Table 1 Plasmid-based surface display of functional elements in Lactobacilli
From: Metabolic engineering of Lactobacilli spp. for disease treatment
Strains | Strategy | Expression mode | Mechanisms | Diseases | Treating effect | Experimental model | Reference |
|---|---|---|---|---|---|---|---|
Displaying vaccines for treating virus infection | |||||||
L. plantarum CGMCC 1.557 | Surface displaying the codon-optimized SARS-CoV-2Â S protein | Plasmid-based | Antigen presentation for SARS-CoV-2 | Against SARS-CoV-2 | Â | Â | [11] |
L. acidophilus NCFM | Surface display of HIV-1 Gag and Salmonella enterica Serovar Typhimurium FliC as adjuvant | Plasmid-based | Inducing the antigen-specific IgA production and stimulating the IFN-γ-producing cells | Against HIV |  | Female BALB/c mice | [12] |
L. casei BLS | Surface displaying HPV type 16 E7 protein (HPV16 E7) with the poly-γ-glutamic acid synthetase complex A (PgsA) of Bacillus subtilis (chungkookjang) served as anchoring motif | Plasmid-based | Inducing the E7-specific serum IgG and mucosal IgA productions | Against HPV16 E7-based cervical cancer | For the C57BL/6 mice that immunized with HPV16 E7-displaying strain, the mean log titer of the serum IgG was increased from 1.24 ± 0.24 to 3.15 ± 0.02 after the first oral vaccination; The E7-specific lymphocyte proliferative response was increased from 7.8 ± 0.9 to 11.0 ± 1.4; The E7-specific cytotoxic T lymphocyte (CTL) response was increased from 21 ± 5 to 510 ± 36 spot-forming cells (SFC)/106 cells. For the TC-1 mouse tumor model, the survival rate of the recombinant HPV16 E7-displaying strain-immunized group was increased from 0–50% | Female C57BL/6 mice and the mice challenged with TC-1 cells | [13] |
L. casei strain 525 | Surface displaying HPV type 16 E7 protein (HPV16 E7) | Plasmid-based | Inducing E7-specific mucosal immunity | Against HPV16 E7-based cervical cancer | Â | Female SPF C57BL/6 (H-2b) mice | [14] |
L. casei strain 525 | Surface displaying HPV type 16 E7 protein (HPV16 E7) | Plasmid-based | Inducing E7-specific mucosal immunity | Against HPV16 E7-based cervical cancer | 70% of the CIN3 patients experienced a pathological down-grade to CIN2 at week 9 | Cervical intraepithelial neoplasia grade 3 (CIN3) patients | [15] |
L. plantarum CGMCC 1.557 | Surface displaying the truncated and codon-optimized viral glycoprotein 5 (GP5) of PRRSV | Plasmid-based | Antigen presentation for PRRSV | Against PRRSV | Â | Â | [16] |
L. plantarum NC8 | Surface displaying the spike antigen of TGEV | Plasmid-based | Inducing cellular, mucosal, and humoral immunity | Against porcine TGEV | Inducing high expression levels of B7 molecules on DCs, as well as high levels of IgG, secretory IgA, and IFN-γ and IL-4 cytokines compared with the control group | SPF mice | [17] |
L. casei ATCC 393 | Surface displaying the core neutralizing epitope (COE) antigen of PEDV conjugated with M cell targeting peptide Co1 (adjuvant) | Plasmid-based | Inducing higher anti-PEDV serum IgG and mucosal SIgA antibody responses | Against PEDV | The mice that orally immunized the recombinant strain could induce the serum IgG antibody response to exhibit stronger PEDV-neutralizing activity (1:24) than and control groups (< 1:2). Moreover, this recombinant strain-induced SIgA antibody response elicited stronger anti-PEDV neutralizing activity (1:20) than the control group (< 1:2) | Female SPF BALB/c mice | [18] |
L. casei ATCC 393 | Surface displaying the core neutralizing epitope (COE) antigen of PEDV conjugated with the M cell-targeting peptide (Col) and dendritic cell-targeting peptide (DCpep) | Plasmid-based | Inducing the anti-PEDV mucosal, humoral, and cellular immune responses | Against PEDV | Providing stronger PEDV-neutralizing ability (1:36) than the control group (< 1:2) | Female SPF BALB/c mice | [19] |
L. casei ATCC 393 | Surface displaying the D antigenic site of the TGEV spike (S) protein and core neutralizing epitope of PEDV S protein | Plasmid-based | Increasing the levels of anti-PEDV and anti-TGEV serum immunoglobulin G (IgG) and mucosal secreted immunoglobulin A (SIgA) antibodies; strengthening the proliferation levels of lymphocytes | Against TGEV and PEDV | Â | BALB/c mice | [20] |
L. plantarum HA33-1 | Surface displaying CSFV E2 protein in conjunction with thymosin α-1 | Plasmid-based | Inducing protective immune responses by eliciting the IgA-based mucosal, IgG-based humoral, and CTL-based cellular immune responses | Against CSFV |  | CSFV infected pigs | [21] |
L. plantarum ZN3 | Surface displaying the H1N1 HA1 protein that fused to DCpep and the M cell-targeting peptide | Plasmid-based | Inducing mucosal, cellular and systemic immune responses | Against swIAV | For oral administration, the survival rate of H1N1 virus-challenged mice was increased from 0–60%; For intranasal administration, the survival rate of H1N1 virus-challenged mice was increased from 0–100% | BALB/c mice inoculated intranasally with H1N1 and H3N2 | [22] |
L. plantarum NC8 | Surface displaying viral 3M2e-HA2 | Plasmid-based | Increasing the mucosal and systemic immune responses | Against AIV | The survival rate of the H9N2-challenged mice that immunized with the recombinant strain were increased from 0–80% | BALB/c mice challenged with mouse-adapted H9N2 AIV or H1N1 influenza virus | [23] |
L. casei L525 | Surface displaying the hemagglutinin 1 (HA1) subunit of the A/Aquatic bird/Korea/W81/2005 (H5N2) that fused with the Bacillus subtilis poly γ-glutamic acid synthetase A (pgsA) | Plasmid-based | Increasing the HA1-specific serum IgG, mucosal IgA and neutralizing antibodies | Against AIV | For the oral and intranasal administration, the survival rate of H5N2 virus-challenged mice was increased from 0–100% | Mice challenged with homologous mouse-adapted H5N2 virus | [24] |
L. plantarum | Surface displaying the VP2 protein of IBDV | Plasmid-based | Inducing humoral and cellular immune responses | Against vvIBDV | The survival rate of the vvIBDV-challenged chickens were increased from 0–100% | Chickens challenged with vvIBDV | [25] |
L. plantarum NC8 | Surface displaying the Gp85 protein of ALV-J | Plasmid-based | Inducing the cellular, humoral, and mucosal immunity responses | Against avian leukosis | The survival rate of the ALV-J-challenged chickens were increased significantly | Chickens that intramuscular injected with ALV-J HB2010001 | [26] |
L. plantarum HA33-1 | Surface displaying the glycoprotein (G) of SVCV and ORF81 protein of KHV | Plasmid-based | Increasing the levels of immunoglobulin M (IgM) | Against SVCV and KHV | Providing effective protection to the vaccinated carps (71% protection) and koi (53% protection) at day 65 post challenge | Cyprinus carpio that oral administrated with SVCV; koi that oral administrated with KHV | [27] |
Displaying vaccines for treating parasites infection | |||||||
L. plantarum NC8 | Surface displaying EtMic2 | Plasmid-based | Antigen presentation for Eimeria tenella | Against chicken coccidiosis | The lesion scores of cecum was decreased from 3.75 ± 0.520 to 2.30 ± 0.506; The OPG (×106) was decreased from 1.44 ± 0.02 to 0.71 ± 0.04; The ACI was increased from 74.93 to 145.15 | Chickens challenged with E. tenella sporulated oocysts | [28] |
L. plantarum NC8 | Surface displaying EtMic2 and AMA1 | Plasmid-based | Antigen presentation for Eimeria tenella | Against chicken coccidiosis | The BWG of E. tenella-challenged chicken was increased from 210.50 ± 16.16 g to 313.71 ± 6.60 g; The lesion scores in cecum were decreased from 3.83 ± 0.41 to 2.00 ± 0.63; The oocyst output (×105) was decreased from 9.50 ± 3.03 to 3.56 ± 1.30 | Chickens challenged with E. tenella sporulated oocysts | [29] |
L. Plantarum NC8 | Surface displaying SO7 that fused to DCpep | Plasmid-based | Dendritic cell-targeting antigen presentation for Eimeria tenella | Against chicken coccidiosis | The body weight gains and serum antibody responses were increased in the E. tenella-challenged chicken, while the fecal oocyst shedding and pathological damage in cecum were decreased | Chickens challenged with E. tenella sporulated oocysts | [30] |
L. plantarum NC8 | Surface displaying eukaryotic initiation factor U6L5H2 | Plasmid-based | Producing higher levels of specific cecal SIgA, serum IgG, transcription of cytokines IFN-γ and IL-2, and lymphocyte proliferation | Against chicken coccidiosis | The body weight gain of E. tenella-challenged chicken was increased from 83.32 ± 3.28 g to 101.57 ± 2.02 g; The average lesion score was decreased from 2.90 ± 0.42 to 1.79 ± 0.31; The oocyst output (×105) was decreased from 5.37 ± 0.43 to 1.35 ± 0.18; The ACI was increased from 109.90 to 168.28 | Chickens challenged with E. tenella sporulated oocysts | [31] |
L. plantarumNC8 | Surface displaying gp43 and nudix hydrolase (TsNd) of Trichinella spiralis | Plasmid-based | Inducing higher levels of specific humoral, mucosal, and cellular immune responses | Against trichinellosis | A 75.67% reduction of adult worms (AW) at 7 days post-infection (dpi) and 57.14% reduction of muscle larva (ML) at 42 dpi were observed in the larval-challenged mice | BALB/c mice challenged with infectious T. spiralis | [32] |
Displaying vaccines for treating pathogens infection | |||||||
L. plantarum WCFS1 | Surface displaying Ag85B and ESAT-6 (AgE6) | Plasmid-based | Inducing specific immune responses | Against tuberculosis (TB) | Inducing antigen-specific proliferative responses in lymphocytes purified from TB-positive donors; Inducing immune responses in mice after nasal or oral immunization | C57BL/6 BomTac mice | [33] |
L. casei ATCC 393 | Surface displaying the toxoid of C. perfringens α-toxin | Plasmid-based | Eliciting mucosal, humoral, and cellular immunity to neutralize the natural α-toxin of C. perfringens | Against C. perfringens infection | Improve the survival rates of C. perfringens-challenged mice from 0–90% | SPF BALB/c mice challenged with C. perfringens natural α-toxin and C. perfringens type A | [34] |
L. casei ATCC 393 | Surface displaying the NetB toxin of C. perfringens | Plasmid-based | Inducing high anti-toxin antibody response | Against C. perfringens infection | Â | Chickens that orally inoculated with virulent C. perfringens | [35] |
L. casei ATCC 393 | Surface displaying the C-terminal domain of α-toxin of C. perfringens | Plasmid-based | Inducing specific serum anti-α antibodies | Against C. perfringens infection | The mean body weight changes of the recombinant strain-immunized chickens (35.61%) were higher than that of the non-vaccinated chickens (24.13%) | Ross 308 broiler chickens challenged with C. perfringens CP58 | [36] |
L. crispatus N-11 | Surface displaying the α-β2-ε-β1 toxoid of C. perfringens | Plasmid-based | Stimulating the mucosal, cellular, and humoral immunity | Against the toxins of C. perfringens | The specific secretory IgA (SIgA) and IgY antibodies in the serum and intestinal mucus and the serum concentration of IFN-γ, lL-2, IL-4, IL-10, IL-12, and IL-17 were increased significantly in the recombinant strain-immunized group | Chickens challenged with the natural α-β2-ε-β1 toxin combined with C. perfringens type A and type B pathogenic bacteria | [37] |
L. gasseri NM713 | Surface displaying the conserved region of streptococcal M6 protein (CRR6) | Plasmid-based | Inducing specific systemic (IgG) and mucosal (IgA) immune responses against the streptococcal M6 antigen | Against the S. pyogenes infection | The mice that orally administered with the recombinant strain showed lower streptococcal infection (10%) and mortality (3.3%) rate as compared to the control group | Seven-weeks old mice that challenged with S. pyogenes | [38] |
L. casei CC16 | Surface displaying the Aha1 of A. veronii fused the cholera toxin B subunit (CTB) as adjuvant | Plasmid-based | Stimulating the humoral and cellular immunity | Against the A. veronii infection | The survival rate of A. veronii-challenged carp was increased from 0–64.29% | Cyprinus carpio that intraperitoneally injected with A. veronii | [39] |
L. casei CC16 | Surface displaying the Aha1 of A. veronii fused the E. coli intolerant enterotoxin B subunit (LTB) | Plasmid-based | Inducing the expression of various immune enzymes in the humoral immunity of carp and increasing the cytokine levels | Against the A. veronii infection | The survival rate of A. veronii-challenged carp was increased from 0–60.71% | Common carp that intraperitoneally injected with A. veronii TH0426 | [40] |
L. casei CC16 | Surface displaying the SH type VI pili B (MshB) from A. veronii as an antigen and cholera toxin B subunit (CTB) as a molecular adjuvant | Plasmid-based | Stimulating the production of high levels of serum-specific immunoglobulin M (IgM) and enhancing the non-specific immunity | Against the A. veronii infection | The survival rate of A. veronii-challenged carp was increased from 0–60% | Crucian carp that intraperitoneally injected with A. veronii | [41] |
L. casei ATCC 393 | Surface displaying the outer membrane protein K (OmpK) of V. mimicus as an antigen, and cholera toxin B subunit (CTB) as a molecular adjuvant | Plasmid-based | Inducing the humoral and cellular immunity | Against the V. mimicus infection | The survival rate of the recombinant strain-immunized Carassius auratus was higher than the control group | C. auratus challenged with V. mimicus | [42] |
L. plantarum NC8 | Surface displaying the FomA protein of F. nucleatum | Plasmid-based | Increasing the mouse-specific humoral immunity and eliciting the mucosal and T cell-mediated immune responses | Against IBD | Decreasing the mortality rate and body weight loss | F. nucleatum-and DSS-induced IBD mice | [43] |
L. reuteri WXD171 | Surface displaying the iron-regulated surface determinant protein B (IsdB) of S. aureus | Plasmid-based | Inducing the mucosal responses in gut-associated lymphoid tissues | Against the S. aureus infection | Improving the survival rate of S. aureus-challenged mice from 10–70% | Mouse model of S. aureus-induced pulmonary, skin, and systemic infection. | [44] |
L. casei ATCC 393 | Surface displaying the outer membrane protein OMP19 of Brucella species | Plasmid-based | Providing a very good general and mucosal immune responses | Against brucellosis | The mice that orally immunized with OMP19-displaying strain showed higher degrees of protection (15-fold reduction of B. abortus 544 in spleen) as compared to the control group | BALB/c mice challenged intraperitoneally with the virulent B. abortus 544 | [45] |
Displaying functional elements for the intestinal exclusion of viruses and pathogens | |||||||
L. rhamnosus GG | Surface displaying IgG-binding domain of protein G | Plasmid-based | Capture rotavirus via hyperimmune bovine colostrum antibodies (HBC-IgG) | Against rotavirus (RRV) | The combination usage of HBC antibodies and this engineered strain was more effective (10 to 100-fold increase) in reducing the prevalence, severity, and duration of diarrhea | Mouse RRV infection model | [46] |
L. paracasei BL23 | Surface displaying rotavirus proteins 1 and 3 (ARP1 and ARP3) | Plasmid-based | Capture rotavirus by anti-rotavirus proteins | Against rotavirus | Â | Â | [47] |
L. casei ATCC 334 | Surface displaying the Listeria adhesion protein (LAP) from a non-pathogenic Listeria (L. innocua) and a pathogenic Listeria (L. monocytogenes) | Plasmid-based | Excluding L. monocytogenes competitively by occupying the surface presented LAP receptor, heat shock protein 60 | Against the L. monocytogenes infection | The number L. monocytogenes cells that adhered to the intestine were 100-fold lower in the mice that treated with the recombinant strain; At 10 days post the L. monocytogenes challenge, the surviving rate of the recombinant strain-treated mice (~ 92%) was higher than the control group (60%) | Female mice (A/J: 6–8 weeks of age) challenged with Listeria monocytogenes F4244 | [48] |
L. casei ATCC 344 | Surface displaying internalins A and B (inlAB) of L. monocytogenes | Plasmid-based | Inhibiting the adhesion, invasion and transcellular passage of L. monocytogenes | Against L. monocytogenes infection | Reducing the adhesion of L. monocytogenes by 50-53.6% at 16 and 24Â h, far more than that of the control group (8%) | Caco-2 cells | [49] |
Displaying pharmaceutical compounds and enzyme | |||||||
L. plantarum NC8 | Surface displaying murine IL-10 | Plasmid-based | Anti-inflammation | Against Th1 Responses of RAW264.7 Cells Stimulated with Poly(I: C) or LPS | Reducing the Poly(I: C)- or LPS-induced Th1 responses in RAW264.7 cells and decreasing the expression of TNF-α, IFN-γ, IL-1β, and IL-6 | RAW264.7 cells stimulated with Poly(I: C) or LPS | [50] |
L. plantarum NC8 | Surface displaying the porcine IFN-λ3 | Plasmid-based | Inhibiting the replication of PEDV and TGEV | Against TGEV and PEDV | Reducing the prevalence of PEDV and TGEV viruses by 53% and 59%, respectively | Intestinal porcine epithelial cell line J2 (IPEC-J2) that inoculated with PEDV strain CV777 or TGEV strain SY | [51] |
L. reuteri CGMCC1.3264 | Surface displaying lactonohydrolase | Plasmid-based | Degrading zearalenone | Against fungal mycotoxins zearalenone | This engineered strain was capable of hydrolyzing 2.5Â mg/kg of ZEN-contaminated corn within 4Â h | Â | [52] |