Immunization with cell‐free‐generated vaccine protects from Porphyromonas gingivalis‐induced alveolar bone loss

Abstract

Introduction Periodontal diseases (PD) are complex oral inflammatory diseases initiated by keystone bacteria such as Porphyromonas gingivalis. A vaccine for PD is desirable as clinical treatment involves protracted maintenance strategies aimed to retain dentition. Although prior immunization approaches targeting P. gingivalis have reported variable success in limiting facets of disease such as oral bone loss, it remains that a vaccine for this disease may be attainable. Aim To investigate cell‐free protein synthesis (CFPS) as a platform to produce vaccinable targets suitable for efficacy testing in a P. gingivalis‐induced murine oral bone loss model. Materials and Methods Recombinantly generated P. gingivalis minor fimbriae protein (Mfa1), RgpA gingipain hemagglutinin domain 1 (HA1), and RgpA gingipain hemagglutinin domain 2 (HA2) were combined in equivalent doses in adjuvants and injected intramuscularly to immunize mice. Serum levels of protein‐specific antibody were measured by ELISA, and oral bone levels were defined by morphometrics. Results Recombinantly generated P. gingivalis proteins possessed high fidelity to predicted size and elicited protein‐specific IgG following immunization. Importantly, immunization with the vaccine cocktail protected from P. gingivalis elicited oral bone loss. Conclusion These data verify the utility of the CFPS technology to synthesize proteins that have the capacity to serve as novel vaccines.

Periodontal Vaccines-A systematic Review

“…[38] Very recently, Huang et al have used a novel technique of cell free protein synthesis as a platform to produce vaccine candidates. They were able to recombinantly generate a cocktail of P.gingivalis proteins that were capable of eliciting a high serum IgG response as well as were capable of protecting the mice from progressive bone loss [39].…”

A Porphyromonas gingivalis Capsule-Conjugate Vaccine Protects From Experimental Oral Bone Loss

“…For all studies, we used P. gingivalis strain A7436, grown anaerobically as previously described [ 21 ]. In brief, bacteria were plated from freezer stocks on brain-heart infusion blood agar supplemented with yeast extract, and harvested plate growth was then cultured in BHI-YE broth to a late log phase.…”

“…To define oral bone loss in response to P. gingivalis and the impact of vaccination, we used the murine oral bone loss model of Baker as we have done previously [ 20 , 21 , 51 ]. Studies using live animals were conducted in accordance with U. Florida IACUC-approved protocols.…”

“…gingivalis is a Gram-negative anaerobic bacterium that possesses an array of molecules, including fimbriae, two types of lipopolysaccharide, gingipains, and a polysaccharide capsule that contribute to the overall virulence potential of this organism [9][10][11][12][13][14][15]. Several groups have targeted these molecules as vaccine candidates and have had varying degrees of success [16][17][18][19][20][21]. It is known that P. gingivalis strains display a broad range of heterogeneity; however, encapsulated strains are associated with highly invasive infection [9,22].…”

Periodontal Vaccine-Armour against Periodontitis

“…Of late, distinct literature is coming forth that highlights the ongoing efforts to develop an effective, biocompatible and practical mode of periodontal vaccination. Based on the rationale that periodontal diseases are complex oral inflammatory diseases initiated by keystone bacteria, Huang, et al [28] investigated cellfree protein synthesis as a platform to produce vaccinable targets suitable for efficacy testing in a P. gingivalisinduced murine oral bone loss model. They reported recombinantly generated P. gingivalis minor fimbriae protein, RgpA gingipain hemagglutinin domain 1, and RgpA gingipain hemagglutinin domain 2, possessed high fidelity to predicted size and elicited protein-specific IgG following immunization.…”