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Protein subunit vaccines of parainfluenza type 3 virus: immunogenic effect in lambs and mice.

Protein subunit vaccines were prepared from a mixture of the haemagglutinin (HN) and fusion (F) glycoproteins of parainfluenza type 3 virus (PI-3). The glycoproteins were isolated in three different forms and characterized by their sedimentation coefficients: 30S protein micelles (a complex of several HN and F glycoproteins devoid of detergent and lipid), 18S protein-TX complexes (a complex of several glycoproteins containing the detergent Triton X-100), and 4S protein-TX complexes (probably monomers of the glycoproteins complexed to Triton X-100). These preparations were tested as vaccines in mice and lambs. The immune response in the mice was assayed both in the serum and in extracts from the lungs using an ELISA technique. Both of the multimeric complexes were highly immunogenic. The 30S protein micelles induced a high antibody response after two injections with either 10 or 1 microgram protein. The serum IgG titres reached levels of about 90 micrograms/ml and 40 micrograms/ml respectively. Similar titres were reached with the 18S protein-TX complexes. After two injections of either the 30S or the 18S complexes IgA antibody responses were detected in the lung extracts. The 4S protein-TX complexes were poor immunogens and induced low antibody responses in mice. The lambs were vaccinated with the 30S protein micelles, and the immune response was evaluated serologically and in challenge experiments. The 30S protein micelles in an oil adjuvant induced detectable serum antibody titres as well as protective immunity against the pneumonia caused by the PI-3 virus.

Animals↗

Antibody responses and HIV-1 viral load in HIV-1-seropositive subjects immunised with either the MF59-adjuvanted influenza vaccine or a conventional non-adjuvanted subunit vaccine during highly active antiretroviral therapy.

OBJECTIVE: To study immunological and virological parameters in HIV-1-seropositive adults treated with highly active antiretroviral therapy (HAART) for at least 7 months after immunisation with MF59-adjuvanted (FLUAD, Chiron, Siena, Italy) or with non-adjuvanted (AGRIPPAL, Chiron) trivalent influenza vaccine. DESIGN: Blood samples, collected before and after vaccination, were analysed for the presence of antibodies against the vaccine antigens, for number of CD4+ T lymphocytes and HIV-1 RNA levels. RESULTS: Forty-four volunteers received FLUAD and 40 AGRIPPAL influenza vaccine. Thirty days after vaccination both adjuvanted and non-adjuvanted vaccines induced significant increases of anti-influenza virus antibodies. However, antibody titres found in volunteers receiving adjuvanted vaccine were in general significantly higher when compared with those found in the non-adjuvanted vaccine group. The requirements of the European Commission of influenza vaccine for a non-elderly adult population were always met by recipients of the adjuvanted vaccine, even in those with the lowest CD4+ cell counts (<200 cells/mmc). The subjects receiving the non-adjuvanted vaccine failed to met these requirements. The CD4+ T lymphocytes and plasma HIV-1 RNA levels remained stable in the long term, both in people receiving adjuvanted or non-adjuvanted vaccine. CONCLUSION: MF59-adjuvanted influenza induced a significant higher immune responses as compared with conventional vaccine in HIV-seropositive HAART-treated patients. Both vaccines were safe regarding HIV RNA viral replication and loss of CD4+ T lymphocytes.

Adjuvants, Immunologic↗

Antigen discovery and delivery of subunit vaccines by nonliving bacterial ghost vectors.

The bacterial ghost (BG) platform system is a novel vaccine delivery system endowed with intrinsic adjuvant properties. BGs are nonliving Gram-negative bacterial cell envelopes which are devoid of their cytoplasmic contents, yet maintain their cellular morphology and antigenic structures, including bioadhesive properties. The main advantages of BGs as carriers of subunit vaccines include their ability to stimulate a high immune response and to target the carrier itself to primary antigen-presenting cells. The intrinsic adjuvant properties of BGs enhance the immune response to target antigens, including T-cell activation and mucosal immunity. Since native and foreign antigens can be carried in the envelope complex of BGs, combination vaccines with multiple antigens of diverse origin can be presented to the immune system simultaneously. Beside the capacity of BGs to function as carriers of protein antigens, they also have a high loading capacity for DNA. Thus, loading BGs with recombinant DNA takes advantage of the excellent bioavailability for DNA-based vaccines and the high expression rates of the DNA-encoded antigens in target cell types such as macrophages and dendritic cells. There are many spaces within BGs including the inner and outer membranes, the periplasmic space and the internal lumen which can carry antigens, DNA or mediators of the immune response. All can be used for subunit antigen to design new vaccine candidates with particle presentation technology. In addition, the fact that BGs can also carry piggyback large-size foreign antigen particles, increases the technologic usefulness of BGs as combination vaccines against viral and bacterial pathogens. Furthermore, the BG antigen carriers can be stored as freeze-dried preparations at room temperature for extended periods without loss of efficacy. The potency, safety and relatively low production cost of BGs offer a significant technical advantage over currently utilized vaccine technologies.

Adjuvants, Immunologic↗

Naturally occurring V1-env region variants mediate simian immunodeficiency virus SIVmac escape from high-titer neutralizing antibodies induced by a protective subunit vaccine.

Macaques which developed high-titer neutralizing antibodies (htNAb) after immunization with a virion-derived oligomeric envelope glycoprotein subunit vaccine were protected against a homologous simian immunodeficiency virus SIVmac challenge. Here we demonstrate that the htNAb could be overcome by V1-env region variants isolated ex vivo from an SIVmac-infected macaque. The results further suggest that the development of V1-env region neutralization escape mutants is also necessary for survival of the virus in infected macaques. The immunological capacity of a single variable region to induce neutralizing antibodies in vaccinated and infected macaques initiate new ideas for a successful vaccine strategy.

Amino Acid Sequence↗

Recombinant subunit vaccines: potentials and constraints.

Recombinant DNA techniques have enormous potential for the development of inexpensive, safe and efficacious vaccines for the aquaculture industry. Aside from rationally attenuated pathogens, two broad categories of recombinant vaccines have been described. The first are so-called "vectored" vaccines, which consist of either viral or plasmid expression vectors harbouring genes for protective antigens from a given pathogen. The second are recombinant subunit antigens produced using heterologous protein expression systems. Less onerous from a regulatory standpoint, recombinant subunit vaccines generate strong antibody responses in recipient animals when administered parenterally with adjuvant-containing formulations. This approach is nevertheless constrained by the fact that low-cost systems for protein expression (especially E. coli, but also yeast) often generate misfolded or incorrectly processed membrane antigens that fail to protect, while more complex insect and mammalian tissue culture cells are prohibitively expensive from a production standpoint. Furthermore, subunit antigens generate less than optimal mucosal and cytotoxic T-cell responses, the last two of which are especially important for the clearance of intracellular pathogens. Fortunately, these constraints are now being overcome through the use of new generation adjuvants and delivery systems that enhance immunogenicity, as well as new expression systems for the production of viral and protozoan membrane antigens.

Adjuvants, Immunologic↗

The new generation of recombinant viral subunit vaccines.

There have been two major focal points for the development of recombinant viral subunit vaccines. Expression strategies for the candidate antigens have focused increasingly on producing more natural antigenic structures, for example, virus-like particles. Also, new adjuvants have been used in order to increase immunogenicity.

Animals↗

[Treatment of recurrent herpes labialis using a herpes simplex Type-I subunit vaccine. A prospective randomized double-blind multicenter study].

A herpes simplex type I subunit vaccine developed by Behring-Werke (Marburg/Lahn, FRG) was compared with placebo for therapeutic effectiveness in the treatment of recurrent orolabial herpes simplex in a multicentre study involving the major hospitals in Frankfurt/Main (n = 34), Hamburg (n = 18), Giessen (n = 17) und Kassel (n = 14). The effectiveness of the vaccine was judged on the reduction in the frequency of recurrences. Within the period of the 8-month study there was no statistically significant difference in the frequency of recurrences with vaccine or placebo. Both placebo and verum led to a significant decrease in the number of recurrences of orolabial herpes simplex, but the effect of placebo was slightly more pronounced.

Adult↗

A simple novel procedure for preparation of herpes simplex virus subunit vaccine.

A simple procedure for preparation of herpes simplex virus type 1 (HSV-1) subunit vaccine is described. The method is based on treatment of virus-infected cells with a nonionic detergent, removal of cell nuclei by low speed centrifugation, separation of viral nucleocapsids by high speed centrifugation through a sucrose cushion and, finally, precipitation of viral and cellular glycoproteins and proteins with ammonium sulphate (AS). Unlike to acetone precipitation, affinity chromatography on lectins or hydroxylapatite chromatography, AS precipitation repeatedly yielded the best and most reliable results as judged by relative protein content, antigenicity and immunogenicity of the final vaccine product.

Ammonium Sulfate↗

A study of acute respiratory disease in the community of Port Chalmers. III. Efficacy of influenza virus subunit vaccines in 1973, 1974 and 1975.

An evaluation of the efficacy of influenza virus subunit vaccine was undertaken during a study of acute respiratory disease in the semi-isolated community of Port Chalmers, New Zealand. In 1973, the administration of a vaccine containing A/England/42/72(H3N2) and B/Roma/1/67 stains was found to produce HI antibody titres greater than or equal to 1:40 to the A and B components in 50 percent of 32 subjects. There was no significant NI antibody response to the A component. During an epidemic of A/Port Chalmers/I/73(H3N2) occurring three to four months after vaccination, vaccinees were not protected from clinical infection. Sixty and ninety-nine subjects received vaccine containing A/Port Chalmers/1/73 and B/Hong Kong/8/73 in 1974 and 1975 respectively. In 1974 all 60 subjects received a second dose of vaccine which was shown to have little effect on the distribution of HI antibody titres. The benefits of annual vaccination in this general practice are unclear.

Adolescent↗

Immunogenicity and safety of respiratory syncytial virus subunit vaccine in seropositive children 18-36 months old.

Twenty-six children (aged 18-36 months) previously hospitalized for respiratory syncytial virus (RSV) infection were randomized to receive 50 micrograms of an RSV subunit vaccine composed primarily of F glycoprotein or saline placebo by intramuscular injection. Serum was obtained at entry and at 1 and 6 months after vaccination for detection of antibody to F glycoprotein and G glycoprotein of subtypes A (Ga) or B (Gb) and of neutralizing antibody (nAb). At 1 month, by comparing the baseline values, vaccinees had statistically significant increases in geometric mean antibody titer (GMT) of more than fourfold to F (P = .0001), Ga (P = .0001), Gb (P = .003), and nAb (P = .009). No differences in GMT were observed between F protein vaccine and placebo recipients at entry, nor between placebo recipients at entry and 1 month. RSV infections were identified in 7 placebo recipients (4 by both viral identification and seroconversion, 3 by seroconversion alone). No vaccine recipient had RSV infection documented in the 6 months after vaccination (P = .003). There were no significant vaccine-related side effects, and no evidence of enhanced respiratory illnesses was observed. The subunit F protein vaccine appears safe and immunogenic and may prevent infection in healthy children primed by prior RSV infection.

Antibodies, Viral↗

Influence of administration dose and route on the immunogenicity and protective efficacy of BBG2Na, a recombinant respiratory syncytial virus subunit vaccine candidate.

The immunogenicity and protective efficacy of BBG2Na, a novel recombinant respiratory syncytial virus subunit vaccine candidate, was assessed in BALB/c mice under various conditions of dose, administration route and number of immunisations. A single intra-peritoneal (i.p.) dose of 2 microg, or two doses of 0.2 microg, were sufficient to induce elevated RSV-A serum antibodies and sterilising lung protective immunity. Serum antibody titres were significantly boosted following second immunisations, but not a third. Of three routes of immunisation, i.p. induced the highest RSV-A antibody titres, followed in efficacy by the intra-muscular (i. m.) and subcutaneous (s.c.) routes. Nonetheless, all three routes induced comparable and sterilising lung protection. In contrast, upper respiratory tract protection was observed only after i.p. vaccination, although significant viral titre reductions were evident following i.m. or s.c. immunisations. Interestingly, Pepscan analyses indicated that antibody epitope usage was highest in i.p. and lowest in i.m. immunised mice, respectively. Nonetheless, all routes resulted in antibody responses to known lung protective epitopes (protectopes). Thus, the prevention of serious lower respiratory tract disease, the principle goal of a RSV vaccine, but not URT infection, is dose dependent but unlikely to be influenced by the route of BBG2Na administration.

Animals↗

Development of a single-shot subunit vaccine for HIV-1. 5. programmable in vivo autoboost and long lasting neutralizing response.

The subunit vaccine for HIV-1, recombinant glycoprotein 120 (rgp120), was used as a model antigen to evaluate the potential for a pulsatile single immunization vaccine formulation consisting of poly(lactic-co-glycolic) acid (PLGA) microspheres. We designed rgp120 PLGA microsphere formulations that provide a pulse of rgp120 at 1 to 6 months (depending on the polymer) after administration, mimicking another immunization. In these studies, the in vitro pulse of rgp120 correlated well with the observed in vivo autoboost as measured by an increase in anti-gp120 antibodies in guinea pigs. The immune response to the rgp120 PLGA microsphere formulations was increased by adding the soluble form of the saponin-derived adjuvant, QS-21. The use of small microspheres, however, did not increase the humoral response to rgp120. A single immunization with rgp120 PLGA microspheres resuspended in soluble rgp120 and QS-21 elicited neutralizing antibody titers that were comparable to titers obtained from two immunizations of rgp120 and QS-21 at the same total dose. Administration of rgp120 PLGA microspheres in baboons resulted in high, long-lasting neutralizing antibody titers that were greater than repeated immunizations with soluble rgp120 and QS-21. These studies also indicated that a continuous release of QS-21 at the injection site may provide a greater immune response than a bolus injection. Overall, this work demonstrated that PLGA microsphere formulations may be designed to provide in vivo pulses of an antigen eliminating the need for repeated immunizations.

AIDS Vaccines↗

Characterization of canine humoral immune responses to outer surface protein subunit vaccines and to natural infection by Lyme disease spirochetes.

Canine antibody responses to Lyme disease subunit vaccines and to natural borrelial infection were investigated. Vaccines were formulated with QS21 and outer surface proteins A (OspA) and B (OspB) derived from Borrelia burgdorferi B31. Vaccines containing QS21 and the lipoproteins gave 4-fold higher IgG1 and 8-fold higher IgG2 antibody responses than without QS21. Antisera to lipidated OspA or OspB vaccines containing QS21 had high antiborrelial activity against isolates B31 and CA-2-87, similar to those with a vaccine containing both OspA and OspB. Only the combination vaccine induced antiborrelial activity against heterologous isolates 24008 Fr and Borrelia garinii G25. Nonlipidated OspA- and OspB-based vaccines with QS21 elicited lower antibody and antiborrelial activity than did lipidated OspA and OspB vaccines; 49% of naturally exposed dogs had low titers to OspA or OspB. Thus, vaccines using lipidated OspA, OspB, and QS21 could induce higher antiborrelial activity than did natural exposure.

Adjuvants, Immunologic↗

Immunological and virological analyses of persons infected by human immunodeficiency virus type 1 while participating in trials of recombinant gp120 subunit vaccines.

We have studied 18 participants in phase I/II clinical trials of recombinant gp120 (rgp120) subunit vaccines (MN and SF-2) who became infected with human immunodeficiency virus type 1 (HIV-1) during the course of the trials. Of the 18 individuals, 2 had received a placebo vaccine, 9 had been immunized with MN rgp120, and seven had been immunized with SF-2 rgp120. Thirteen of the 18 infected vaccinees had received three or four immunizations prior to becoming infected. Of these, two were placebo recipients, six had received MN rgp120, and five had received SF-2 rgp120. Only 1 of the 11 rgp120 recipients who had multiple immunizations failed to develop a strong immunoglobulin G antibody response to the immunogen. However, the antibody response to rgp120 was transient, typically having a half-life of 40 to 60 days. No significant neutralizing activity against the infecting strain was detected in any of the infected individuals at any time prior to infection. Antibody titers in subjects infected despite vaccination and in noninfected subjects were not significantly different. Envelope-specific cytotoxic T-lymphocyte responses measured after infection were infrequent and weak in the nine vaccinees who were tested. HIV-1 was isolated successfully from all 18 individuals. Sixteen of these strains had a non-syncytium-inducing (NSI) phenotype, while two had a syncytium-inducing (SI) phenotype. NSI strains used the CCR5 coreceptor to enter CD4+ cells, while an SI strain from one of the vaccinees also used CXCR4. Viruses isolated from the blood of rgp120 vaccinees were indistinguishable from viruses isolated from control individuals in terms of their inherent sensitivity to neutralization by specific monoclonal antibodies and their replication rates in vitro. Furthermore, genetic sequencing of the env genes of strains infecting the vaccinees did not reveal any features that clearly distinguished these viruses from contemporary clade B viruses circulating in the United States. Thus, despite rigorous genetic analyses, using various breakdowns of the data sets, we could find no evidence that rgp120 vaccination exerted selection pressure on the infecting HIV-1 strains. The viral burdens in the infected rgp120 vaccine recipients were also determined, and they were found to be not significantly different from those in cohorts of placebo-vaccinated and nonvaccinated individuals. In summary, we conclude that vaccination with rgp120 has had,to date, no obvious beneficial or adverse effects on the individuals we have studied.

AIDS Vaccines↗

Early studies on a candidate intracellular subunit vaccine NFU.Ac.HIV[JM] for prevention and/or modification of HIV-related disease.

A candidate HIV subunit vaccine NFU.Ac.HIV[JM] was prepared by detergent and formaldehyde treatment of HIV-infected JM cells. The preparation contained HIV polypeptides at 200, 118, 70, 41 and 24 kD, with reactivity by immunoblotting of approximately fifteen virus-specific polypeptides including polypeptides at 119, 55, 41 and 24 kD, which may represent gp 120, p55, gp41 and p24, respectively. The content of gp120 was estimated to be 5 micrograms/ml. The vaccine was immunogenic in a rabbit, inducing neutralising antibody and reactivity by ELISA and by immunoblotting against a number of HIV polypeptides including those of molecular weights 24, 41, 55 nd 120 kD. The vaccine contained no infectious HIV or reverse transcriptase enzyme activity.

AIDS Vaccines↗

Introductory evaluation of an oral, killed whole cell enterotoxigenic Escherichia coli plus cholera toxin B subunit vaccine in Egyptian infants.

BACKGROUND: We conducted the first trial to assess the safety and immunogenicity of an oral, killed enterotoxigenic Escherichia coli plus cholera toxin B-subunit vaccine in children <2 years old. METHODS: Three doses of vaccine or killed E. coli K-12 control were given at 2-week intervals to 64 Egyptian infants, 6 to 18 months old, in a randomized, double blind manner. Adverse events were monitored for 3 days after each dose. Blood was collected before immunization and 7 to 10 days after each dose to assess vaccine-specific serologic responses. RESULTS: There was no statistically significant intergroup difference in the percentage of subjects reporting the primary safety endpoint (diarrhea or vomiting) after the first (31%, vaccine; 30%, control) or third (14%, vaccine; 18%, control) dose, whereas there was a trend toward greater reporting in the vaccine group after Dose 2 (36%, vaccine; 18%, control; P = 0.052). The percentage of children showing IgA seroconversion after any dose was higher in the vaccine than the control group for recombinant cholera toxin B-subunit (97% vs. 46%), colonization factor antigen I (61% vs. 18%) and coli surface antigen 4 (39% vs. 4%) (P < 0.001 for each comparison). IgG seroconversion rates in the vaccine and control groups were 97 and 21% to recombinant cholera toxin B-subunit (P < 0.001), 64 and 29% for colonization factor antigen I (P < 0.01), 53 and 21% for coli surface antigen 2 (P < 0.05) and 58 and 4% for coli surface antigen 4 (P < 0.001), respectively. The third vaccine dose was followed by augmented IgG antitoxin titers. CONCLUSION: The oral enterotoxigenic E. coli vaccine was safe and immunogenic in this setting in Egyptian infants.

Adjuvants, Immunologic↗