PspA, a protection-eliciting pneumococcal protein: immunogenicity of isolated native PspA in mice

Briles, David E.; King, Janice; Gray, Mary Ann; McDaniel, Larry S.; Swiatlo, Edwin; Benton, Kimberly A.

doi:10.1016/0264-410x(96)82948-3

Cited by 127 publications

(117 citation statements)

References 48 publications

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“…DAP was added (50 g/mL) for the growth of Asd Ϫ strains (27). S. pneumoniae WU2 was cultured on brain heart infusion (BHI) agar containing 5% sheep blood or in Todd-Hewitt broth plus 0.5% yeast extract (25). Plasmid pYA3493 is an expression vector for constructing antigen fusions to the first 23 aa of ␤-lactamase (17).…”

Section: Methodsmentioning

confidence: 99%

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Evaluation of new generationSalmonella entericaserovar Typhimurium vaccines with regulated delayed attenuation to induce immune responses against PspA

Wang

Scarpellini

et al. 2009

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

112

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Increasing the immunogenicity to delivered antigens by recombinant attenuated Salmonella vaccines (RASV) has been the subject of intensive study. With this goal in mind, we have designed and constructed a new generation of RASV that exhibit regulated delayed attenuation. These vaccine strains are phenotypically wild type at the time of immunization and become attenuated after colonization of host tissues. The vaccine strains are grown under conditions that allow expression of genes required for optimal invasion and colonization of host tissues. Once established in the host, these virulence genes are turned off, fully attenuating the vaccine strain. In this study, we compared 2 of our newly developed regulated delayed attenuation Salmonella enterica serovar Typhimurium strains 9088 and 9558 with the ⌬cya ⌬crp ⌬asd strain 8133, for their abilities to express and present a secreted form of the ␣-helical region of pneumococcal surface protein A (PspA) to the mouse immune system. All 3 strains induced high levels of serum antibodies specific for PspA as well as to Salmonella antigens in orally immunized mice. However, both RASVs expressing delayed attenuation elicited significantly greater anti-PspA immune responses, including serum IgG and T cell secretion of IL-4 and IFN-␥, than other groups. Also, vaccination with delayed attenuation strains resulted in a greater degree of protection against Streptococcus pneumoniae challenge than in mice vaccinated with 8133 (71-86% vs. 21% survival, P < 0.006). Together, the results demonstrate that the regulated attenuation strategy results in highly immunogenic antigen delivery vectors for oral vaccination.Streptococcus pneumoniae ͉ bacterial vectors G enerating a safe and immunogenic vaccine strain is the biggest challenge in the development of live Salmonella vaccines (1). An ideal Salmonella vaccine strain should exhibit wild-type abilities to withstand stresses (enzymatic, acid, osmotic, ionic, etc.) and host defenses (bile, antibacterial peptides, etc.) encountered after oral or intranasal immunization, and should exhibit wild-type ability to colonize and invade host lymphoid tissues while remaining avirulent. Various attenuated Salmonella strains have been used as live vaccines to induce mucosal and systemic immunity against either the carrier itself or to a vectored antigen (2). Salmonella vaccine strains typically carry defined deletion mutations affecting either metabolic functions or virulence factors (3). Various attenuating mutations have been investigated in the pursuit to develop optimal immune responses (4, 5). Many attenuating mutations were found to either reduce Salmonella survival due to host-induced stresses and/or reduce colonization of lymphoid effector tissues leading to less than ideal immunogenicity (6, 7). To circumvent these problems, we explored ways to achieve regulated delayed attenuation in vivo (8, 9) to create vaccine strains that are phenotypically wild-type at the time of immunization and become attenuated after colonization of host tissues.O...

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Section: Methodsmentioning

confidence: 99%

“…4). S. pneumoniae WU2 produces PspA that is cross-reactive with PspA Rx1 (25). Immunization with any of the pspA-expressing strains provided significant protection against challenge (P Ͻ 0.01).…”

Section: Antigen-specific Stimulation Of Il-4 or Ifn-␥ Productionmentioning

confidence: 99%

Evaluation of new generationSalmonella entericaserovar Typhimurium vaccines with regulated delayed attenuation to induce immune responses against PspA

Wang

Scarpellini

et al. 2009

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

112

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“…Bacterial cultures were pelleted by centrifugation and treated for 20 min at room temperature with 2% choline chloride (Sigma-Aldrich) in PBS containing 0.25 M NaCl to release CBPs from the bacterial cell wall (8). The resulting CBP-depleted bacteria (R36Ach) were washed by centrifugation, heat killed, and stored as indicated previously.…”

Section: Preparation Of R36a Depleted Of Choline-binding Proteins (Cbp)mentioning

confidence: 99%

“…8), radically changes the nature of the in vivo IgG anti-PPS14 response relative to both purified capsular PPS14 as well as intact Pn. Thus, the IgG anti-PPS14 response to soluble PPS14-PspA conjugate is largely similar to the IgG anti-PspA responses to both conjugate and intact Pn14, including prolonged primary kinetics of induction and the generation of a CD4 ϩ T cell-dependent, PPS14-specific IgG memory response (7,9).…”

mentioning

confidence: 99%

Parameters Underlying Distinct T Cell-Dependent Polysaccharide-Specific IgG Responses to an Intact Gram-Positive Bacterium versus a Soluble Conjugate Vaccine

Colino

Chattopadhyay

Sen

et al. 2009

The Journal of Immunology

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IgG anti-polysaccharide (PS) responses to both intact Streptococcus pneumoniae (Pn) and PS conjugate vaccines are dependent on CD4+ T cells, B7-dependent costimulation, and CD40-CD40-ligand interactions. Nevertheless, the former response, in contrast to the latter, is mediated by an ICOS-independent, apoptosis-prone, extrafollicular pathway that fails to generate PS-specific memory. We show that pre-existing PS-specific Igs, the bacterial surface or particulation, selective recruitment of B cell subsets, or activation and recruitment of Pn protein-specific CD4+ T cells do not account for the failure of Pn to generate PS-specific IgG memory. Rather, the data suggest that the critical factor may be the lack of covalent attachment of PS to protein in intact Pn, highlighting the potential importance of the physicochemical relationship of PS capsule with the underlying bacterial structure for in vivo induction of PS-specific Igs.

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“…The pneumococcal surface protein A (PspA) is highly immunogenic in mice and humans (18,19). In previous studies, mice orally immunized with an S. typhimurium vaccine strain expressing the ␣-helical domain of S. pneumoniae strain Rx1 PspA generated PspA-specific immune responses and were protected against challenge with virulent S. pneumoniae (20).…”

mentioning

confidence: 99%

Regulated programmed lysis of recombinant Salmonella in host tissues to release protective antigens and confer biological containment

Kong

Wanda

Zhang

et al. 2008

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

129

185

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We have devised and constructed a biological containment system designed to cause programmed bacterial cell lysis with no survivors. We have validated this system, using Salmonella enterica serovar Typhimurium vaccines for antigen delivery after colonization of host lymphoid tissues. The system is composed of two parts. The first component is Salmonella typhimurium strain 8937, with deletions of asdA and arabinose-regulated expression of murA, two genes required for peptidoglycan synthesis and additional mutations to enhance complete lysis and antigen delivery. The second component is plasmid pYA3681, which encodes arabinoseregulated murA and asdA expression and C2-regulated synthesis of antisense asdA and murA mRNA transcribed from the P22 P R promoter. An arabinose-regulated c2 gene is present in the chromosome. 8937(pYA3681) exhibits arabinose-dependent growth. Upon invasion of host tissues, an arabinose-free environment, transcription of asdA, murA, and c2 ceases, and concentrations of their gene products decrease because of cell division. The drop in C2 concentration results in activation of P R, driving synthesis of antisense mRNA to block translation of any residual asdA and murA mRNA. A highly antigenic ␣-helical domain of Streptococcus pneumoniae Rx1 PspA was cloned into pYA3681, resulting in pYA3685 to test antigen delivery. Mice orally immunized with 8937(pYA3685) developed antibody responses to PspA and Salmonella outer membrane proteins. No viable vaccine strain cells were detected in host tissues after 21 days. This system has potential applications with other Gram-negative bacteria in which biological containment would be desirable.programmed cell lysis ͉ rPspA ͉ Rx1

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PspA, a protection-eliciting pneumococcal protein: immunogenicity of isolated native PspA in mice

Cited by 127 publications

References 48 publications

Evaluation of new generationSalmonella entericaserovar Typhimurium vaccines with regulated delayed attenuation to induce immune responses against PspA

Evaluation of new generationSalmonella entericaserovar Typhimurium vaccines with regulated delayed attenuation to induce immune responses against PspA

Parameters Underlying Distinct T Cell-Dependent Polysaccharide-Specific IgG Responses to an Intact Gram-Positive Bacterium versus a Soluble Conjugate Vaccine

Regulated programmed lysis of recombinant Salmonella in host tissues to release protective antigens and confer biological containment

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