Intranasal Immunization with Killed Unencapsulated Whole Cells Prevents Colonization and Invasive Disease by Capsulated Pneumococci

Malley, Richard; Lipsitch, Marc; Stack, Anne M.; Saladino, Richard A.; Fleisher, Gary; Pelton, Stephen I.; Thompson, Claudette M.; Briles, David E.; Anderson, Patrick L.

doi:10.1128/iai.69.8.4870-4873.2001

Cited by 168 publications

(202 citation statements)

References 24 publications

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“…In previous experiments (ref. 41 and data not shown), we noted that ethanol-but not heat-killed whole-cell pneumococcal preparations administered intranasally with an adjuvant (cholera toxin) were immunogenic and protected rats against sepsis. These findings raised the possibility that heat treatment but not ethanol treatment had inactivated an important immunogenic component(s) of whole pneumococci.…”

Section: Ethanol-killed But Not Heat-killed Pneumococci Stimulate Hekmentioning

confidence: 94%

Recognition of pneumolysin by Toll-like receptor 4 confers resistance to pneumococcal infection

Malley

Henneke

Morse

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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630

583

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Streptococcus pneumoniae is one of the leading causes of invasive bacterial disease worldwide. Fragments of the cell wall and the cytolytic toxin pneumolysin have been shown to contribute substantially to inflammatory damage, although the interactions between pneumococcal components and host-cell structures have not been elucidated completely. Results of a previous study indicated that cell-wall components of pneumococci are recognized by Toll-like receptor (TLR)2 but suggested that pneumolysin induces inflammatory events independently of this receptor. In this study we tested the hypothesis that pneumolysin interacts with surface proteins of the TLR family other than TLR2. We found that pneumolysin stimulates tumor necrosis factor-␣ and IL-6 release in wild-type macrophages but not in macrophages from mice with a targeted deletion of the cytoplasmic TLR-adapter molecule myeloid differentiation factor 88, suggesting the involvement of the TLRs in pneumolysin recognition. Purified pneumolysin synergistically activated macrophage responses together with preparations of pneumococcal cell walls or staphylococcal peptidoglycan, which are known to activate TLR2. Furthermore, when compared with wild-type macrophages, macrophages from mice that carry a spontaneous mutation in TLR4 (P712H) were hyporesponsive to both pneumolysin alone and the combination of pneumolysin with pneumococcal cell walls. Finally, these TLR4-mutant mice were significantly more susceptible to lethal infection after intranasal colonization with pneumolysin-positive pneumococci than were control mice. We conclude that the interaction of pneumolysin with TLR4 is critically involved in the innate immune response to pneumococcus.

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Section: Ethanol-killed But Not Heat-killed Pneumococci Stimulate Hekmentioning

confidence: 94%

Recognition of pneumolysin by Toll-like receptor 4 confers resistance to pneumococcal infection

Malley

Henneke

Morse

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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630

583

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“…WCV was derived from strain RX1AL Ϫ , a capsule-and autolysin-negative mutant, and prepared as described in ref. 9. The final vaccine mixture contained 10 8 (killed) cfu of this strain plus 1 g of CT (List Biological Laboratories, Campbell, CA) per 10-l dose.…”

Section: Methodsmentioning

confidence: 99%

“…To assess the role of capsular serotype in protection against nasopharyngeal carriage, groups of 20 C57BL͞6J mice (female, age 6 weeks, The Jackson Laboratories) were randomized by cage to receive Ϸ10 6 cfu of TIGR4:6B 4 , TIGR4:7F 4 , TIGR4:14 4 , or saline control, as described in ref. 9. Inoculations were given three times at weekly intervals.…”

Section: Methodsmentioning

confidence: 99%

“…TIGR4J is acapsular; the other strains express serotype 6B, 7F, or 14 capsules, respectively (12). Strain 0603 is a serotype 6B clinical strain (9). Frozen midlog-phase aliquots were thawed and diluted to Ϸ10 6 colonyforming units (cfu) per 10 l of intranasal inoculum for challenge.…”

Section: Methodsmentioning

confidence: 99%

“…The age-specific incidence of pneumococcal disease in humans declines simultaneously and in a parallel fashion for a wide range of serotypes and well before natural acquisition of anticapsular Abs (8), suggesting a common and probably capsular serotypeindependent mechanism of protection. Additionally, we have previously shown that a vaccine made from killed, unencapsulated pneumococci whole-cell vaccine (WCV) given intranasally to mice with cholera toxin (CT) as an adjuvant prevents colonization by pneumococci of various capsular serotypes (9,10). More recently, McCool and Weiser (11) reported that clearance of carriage of a recently acquired pneumococcal strain in mice could occur independently of Ab, further supporting the possibility that other components of the immune response (whether innate or acquired, specific or not) may play an important role in this process, but the mechanisms for this protection have not been elucidated to date.…”

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confidence: 99%

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CD4⁺T cells mediate antibody-independent acquired immunity to pneumococcal colonization

Malley

Trzciński

Srivastava

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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Acquired immunity to Streptococcus pneumoniae (pneumococcus) has long been assumed to depend on the presence of anticapsular antibodies. We found, however, that colonization with live pneumococci of serotypes 6B, 7F, or 14 protected mice against recolonization by any of the serotypes and that protection from acquisition of a heterologous or homologous strain did not depend on anticapsular antibody. Further, intranasal immunization by live pneumococcal colonization or by a killed, nonencapsulated wholecell vaccine protected antibody-deficient mice against colonization, suggesting independence of antibodies to any pneumococcal antigens. Protection by intranasal immunization with whole-cell vaccine was completely abrogated in T cell-deficient mice, and in mice that were congenitally deficient in CD4 ؉ T cells or depleted of these cells at the time of challenge. In contrast, mice congenitally deficient in, or depleted of, CD8 ؉ T cells were fully protected. Protection in this model was observed beyond 2 months after immunization, arguing against innate or nonspecific immune mechanisms. Thus, we find that immunity to pneumococcal colonization can be induced in the absence of antibody, independent of the capsular type, and this protection requires the presence of CD4 ؉ T cells at the time of challenge.Streptococcus pneumoniae ͉ cell-mediated immunity ͉ vaccine A lmost 1 million children in the developing world die of infections due to Streptococcus pneumoniae (pneumococcus) each year (1). Pneumococcus is considered an ''extracellular'' bacterial pathogen, i.e., it is killed upon ingestion by phagocytic cells. Ingestion is facilitated by antibody (Ab) to its capsular polysaccharides (PS), of which there are at least 90 different serotypes. The two existing pneumococcal vaccines are based on injected mixtures of PS. Plain (unconjugated) PS vaccine contains 23 serotypes but is not efficacious in children Ͻ2 years old and therefore fails to protect those at highest risk. Protein-conjugated PS contains seven serotypes and protects infants (2), but it is difficult to manufacture (resulting in repeated shortages), is expensive, needs refrigeration, requires multiple injections, and does not include many of the capsular serotypes that cause pneumococcal disease in the developing world. Furthermore, serotype replacement, whereby pneumococcal serotypes not included in the conjugate vaccine become more prevalent causes of colonization and disease, has already been observed in clinical trials (3) and in epidemiologic studies (4) after implementation of conjugate vaccine immunization programs. Therefore, despite the success of the capsule-based vaccines, alternative strategies are urgently needed.The success of serum therapy (passive transfer of anticapsular Ab from hyperimmune animals) and the efficacy of PS and PS-protein conjugate vaccines have clearly demonstrated that anticapsular Ab alone is sufficient to treat or prevent pneumococcal disease. Furthermore, studies in animals (5) and humans (6) clearly demonstrate that antica...

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Process intensification for production of Streptococcus pneumoniae whole‐cell vaccine

Campos

Cardoso

Fratelli

et al. 2020

Biotech & Bioengineering

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The available pneumococcal conjugate vaccines provide protection against only those serotypes that are included in the vaccine, which leads to a selective pressure and serotype replacement in the population. An alternative low‐cost, safe and serotype‐independent vaccine was developed based on a nonencapsulated pneumococcus strain. This study evaluates process intensification to improve biomass production and shows for the first time the use of perfusion‐batch with cell recycling for bacterial vaccine production. Batch, fed‐batch, and perfusion‐batch were performed at 10 L scale using a complex animal component‐free culture medium. Cells were harvested at the highest optical density, concentrated and washed using microfiltration or centrifugation to compare cell separation methods. Higher biomass was achieved using perfusion‐batch, which removes lactate while retaining cells. The biomass produced in perfusion‐batch would represent at least a fourfold greater number of doses per cultivation than in the previously described batch process. Each strategy yielded similar vaccines in terms of quality as evaluated by western blot and animal immunization assays, indicating that so far, perfusion‐batch is the best strategy for the intensification of pneumococcal whole‐cell vaccine production, as it can be integrated to the cell separation process keeping the same vaccine quality.

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Intranasal Immunization with Killed Unencapsulated Whole Cells Prevents Colonization and Invasive Disease by Capsulated Pneumococci

Cited by 168 publications

References 24 publications

Recognition of pneumolysin by Toll-like receptor 4 confers resistance to pneumococcal infection

Recognition of pneumolysin by Toll-like receptor 4 confers resistance to pneumococcal infection

CD4⁺T cells mediate antibody-independent acquired immunity to pneumococcal colonization

Process intensification for production of Streptococcus pneumoniae whole‐cell vaccine

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Intranasal Immunization with Killed Unencapsulated Whole Cells Prevents Colonization and Invasive Disease by Capsulated Pneumococci

Cited by 168 publications

References 24 publications

Recognition of pneumolysin by Toll-like receptor 4 confers resistance to pneumococcal infection

Recognition of pneumolysin by Toll-like receptor 4 confers resistance to pneumococcal infection

CD4+T cells mediate antibody-independent acquired immunity to pneumococcal colonization

Process intensification for production of Streptococcus pneumoniae whole‐cell vaccine

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CD4⁺T cells mediate antibody-independent acquired immunity to pneumococcal colonization