, including modified immunogens, adjuvants and modes of administration have been designed, which hold promise for human testing. Clinical trials using a safer vaccine, which is potent enough to elicit a robust antibody response in the absence of encephalitis may prove effective in mitigating progressive neurodegeneration seen in AD. If so, Abeta vaccination could supplant current symptomatic treatment and represent one of the first therapeutic options for AD based on the amyloid cascade hypothesis.

Alzheimer Disease↗

Vaccine development against Theileria parasite.

Bovine piroplasmosis caused by Theileria sergenti is a major cause of economical loss in grazing cattle in Japan. We found that parasite stocks and isolates consist of genetically and antigenically mixed population. To differentiate parasite populations bearing 2 allelic forms of p32, an immunodominant piroplasm surface protein, 2 sets of oligonucleotide primers were designed to amplify either of the 2 alleles by polymerase chain reaction (PCR). By using this allele-specific PCR, we found that the majority of T. sergenti-infected calves in Japan harbored mixed parasite populations with C and I type parasites. Amino acid sequence of p32 contains Lys-Glu-Lys (KEK) motif which is one of tripeptide necessary for malaria parasite to invade erythrocytes. We produced 2 vaccine candidates, recombinant baculovirus p32 and synthetic peptide containing KEK motif. Immunization of either recombinant p32 or synthetic peptide containing a KEK sequence with adjuvant resulted in low parasitemia and reduced the clinical symptoms compared to control calves. Interestingly, parasites with a p32 allelic form corresponding to one used as the immunogen were suppressed. Therefore, a cocktail vaccine containing KEK peptides derived from C and I type parasites is desired for control Theileria parasite infection in Japan.

Amino Acid Sequence↗

Host response and vaccine development to Helicobacter pylori infection.

Studies in both humans and animals demonstrate that H. pylori is capable of illiciting an innate response that in part is regulated by the genetic makeup of the host. These innate responses includes stimulating immune effector mechanisms at the cellular and biochemical level resulting in the influx of neutrophils into the lamina propria and have even been shown to modify gastric acid secretion. The availability of good animal models of chronic Helicobacter infection has also allowed investigators to begin to examine how the adaptive host immune response prevents and/or exacerbates Helicobacter-induced gastroduodenal disease. The experimental H. felis/mouse model has been utilized by a number of laboratories to investigate mechanisms of host defense against chronic Helicobacter infection. This model and the more recently developed H. pylori rodent model has not only allowed investigators to confirm the feasibility of immunotherapy to prevent and/or cure Helicobacter infection but also to begin to examine how the host immune response prevents and/or exacerbates Helicobacter-induced gastroduodenal disease. Based on these studies a hypothesis is emerging that suggests that protection and/or cure from Helicobacter infection is mediated primarily by an upregulated cellular immune response which may act via an antibody independent mechanism. Paradoxically, following natural infection with H. pylori, a component of the cellular immune response also promotes chronic gastric inflammation without clearance of the organism. The recent development of reliable and reproducible H. pylori/rodent models of disease and the availability of numerous inbred strains, transgenic and knockout animals, will allow investigators to continue to explore the role the host cellular and humoral immune response plays in promoting or preventing this infection.

Adjuvants, Immunologic↗

Analysis of antibody response in humans to the type A OspC loop 5 domain and assessment of the potential utility of the loop 5 epitope in Lyme disease vaccine development.

The OspC protein of Borrelia burgdorferi is an immunodominant antigen. Here we demonstrate that the loop 5 domain of type A OspC is surface exposed, elicits bactericidal antibody in mice, and is antigenic in humans. The data suggest that loop 5 may be suitable for inclusion in a polyvalent, chimeric OspC vaccinogen.

Amino Acid Sequence↗

Analysis of MHC-presented peptides: applications in autoimmunity and vaccine development.

T cells recognize antigenic peptides presented on the cell surface by major histocompatibility complex (MHC) molecules. Here, Roman Chicz and Robert Urban describe the features of peptides bound to MHC molecules and the mechanism by which these surface proteins bind diverse peptide ligands with high affinity. In addition, they discuss the application of new technologies to the identification of MHC-associated peptides.

Animals↗

Future vaccine development at NICHD.

Using published data and the results of our studies, we hypothesized that a critical level of serum IgG antibodies to the surface structures of invasive pathogens (capsular polysaccharides of Haemophilus influenzae type b, pneumococcus, meningococcus, Salmonella typhi, Escherichia coli, and Staphylococcus aureus, the O-specific polysaccharide LPS domain of the LPS of Shigella, non-typhoidal Salmonella, and E. coli, and the capsular polypeptide of Bacillus anthraces) confer immunity to these pathogens. Covalent attachment to a protein increases their immunogenicity and bestows T-cell properties to these antigens. We have also shown that a critical level of serum IgG antibodies to pertussis toxin alone induces immunity on both an individual and on a community basis (herd immunity) to Bordetella pertussis. It is likely that all the above conjugates and pertussis toxoid will be incorporated into vaccines for routine infant immunization.

Bacterial Infections↗

Malaria in 1998: advances in diagnosis, drugs and vaccine development.

Malaria threatens almost half the world's population. Although a concerted worldwide effort might control this disease, the reality is that inadequate and ineffective laboratory diagnosis and treatment conspire in the yearly deaths of 2,000,000 children from malaria. The failure to institute and implement safeguards to maintain the efficacy of new antimalarial drugs is likely to accelerate the emergence of untreatable malaria, creating an ominous parallel to tuberculosis. There is hope. Field expedient, simple and affordable malaria diagnostics are at hand. Artemisinin derivatives remain surprisingly effective against the multiply drug-resistant falciparum malaria of southeast Asia, despite the widespread and unregulated use of these agents. Human trials have begun of WR238605, a promising primaquine replacement that has prophylactic, treatment and transmission-blocking potential. After demonstrating unprecedented protection against homologous challenge, RTS,S, a new sporozite-based malaria vaccine candidate, is now in field trials.

Journal Article↗

Immune response to group A streptococcal C5a peptidase in children: implications for vaccine development.

The group A streptococcal C5a peptidase (SCPA) is a major surface virulence protein that facilitates the establishment of local infection by group A streptococci (GAS). We measured the human immune response to SCPA, using a standardized indirect enzyme-linked immunosorbent assay. Paired acute and convalescent serum samples from children with GAS-associated pharyngitis were assayed, and a strong immune response to SCPA was demonstrated that was independent of the infecting M type and the age of the patient. Western blot analysis of bacterial extracts revealed that all tested M types expressed SCPA. The immune response to SCPA correlated with the anti-streptolysin O and anti-DNase B responses. These data confirm the immunogenicity of SCPA in humans. Previous knowledge of SPCA's role in virulence, its highly conserved nature, and the results of mouse protection studies make SCPA an ideal vaccine candidate for the prevention of GAS disease.

Acute Disease↗

Vaccine development for group A streptococcus infections and associated disease.

Group A streptococcus (GAS) is responsible for a number of diseases ranging from uncomplicated pharyngitis through to life-treating invasive and post-infectious diseases such as necrotizing fasciitis and rheumatic heart disease. GAS associated diseases occur globally and are serious problems in many developing nations and indigenous populations of many developed nations. This, and the resurgence in industrialized countries, and increased virulence of GAS in the 1980s highlight the need of cost-effective control strategies. Here we highlight the GAS diseases that are still a problem in many populations and discuss potentially useful strategies to combat GAS infections and disease.

Adjuvants, Immunologic↗

Involvement of T cells in malaria immunity: implications for vaccine development.

T cells are critical for immunity to malaria, not only because they function as helper cells for an antibody response, but also because they serve as effector cells. Such cellular immunity is directly implicated in protection from sporozoites and plays an important role in protection from blood-stage parasites. It also can block transmission of malaria from mammalian host to the mosquito. Both CD8 and CD4 effector cells have important roles. The parasite's defence from immune attack, however, is designed to minimize activation of T cells. Thus, there appears to be limitation of the number of T sites within many malaria proteins and variation within these limited sites. Homology to host proteins and resultant immune-escape due to tolerance may be another mechanism. These parasite defence mechanisms highlight both the importance of T-cell immunity in malaria and the challenge of designing effective vaccines to stimulate T cells.

Animals↗

Peptide sequences with strong stimulatory activity for lymphoid cells: implications for vaccine development.

Seven peptides derived from the bacterial major outer-membrane protein TraT were synthesized and then tested in lymphoproliferative assays using lymphoid cells from a variety of animals that had been immunized with the native TraT molecule in saline. A hierarchical pattern of responsiveness to the peptides was observed in the four animal species studied and in particular three of the peptides (T2, T4 and T6) showed very strong responses in all species. The 'universality' of the TraT-derived peptides was confirmed by studying the responsiveness of lymphoid cells obtained from the peripheral blood of twenty clinically normal human donors. Thus, following a secondary in vitro immunization with TraT-pulsed human peripheral blood mononuclear cells, responsiveness to TraT and to the TraT-derived peptides was observed in the cultures derived from all twenty donors. Taken together, our findings imply that the putative T-cell epitope peptides (T2, T4 and T6) could be employed as carriers in subunit vaccines and thereby help to overcome the unresponsiveness observed in animals and humans as a result of MHC restriction.

Adjuvants, Immunologic↗

Use of liposomes as an immunopotentiating delivery system: in perspective of vaccine development.

Liposomes have been widely used to deliver antigens to the antigen-presenting cells (APCs) and also to modify their immunological behaviour in model animals. We recently demonstrated the potential of yeast lipid liposomes to undergo membrane-membrane fusion with cytoplasmic membrane of the target cells. Interestingly, studies in the present report revealed that antigen encapsulated in yeast lipid liposomes could be successfully delivered simultaneously into the cytosolic as well as endosomal processing pathways of APCs, leading to the generation of both CD4+ T helper and CD8+ cytotoxic T cells. In contrast, encapsulation of same antigen in egg phosphatidyl-choline (PC) liposomes, just like its free form, has inefficient access to the cytosolic pathway of major histocompatibility complex (MHC) I dependent antigen presentation and failed to generate antigen specific CD8+ cytotoxic T-cell response. However, both egg PC as well as yeast lipid liposomes have elicited strong antigen specific antibody responses in immunized animals. These results imply usage of liposome encapsulated antigen as potential candidate vaccine capable of eliciting both cell mediated as well as humoral immune responses.

Animals↗

Elevation of immune activation in kenyan women is associated with alterations in immune function: implications for vaccine development.

The infectious burden leading to immune activation can vary between different populations and lead to various immune dysfunctions. We compared the effect of immune activation on apoptosis and T cell function in HIV uninfected individuals from Nairobi, Kenya (n=34), and Winnipeg, Canada (n=10). Women from Nairobi had a significantly greater number of CD8+ T cells expressing the activation markers CD38 and HLA DR. Kenyan women also had significantly higher levels of CTLA-4+ CD4 and CD8+ T cells, and reduced levels of CD28+ CD8+ cells. Levels of CD95+ CD4+ T cells were higher in Kenyan women and, correspondingly, showed higher levels of spontaneous apoptosis. Kenyan women also demonstrated hyper-responsiveness to T cell activation as assessed by interferon gamma production. This study demonstrates that in a population of Kenyan women with high levels of T cell activation, there were also elevated levels of T cell apoptotic death and hyper-responsiveness. These differences may influence the efficacy of immune responses to pathogens and must be considered when testing candidate vaccines.

Apoptosis↗