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PubMed1991

Algammulin (gamma inulin/alum hybrid adjuvant) has greater adjuvanticity than alum for hepatitis B surface antigen in mice.

Algammulin is a new vaccine adjuvant comprising a stable suspension of 1-2 microns ovoids of the immune stimulant gamma inulin in which alum (Alhydrogel) is embedded as a protein carrier. Adjuvanticity tests in mice with Algammulin show that the presence of gamma inulin on the alum particles has synergistically enhanced their adjuvanticity for low doses of hepatitis B surface antigen. The primary-response titres of HBsAg-specific antibody from a given low dose of alum injected as Algammulin were 3- to 5.6-fold greater than those from the same alum dose injected as free alum. This corresponds closely with more extensive previous work using keyhole limpet haemocyanin as antigen.

Adjuvants, Immunologic↗
PubMed1989

Mechanism of lung injury in cotton rats immunized with formalin-inactivated respiratory syncytial virus.

Respiratory syncytial virus (RSV) seronegative cotton rats were immunized intramuscularly at four and eight weeks of age with either formalin-inactivated, alum-precipitated RSV grown in HEp-2 cell tissue cultures or virus-free HEp-2 cell tissue cultures similarly prepared. Sham-immunized animals served as controls. At 12 weeks of age, all animals were challenged with 6 x 10(5) plaque forming units of live RSV via the intranasal route. Animals were killed at predetermined days and evaluated for RSV antibody, virus replication and pulmonary histopathology. Of animals immunized with inactivated-RSV, 88% developed neutralizing antibody to RSV. Virus replication in the lungs of such animals was significantly reduced compared with tissue-culture-immunized animals. Surprisingly, however, both groups exhibited pulmonary histopathology, characterized by polymorphonuclear and mononuclear interstitial infiltrates. The virus-immunized animals manifested a more severe inflammatory reaction that reached a peak earlier than the virus-free, tissue-culture-immunized control group. In contrast, sham-immunized animals, when infected with live RSV for the first time, developed little or no pulmonary histopathology. The data suggest that the pathogenesis of pulmonary injury during natural RSV infection in the immunized host is due primarily to prior sensitization to the virus. In the animal model, sensitization to non-viral tissue culture components also contributes to lung injury.

Alum Compounds↗
PubMed1991

Algammulin, a new vaccine adjuvant comprising gamma inulin particles containing alum: preparation and in vitro properties.

Crystallization of inulin with alum forms a fine (1-2 micron) suspension of electron-dense ovoids; the alum is embedded in inulin particles, which are then converted to the immune stimulant polymorphic form, gamma inulin. This very stable hybrid preparation is termed Algammulin. Preferred conditions for its preparation are described. The alum still adsorbs protein. Gamma inulin is equally able to activate the alternative pathway of complement in vitro whether free or combined as Algammulin. Gamma inulin, either free or combined as Algammulin, dissolved on heating over a narrow temperature range that can be used to characterize the polymorphic form of the inulin.

Adjuvants, Immunologic↗
PubMed1991

The adjuvanticity of Algammulin, a new vaccine adjuvant.

Algammulin is a suspension of 1-2 microns ovoids of the immune stimulant gamma inulin (g-IN) in which alum is embedded as a carrier for protein or other anionic antigens. Tests for specific IgG and seroconversion responses in mice immunized with keyhole limpet haemocyanin (KLH) show that the presence of g-IN on the alum has increased its adjuvanticity 6- to 17-fold (p less than 0.001), and thus has a synergistic effect. A mixture of alum and g-IN was no more active than alum alone. Low KLH doses at borderline seroconversion levels (0.1-1 microgram/mouse) allow comparison with a vaccine situation. The improved Algammulin responses extended to 58 days after primary doses and to memory recall after boost at 65 days. At 1 mg Algammulin i.p. in primary and secondary doses the anti-KLH IgG responses were equal to those from Freund's complete adjuvant. The g-IN on the alum increased all responses tested (IgG 1, 2a, 2b, 3 and total IgG, IgM, IgA and IgE), but changed the emphasis from that of alum (favouring mostly IgG 2a, IgG 2b and IgA).

Adjuvants, Immunologic↗
PubMed1992

Parameters affecting the immunogenicity of microencapsulated tetanus toxoid.

The immunogenic potential of tetanus toxoid (TT) was compared when either adsorbed to aluminium hydroxide (TT-alum) or entrapped in microparticles consisting of poly(D,L-lactic-co-glycolic acid) (PLA:PGA, 55:45) derived polymers. Furthermore, the effect of administering the microparticles in an aqueous buffer or water-in-oil emulsion on the TT immunogenicity was also investigated. When mice were immunized with the different formulations, similar levels of anti-TT antibodies were observed during the primary IgG response. The choice of the carrier seemed to play an important role for both the level and maintenance of the secondary IgG response, attained as a consequence of a booster immunization with TT-alum. The strongest secondary antibody response was obtained by priming with TT-containing microparticles, resuspended in water-in-oil emulsions. As expected, incomplete Freund's adjuvant (IFA) proved to be a more potent adjuvant than peanut oil, whereas resuspension of the microparticles in aqueous solution induced a relatively less efficient antibody response. Overall, microencapsulated TT primed the mice more effectively, since the secondary antibody response was higher and persisted longer compared with TT-alum priming. These results indicate that in addition to TT maintaining its antigenicity after microencapsulation, the microparticles also potentiate its immunogenic properties. This approach should prove very useful for designing more effective vaccines.

Adjuvants, Immunologic↗
PubMed1993

Adhesion-mediated enhancement of the adjuvant activity of alum.

Alum, the only adjuvant currently licensed for use in humans, fails to adsorb influenza virus haemagglutinin (BHA) and is a poor adjuvant for this antigen. A specific monoclonal anti-BHA antibody adsorbed to alum promoted adhesion of the antigen to the adjuvant. The 'alum-anti-BHA-BHA' complex was found to be 1500-fold more immunogenic in mice and 5-fold more immunogenic in rabbits than a mixture of alum and BHA lacking the anti-BHA antibody. The biotin-binding protein, avidin, also mediated adsorption of biotinylated BHA to alum, and enhanced its immunogenicity to a comparable extent. These results indicate that the adjuvant activity of alum can be markedly enhanced by promoting a physical association between the antigen and the adjuvant.

Adhesiveness↗
PubMed1993

A self-associating hepatitis B surface antigen-derived peptide that is immunogenic in alum.

We previously described an oligomeric synthetic peptide derived from the hepatitis B surface antigen that displayed a limited tendency to form self-associating macromolecular structures in solution. Here it is demonstrated that amino-terminal myristylation of this peptide results in near quantitative aggregation of the oligomeric peptide. The myristylated peptide is highly immunogenic when used in conjunction with alum as adjuvant in both the rabbit and rhesus monkey models. The antibody response generated by peptide also cross-reacted with native antigen and was long-lasting. Collectively the results described in this and previous reports offer an attractive new approach for generating immunogenic peptide mimetics of conformational epitopes that may find application as vaccines.

Alum Compounds↗
PubMed1995

Effect of DETOX as an adjuvant for melanoma vaccine.

The identification of effective adjuvants is critical for tumor vaccine development. Towards this end, we examined whether the immunogenicity of a melanoma vaccine could be potentiated by DETOX, an adjuvant consisting of monophosphoryl lipid A (MPL) and purified mycobacterial cell-wall skeleton (CWS). Nineteen patients with resected stage III melanoma were immunized with a polyvalent melanoma antigen vaccine (40 micrograms) admixed with DETOX, q3 wks x 4. Seven patients received vaccine + low-dose DETOX (10 micrograms MPL + 100 micrograms CWS) and 12 received vaccine + high-dose DETOX (20 micrograms MPL + 200 micrograms CWS). A non-randomized control group of 35 patients was treated similarly with 40 micrograms vaccine + alum. One week after the fourth vaccine immunization, melanoma antibodies were increased over baseline in 7/7 (100%) patients treated with vaccine + low-dose DETOX, 8/12 (67%) patients treated with vaccine + high-dose DETOX, and in 4/19 (21%) of vaccine + alum patients. For the entire DETOX group, the antibody response rate was 15/19 (79%) compared 4/19 (21%) in the alum group (p < 0.001). In contrast, a strong delayed-type hypersensitivity (DTH) response (> or = 15 mm increase in DTH response over baseline) was induced in 50% of the entire DETOX group versus in 47% of the alum group. Median disease-free (DF) survival for the entire DETOX group was 17.8 months compared with 32.1 months in the alum group (p < 0.05). In conclusion, DETOX markedly potentiated antibody but had little effect on DTH responses to melanoma vaccine immunization. It did not appear to improve disease-free survival in comparison to alum in this non-randomized study.

Adjuvants, Immunologic↗
PubMed1995

The influence of gamma inulin and Algammulin on the immune response in sheep to a recombinant antigen of Taenia ovis.

Gamma inulin and Algammulin, two new adjuvants, were examined and compared with alum and Freund's Complete/Incomplete Adjuvant (FCA/FIA), for potentiation of cell-mediated immunity (CMI) and humoral immunity in sheep to a recombinant Taenia ovis antigen. The ability to protect sheep when challenged with live T. ovis eggs was also assessed. The results showed that gamma inulin and Algammulin induced a CMI response which was comparable to the FCA/FIA and alum groups and significantly higher than the control saline group. While gamma inulin, Algammulin and alum performed similarly and induced a significantly higher humoral immune response than the saline group. FCA/FIA elicited a much higher humoral immune response. Algammulin did not show the synergistic effect seen in mice and performed similarly to gamma inulin and alum alone. All the adjuvant groups induced significantly higher IgG1 and IgG2 levels than the saline group and they all favoured IgG1 production. When the sheep were challenged with live T. ovis eggs, at 25 weeks after primary immunization, the only group to show significant protection was the one which received FCA/FIA.

Adjuvants, Immunologic↗
PubMed1995

Humoral and cellular responses induced by intradermally administered cytokine and conventional adjuvants.

Serum antibody responses to the model protein antigen avidin were monitored in sheep following intradermal injection of avidin formulated with a range of commercially available and experimental adjuvants, including muramyl dipeptide (MDP), aluminium hydroxide gel (alum), recombinant ovine interleukin1 beta (rovIL-1 beta), rovIL-1 beta + alum, Quil A + alum or Emulsigen Plus. The highest antibody responses were recorded for animals immunised with avidin in rovIL-1 beta + alum, Quil A + alum or Emulsigen Plus, with moderate responses resulting from use of rovIL-1 beta or alum alone as adjuvants. Lower antibody responses to avidin were recorded when avidin was administered alone or with MDP. Delayed-type hypersensitivity (DTH) responses to avidin indicated that the most pronounced cellular response occurred in animals immunised with rovIL-1 beta + alum. Local cellular changes induced after primary and secondary intradermal injections indicated that distinct patterns of cellular recruitment were induced by the different adjuvants. Avidin with MDP resulted in an elevation of CD4+ T cells in the upper dermis while Emulsigen Plus induced an infiltration of large numbers of neutrophils throughout the dermis and reticular layers. CD4+, CD8+ and gamma delta + T cells increased in number and were found evenly distributed throughout these regions. Alum-based adjuvants resulted in the development of distinct cellular accumulations comprising primarily CD4+ T cells and CD45R + B cells arranged in distinct foci in the reticular layer. These cells were strongly class II positive as were the majority of macrophage like cells surrounding the foci. Staining for factor VIII related antigen indicated the presence of endothelial venules in the T and B cell foci and surrounding tissues.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1994

Immunization of cottontop tamarins and rabbits with a candidate vaccine against the Epstein-Barr virus based on the major viral envelope glycoprotein gp340 and alum.

The Epstein-Barr virus (EBV) is associated with a range of life-threatening diseases in humans. Development of an effective vaccine has therefore been an important objective. One problem in the development of a subunit vaccine for human administration is the selection of a satisfactory adjuvant since the only one currently licensed for human use is alum, although this is not considered to be very effective. The present study demonstrated that a subunit vaccine composed of the EBV envelope glycoprotein gp340 with alum as the adjuvant did elicit protective immunity against EBV-induced lymphoma in three out of five cottontop tamarins. Furthermore, rabbits immunized with gp340/alum developed the same range of antibody responses as rabbits immunized with gp340/SAF-1, an experimental adjuvant claimed to be more effective than alum. Therefore, these results indicate that alum should be evaluated as an adjuvant as part of a human trial of a gp340-based subunit vaccine.

Adjuvants, Immunologic↗
PubMed1994

Immunogenicity of an alum-adsorbed synthetic multiple-antigen peptide based on B- and T-cell epitopes of the Plasmodium falciparum CS protein: possible vaccine application.

Multiple-antigen peptides (MAPs), containing B- and T-cell epitopes of the Plasmodium falciparum circumsporozoite (CS) protein, have been designed to overcome the limitations of first-generation peptide vaccines caused by low epitope density, carrier toxicity and the lack of parasite-derived T-cell epitopes. The immunogenicity of a P. falciparum MAP construct (T1B4), containing four copies of the 5' repeat cell T epitope (T1) combined with the 3' repeat epitope (NANP)3, has been examined using different adjuvant formulations. Mice immunized intraperitoneally or subcutaneously with (T1B)4 in alum, a formulation suitable for human vaccines, developed high anti-peptide and anti-sporozoite antibody titres, comparable with those obtained with Freund's adjuvant. The MAP/alum formulation also elicited a strong anamnestic antibody response in sporozoite-primed mice, raising the possibility of using a MAP/alum vaccine to increase the low anti-sporozoite antibody levels of people living in malaria-endemic areas.

Adjuvants, Immunologic↗
PubMed1995

Virosomes as carriers for combined vaccines.

Immunopotentiating reconstituted influenza virosomes (IRIV) are liposomes which carry the two glycoproteins of the influenza virus on their surface. A hepatitis A vaccine using IRIV as carrier has very good immunogenicity and is well tolerated. The objective of this study was to produce a fivefold combined vaccine against hepatitis A and B, diphtheria, tetanus and influenza A/B, and to show that in principle IRIVs can serve as carriers for multiple antigens which have good immunogenicity and are well tolerated. A total of four studies were carried out. Either the combined vaccine or the corresponding adequately tested alum-adsorbed single vaccines were tested for reactogenicity and immunogenicity in young adults. A hepatitis A and B combination on an IRIV base showed the same immunogenicity and toleration as the single vaccines. However, with the simultaneous coupling of all five vaccines on the same IRIV or the binding of Di-Te and HAV on different IRIVs there was a suppression of the humoral immune response against HAV (p = 0.03). The possibility that epitope-specific suppression had occurred could be ruled out. The suppression of the response against HAV could be circumvented by halving the quantity of Di-Te antigen in the combined vaccine so as to avoid antigenic competition. Surprisingly, the immunogenicity of Di-Te vaccination in the combination proved superior to that of a separate vaccination. All vaccinations were well tolerated.

Adult↗
PubMed1995

Interferon-gamma inductive effect of liposomes as an immunoadjuvant.

Adjuvant effect of liposomes was compared to that of aluminium hydroxide (Alum) using ovalbumin (OVA) as a model antigen, and the difference in adjuvanticity of liposomes from Alum has been evaluated on the basis of cytokine production. Both adjuvants enhanced the IgG levels, but a remarkable difference was observed in the production of IgG subclass; Alum enhanced IgG1 levels, and liposomes enhanced IgG2a, IG2b, and IG3 levels. Further, Alum enhanced antigen-specific IgE levels, whereas liposomes did not. To clarify the difference in adjuvant effect of these adjuvants, secretion of cytokines, especially interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) which regulate IgE production, was estimated ex vivo. Overnight culture of splenic cells obtained from mice immunized with liposomes encapsulating OVA elicits IFN-gamma secretion, but not IL-4 secretion. On the other hand, the production of both cytokines was elevated by the immunization with OVA-Alum complex. The results indicate that the difference of adjuvant activity between liposomes and Alum may come from the difference in the secretion of IL-4 and that, consequently, a different class of antibody response is developed. More importantly, negatively charged liposomes containing phosphatidylserine remarkably promoted IFN-gamma secretion compared to neutral liposomes.

Adjuvants, Immunologic↗
PubMed1995

HIV-1 recombinant gp160 vaccine induced antibodies in serum and saliva. The NIAID AIDS Vaccine Clinical Trials Network.

As part of a phase I safety and immunogenicity trial of a vaccinia-expressed HIV-1 recombinant gp160 (rgp160) candidate vaccine, we measured serum and saliva antibody responses in low risk, uninfected volunteers. Six healthy adult volunteers received 50 micrograms doses of rgp160 vaccine adjuvanted in alum and deoxycholate at months 0, 1, 6, and 12. A 200 micrograms rgp160 immunization was given to four volunteers at 18 months. The vaccine induced anti-envelope glycoprotein IgG and IgA serum antibodies in all six volunteers. Saliva antibodies to envelope glycoprotein appeared in some volunteers at certain timepoints. Three volunteers appeared to transiently develop vaccine-induced secretory IgA antibody to envelope glycoprotein in whole saliva.

AIDS Vaccines↗
PubMed1995

Immune response of mice to immunization with subunit influenza A vaccine in DTP vaccine.

Experiments were carried out to examine the initial feasibility of immunizing infants and children with inactivated influenza virus vaccine combined with diphtheria-tetanus-pertussis (DTP) vaccine. Groups of mice were immunized with saline vaccine or vaccine mixed with DTP: three doses of vaccine were given 3 weeks apart, and the antibody response and resistance to challenge infection were tested 3 weeks after the 1st and 3rd immunizations. The results showed that the antibody response and immunity to challenge virus infection were significantly greater for mice given vaccine in DTP than for mice given saline vaccine alone. Comparison of the response to graded doses of vaccine in saline or DTP indicated that vaccine in DTP was > 250-fold more effective in inducing serum antibody and protection than saline vaccine alone. The enhancing activity of DTP was significant for the alum component alone; however, most of the adjuvant effect was from the antigen components of the DTP vaccine. The results suggest that immunization against influenza in infants and young children could be achieved by combining small amounts of influenza antigen with DTP vaccines; however, the present results have been obtained in mice, and, since the responses to vaccines and adjuvants vary from species to species, the present results cannot be used to indicate similar results in human volunteers. The results indicate the potential value of an immunization procedure which should be tested in volunteers and which could provide a simple strategy for the immunization of at-risk infants and children against influenza.

Adjuvants, Immunologic↗
PubMed1996

The effects of adjuvants on CTL induction by V3:Ty-virus-like particles (V3-VLPs) in mice.

We have previously described the generation of HIV-1 V3-specific cytotoxic T-lymphocytes (CTL) responses in BALB/c (H-2d) mice following immunization with Ty-virus-like particles carrying the V3 loop of gp120 (V3-VLPs) without adjuvant. In this study the effects of various adjuvants on CTL induction by V3-VLPs was examined. Mice immunized with V3-VLPs formulated in aqueous-based adjuvants, Detox, gamma-inulin, galactosaminylmuramyl dipeptide and Chemivax generated V3-specific CTL responses, although at reduced levels when compared to the no adjuvant group. V3-VLPs prepared in Alhydrogel, algamulin or as an oil emulsion in SAF-MF failed to generate V3-specific CTL responses. The mechanism whereby alum prevented the induction of a CTL response was investigated further. Immunization with V3-VLPs prepared in non-saturating doses of alum or alum plus EDTA primed for strong CTL responses, indicating that free VLPs do, but alum-bound VLPs do not enter the MHC class I processing pathway of antigen-presenting cells (APCs). Furthermore, V3-VLPs with very low doses of alum led to an enhancement of the CTL response. The formulation of hybrid Ty-VLPs in oil based or precipitating adjuvants, therefore, inhibits access to the MHC class I processing pathway of APCs. The intact particulate structure of hybrid VLPs is therefore strictly necessary for CTL induction.

Adjuvants, Immunologic↗