Characterization of the cattle serum antibody responses against TEM β-lactamase and the nonimmunogenicEscherichia coliheat-stable enterotoxin (STal)

Abstract

In order to test the use of a subunit recombinant vaccine for its capacity to induce antibodies against the nonimmunogenic heat-stable enterotoxin STa from Escherichia coli and the TEM-1 beta-lactamase, cattle were immunized with a hybrid protein created by insertion of the STa sequence at position 197 of the TEM-1 beta-lactamase. Specific anti-STa IgG and IgG1 antibodies were detected at low levels, while no IgG2 antibodies were detected. In contrast, high levels of the different anti-TEM IgG subtypes were detected in cattle sera. In addition, beta-lactamase activity was inhibited by the sera. The presence of antibodies against STa and TEM-1 beta-lactamase was assessed in sera from 366 cattle taken from the field. No significant level of IgGs against the toxin or the TEM-1 was detected. A comparison of the antibody level between the immunized and the nonimmunized animals clearly demonstrated that STa was not able to induce a significant level of antibodies in the vaccinated animals. In contrast, a strong antibody response against TEM-1 beta-lactamase was demonstrated.

Antibiotic Resistance Determinant-Focused Acinetobacter baumannii Vaccine Designed Using Reverse Vaccinology

“…It is clear that many valuable results have been identified from our bioinformatics study. It is noted that many published experimental studies [14,22,46,73,81] have experimentally verified many of our predictions. Considering that we did not use these published results ahead of our bioinformatics predictions, these experimental evidences have provided some proof-of-concept evaluations for our analysis.…”

“…Interestingly, our Vaxign predicted PER-1 as a cytoplasmic protein with an adhesin probability =0.378. Given previous reports of β-lactamases being feasible vaccine candidates [14,15,81] and our Vaxign analysis of the differences among β-lactamases, we would recommend that A. baumannii CTX-M-2, -5, and -43 enzymes be better β-lactamase vaccine candidates, which could potentially stimulate strong immune responses that not only reduce antibiotic resistance but also prevent MDR bacterial invasion and infection.…”

“…Previous reports show that β-lactamases can be used as vaccine candidates to stimulate protective immunity against antibiotic-resistant pathogens [14,15,81]. Ciofu et al showed that animals immunized with the AmpC β-lactamase protein could induce strong neutralizing antibody responses and demonstrated a synergistic effect with ceftazidime treatment of resistant P. aeruginosa in a rat model of chronic lung infection [14].…”

“…Ciofu [15] also found that a rat with chronic lung infection immunized with purified chromosomal β-lactamase showed significantly lower bacterial load and reduced lung pathology compared to non-immunized rats. Zervosen et al [81] generated a subunit recombinant TEM-1 β-lactamase by insertion of a heat-stable enterotoxin sequence at position 197 of the TEM-1, and immunization of cattle with this hybrid β-lactamase protein resulted in high levels of the anti-TEM IgG in cattle sera that inhibited β-lactamase activity. These results support the feasibility of developing a preventive or therapeutic β-lactamase vaccine that would induce neutralizing antibodies to inhibit the activity of β-lactamase.…”

Class A -Lactamases as Versatile Scaffolds to Create Hybrid Enzymes: Applications from Basic Research to Medicine

“…Such hybrid proteins can be designed for a range of applications such as (i) the creation of bifunctional hybrid proteins [8–11], (ii) the production, purification, and characterization of proteins otherwise difficult to express [10, 12, 13], (iii) the determination of the epitope of a specific antibody at the surface of an antigen [8, 11], (iv) the generation of antibodies against specific antigens or fragments thereof [9, 13–15], and (v) the investigation of fundamental aspects of structure, stability, and function of proteins [10, 12, 16]. …”