Comparative immunogenicity of M. hyopneumoniae NrdF encoded in different expression systems delivered orally via attenuated S. typhimurium aroA in mice

Chen, Austen Y.; Fry, Scott; Forbes-Faulkner, Judy; Daggard, Grant; Mukkur, T.K.S.

doi:10.1016/j.vetmic.2005.12.009

Cited by 19 publications

(13 citation statements)

References 21 publications

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“…P97 is a proteolytically processed adhesin protein capable of binding swine cilia (11,53). NrdF has been extensively studied as a putative vaccine candidate, and previous vaccination studies have demonstrated that NrdF can invoke Th1 and Th2 immune responses and improve the average daily weight gain of swine challenged with M. hyopneumoniae (7,16,17).…”

Section: Resultsmentioning

confidence: 99%

“…M. hyopneumoniae is a pathogen of swine that colonizes the ciliated epithelial cells of the respiratory tract and causes significant economic losses (11). Adh and NrdF have both been previously studied in vaccination experiments when expressed from plasmid-based systems in S. enterica serovar Typhimurium aroA (7,8,15,16,17). Oral vaccination with NrdF-expressing S. enterica serovar Typhimurium aroA has resulted in significant immunoglobulin A (IgA) responses in murine lungs (16), increased murine splenocyte NrdF-specific gamma interferon (IFN-␥) production (7), and primed the porcine respiratory tract for an NrdFspecific secretory IgA response (17).…”

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confidence: 99%

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Development of Non-Antibiotic-Resistant, Chromosomally Based, Constitutive and Inducible Expression Systems for aroA -Attenuated Salmonella enterica Serovar Typhimurium

et al. 2009

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Live-vaccine delivery systems expressing two model antigens from Mycoplasma hyopneumoniae, F2 P97 (Adh) and NrdF, were constructed using Salmonella enterica serovar Typhimurium aroA (STM-1), and immunogenicity in mice was evaluated. Recombinant plasmid-based expression (PBE) and chromosomally based expression (CBE) systems were constructed. The PBE system was formed by cloning both antigen genes into pJLA507 to create an operon downstream of temperature-inducible promoters. Constitutive CBE was achieved using a promoter-trapping technique whereby the promoterless operon was stably integrated into the chromosome of STM-1, and the expression of antigens was assessed. The chromosomal position of the operon was mapped in four clones. Inducible CBE was obtained by using the in vivo-induced sspA promoter and recombining the expression construct into aroD. Dual expression of the antigens was detected in all systems, with PBE producing much larger quantities of both antigens. The stability of antigen expression after in vivo passage was 100% for all CBE strains recovered. PBE and CBE strains were selected for comparison in a vaccination trial. The vaccine strains were delivered orally into mice, and significant systemic immunoglobulin M (IgM) and IgG responses against both antigens were detected among all CBE groups. No significant immune response was detected using PBE strains. Expression of recombinant antigens in S. enterica serovar Typhimurium aroA from chromosomally located strong promoters without the use of antibiotic resistance markers is a reliable and effective method of inducing a significant immune response.The use of live, attenuated bacteria as vaccine delivery systems for heterologous antigens has been extensively studied. In particular, attenuated Salmonella strains have been modified to express a wide range of antigens from bacterial, parasitic, and viral sources (reviewed in references 20, 29, and 41). After oral administration, Salmonella can penetrate the Peyer's patches via M cells and colonize the mesenteric lymph nodes, which contain various antigen-presenting cells (reviewed in reference 5). This can generate a range of immune responses, including systemic and mucosal responses at local and distal sites. Other advantages of using attenuated Salmonella include the ease of oral administration, which bypasses the need for needle administration; increased antigen presentation due to the use of a live-vector delivery system, the induction of both Th1 and Th2 immune responses; and the wide range of attenuated Salmonella and recombinant vectors available to researchers (19,20,51).However, there are a number of issues to overcome. Most methods for the expression of heterologous antigens in Salmonella use plasmids to express the antigenic proteins. This can have several drawbacks. The stable maintenance of the expression plasmid in vivo can be difficult to achieve. Tightly regulated promoters are often used to increase plasmid stability, and several in vivo-inducible promoters have delivered promising res...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Development of Non-Antibiotic-Resistant, Chromosomally Based, Constitutive and Inducible Expression Systems for aroA -Attenuated Salmonella enterica Serovar Typhimurium

et al. 2009

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“…COS-7 cells were transfected with pcDNA3.1/ptS1, pcDNA3.1/ptS1.13L.129G or pcDNA3.1 vector (negative control) using Lipofectamine 2000 (Invitrogen) as described previously (Chen et al, 2006). Expression of rPTS1 and rPTS1.13L.129G was detected by Western blotting with an alkaline phosphatase-labelled anti-V5 antibody (Invitrogen) specific for the V5 epitope, which was fused to the C terminus of the recombinant S1.…”

Section: Methodsmentioning

confidence: 99%

“…His-tagged rPTS1.13L.129G was purified by IMAC using Ni-NTA resin (Qiagen) as described previously (Chen et al, 2006). A 100 ml culture of XL10-Gold E. coli harbouring the pTrcHis2/ptS1.13L.129G plasmid was grown in LB broth to OD 600 0.7 and then induced with 1 mM IPTG for 4 h at 37 uC with shaking at 200 r.p.m.…”

Section: Methodsmentioning

confidence: 99%

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Parenteral immunization of mice with a genetically inactivated pertussis toxin DNA vaccine induces cell-mediated immunity and protection

Fry

Chen

Daggard

et al. 2008

Journal of Medical Microbiology

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The immunogenicity and protective efficacy of a DNA vaccine encoding a genetically inactivated S1 domain of pertussis toxin was evaluated using a murine respiratory challenge model of Bordetella pertussis infection. It was found that mice immunized via the intramuscular route elicited a purely cell-mediated immune response to the DNA vaccine, with high levels of gamma interferon (IFN-c) and interleukin (IL)-2 detected in the S1-stimulated splenocyte supernatants and no serum IgG. Despite the lack of an antibody response, the lungs of DNA-immunized mice were cleared of B. pertussis at a significantly faster rate compared with mock-immunized mice following an aerosol challenge. To gauge the true potential of this S1 DNA vaccine, the immune response and protective efficacy of the commercial diphtheria-tetanus-acellular pertussis (DTaP) vaccine were included as the gold standard. Immunization with DTaP elicited a typically strong T-helper (Th)2-polarized immune response with significantly higher titres of serum IgG than in the DNA vaccine group, but a relatively weak Th1 response with low levels of IFN-c and IL-2 detected in the supernatants of antigen-stimulated splenocytes. DTaP-immunized mice cleared the aerosol challenge more efficiently than DNA-immunized mice, with no detectable pathogen after day 7 post-challenge.

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Development of Mycoplasma hyopneumoniae Recombinant Vaccines

2016

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Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia (EP), a disease that affects swine production worldwide. Vaccination is the most cost-effective strategy for the control and prevention of the disease. Research using genome-based approach has the potential to elucidate the biology and pathogenesis of M. hyopneumoniae and contribute to the development of more effective vaccines. Here, we describe the protocol for developing M. hyopneumoniae recombinant vaccines using reverse vaccinology approaches.

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Comparative immunogenicity of M. hyopneumoniae NrdF encoded in different expression systems delivered orally via attenuated S. typhimurium aroA in mice

Cited by 19 publications

References 21 publications

Development of Non-Antibiotic-Resistant, Chromosomally Based, Constitutive and Inducible Expression Systems for aroA -Attenuated Salmonella enterica Serovar Typhimurium

Development of Non-Antibiotic-Resistant, Chromosomally Based, Constitutive and Inducible Expression Systems for aroA -Attenuated Salmonella enterica Serovar Typhimurium

Parenteral immunization of mice with a genetically inactivated pertussis toxin DNA vaccine induces cell-mediated immunity and protection

Development of Mycoplasma hyopneumoniae Recombinant Vaccines

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