Recent Trends and Strategies for Targeting M – Cells via Oral Vaccine against Hepatitis B: A Review

Orally administered fungal vaccines show promise for the prevention of infectious diseases. Edible mushrooms are deemed appropriate hosts to produce oral vaccines due to their low production cost and low risk of gene contamination. However, their low expression level of antigens has limited the potential development of oral vaccines using mushrooms. The low expression level might result from impurity of the transgenic mycelia since dikaryotic mycelia are commonly used as transformation materials. In this study, stable transgenic hepatitis B virus surface antigen (HBsAg) in Flammulina velutipes transformants was obtained by Agrobacterium-mediated transformation, followed by fruiting and basidiospore mating. The formation of HBsAg was detected by western blot analysis. The expression levels of HBsAg in transgenic F. velutipes fruiting bodies were (129.3±15.1), (110.9±1.7) and (161.1±8.5) ng/g total soluble protein. However, the values may be underestimated due to incomplete protein extraction. Two of the four pigs in the experimental group produced positive anti-HBsAg-specific IgG after being fed the HBsAg transgenic F. velutipes fruiting bodies for 20 weeks, while no anti-HBsAg antibody was detected in the control group. One of the positive pigs had HBsAg titres of 5.36 and 14.9 mIU/mL in weeks 10 and 14, respectively, but expression faded thereafter. The other positive pig displayed HBsAg titres of 9.75, 17.86 and 39.87 mIU/mL in weeks 14, 18 and 20, respectively. The successful immunogenicity in pigs fed transgenic F. velutipes fruiting bodies demonstrated the potential of using the fungus as an oral vaccine.

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M-cell Targeting Acid-resistant Oral Vaccine Delivery for Immunization Against Hepatitis B Infection Using Cationic Solid Lipid Nanoparticles

10.21203/rs.3.rs-904521/v1 ◽

2021 ◽

Author(s):

Surendra Saraf ◽

Rudra Narayan Sahoo ◽

Shailesh Jain ◽

Subrata Mallick

Keyword(s):

Hepatitis B ◽

Solid Lipid Nanoparticles ◽

Oral Vaccine ◽

Vaccine Delivery ◽

Lipid Nanoparticles ◽

Hepatitis B Infection ◽

Cell Targeting ◽

M Cell ◽

Promising Alternative ◽

Solid Lipid

Abstract Background: Viral infection caused by Hepatitis B is transmitted by permucosal or parenteral exposure and also one of the prime causes of hepatocellular carcinoma and liver cirrhosis. Objectives: M-cell targeting acid-resistant oral vaccine delivery have been formulated for immunization against Hepatitis B infection. Methods: Cationic solid lipid nanoparticles (cSLNs) were prepared utilizing solvent injection technique. Hepatitis B surface antigen (HBsAg) loaded alginate coated cSLNs were anchored with lipopolysaccharide (LPS). SDS-PAGE was performed to evaluate acid degradation protection of prepared formulation. Induction of immunity produced by prepared nanoparticle for Hepatitis B was determined on female Balb/c mice followed by ELISA assays for assessing anti-HBsAg IgG/IgA antibodies in mucosal fluids. Results: Sustained release of HBsAg (60.66 %) has been exhibited from alginate coated cSLNs in comparison to cSLNs without alginate coating (97.72 %) after 48 h. The production of anti-HBs titer in intestinal, salivary and vaginal secretions was 3.41 IU/ml, 3.1 IU/ml and 2.51 IU/ml respectively in comparison to the control group. Integrity of the M-cells has been maintained after binding with SLN, and oral administration delivered the antigen to the desired site of gut. Conclusion: It was found effective in producing antibodies in mucosal immunization against Hepatitis B virus. So, this formulation could be used as a promising alternative preexisting vaccine to prevent Hepatitis B infection.

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Keynote review: Intestinal Peyer's patch M cells and oral vaccine targeting

Drug Discovery Today ◽

10.1016/s1359-6446(05)03536-1 ◽

2005 ◽

Vol 10 (17) ◽

pp. 1145-1157 ◽

Cited By ~ 137

Author(s):

David J. Brayden ◽

Mark A. Jepson ◽

Alan W. Baird

Keyword(s):

Oral Vaccine ◽

M Cells ◽

Peyer’S Patch ◽

Peyer's Patch

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Freeze-Drying of Plant Tissue Containing HBV Surface Antigen for the Oral Vaccine against Hepatitis B

BioMed Research International ◽

10.1155/2014/485689 ◽

2014 ◽

Vol 2014 ◽

pp. 1-10 ◽

Cited By ~ 20

Author(s):

Marcin Czyż ◽

Radosław Dembczyński ◽

Roman Marecik ◽

Justyna Wojas-Turek ◽

Magdalena Milczarek ◽

...

Keyword(s):

Hepatitis B ◽

Surface Antigen ◽

Freeze Drying ◽

Elevated Temperatures ◽

Oral Vaccine ◽

Leaf Tissue ◽

Residual Moisture ◽

Small Surface ◽

Process Step ◽

Freeze Dried

The aim of this study was to develop a freeze-drying protocol facilitating successful processing of plant material containing the small surface antigen of hepatitis B virus (S-HBsAg) while preserving its VLP structure and immunogenicity. Freeze-drying of the antigen in lettuce leaf tissue, without any isolation or purification step, was investigated. Each process step was consecutively evaluated and the best parameters were applied. Several drying profiles and excipients were tested. The profile of 20°C for 20 h for primary and 22°C for 2 h for secondary drying as well as sucrose expressed efficient stabilisation of S-HBsAg during freeze-drying. Freezing rate and postprocess residual moisture were also analysed as important factors affecting S-HBsAg preservation. The process was reproducible and provided a product with VLP content up to 200 µg/g DW. Assays for VLPs and total antigen together with animal immunisation trials confirmed preservation of antigenicity and immunogenicity of S-HBsAg in freeze-dried powder. Long-term stability tests revealed that the stored freeze-dried product was stable at 4°C for one year, but degraded at elevated temperatures. As a result, a basis for an efficient freeze-drying process has been established and a suitable semiproduct for oral plant-derived vaccine against HBV was obtained.

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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.

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Recent trends in acute hepatitis B in Japan

Kanzo ◽

10.2957/kanzo.47.419 ◽

2006 ◽

Vol 47 (9) ◽

pp. 419-424 ◽

Cited By ~ 2

Author(s):

Fuminaka Sugauchi ◽

Masashi Mizokami

Keyword(s):

Hepatitis B ◽

Acute Hepatitis ◽

Acute Hepatitis B ◽

Recent Trends

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Structural and functional characterization of a maize‐derived hepatitis B surface antigen (HBsAg) for the development of an oral vaccine

The FASEB Journal ◽

10.1096/fasebj.29.1_supplement.571.22 ◽

2015 ◽

Vol 29 (S1) ◽

Author(s):

Shweta Shah ◽

Celine Hayden ◽

John Howard ◽

A Rao

Keyword(s):

Hepatitis B ◽

Surface Antigen ◽

Hepatitis B Surface Antigen ◽

Oral Vaccine ◽

Functional Characterization

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Cloning, expression, purification and interaction evaluation of 30 amino acids in C terminus of Clostridium perfringens toxin with its receptor Claudin-4

Science and Technology Development Journal - Natural Sciences ◽

10.32508/stdjns.v2i4.809 ◽

2019 ◽

Vol 2 (4) ◽

pp. 47-55

Author(s):

Quang Kien Huynh ◽

An Hoang Nguyen ◽

Quynh Thi Mong Pham ◽

Hoan Phuoc Khai Nguyen ◽

Hieu Van Tran

Keyword(s):

Amino Acids ◽

Fusion Protein ◽

Clostridium Perfringens ◽

Field Effect Transistors ◽

Oral Vaccine ◽

Soluble Form ◽

M Cells ◽

Gastrointestinal Infections ◽

E Coli ◽

C Terminus

Oral vaccine is a strategy being the most interested about treatments of gastrointestinal infections because of many great benefits outweigh conventional injection vaccines. In order to resolve the dispersion of antigens in gastrointestinal surfaces, the immunological tolerance and also be capable to stimulate immune responses effectively, M cells are targeted for antigens delivery. A number of researches reported that 30 amino acids in C terminus of Clostridium perfringens toxin (CPE30) have a high affinity to Claudin-4 receptor presenting on M cells. It is highly indispensable to produce a resource for assessing of CPE30 binding ability so cpe30 gene was cloned into the pET-gfp plasmid by two restriction enzymes BamHI and NdeI on the E. coli DH5α strain. The expression and confirmation of the fusion protein CPE30-GFP which was induced by IPTG in E. coli BL21 (DE3) strain and assessed by SDS-PAGE and Western blot with 6xHis Taq antibody demonstrated that there was the over expression of CPE30 GFP fusion protein in the cytoplasm, mainly in the soluble form. Finally, CPE30-GFP was purified which the purity was approximately 92.3%. In vitro protein interaction measurement using silicon nanowire field-effect transistors (SiNW FETs) showed that CPE30-GFP had a good binding affinity with its receptor Claudin-4 (R4). This result laid the groundwork for the CPE30 interaction study with the M cell in vivo.

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Immunogenicity of Oral Vaccine Candidate in Recombinant Lactococcus lactis Expressing HBcAg and IFNα-2b for Hepatitis B Prevention

10.21203/rs.3.rs-198617/v1 ◽

2021 ◽

Author(s):

Apon Zaenal Mustopa ◽

Lita Meilina ◽

Sri Budiarti ◽

Huda Shalahudin Darusman ◽

Lita Triratna ◽

...

Keyword(s):

Immune Response ◽

Hepatitis B ◽

Vaccine Candidate ◽

Oral Vaccine ◽

Adaptive Immune System ◽

Control Group ◽

Interferon Α ◽

Single Treatment ◽

In Vivo Test

Abstract Background: Hepatitis B is a liver inflammation caused by virus infection leading to acute and chronic conditions. Antigenic compound of HBcAg could induce specific T and B cell that generating high immunity response rather than HBsAg. Therapeutic agent for hepatitis and cancer treatment which has been approved by USFDA is Interferon α-2b (IFNα-2b). This type of class I cytokine plays a role in inhibiting viral replication and modulating adaptive immune system. Objectives: In this study, we analyzed immunogenicity of recombinant HBcAg and IFNα-2b expressed in L. lactis NZ3900. Methods: In vivo test was carried out using female Balb/c mice. Antibody responses for oral immunization were quantified as total IgG using ELISA on days 21, 35 and 51. Safety compound of this oral vaccine candidate was also described by liver and spleen condition after immunization. The differential leukocyte counting was performed to confirm the inflammatory process. Results: Post immunization with L. lactis recombinant strains could induce optimum total humoral immune responses on day 35, with IgG concentration at 4.96±1.03 mg/mL. Single treatment with HBcAg was more potent in inducing immune response (IgG) rather than HBcAg-IFNa-2b combination. Immunization for 51 days could not alleviate animal body weight in each group. The maintaining IgG production until 51 days was just because lymphocytes activities persisted above 70%. The lymphocytes number which achieved 76.3% (P2) and 78.3% (P3) compared to Control group with 62%. Conclusion: Single treatment with recombinant HBcAg expressed in L. lactis NZ3900 was better inducing IgG production and maintaining for 51 days. This result suggested, L. lactis recombinant strain can be as potential vaccine candidate to induce immune response protecting from hepatitis B virus. Moreover, no organ damage was found in liver and spleen of Balb/c mice.

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