Immunogenicity of Recombinant-Deficient Lactobacillus casei with Complementary Plasmid Expressing Alanine Racemase Gene and Core Neutralizing Epitope Antigen against Porcine Epidemic Diarrhea Virus

Li, Fengsai; Wang, Xiaona; Fan, Xiaolong; Sui, Ling; Zhang, Hailin; Li, Yue; Zhou, Han; Wang, Li; Qiao, Xin; Tang, Lijie; Li, Yijing

doi:10.3390/vaccines9101084

Cited by 6 publications

(4 citation statements)

References 49 publications

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“…Li et al applied alanine racemase-deficient L. casei W56 as the host strain and a complementary plasmid as a vector to express the neutralizing antigen COE of PEDV. After the oral immunization of mice, both SIgA and IgG levels increased significantly (Li et al 2021 ). In this study, the protective antigen S1 gene of PEDV was inserted into the genome of the strain △ Alr HLJ-27 to construct the mutant strain S1/ △ Alr HLJ-27 .…”

Section: Discussionmentioning

confidence: 99%

Assessing immunogenicity of CRISPR-NCas9 engineered strain against porcine epidemic diarrhea virus

Li,

Zhao,

Sui

et al. 2024

Appl Microbiol Biotechnol

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Porcine epidemic diarrhea (PED) caused by porcine epidemic diarrhea virus (PEDV), is an acute and highly infectious disease, resulting in substantial economic losses in the pig industry. Given that PEDV primarily infects the mucosal surfaces of the intestinal tract, it is crucial to improve the mucosal immunity to prevent viral invasion. Lactic acid bacteria (LAB) oral vaccines offer unique advantages and potential applications in combatting mucosal infectious diseases, making them an ideal approach for controlling PED outbreaks. However, traditional LAB oral vaccines use plasmids for exogenous protein expression and antibiotic genes as selection markers. Antibiotic genes can be diffused through transposition, transfer, or homologous recombination, resulting in the generation of drug-resistant strains. To overcome these issues, genome-editing technology has been developed to achieve gene expression in LAB genomes. In this study, we used the CRISPR-NCas9 system to integrate the PEDV S1 gene into the genome of alanine racemase-deficient Lactobacillus paracasei △Alr HLJ-27 (L. paracasei △Alr HLJ-27) at the thymidylate synthase (thyA) site, generating a strain, S1/△Alr HLJ-27. We conducted immunization assays in mice and piglets to evaluate the level of immune response and evaluated its protective effect against PEDV through challenge tests in piglets. Oral administration of the strain S1/△Alr HLJ-27 in mice and piglets elicited mucosal, humoral, and cellular immune responses. The strain also exhibited a certain level of resistance against PEDV infection in piglets. These results demonstrate the potential of S1/△Alr HLJ-27 as an oral vaccine candidate for PEDV control. Key points • A strain S1/△Alr HLJ-27 was constructed as the candidate for an oral vaccine. • Immunogenicity response and challenge test was carried out to analyze the ability of the strain. • The strain S1/△Alr HLJ-27 could provide protection for piglets to a certain extent.

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Section: Discussionmentioning

confidence: 99%

Assessing immunogenicity of CRISPR-NCas9 engineered strain against porcine epidemic diarrhea virus

Li,

Zhao,

Sui

et al. 2024

Appl Microbiol Biotechnol

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“…Studies have confirmed that utilizing LAB to develop an oral vaccine to deliver foreign antigens can help induce the production of IgA antibodies in animals. Li et al used auxotrophic L. casei and its complementary plasmid to express the non-toxic antigen of Porcine Epidemic Diarrhea Virus (PEDV), and the results showed that it could stimulate the production of sIgA and IgG in mice [ 35 ]. Therefore, we endeavored to test the immunogenicity of pPG-E-α-β2-ε-β1/ L. crispatus N-11 in chickens, immunized orally using ELISA and immunohistochemistry.…”

Section: Discussionmentioning

confidence: 99%

Oral Immunization of Chickens with Probiotic Lactobacillus crispatus Constitutively Expressing the α-β2-ε-β1 Toxoids to Induce Protective Immunity

Khan

Huang

et al. 2022

Vaccines

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Clostridium perfringens (C. perfringens) is a bacterium that commonly causes zoonotic disease. The pathogenicity of C. perfringens is a result of the combined action of α, β, and ε exotoxins. In this study, Lactobacillus crispatus (pPG-T7g10/L. crispatus) expressing the main toxoids of C. perfringens, α, ε, β1, and β2, with EGFP-labeling, was constructed, and the protective effect was estimated in chickens. The α-β2-ε-β1 toxoid was constitutively expressed for confirmation by laser confocal microscopy and western blotting, and its immunogenicity was analyzed by enzyme-linked immunosorbent assay (ELISA) and immunohistochemical assays. After booster immunization, the probiotic vaccine group showed significantly higher levels (p < 0.05) of specific secretory IgA (sIgA) and IgY antibodies in the serum and intestinal mucus. Furthermore, the levels of cytokines, including interferon (IFN)-γ, interleukin (lL)-2, IL-4, IL-10, IL-12, and IL-17, and the proliferation of spleen lymphocytes in chickens orally immunized with pPG-E-α-β2-ε-β1/L. crispatus increased significantly. Histopathological observations showed that the intestinal pathological changes in chickens immunized with pPG-E-α-β2ε-β1/L. crispatus were significantly alleviated. These data reveal that the probiotic vaccine could stimulate mucosal, cellular, and humoral immunity and provide an active defense against the toxins of C. perfringens, suggesting a promising candidate for oral vaccines against C. perfringens.

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“…Since the first report on the use of lactic acid bacteria as a carrier for vaccine antigens, many studies have been published ( 12 ). Attempts have been made to use recombinant LAB strains in immunoprophylaxis against gastrointestinal pathogens ( Salmonella , Helicobacter , Yersinia ), bacteria that cause respiratory infections ( Streptococcus pneumoniae ) ( 5 ), as well as against numerous viruses such as SARS-CoV-2 ( 13 ), Human Papilloma Virus ( 14 ), rotavirus ( 15 ), Porcine Epidemic Diarrhea Virus ( 16 , 17 ), and subgroup J Avian Leukosis Virus ( 18 ). Various strategies have been developed to efficiently express genes encoding heterologous antigens in LAB strains, mainly of the genera Lactococcus and Lactobacillus .…”

Section: Introductionmentioning

confidence: 99%

Genomic and transcriptomic analysis of Ligilactobacillus salivarius IBB3154—in search of new promoters for vaccine construction

Kobierecka,

Wyszyńska,

Aleksandrzak-Piekarczyk

et al. 2023

Microbiol Spectr

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Transcriptomic analysis of the genome sequenced Ligilactobacillus salivarius strain IBB3154 grown at two different temperatures (37°C vs 42°C) identified differentially expressed genes involved in metabolic pathways, osmoregulation, and surface protein expression. Two highly expressed genes, sasA1 and sasA2 , which encode cell wall-anchored proteins belonging to the serine-rich repeat protein group, were found to be temperature-inducible. Moonlighting proteins with various functions, such as glyceraldehyde 3-phosphate dehydrogenase, fructose-bisphosphate aldolase, elongation factor Tu, and enolase, were highly expressed at both temperatures. The efficiency of promoters has been confirmed by the β-glucuronidase activity test; however, temperature dependence was not detected. We also found that the P sasA1 promoter retained its activity in the presence of bile salts. Knowledge of promoters that are highly active in L. salivarius cells can be used to produce strains that are carriers of immunogenic proteins. IMPORTANCE The genome of the strain Ligilactobacillus salivarius IBB3154 was sequenced, and transcriptome analysis was carried out at two different temperatures, allowing the determination of gene expression levels in response to environmental changes (temperature). Genes with higher expression at 42°C were identified. The use of a reporter gene (β- glucuronidase) did not confirm the transcriptomic results; it was found that the promoters of the genes sasA1 and sasA2 were active in the presence of bile salts. This opens up new opportunities for the overexpression of genes of other bacterial species in Ligilactobacillus cells in the intestinal environment.

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Immunogenicity of Recombinant-Deficient Lactobacillus casei with Complementary Plasmid Expressing Alanine Racemase Gene and Core Neutralizing Epitope Antigen against Porcine Epidemic Diarrhea Virus

Cited by 6 publications

References 49 publications

Assessing immunogenicity of CRISPR-NCas9 engineered strain against porcine epidemic diarrhea virus

Assessing immunogenicity of CRISPR-NCas9 engineered strain against porcine epidemic diarrhea virus

Oral Immunization of Chickens with Probiotic Lactobacillus crispatus Constitutively Expressing the α-β2-ε-β1 Toxoids to Induce Protective Immunity

Genomic and transcriptomic analysis of Ligilactobacillus salivarius IBB3154—in search of new promoters for vaccine construction

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