Immunogenicity and protective efficacy against Treponema pallidum in New Zealand rabbits immunized with plasmid DNA encoding flagellin

Abstract

Plasmid DNA encoding flagellin FlaB3 was used as a vaccination candidate for the evaluation of immunogenicity and protection against Treponema pallidum subsp. pallidum dissemination. First, intramuscular injection of the flagellin encoded by the plasmid DNA into New Zealand rabbits elicited both humoral and cellular immune responses. Total IgG production increased in response to flagellin. In addition, serum IFN-γ secretion and CD8+ cells were substantially greater in the rabbits immunized with the plasmid encoding flagellin FlaB3 than those in the rabbits immunized with recombinant flagellin. The flagellin encoded by the plasmid DNA induced significant upregulation of serum IL-6 and IL-8 compared to that of the control rabbits. Subsequently, intradermal challenge of the vaccinated New Zealand rabbits with 1 × 107T. pallidum resulted in a significant reduction of the bacterial organ burden in the blood, liver, spleen, and testicles in the flagellin plasmid DNA-vaccinated rabbits. Furthermore, the histopathological analysis demonstrated that the rabbits immunized with the plasmid DNA-encoded flagellin (FlaB3) showed better immune protection. These findings provide evidence that plasmid DNA-encoded flagellin (FlaB3) may be useful as a potential immunization route for future development of a vaccine to inhibit T. pallidum dissemination in related animals.

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“…In fact, it has also been used in the vaccine and bacterial resistance fields. For example, Lithgow Karen et al utilized real‐time PCR to detect the quantity of T. pallidum in New Zealand rabbits, and this method provided the precise immune effect of recombinational proteins Tp0751 and FlaB3. Although T. pallidum has been sensitive to penicillin since it first appeared, we should continue to be vigilant at drug‐resistant strains because of antibiotic abuse and gene mutations.…”

“…In fact, it has also been used in the vaccine and bacterial resistance fields. For example, Lithgow Karen et al34,35 utilized real-time PCR to detect the quantity of T. pallidum in New Zealand rabbits, and this method provided…”

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“…Total RNA was extracted using TRIzol (Beyotime, China) and then converted to cDNA using BeyoRT II First Strand cDNA synthesis kit (RNase H minus). Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) was performed using the SYBR Green mix (Qiagen, Shanghai, China) in the LightCycle 96 apparatus (Roche, Basel, Switzerland) (19). The primers used are shown in Table 1.…”

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“…Новым подходом к созданию вакцины было использование инактивированной Borrelia burgdorferi в качестве эффективного носителя для доставки антигенов и индукции специфического ответа макроорганизма на антигены T. pallidum, экспрессируемые генами tp0897 (tprK) и tp0435 [11]. В другом исследовании в качестве вакцины-кандидата для оценки иммуногенности и защиты от распространения возбудителя использовался флагеллин FlaB3, кодируемый плазмидной ДНК [12]. У кроликов, вакцинированных флагеллином FlaB3, кодируемым плазмидной ДНК, при последующем внутрикожном заражении наблюдалось значимое снижение бактериальной нагрузки в крови, печени, селезенке и яичках.…”