Cell Wall Anchoring of the 37-Kilodalton Oncofetal Antigen by
            <i>Lactobacillus plantarum</i>
            for Mucosal Cancer Vaccine Delivery

Fredriksen, Lasse; Mathiesen, Geir; Sioud, Mouldy; Eijsink, Vincent G. H.

doi:10.1128/aem.01031-10

Cited by 55 publications

(74 citation statements)

References 20 publications

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“…for protein secretion and anchoring (13,26). These vectors were constructed in order to complement the acm2 mutant with either native Acm2 or an N-terminally truncated version of Acm2 for cytoplasmic expression.…”

Section: Methodsmentioning

confidence: 99%

The Major Autolysin Acm2 from Lactobacillus plantarum Undergoes Cytoplasmic O-Glycosylation

et al. 2012

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The major autolysin Acm2 from the probiotic strain Lactobacillus plantarum WCFS1 contains high proportions of alanine, serine, and threonine in its N-terminal so-called AST domain. It has been suggested that this extracellular protein might be glycosylated, but this has not been experimentally verified. We used high-resolution liquid chromatography-tandem mass spectrometry (LC-MS/MS) to study the possible occurrence of glycans on peptides generated from lactobacillary surface proteins by protease treatment. This approach yielded five glycopeptides in various glycoforms, all derived from the AST domain of Acm2. All five glycopeptides contained the hydroxy-amino acids serine and threonine, suggesting that Acm2 is O-glycosylated. By using lectin blotting with succinylated wheat germ agglutinin, and by comparing the wild-type strain with an Acm2-negative derivative (NZ3557), we found that the attached N-acetylhexosamines are most likely N-acetylglucosamines (GlcNAc). NZ3557 was further used as a genetic background to express an Acm2 variant lacking its secretion signal, resulting in intracellular expression of Acm2. We show that this intracellular version of Acm2 is also glycosylated, indicating that the GlcNAc modification is an intracellular process.

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Section: Methodsmentioning

confidence: 99%

The Major Autolysin Acm2 from Lactobacillus plantarum Undergoes Cytoplasmic O-Glycosylation

et al. 2012

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“…lactis D17 ( Table 1). The recombinant plasmids were transformed into Lactobacillus by electroporation as previously described (8).…”

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confidence: 99%

Mucosal and Systemic Immune Responses Induced by Recombinant Lactobacillus spp. Expressing the Hemagglutinin of the Avian Influenza Virus H5N1

Wang

Gao

et al. 2012

Clin Vaccine Immunol

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To develop a safe, effective, and convenient vaccine for the prevention of highly pathogenic avian influenza (HPAI), we have successfully constructed two recombinant lactobacillus strains (LA4356-pH and DLD17-pH) that express the foreign HPAI virus protein hemagglutinin 1 (HA 1 ). The mucosal and systemic immune responses triggered by these two recombinant lactobacilli following oral administration to BALB/c mice were evaluated. The results showed that both LA4356-pH and DLD17-pH could significantly increase the specific anti-HA 1 IgA antibody level in the mucosa and the anti-HA 1 IgG level in serum, as well as stimulating the splenic lymphocyte proliferative reaction through increased expression of interleukin-4 (IL-4). Compared with LA4356-pH, DLD17-pH was more effective at inducing systemic and mucosal immune responses, with higher anti-HA 1 -specific IgA and IgG levels. Therefore, DLD17-pH could be a promising oral vaccine candidate against HPAI. Highly pathogenic avian influenza (HPAI) virus (H5N1) is a threat to the world's poultry industry. As a zoonotic agent, this virus also has the potential to cause a human pandemic (29). Current vaccination (by intramuscular immunization) against HPAI has succeeded in reducing morbidity and mortality in poultry. However, the intense stress caused by injection could hamper the animal's growth (34). Thus, many investigators are pursuing more convenient and economical avenues for construction of new vaccine candidates, such as recombinant subunit vaccines using baculovirus (28), plasmid DNA (3), or replication-incompetent adenovirus (rAd) (26, 27) vectors. The H5N1 strain can infect animals through their respiratory and intestinal tracts (10). Because there are many mucosa-associated lymphoid tissues underneath the epithelia of the respiratory and intestinal tracts, protective HPAI virus antigens may induce an effective mucosal immune response to prevent the invasion of HPAI virus if they can be transported easily to these tissues. Thus, developing mucosal vaccine candidates based on these considerations is a feasible strategy.Mucosal vaccines, which are administered mainly orally or intranasally, cause less stress in the animal and have been the subject of growing interest due to the advantages that they offer over conventional parenteral vaccines. The biggest advantage is the stimulation of mucosal immune responses (15,24). Lactobacillus is a genus of Gram-positive facultative anaerobic or microaerophilic rod-shaped bacteria. These beneficial bacteria normally live in the digestive, urinary, and genital systems without causing disease. Their abilities to transit through the stomach intact and to associate closely with the intestinal epithelium, combined with their immunomodulatory properties, have made Lactobacillus spp. attractive candidates as live vehicles for the delivery of immunogens to the intestinal mucosa (6, 12). It was recently shown that specific Lactobacillus species can induce inflammatory responses against infection, increase IgA production, activate mo...

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“…35 The parental vectors used for cloning were pSIP401 derivatives that had previously been modified for C-terminal cell wall anchoring of the heterologously expressed protein of interest. 23 The NY-ESO-1 sequence previously optimized for secretion 36 was codon-optimized for expression in L. plantarum WCFS1 and ordered from Genscript (Piscataway, NJ, USA) in a pUC57-vector (pUC57-NY-ESO). The NY-ESO-1 gene fragment was amplified using pUC57-NY-ESO as template with the primer pair NYESOF (TGCTTCATCAGTCGACCAAGCG-GAAGGCCGA) and NYESOcwa3R (CTGGTTGGCT ACGCGTTCGCCGTTGAC) (restriction sites in italics).…”

Section: Cloningmentioning

confidence: 99%

“…The pUC57-Gal-1 vector was digested with SalI and EcoRI and ligated into the SalI/EcoRI-digested pLp_3050sNucA, 40 yielding pLp_3050sGal-1. The Gal-1 anchoring construct was made by amplifying the 3050sGal-1 fragment (Gal-1 fused to the Lp_3050 signal peptide) from pLp_3050sGal-1 using the primer pair 3050F (GGAGTATGATTCATATGAAAAAATTTAACT-TTAAAACCATGTT) and Galcwa2R (GTTCAGTGACAC-GCGTATCAAACGCCACACACTTAATC) and subsequently in-fusion cloned into the NdeI/MluI digested pLp_0373sO-FAcwa2 plasmid, 23 yielding pLp_3050sGal-1cwa2. The cell wall anchor referred to as cwa2 is similar to cwa3, described above, but the anchor sequence consist of 194 residues.…”

Section: Cloningmentioning

confidence: 99%

“…We previously reported that the 37 kDa immunogenic oncofetal antigen (OFA) expressed on the cell surface of L. plantarum can induce antibody response. 23 Encouraged by this finding, in this study we investigated whether L. plantarum expressing NY-ESO-1 can induce specific immunity in mice. As controls for immunogenicity, we included 2 conserved proteins between human and mouse, HSP-27 and galectin-1 (Gal-1).…”

Section: Introductionmentioning

confidence: 99%

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RecombinantLactobacillus plantaruminduces immune responses to cancer testis antigen NY-ESO-1 and maturation of dendritic cells

Mobergslien

Vasovič

Mathiesen

et al. 2015

Human Vaccines & Immunotherapeutics

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Given their safe use in humans and inherent adjuvanticity, Lactic Acid Bacteria may offer several advantages over other mucosal delivery strategies for cancer vaccines. The objective of this study is to evaluate the immune responses in mice after oral immunization with Lactobacillus (L) plantarum WCFS1 expressing a cell-wall anchored tumor antigen NY-ESO-1. And to investigate the immunostimulatory potency of this new candidate vaccine on human dendritic cells (DCs). L. plantarum displaying NY-ESO-1 induced NY-ESO-1 specific antibodies and T-cell responses in mice. By contrast, L. plantarum displaying conserved proteins such as heat shock protein-27 and galectin-1, did not induce immunity, suggesting that immune tolerance to self-proteins cannot be broken by oral administration of L. plantarum. With respect to immunomodulation, immature DCs incubated with wild type or L. plantarum-NY-ESO-1 upregulated the expression of co-stimulatory molecules and secreted a large amount of interleukin (IL)-12, TNF-a, but not IL-4. Moreover, they upregulated the expression of immunosuppressive factors such as IL-10 and indoleamine 2,3-dioxygenase. Although L. plantarum-matured DCs expressed inhibitory molecules, they stimulated allogeneic T cells invitro. Collectively, the data indicate that L. plantarum-NY-ESO-1 can evoke antigen-specific immunity upon oral administration and induce DC maturation, raising the potential of its use in cancer immunotherapies.

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Cell Wall Anchoring of the 37-Kilodalton Oncofetal Antigen by Lactobacillus plantarum for Mucosal Cancer Vaccine Delivery

Cited by 55 publications

References 20 publications

The Major Autolysin Acm2 from Lactobacillus plantarum Undergoes Cytoplasmic O-Glycosylation

The Major Autolysin Acm2 from Lactobacillus plantarum Undergoes Cytoplasmic O-Glycosylation

Mucosal and Systemic Immune Responses Induced by Recombinant Lactobacillus spp. Expressing the Hemagglutinin of the Avian Influenza Virus H5N1

RecombinantLactobacillus plantaruminduces immune responses to cancer testis antigen NY-ESO-1 and maturation of dendritic cells

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