Production and Targeting of the<i>Brucella abortus</i>Antigen L7/L12 in<i>Lactococcus lactis</i>: a First Step towards Food-Grade Live Vaccines against Brucellosis

Ribeiro, Lucilene Arilho; Azevedo, Vasco; Loir, Yves Le; Oliveira, Sérgio C.; Dièye, Yakhya; Piard, Jean-Christophe; Gruss, Alexandra; Langella, Philippe

doi:10.1128/aem.68.2.910-916.2002

Cited by 127 publications

(113 citation statements)

References 51 publications

Supporting

Mentioning

104

Contrasting

Unclassified

Order By: Relevance

“…In the last decade, several studies have used L. lactis as an antigen delivery vehicle to develop safe, live vaccines (Robinson et al, 1997;Steidler et al, 1998;Chatel et al, 2001;Enouf et al, 2001;Bermú dez-Humarán et al, 2002;Ribeiro et al, 2002;Xin et al, 2003). In these studies, different transcriptional and translational lactococcal signals have been used to express antigens.…”

Section: Resultsmentioning

confidence: 99%

An inducible surface presentation system improves cellular immunity against human papillomavirus type 16 E7 antigen in mice after nasal administration with recombinant lactococci

Bermúdez‐Humarán

Cortes-Perez

Loir

et al. 2004

Journal of Medical Microbiology

Self Cite

133

View full text Add to dashboard Cite

Human papillomavirus type 16 (HPV-16) is the major causative agent of cervical cancer. To date, vaccine strategies against HPV-16 are based on the ability of the E7 oncoprotein to elicit an immune response against this virus. In this study, the use of an inducible or a constitutive system to produce the HPV-16 E7 protein in Lactococcus lactis, a non-pathogenic and non-invasive Gram-positive bacterium, was compared. The highest E7 production was obtained with the inducible system. When mice were immunized intranasally with recombinant lactococci expressing either inducible or constitutive E7, an antigen-specific cellular response (i.e. secretion of IL2 and IFN-ª cytokines) was evoked and was substantially higher in mice receiving L. lactis expressing E7 with the inducible system. As bacterial antigen location may influence the immune response, recombinant L. lactis strains that produced E7 in three cellular locations, intracellular, secreted or cell-wall-anchored were evaluated. The highest immune response was elicited by administration of L. lactis producing an inducible cell-wall-anchored form of E7 protein. These promising results represent a step towards the development of a new, safe mucosal vector to treat HPV-related cervical cancer. INTRODUCTIONEpidemiological data have clearly shown that human papillomavirus type 16 (HPV-16) infection is the main aetiological factor for cervical cancer (CxCa) (Furumoto & Irahara, 2002). Worldwide, $400 000 women die annually from CxCa (Parkin et al., 1999). A prophylactic and/or therapeutic vaccine against this virus is thus a priority to prevent or to treat, respectively, CxCa. A prophylactic vaccine based on highly purified virus-like particles has recently been successfully used in trials in women, with a significant reduction observed in the incidence of both HPV-16 infection and related CxCa (Koutsky et al., 2002). However, such vaccines could probably not be used therapeutically in alreadyinfected patients because the virion capsid proteins are not detected in CxCa. The HPV-16 E7 protein, constitutively produced in cervical carcinomas, is required for the transformation process (Baker et al., 1987;Bedell et al., 1987;Dyson et al., 1989;Tanaka et al., 1989) and is considered a good antigen candidate for the development of a therapeutic vaccine against CxCa.Several studies have investigated the use of bacteria as E7 antigen delivery vehicles to elicit an immune response against HPV-16. In these studies, the vectors used were attenuated strains of pathogenic bacteria such as Salmonella and Mycobacterium spp. (Londoño et al., 1996; Jabbar et al., 2000). Although these recombinant strains elicited immune responses, invasiveness of the vectors and risks of reversion to pathogenicity limit their use in vulnerable groups such as immunocompromised patients or children. There is thus a need for the development of a new generation of safe delivery vehicles.Lactic acid bacteria (LAB) are promising candidates as safe vehicles for in vivo delivery of antigens. Compared with att...

show abstract

Section: Resultsmentioning

confidence: 99%

An inducible surface presentation system improves cellular immunity against human papillomavirus type 16 E7 antigen in mice after nasal administration with recombinant lactococci

Bermúdez‐Humarán

Cortes-Perez

Loir

et al. 2004

Journal of Medical Microbiology

Self Cite

133

View full text Add to dashboard Cite

show abstract

“…The only approved mucosal vaccines are pathogens (49), some of which may pose a risk of infection, particularly to children or immunocompromised individuals (36,39,46). Therefore, recent efforts to develop mucosal vaccine delivery systems have included ones that are not pathogenic, such as liposomes or microparticles (10,50), commensal bacteria (19,20,32,34,39,45,46), or noninvasive, noncommensal bacteria (11,18,31,42).…”

Section: Discussionmentioning

confidence: 99%

“…It is generally recognized as safe by the U.S. Food and Drug Administration. Many heterologous proteins have been expressed in L. lactis (11,12,18,23,31,42,53), and immunization with these strains elicits immune responses specific to heterologous antigens (43,52,55). However, we are aware of only one report (55) that shows that mucosal immunization with a lactococcal vaccine can reduce infection.…”

mentioning

confidence: 91%

Mucosal Vaccine Made from Live, RecombinantLactococcus lactisProtects Mice against Pharyngeal Infection withStreptococcus pyogenes

2004

View full text Add to dashboard Cite

A novel vaccine (LL-CRR) made from live, nonpathogenic Lactococcus lactis that expresses the conserved C-repeat region (CRR) of M protein from Streptococcus pyogenes serotype 6 was tested in mice. Nasally vaccinated mice produced CRR-specific salivary immunoglobulin A (IgA) and serum IgG. Subcutaneously vaccinated mice produced CRR-specific serum IgG but not salivary IgA. A combined regimen produced responses similar to the salivary IgA of nasally vaccinated mice and serum IgG of subcutaneously vaccinated mice. Mice vaccinated nasally or with the combined regimen were significantly protected against pharyngeal infection following a nasal challenge with S. pyogenes M serotype 14. Mice vaccinated subcutaneously were not protected against pharyngeal infection. Mice in all three LL-CRR vaccination groups were significantly protected against the lethal effects of S. pyogenes. Only 1 of 77 challenged mice that were vaccinated with LL-CRR died, whereas 60 of 118 challenged mice that were vaccinated with a control strain or phosphatebuffered saline died. In conclusion, mucosal vaccination with LL-CRR produced CRR-specific salivary IgA and serum IgG, prevented pharyngeal infection with S. pyogenes, and promoted survival.

show abstract

“…Expression of heterologous protein was described by Ribeiro et al, [7], we followed the same for OmpA expression with minor modification in nisin concentration i.e. 10 ng/ml.…”

Section: Ompa Expressionmentioning

confidence: 99%

Factors Affecting Inducible Expression of Outer Membrane Protein A (OmpA) of Shigella dysenteriae Type-1 in Lactococcus lactis Using Nisin Inducible Controlled Expression (NICE)

et al. 2015

View full text Add to dashboard Cite

Potential use of Lactococcus lactis (L. lactis) as a heterologous protein expression host as well as for delivery of multiple therapeutic proteins has been investigated extensively using Nisin Inducible Controlled Expression (NICE) system. Optimum inducible expression of heterologous protein by NICE system in L. lactis depends on multiple factors. To study the unexplored role of factors affecting heterologous protein expression in L. lactis using NICE, the present study outlines the optimization of various key parameters such as inducer concentration, host's proteases and precipitating agent using Outer membrane protein A (OmpA). For efficient expression and secretion of OmpA, pSEC:OmpA vector was successfully constructed. To circumvent the troubles encountered during detection of expressed OmpA, the precipitating agent was switched from TCA to methanol.Nevertheless, detection was achieved accompanied by degraded protein products. Speculating the accountability of observed degradation at higher inducer concentration, different nisin concentrations were evaluated. Lower nisin concentrations were found desirable for optimum expression of OmpA. Consistently observed degradation was eliminated by incorporation of protease inhibitor cocktail which inhibits intracellular proteases and expression in VEL1153 (NZ9000 DhtrA) strain which inhibits extracellular protease leading to optimum expression of OmpA. Versatility and complexity of NICE system in L. lactis requires fine-tuning of target protein specific parameters for optimum expression.

show abstract

Production and Targeting of theBrucella abortusAntigen L7/L12 inLactococcus lactis: a First Step towards Food-Grade Live Vaccines against Brucellosis

Cited by 127 publications

References 51 publications

An inducible surface presentation system improves cellular immunity against human papillomavirus type 16 E7 antigen in mice after nasal administration with recombinant lactococci

An inducible surface presentation system improves cellular immunity against human papillomavirus type 16 E7 antigen in mice after nasal administration with recombinant lactococci

Mucosal Vaccine Made from Live, RecombinantLactococcus lactisProtects Mice against Pharyngeal Infection withStreptococcus pyogenes

Factors Affecting Inducible Expression of Outer Membrane Protein A (OmpA) of Shigella dysenteriae Type-1 in Lactococcus lactis Using Nisin Inducible Controlled Expression (NICE)

Contact Info

Product

Resources

About