Immunogenicity of Endolysin PlyC

Abstract

Endolysins are bacteriolytic enzymes derived from bacteriophages. They represent an alternative to antibiotics, since they are not susceptible to conventional antimicrobial resistance mechanisms. Since non-human proteins are efficient inducers of specific immune responses, including the IgG response or the development of an allergic response mediated by IgE, we evaluated the general immunogenicity of the highly active antibacterial enzyme, PlyC, in a human population and in a mouse model. The study includes the identification of molecular epitopes of PlyC. The overall assessment of potential hypersensitivity to this protein and PlyC-specific IgE testing was also conducted in mice. PlyC induced efficient IgG production in mice, and the molecular analysis revealed that PlyC-specific IgG interacted with four immunogenic regions identified within the PlyCA subunit. In humans, approximately 10% of the population demonstrated IgG reactivity to the PlyCB subunit only, which is attributed to cross-reactions since this was a naïve serum. Of note, in spite of being immunogenic, PlyC induced a normal immune response, without hypersensitivity, since both the animals challenged with PlyC and in the human population PlyC-specific IgE was not detected.

Experimental Identification of Cross-Reacting IgG Hotspots to Predict Existing Immunity Evasion of SARS-CoV-2 Variants by a New Biotechnological Application of Phage Display

“…These oligonucleotides were used to create a phage display library using the T7Select 415-1 Cloning Kit (Merck Millipore, Burlington, USA). Immunoprecipitation of the library was performed in accordance with a previously published protocol [ 7 , 11 ]. Briefly, the phage library was amplified in a standard culture as described in the manufacturer’s manual and then purified by hollow fiber dialysis against a Phage Extraction Buffer (20 mM Tris-HCl, 100 mM NaCl, 6 mM MgSO 4 , pH 8.0).…”

Bacteriophage vB_SepP_134 and Endolysin LysSte_134_1 as Potential Staphylococcus-Biofilm-Removing Biological Agents

“…Phage enzymes capable of destroying bacterial cell walls have advantages over bacteriophages [ 21 , 22 , 23 ]. These lytic enzymes have a wider host spectrum than their parental phages and bacteria do not acquire resistance to them; in addition, these lytic proteins demonstrate clear bioavailability and pharmacokinetics [ 24 ]. Importantly, standardized biotechnological methods can be used for their production.…”

“…Their CBD can be C-terminal or it can be located between two EADs. The potential efficacy of the endolysins of Staphylococcus phages has been demonstrated [ 24 , 28 , 29 , 30 , 31 , 32 ].…”

Bacteriophage St_134 and its Endolysin as a Potential Staphy-lococcus Biofilm-Removing Biological Agent

“…Phage enzymes capable of destroying bacterial surfaces have advantages over bacteriophages [14][15][16][17]. The phage lysins have a wider host spectrum and low bacterial resistance; they demonstrate clear bioavailability and pharmacokinetics [18]. Importantly, standardized biotechnological methods can be used for their production.…”

“…Notably, the pentaglycine interpeptide bridge is a unique component of most Staphylococcus strains that can be digested by the specific peptidoglycan hydrolases [25]. Last years, efficacy of endolysins of phages of Gram-positive bacteria as antibacterial drugs has been demonstrated [18,[26][27][28][29][30].…”