Oral Immunization with Recombinant Lactobacillus plantarum Induces a Protective Immune Response in Mice with Lyme Disease

ABSTRACT Mucosal immunization is advantageous over other routes of antigen delivery because it can induce both mucosal and systemic immune responses. Our goal was to develop a mucosal delivery vehicle based on bacteria generally regarded as safe, such as Lactobacillus spp. In this study, we used the Lyme disease mouse model as a proof of concept. We demonstrate that an oral vaccine based on live recombinant Lactobacillus plantarum protects mice from tick-transmitted Borrelia burgdorferi infection. Our method of expressing vaccine antigens in L. plantarum induces both systemic and mucosal immunity after oral administration. This platform technology can be applied to design oral vaccine delivery vehicles against several microbial pathogens.

Download Full-text

Antigen Delivery to Mucosa-Associated Lymphoid Tissues Using Liposomes as a Carrier

Bioscience Reports ◽

10.1023/a:1020103109483 ◽

2002 ◽

Vol 22 (2) ◽

pp. 355-369 ◽

Cited By ~ 45

Author(s):

Fan Zhou ◽

Marian R. Neutra

Keyword(s):

Oral Vaccine ◽

Lymphoid Tissues ◽

M Cells ◽

Antigen Delivery ◽

Mucosal Immunization ◽

Oral Vaccines ◽

Oral Vaccination ◽

Particulate Form ◽

Delivery Vehicles ◽

Antigen Carrier

Oral vaccination requires an antigen delivery vehicle to protect the antigen and to enhance translocation of the antigen to the mucosa-associated lymphoid tissue. A variety of antigen delivery vehicles including liposomes have been studied for mucosal immunization. The advantages of liposome formulations are their particulate form and the ability to accommodate immunomodulators and targeting molecules in the same package. Many conventional liposomes are variably unstable in acids, pancreatic juice and bile. Nevertheless, carefully designed liposomes have demonstrated an impressive efficacy in inducing mucosal IgA responses, compared to free antigens and other delivery vehicles. However, liposomes as an oral vaccine vehicle are not yet optimized. To design liposomes that are stable in the harsh intestinal environment and are efficiently taken up by the M cells remains a challenge. This review summarizes recent research efforts using liposomes as an antigen carrier for oral vaccines with practical attention to liposome designs and interaction with the M cells.

Download Full-text

Gut Adhesive Bacillus subtilis Spores as a Platform for Mucosal Delivery of Antigens

Infection and Immunity ◽

10.1128/iai.01255-13 ◽

2014 ◽

Vol 82 (4) ◽

pp. 1414-1423 ◽

Cited By ~ 23

Author(s):

Milene Tavares Batista ◽

Renata D. Souza ◽

Juliano D. Paccez ◽

Wilson B. Luiz ◽

Ewerton L. Ferreira ◽

...

Keyword(s):

Bacillus Subtilis ◽

Streptococcus Mutans ◽

Sublingual Administration ◽

Antigen Delivery ◽

Content Type ◽

Mucosal Delivery ◽

Abiotic Surfaces ◽

Bacterial Adhesins ◽

Vaccine Antigens

ABSTRACTBacillus subtilisspores have been used as safe and heat-resistant antigen delivery vectors. Nonetheless, the oral administration of spores typically induces weak immune responses to the passenger antigens, which may be attributed to the fast transit through the gastrointestinal tract. To overcome this limitation, we have developedB. subtilisspores capable of binding to the gut epithelium by means of expressing bacterial adhesins on the spore surface. The resulting spores bound toin vitrointestinal cells, showed a longer transit through the mouse intestinal tract, and interacted with Peyer's patch cells. The adhesive spores increased the systemic and secreted antibody responses to theStreptococcus mutansP1 protein, used as a model antigen, following oral, intranasal, and sublingual administration. Additionally, P1-specific antibodies efficiently inhibited the adhesion of the oral pathogenStreptococcus mutansto abiotic surfaces. These results support the use of gut-colonizingB. subtilisspores as a new platform for the mucosal delivery of vaccine antigens.

Download Full-text

Lactic acid bacteria as antigen delivery vehicles for oral immunization purposes

International Journal of Food Microbiology ◽

10.1016/s0168-1605(98)00048-8 ◽

1998 ◽

Vol 41 (2) ◽

pp. 155-167 ◽

Cited By ~ 129

Author(s):

Peter H Pouwels ◽

Rob J Leer ◽

Michael Shaw ◽

Marie-Joan Heijne den Bak-Glashouwer ◽

Frans D Tielen ◽

...

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Oral Immunization ◽

Antigen Delivery ◽

Delivery Vehicles

Download Full-text

Mucosal immune responses induced by oral administration recombinant Bacillus subtilis expressing the COE antigen of PEDV in newborn piglets

Bioscience Reports ◽

10.1042/bsr20182028 ◽

2019 ◽

Vol 39 (3) ◽

Cited By ~ 6

Author(s):

Jialu Wang ◽

Lulu Huang ◽

Chunxiao Mou ◽

En Zhang ◽

Yongheng Wang ◽

...

Keyword(s):

Bacillus Subtilis ◽

Immune Responses ◽

Oral Administration ◽

Oral Vaccine ◽

Oral Immunization ◽

Economic Losses ◽

Antigen Delivery ◽

Newborn Piglets ◽

Mucosal Immune Responses ◽

Recombinant Bacillus Subtilis

Abstract Porcine epidemic diarrhea (PED) is a highly contagious disease in newborn piglets and causes substantial economic losses in the world. PED virus (PEDV) spreads by fecal–oral contact and can be prevented by oral immunization. Therefore, it is necessary to develop an effective oral vaccine against PEDV infection. Currently, Bacillus subtilis as recombinant vaccine carrier has been used for antigen delivery and proved well in immune effect and safety. The present study evaluated the immunogenicity of recombinant Bacillus subtilis (B. subtilis-RC) in piglets via oral administration. After oral immunization in piglets, B. subtilis-RC significantly increased the local mucosal immune responses. Oral administration with B. subtilis-RC significantly improved the level of specific mucosal immunoglobulin A (IgA) antibodies against PEDV infection, through enlarging the area of Peyer’s patches (PPs) and increasing the number of ileum IgA+ secreting (SIgA) cells. In the meantime, B. subtilis-RC remarkably increased the number of intraepithelial lymphocytes (IELs). We also observed that oral administration of B. subtilis-RC significantly increased CD3+T lymphocytes’ numbers and up-regulated the ratio of CD4+/CD8+ T cells. Furthermore, high titers of specific serum immunoglobulin G (IgG) revealed satisfactory systemic immune response against PEDV infection. In summary, our study demonstrated that oral administration of B. subtilis-RC could trigger a high level of local and systemic immune responses and would be a promising candidate vaccine against PEDV infection in piglets.

Download Full-text

Surface Display of N-Terminally Anchored Invasin by Lactobacillus plantarum Activates NF-κB in Monocytes

Applied and Environmental Microbiology ◽

10.1128/aem.01227-12 ◽

2012 ◽

Vol 78 (16) ◽

pp. 5864-5871 ◽

Cited By ~ 40

Author(s):

Lasse Fredriksen ◽

Charlotte R. Kleiveland ◽

Lene T. Olsen Hult ◽

Tor Lea ◽

Cathrine S. Nygaard ◽

...

Keyword(s):

Lactobacillus Plantarum ◽

Yersinia Pseudotuberculosis ◽

Surface Display ◽

Extracellular Domain ◽

Content Type ◽

Bacterial Surface ◽

Mucosal Surfaces ◽

Vaccine Antigens ◽

Mucosal Vaccines ◽

Delivery Vehicles

ABSTRACTThe probiotic lactic acid bacteriumLactobacillus plantarumis a potential delivery vehicle for mucosal vaccines because of its generally regarded as safe (GRAS) status and ability to persist at the mucosal surfaces of the human intestine. However, the inherent immunogenicity of vaccine antigens is in many cases insufficient to elicit an efficient immune response, implying that additional adjuvants are needed to enhance the antigen immunogenicity. The goal of the present study was to increase the proinflammatory properties ofL. plantarumby expressing a long (D1 to D5 [D1-D5]) and a short (D4-D5) version of the extracellular domain of invasin from the human pathogenYersinia pseudotuberculosis. To display these proteins on the bacterial surface, four different N-terminal anchoring motifs fromL. plantarumwere used, comprising two different lipoprotein anchors, a transmembrane signal peptide anchor, and a LysM-type anchor. All these anchors mediated surface display of invasin, and several of the engineered strains were potent activators of NF-κB when interacting with monocytes in cell culture. The most distinct NF-κB responses were obtained with constructs in which the complete invasin extracellular domain was fused to a lipoanchor. The proinflammatoryL. plantarumstrains constructed here represent promising mucosal delivery vehicles for vaccine antigens.

Download Full-text

Lactic Acid Bacteria as Mucosal Delivery Vehicles

Genetics of Lactic Acid Bacteria ◽

10.1007/978-1-4615-7090-5_9 ◽

2003 ◽

pp. 261-290

Author(s):

Jerry M. Wells ◽

Annick Mercenier

Keyword(s):

Lactic Acid ◽

Lactic Acid Bacteria ◽

Mucosal Delivery ◽

Delivery Vehicles

Download Full-text

Immunogenicity in Swine of Orally Administered Recombinant Lactobacillus plantarum Expressing Classical Swine Fever Virus E2 Protein in Conjunction with Thymosin α-1 as an Adjuvant

Applied and Environmental Microbiology ◽

10.1128/aem.00127-15 ◽

2015 ◽

Vol 81 (11) ◽

pp. 3745-3752 ◽

Cited By ~ 27

Author(s):

Yi-Gang Xu ◽

Xue-Ting Guan ◽

Zhong-Mei Liu ◽

Chang-Yong Tian ◽

Li-Chun Cui

Keyword(s):

Immune Responses ◽

Lactobacillus Plantarum ◽

Classical Swine Fever Virus ◽

Oral Vaccine ◽

Classical Swine Fever ◽

Fever Virus ◽

Economic Losses ◽

E2 Protein ◽

Content Type ◽

Cellular Immune

ABSTRACTClassical swine fever, caused by classical swine fever virus (CSFV), is a highly contagious disease that results in enormous economic losses in pig industries. The E2 protein is one of the main structural proteins of CSFV and is capable of inducing CSFV-neutralizing antibodies and cytotoxic T lymphocyte (CTL) activitiesin vivo. Thymosin α-1 (Tα1), an immune-modifier peptide, plays a very important role in the cellular immune response. In this study, genetically engineeredLactobacillus plantarumbacteria expressing CSFV E2 protein alone (L. plantarum/pYG-E2) and in combination with Tα1 (L. plantarum/pYG-E2-Tα1) were developed, and the immunogenicity of each as an oral vaccine to induce protective immunity against CSFV in pigs was evaluated. The results showed that recombinantL. plantarum/pYG-E2 andL. plantarum/pYG-E2-Tα1 were both able to effectively induce protective immune responses in pigs against CSFV infection by eliciting immunoglobulin A (IgA)-based mucosal, immunoglobulin G (IgG)-based humoral, and CTL-based cellular immune responses via oral vaccination. Significant differences (P< 0.05) in the levels of immune responses were observed betweenL. plantarum/pYG-E2-Tα1 andL. plantarum/pYG-E2, suggesting a better immunogenicity ofL. plantarum/pYG-E2-Tα1 as a result of the Tα1 molecular adjuvant that can enhance immune responsiveness and augment specific lymphocyte functions. Our data suggest that the recombinantLactobacillusmicroecological agent expressing CSFV E2 protein combined with Tα1 as an adjuvant provides a promising strategy for vaccine development against CSFV.

Download Full-text

Alga-Produced Cholera Toxin-Pfs25 Fusion Proteins as Oral Vaccines

Applied and Environmental Microbiology ◽

10.1128/aem.00714-13 ◽

2013 ◽

Vol 79 (13) ◽

pp. 3917-3925 ◽

Cited By ~ 64

Author(s):

James A. Gregory ◽

Aaron B. Topol ◽

David Z. Doerner ◽

Stephen Mayfield

Keyword(s):

Cholera Toxin ◽

Tandem Repeats ◽

Low Cost ◽

Oral Vaccine ◽

Childhood Mortality ◽

Secretory Iga ◽

Content Type ◽

Freeze Dried ◽

Vaccine Antigens ◽

Iga Antibodies

ABSTRACTInfectious diseases disproportionately affect indigent regions and are the greatest cause of childhood mortality in developing countries. Practical, low-cost vaccines for use in these countries are paramount to reducing disease burdens and concomitant poverty. Algae are a promising low-cost system for producing vaccines that can be orally delivered, thereby avoiding expensive purification and injectable delivery. We engineered the chloroplast of the eukaryotic algaChlamydomonas reinhardtiito produce a chimeric protein consisting of the 25-kDaPlasmodium falciparumsurface protein (Pfs25) fused to the β subunit of the cholera toxin (CtxB) to investigate an alga-based whole-cell oral vaccine. Pfs25 is a promising malaria transmission-blocking vaccine candidate that has been difficult to produce in traditional recombinant systems due to its structurally complex tandem repeats of epidermal growth factor-like domains. The noncatalytic CtxB domain of the cholera holotoxin assembles into a pentameric structure and acts as a mucosal adjuvant by binding GM1 ganglioside receptors on gut epithelial cells. We demonstrate that CtxB-Pfs25 accumulates as a soluble, properly folded and functional protein within algal chloroplasts, and it is stable in freeze-dried alga cells at ambient temperatures. In mice, oral vaccination using freeze-dried algae that produce CtxB-Pfs25 elicited CtxB-specific serum IgG antibodies and both CtxB- and Pfs25-specific secretory IgA antibodies. These data suggest that algae are a promising system for production and oral delivery of vaccine antigens, but as an orally delivered adjuvant, CtxB is best suited for eliciting secretory IgA antibodies for vaccine antigens against pathogens that invade mucosal surfaces using this strategy.

Download Full-text

Peptide nanofiber–CaCO3 composite microparticles as adjuvant-free oral vaccine delivery vehicles

Journal of Materials Chemistry B ◽

10.1039/c5tb01623a ◽

2016 ◽

Vol 4 (9) ◽

pp. 1640-1649 ◽

Cited By ~ 13

Author(s):

Joshua D. Snook ◽

Charles B. Chesson ◽

Alex G. Peniche ◽

Sara M. Dann ◽

Adriana Paulucci ◽

...

Keyword(s):

Mucosal Immunity ◽

Oral Vaccine ◽

Vaccine Delivery ◽

Oral Vaccination ◽

Delivery Vehicles ◽

Gi Infections ◽

Mucosal Pathogens

To combat mucosal pathogens that cause gastrointestinal (GI) infections, local mucosal immunity is required which is best achieved through oral vaccination.

Download Full-text