Affinity constants of naturally acquired and vaccine-induced anti-Pseudomonas aeruginosa antibodies in healthy adults and cystic fibrosis patients.

Naturally acquired anti-Pseudomonas aeruginosa antibody fails to afford protection against repeated P. aeruginosa bronchopulmonary exacerbations in cystic fibrosis (CF) patients. In an effort to explain this phenomenon, the titer and affinity constants of serum anti-lipopolysaccharide (LPS) IgG were determined in five study groups: healthy adults before and after immunization with a polyvalent LPS-based vaccine, healthy noncolonized CF patients before and after immunization, nonimmunized CF patients with significantly elevated anti-LPS antibody titers without documented colonization, recently colonized CF patients before and after immunization, and nonimmunized CF patients chronically colonized with P. aeruginosa. Immunization elicited a significant rise in total anti-LPS immunoglobulin levels and affinity constants in both healthy adults and CF patients. Although chronically colonized patients had elevated levels of total anti-LPS antibody, these antibodies possessed affinities at least 100-fold less than those of vaccine-induced antibodies.     

Antibody activity against Pseudomonas aeruginosa in immune globulins prepared for intravenous use in humans

Twenty-seven lots of human immune globulin for intravenous use (IGIV) from seven different producers, including one hyperimmune preparation, were examined for immunologic reactivity and opsonic and protective activity against Pseudomonas aeruginosa. All IGIVs contained hemagglutinating antibodies to seven immunotype-specific lipopolysaccharides of P. aeruginosa (geometric mean titer +/- SE, 14 +/- 3 for nonhyperimmune preparations and 420 for the hyperimmune product) and to exotoxin A (77 +/- 15). All IGIVs tested demonstrated opsonic activity against P. aeruginosa in an in vitro granulocyte-dependent bactericidal assay. All IGIVs conferred dose-dependent protection (hyperimmune more so than nonhyperimmune) against fatal burn-wound infections due to P. aeruginosa in mice. In contrast, single lots of hyperimmune and nonhyperimmune IGIV conferred limited protection against infections due to P. aeruginosa in granulocytopenic mice. These studies indicate the potential prophylactic efficacy of IGIV in human pseudomonas disease, and the possible need for high doses of hyperimmune IGIV in granulocytopenic patients.     

Outer membrane protein F (porin) preparation of Pseudomonas aeruginosa as a protective vaccine against heterologous immunotype strains in a burned mouse model

Outer membrane (OM) protein F (porin) was purified by extraction from polyacrylamide gels of cell envelope proteins of the Pseudomonas aeruginosa PA01 strain. Mice were immunized intramuscularly with 10 micrograms of protein F preparation on days 1 and 14 and then subjected to burn and challenge on day 28. Protein F immunization afforded significant protection above that provided by PA01 lipopolysaccharide (LPS) immunization against subsequent challenge with six of six heterologous LPS immunotype strains of P. aeruginosa. By an ELISA, the murine immune response revealed an IgG titer of 5,120 to protein F by day 30. Immunoblot analysis of antisera from protein F-immunized mice revealed bands with both protein F and protein H of cell envelopes of all immunotypes tested. Active immunization with OM protein H did not, however, afford significant protection to mice in this burned mouse model. These data show the efficacy of OM protein F as a protective vaccine in a murine model representative of human infection.     

Immunologic approaches to blockade of the renin-angiotensin system: a review

Several immunologic approaches to blockade of the renin-angiotensin system (RAS) have been reported, involving most of the proteins and peptides of the biochemical cascade: renin, substrate, angiotensins, and converting enzyme. None as yet has involved blockade of angiotensin II receptors. Earlier and more recent studies used passive transfer of heterologous antibodies or active immunization against RAS proteins and peptides. Passive transfers have been performed with both polyclonal antibodies and now with specific monoclonal immunoglobulins. The latter are better defined in affinity, quantity, and capacity to bind and thus inhibit the biologic activity of the antigen. Active immunization produced long-term blockade of part or all of the biologic activity of the system. The immunopathologic consequences of the use of antibodies raised against a self-antigen could be of interest in defining the predominant site of storage and secretion of the relevant protein and hence the respective roles of different tissues in the production of specific proteins in, for example, the vascular pulmonary bed for converting enzyme and renal arterial tree for renin. In all cases immunologic methods offer in vivo experimental models of short- or long-term RAS blockade that could be compared with pharmacologic methods, such as converting-enzyme inhibition, angiotensin II antagonists, and renin inhibitors.     

Implications of endotoxin contamination in the evaluation of antibodies to lipopolysaccharides in a murine model of gram-negative sepsis

The pharmacological factors involved in lipopolysaccharide (LPS)-induced host resistance against infection were investigated in relation to the problem of endotoxin contamination of preparations of monoclonal antibody to the common structure of endotoxin. When administered prophylactically, purified LPS (as low as 4 ng/kg of mouse body weight) or antibody preparations contaminated with endotoxin (assayed by Limulus amoebocyte lysate test) were protective against lethal challenge with a clinical isolate of Escherichia coli (P less than .01). Antibodies that were nonreactive to LPS were similarly protective (P less than .001) when spiked with low doses of LPS (100 ng/mg of protein) but, as with LPS, were without effect when administered after infection. Endotoxin contamination of core-reactive antibody to LPS is mostly a problem associated with the large-scale production, purification, and concentration of the monoclonal antibody. The efficacy (as reported in studies in animal models of gram-negative bacterial sepsis) of antibody to LPS core is controversial. We suggest that endotoxin contamination is a likely factor in this controversy.     

Application of Viral Vectors for Vaccine Development with a Special Emphasis on COVID-19

Viral vectors can generate high levels of recombinant protein expression providing the basis for modern vaccine development. A large number of different viral vector expression systems have been utilized for targeting viral surface proteins and tumor-associated antigens. Immunization studies in preclinical animal models have evaluated the elicited humoral and cellular responses and the possible protection against challenges with lethal doses of infectious pathogens or tumor cells. Several vaccine candidates for both infectious diseases and various cancers have been subjected to a number of clinical trials. Human immunization trials have confirmed safe application of viral vectors, generation of neutralizing antibodies and protection against challenges with lethal doses. A special emphasis is placed on COVID-19 vaccines based on viral vectors. Likewise, the flexibility and advantages of applying viral particles, RNA replicons and DNA replicon vectors of self-replicating RNA viruses for vaccine development are presented.     

Pseudomonas aeruginosa polysaccharide-tetanus toxoid conjugate vaccine: safety and immunogenicity in humans

A Pseudomonas aeruginosa polysaccharide-tetanus toxoid (Ttxd) conjugate vaccine was produced. Polysaccharide was derived from lipopolysaccharide (LPS) and covalently linked to Ttxd by using carbodiimide with adipic acid dihydrazide as a spacer molecule. The conjugate possessed a relative molecular weight of greater than 350,000 and was nontoxic and nonpyrogenic. The vaccine bound serospecific monoclonal antibodies with an avidity similar to LPS and reacted with murine and human opsonic antibody. The vaccine was immunogenic in rabbits and mice and elicited IgG antibody to both LPS and Ttxd. The vaccine was safe when parenterally administered to humans and evoked only mild, transient reactions. Mean titers of IgG antibody to LPS rose 19-fold after immunization, with 82% of the volunteers responding with a fourfold or greater rise in titer. IgG antibody to LPS evoked after immunization was opsonic and highly effective at preventing fatal experimental burn wound sepsis due to P. aeruginosa.     

Protection against experimental Staphylococcus aureus arthritis by vaccination with clumping factor A, a novel virulence determinant.

The importance of the fibrinogen-binding adhesin clumping factor A (ClfA) in the pathogenesis of Staphylococcus aureus septic arthritis was examined in an animal model. The protective effect of active and passive immunization with ClfA also was investigated in S. aureus infection models. The severity of arthritis was markedly reduced in mice challenged intravenously with a clfA mutant, compared with mice infected with the wild-type strain. Mice immunized with recombinant ClfA and challenged with S. aureus developed less-severe arthritis than did mice immunized with a control antigen. Passive immunization of mice with rat and rabbit anti-ClfA antibodies protected against S. aureus arthritis and sepsis-induced death, indicating that the protection by active immunization is antibody mediated. Taken together, these data strongly suggest that ClfA is a crucial virulence determinant for septic arthritis and an excellent target for the generation of immune therapies directed against S. aureus.     

Use of immunoglobulins in prevention and treatment of infection in critically ill patients: review and critique

The study of the use of standard intravenous immunoglobulin (IVIG) preparations as adjunctive therapy for seriously ill patients is motivated by the need to restore immunoglobulin G depleted because of trauma or surgery and/or by the need to provide patients with specific antibodies to various microorganisms. Whereas no clinical studies have shown that standard IVIG has therapeutic efficacy, some data suggest that its prophylactic use is beneficial. Antisera or IVIG prepared from individuals who are hyperimmunized with the biologically active, highly conserved core portion of the endotoxin of gram-negative bacteria confer variable degrees of protection in animal models and clinical trials. Two clinical trials with use of monoclonal antibodies to core lipopolysaccharide have been completed. Only subsets of patients with gram-negative sepsis were protected by the monoclonal antibodies, but the results of the studies were discrepant in regard to the specific characteristics of patients who benefited from the administration of these antibodies. Further studies will be necessary to establish whether this therapy can be recommended for critically ill patients.     

Immunization with a Pseudomonas aeruginosa elastase peptide reduces severity of experimental lung infections due to P. aeruginosa Or Burkholderia cepacia

Pseudomonas aeruginosa and Burkholderia cepacia produce metalloproteases that effect lung injury. Two epitopes (peptides 15 and 42) previously identified on P. aeruginosa elastase induce the production of antibodies that neutralize protease activity. The effects of immunization with synthetic peptides based on these epitopes on experimental lung infections due to P. aeruginosa or B. cepacia were examined. Rats were immunized with peptides conjugated to keyhole limpet hemocyanin or tetanus toxoid before infection. Immunization with peptide 15 (pep15) resulted in a decrease in total cells and polymorphonuclear leukocytes in bronchoalveolar lavage (BAL) fluid and a 50%-70% decrease in lung histopathologic changes, compared with findings in controls. Immunization with peptide 42 decreased cells in BAL fluid but did not decrease lung pathologic changes. Immunization with pep15 alone was just as effective in protecting against lung injury as immunization with a combination of both peptides. These studies suggest that immunization with pep15 can reduce the severity of lung infections due to P. aeruginosa or B. cepacia.     

IgG subclass antibody responses to alginate from Pseudomonas aeruginosa in patients with cystic fibrosis and chronic P. aeruginosa infection.

Chronic bronchopulmonary infection with alginate-producing, mucoid Pseudomonas aeruginosa is characteristically associated with cystic fibrosis (CF). A significant correlation between the antibody response to alginate and poor lung function has been reported. Enzyme-linked immunosorbent assays were developed for the quantitation of human IgG1, IgG2, IgG3, and IgG4 antibodies to P. aeruginosa alginate. We investigated the pattern of IgG subclass antibodies against P. aeruginosa alginate in serum of patients with CF, others with chronic P. aeruginosa infection, and healthy controls. Healthy controls and patients with CF, before they acquired P. aeruginosa infection, had no or very low titers of antibodies against P. aeruginosa alginate. The latter with chronic infection had significantly higher antibody levels than all others groups, including patients with chronic P. aeruginosa infection but no CF. CF with chronic P. aeruginosa infection led to an inverse correlation between lung function parameters and levels of IgG3 and IgG4. Fifty-seven patients with CF have been followed for an average of 12 years with multiple antibody assays covering the preinfection, early, and late stage of chronic infection. All of them developed IgG1 and IgG3 antibodies to alginate at the start of infection. IgG2 antibodies developed later and showed only a slow increase during the chronic infection. Patients who died had significantly higher IgG2 anti-alginate antibody levels than other investigated groups. Elevated levels of IgG2 and IgG3 antibodies to P. aeruginosa alginate are a sign of poor prognosis in CF.     

Association of systemic immune complexes, complement activation, and antibodies to Pseudomonas aeruginosa lipopolysaccharide and exotoxin A with mortality in cystic fibrosis

The relevance of circulating immune complexes, plasma complement activation, and serum antibodies against discrete antigens of Pseudomonas aeruginosa, to the clinical course in patients with cystic fibrosis (CF) is unknown. We related these factors to outcome in 49 patients with CF colonized by P. aeruginosa, comparing 14 who died of lung disease with 35 survivors of similar age and duration of colonization, as well as 9 uncolonized patients with CF, 24 patients with other bronchorrheic lung disease, and 10 healthy control subjects. The patients with CF colonized by P. aeruginosa who died had a higher incidence of immune complexes than did survivors (71 versus 40%, p less than 0.05). Moreover, C4 activation was highly associated with immune complexes and mortality (p less than 0.001 for each). Those who died also had much higher levels of IgG antibodies to P. aeruginosa lipopolysaccharide (LPS) and exotoxin A than did survivors colonized by P. aeruginosa (p less than 0.005 and p = 0.01, respectively), whereas both groups had similar levels of P. aeruginosa sonicate, elastase, alkaline protease, and endotoxin core antibodies. We conclude that increasing levels of serum IgG antibodies to P. aeruginosa LPS and exotoxin A and the presence of systemic immune complexes and complement activation are associated with poor prognosis in CF, and may provide useful noninvasive markers for studying the possible immunopathogenesis of CF lung disease.     

Studies on the ability of alginate to act as a protective immunogen against infection with Pseudomonas aeruginosa in animals

Most patients with cystic fibrosis become colonized and infected with mucoid Pseudomonas aeruginosa. The major component of the mucoid material has been identified as the polysaccharide alginic acid. The present work was undertaken to determine whether antibody to alginate is protective in a model of chronic lung infection with P. aeruginosa in rats. Bacterial clearance was associated with a rise in titers of antibody to alginate. In a number of animals a rise in antibody titers was not seen, and in fact a decrease was noted at 30 days compared with 10 days. This observation suggested the possibility of immune complex formation due to antigen excess. Evidence for immune complex deposition in tissues was obtained by immunofluorescence studies. Thus antibody to alginate may offer strain-dependent protection against chronic lung infection with P. aeruginosa in rats; however, immune complex formation should be considered as a possible consequence of immunization with alginate.     

Protection against infection with Pseudomonas aeruginosa by passive transfer of monoclonal antibodies to lipopolysaccharides and outer membrane proteins

Experimental infection with Pseudomonas aeruginosa was treated with eight different monoclonal antibodies (MCAs) produced by hybridoma cells obtained through cell fusion of mouse plasmacytoma cells and spleen cells from mice immunized with a virulent strain of P. aeruginosa (Homma serotype 7). Five MCAs bound to lipopolysaccharides (LPSs) specific to serotype 7 or serotypes 2, 7, and 13, whereas the other three MCAs bound with broad specificities to outer membrane protein (OMP) fractions. The MCAs to LPS were highly protective against infection, with 50% protective doses of 0.05-2.5 micrograms of immunoglobulin per mouse. In contrast, the MCAs to OMP were much less protective, with a 50% protective dose range of 10 to greater than 100 micrograms of immunoglobulin per mouse. Most of the MCAs to LPS agglutinated P. aeruginosa cells, but all the MCAs to OMP produced so far have not, although all the MCAs bound well to the cells. Agglutinating MCAs provided better protection than did nonagglutinating MCAs.     

Hypothesis for vaccine development: protective immunity to enteric diseases caused by nontyphoidal salmonellae and shigellae may be conferred by serum IgG antibodies to the O-specific polysaccharide of their lipopolysaccharides.

Immunoprophylaxis for bacterial enteric diseases is hindered because the protective immune mechanism(s) against nontyphoidal salmonellae or shigellae in humans are not established. On the basis of the similarities between the clinical signs, epidemiology, pathogenesis, and pathology of as well as protective immunity to salmonellae and shigellae, we propose that serum IgG antibodies to the O-specific polysaccharide (O-SP) of their lipopolysaccharides (LPSs) will confer protective immunity to these two pathogens. Critical to this notion is that (1) the virulence of these two pathogens requires full expression of their LPS; (2) active or passive immunization with serum IgG O-SP antibodies confers protection of mice against Salmonella typhimurium (there are no comparable data for humans); and (3) in humans, convalescence from shigellosis confers type (O-SP) -specific protective immunity, and indirect evidence shows a correlation between the level of serum LPS antibodies and resistance to shigellosis. We designed conjugate vaccines to elicit high levels of long-lived serum IgG O-SP antibodies to nontyphoidal salmonellae and shigellae to test this hypothesis.     

Controversies in the use of passive immunotherapy for bacterial infections in the critically ill patient

Several preparations of standard immunoglobulins for intravenous use have been tested as adjunctive therapy for bacterial infections in premature neonates and in critically ill adults after major surgery, trauma, and burn. The use of intravenous immunoglobulins in these settings is controversial because the efficacy and cost-effectiveness of this treatment are still not definitively established. Specific preparations of immunoglobulins against Pseudomonas aeruginosa for intramuscular administration have shown promising efficacy, and preparations for intravenous administration are now under investigation. Cross-protection against a wide range of gram-negative infections has been attempted by the administration of antiserum to the core glycolipid of lipopolysaccharide prepared from volunteers immunized with the J5 mutant of Escherichia coli 0111. Treatment with this preparation improved the survival rate of patients with gram-negative bacteremia and, when administered prophylactically to high-risk surgical patients, prevented shock and death related to gram-negative infections. The mechanism of protection of the J5 antiserum is not clearly understood because of our inability to measure the actual protective antibody in polyclonal J5 antiserum. Thus, the preparation of readily available cross-protective hyperimmune immunoglobulins is hampered because there is presently no method of selecting appropriate donors or high-titered plasma pools.     

Vaccine protection against HIV-2 infection in cynomolgus monkeys

The aim of this study was to determine if protection against an infectious human immunodeficiency virus type 2 (HIV-2) challenge could be obtained in cynomolgus macaques by active immunization using whole killed virus vaccine. Four monkeys were immunized with killed HIV-2SBL-6669, two of them with five intramuscular (im) injections of viral preparation containing 100 or 300 micrograms protein emulsified in incomplete Freund's adjuvant (IFA) and the two remaining received four im injections of 25-50 micrograms viral protein in iscoms. Each of the four vaccinated cynomolgus monkeys, along with four unvaccinated controls, were challenged intravenously two weeks after the last booster with approximately 100 animal infectious doses (ID50) of live HIV-2SBL-6669. All four immunized monkeys developed antibodies to HIV-2 envelope and core proteins before challenge exposure to HIV-2, but only the two animals vaccinated with virus in IFA developed detectable neutralizing antibodies. The two monkeys immunized with killed virus in IFA have shown no evidence of infection nine months after challenge with live virus. When blood and lymph node cells from these animals were transfused into naive cynomolgus monkeys, the recipients remained free of infection. In contrast, virus was recovered repeatedly in all nonimmunized animals and in the two animals immunized with iscom-associated viral antigens, which had a low content of envelope gp125 antigen. The demonstration of vaccine-induced protection against HIV-2 in a nonhuman primate raises hope for effective immunization against HIV infections in humans as well.     

Active immunization against nicotine suppresses nicotine-induced dopamine release in the rat nucleus accumbens shell

BACKGROUND: Tobacco smoking is the largest preventable cause of morbidity and premature mortality in the world. Although its medical consequences are well documented, 20-50% of the population even in developed countries remain tobacco smokers. The drugs presently used in smoking cessation have limited efficiency and, therefore, there is a need for alternative and improved treatments. One novel approach in this regard may be provided by immunization against nicotine. OBJECTIVE: The present study in male Wistar rats investigated if active immunization with a novel nicotine immunogen, IP18-KLH, may generate nicotine-selective antibodies and, furthermore, whether this treatment might prevent nicotine from exerting its stimulating effect on the mesolimbic, dopaminergic reward system in the brain. METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to determine the titer of nicotine antibodies in plasma after immunization with IP18-KLH in Freund's adjuvant. Competitive ELISA was used to assess the selectivity of the antibodies. Finally, we used in vivo voltammetry to investigate whether active immunization with IP18-KLH could prevent nicotine-induced dopamine release in the shell of nucleus accumbens (NAC(shell)). RESULTS: The present study shows that active immunization with IP18-KLH generates antibodies that are highly selective for nicotine. Furthermore, immunization with IP18-KLH prevented the nicotine-induced increase in dopamine release in the NAC(shell), a biochemical correlate to the rewarding properties of nicotine. CONCLUSIONS: Active immunization with IP18-KLH prevents a central effect of nicotine that is considered critical for the induction of nicotine dependence. Consequently, active immunization may provide long-term protection against initiation of tobacco dependence, an effect that may prove particularly advantageous in relapse prevention.     

Conformational Masking and Receptor-Dependent Unmasking of Highly Conserved Env Epitopes Recognized by Non-Neutralizing Antibodies That Mediate Potent ADCC against HIV-1

The mechanism of antibody-mediated protection is a major focus of HIV-1 vaccine development and a significant issue in the control of viremia. Virus neutralization, Fc-mediated effector function, or both, are major mechanisms of antibody-mediated protection against HIV-1, although other mechanisms, such as virus aggregation, are known. The interplay between virus neutralization and Fc-mediated effector function in protection against HIV-1 is complex and only partially understood. Passive immunization studies using potent broadly neutralizing antibodies (bnAbs) show that both neutralization and Fc-mediated effector function provides the widest dynamic range of protection; however, a vaccine to elicit these responses remains elusive. By contrast, active immunization studies in both humans and non-human primates using HIV-1 vaccine candidates suggest that weakly neutralizing or non-neutralizing antibodies can protect by Fc-mediated effector function, albeit with a much lower dynamic range seen for passive immunization with bnAbs. HIV-1 has evolved mechanisms to evade each type of antibody-mediated protection that must be countered by a successful AIDS vaccine. Overcoming the hurdles required to elicit bnAbs has become a major focus of HIV-1 vaccine development. Here, we discuss a less studied problem, the structural basis of protection (and its evasion) by antibodies that protect only by potent Fc-mediated effector function.     

Immunotherapeutic approaches against Staphylococcus aureus

Staphylococcus aureus is a major cause of life-threatening infections such as bacteremia and endocarditis. Unfortunately, many strains of this bacterial species have become resistant to certain antibiotics, including methicillin and amoxicillin. These strains are known as methicillin-resistant S. aureus (MRSA). Therefore, the prophylactic and therapeutic potential of antistaphylococcal vaccines is currently being explored with priority. In animal models, (passive) immunization with (antibodies directed against) certain S. aureus surface components, staphylococcal toxins and capsular polysaccharides protects against S. aureus colonization or infection. However, immunization studies performed in humans show less promising results. So far, not a single antistaphylococcal vaccine successfully passed clinical trials. This article focuses on the results that were obtained with immunotherapeutic approaches directed against S. aureus in animal and human studies. In addition, it is discussed whether effective immunization approaches against S. aureus are feasible in humans.