Abstract
We characterized two LysM domains of Limosilactobacillus fermentum, belonging to proteins Acglu (GenBank: KPH22907.1) and Pgb (GenBank: KPH22047.1) and bacterium like particles (BLP) derived from the immunomodulatory strain Lacticaseibacillus rhamnosus IBL027 (BLPs027) as an antigen display platform. The fluorescence protein Venus fused to the novel LysM domains could bind to the peptidoglycan shell of lactobacilli and resisted harsh conditions such as high NaCl and urea concentrations. Acglu with five LysM domains was a better anchor than Pgb baring only one domain. Six-week-old BALB/c mice were nasally immunized with the complex Venus-Acglu-BLPs027 at days 0, 14 and 28. The levels of specific serum IgG, IgG1 and IgG2a and the levels of total immunoglobulins (IgT) and IgA in broncho-alveolar lavage (BAL) were evaluated ten days after the last boosting. Venus-Acglu-BLPs027, nasally administered, significantly increased specific BAL IgT and IgA, and serum IgG levels. In addition, spleen cells of mice immunized with Venus-Acglu-BLPs027 secreted TNF-α, IFN-γ and IL-4 when stimulated ex vivo in a dose-dependent manner. We constructed a Gateway compatible destination vector to easily fuse the selected LysM domain to proteins of interest for antigen display to develop mucosal subunit vaccines.
Highlights
With the use of vaccines that employ genetically modified organisms as carriers
We reported that Lacticaseibacillus rhamnosus IBL027 (Basonym: Lactobacillus rhamnosus) has both intrinsic antiviral immunomodulatory and mucosal adjuvant p roperties[10]
We demonstrated that bacterium-like particles (BLPs) derived from the IBL027 strain (BLPs027) enhance local and systemic, humoral and cellular immune responses when co-administered with a mucosal viral vaccine[11]
Summary
With the use of vaccines that employ genetically modified organisms as carriers. We reported that Lacticaseibacillus rhamnosus IBL027 (Basonym: Lactobacillus rhamnosus) has both intrinsic antiviral immunomodulatory and mucosal adjuvant p roperties[10]. BLPs are obtained by treating LAB with hot acid, resulting in bacterial death, loss of DNA and cytoplasmic proteins, and exposure of the cell wall peptidoglycan[11], which can be used to deliver antigens[12] In this sense, there are some studies where particles derived from Lactococcus lactis (known as GEM, gram-positive enhancer matrix) were used as mucosal delivery vehicles for heterologous antigens of bacterial, viral or parasitic nature[13]. There are some studies where particles derived from Lactococcus lactis (known as GEM, gram-positive enhancer matrix) were used as mucosal delivery vehicles for heterologous antigens of bacterial, viral or parasitic nature[13] These vaccines induce robust and long-lasting adaptive immune responses thanks to an optimal activation of innate immune responses through the Toll-like receptor 2 (TLR2)[14]. The mucosal and systemic immune responses elicited by the experimental vaccine, i.e., BLPs exposing the antigen bound to their peptidoglycan, were studied in intranasally immunized mice
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