Suitability of a recombinant Staphylococcus aureus enterotoxin C bovine variant for immunodiagnostics and therapeutic vaccine development.

Recombinant bovine variant of staphylococcal enterotoxin C (SECbovine), produced as a NH2-terminal histidine hexamer fusion protein (His6-tagged SECbovine), expressed at high levels (25%) in Escherichia coli and affinity purified to homogeneity (99.9%), was tested for its diagnostic and therapeutic potentials. His6-tagged SECbovine is antigenically authentic to native SECbovine across host species, as confirmed by antibody-based capture detection assays using human, mouse, rabbit and chicken hyperimmune sera. His6-tagged SECbovine showed significant T-cell stimulation activity in vitro. His6-tagged SECbovine was immunogenic for IgG in mice (intragastric and intravenous routes) and rabbits (intramuscular and subcutaneous routes), dispensing immunoadjuvant coadministration. The formation of neutralizing antibodies reduced the severity of intoxication symptoms in immunized rabbits. Purified anti-recombinant SECbovine rabbit polyclonal IgG neutralized the pyrexic and diarrhoeagenic effects of native SEC/SED and recombinant SEC, tested by the kitten and rabbit bioassays, respectively.

Prevention of lethal murine candidiasis using HP (2-20), an antimicrobial peptide derived from the N-terminus of Helicobacter pylori ribosomal protein L1.

Peptide HP (2-20), A(2)KKVFKRLEKLFSKIQNDK(20), is a cationic antimicrobial peptide derived from the N-terminus of Helicobacter pylori ribosomal protein 1, HpRpL1. Native peptide HP (2-20) and its synthetic derivatives have been shown in vitro to exhibit potent killing activity against Gram-positive, Gram-negative and yeast cells, thus, making them promising candidates for treatment of polymicrobial infections. However, the therapeutic potential of peptide HP (2-20) or its synthetic derivatives in any animal model of either bacterial or fungal diseases has not yet been investigated. In this study, we demonstrate that synthetic peptide amide HP (2-20), administered in six doses (300microg each; one intraperitoneal dose at the time of the infection, followed by five intravenous doses at 12h intervals) to CBA/J male mice experimentally infected with a lethal inoculum ( [Formula: see text] CFU) of Candida albicans, delayed the onset of disease, suppressed disease progression, and greatly increased survival rate and time (16.6% by day 14), as compared with the untreated infected control mice (100% mortality by day 5). Further, using isotonic buffer systems differing in ionic strength, peptide HP (2-20) was shown in vitro to exhibit an ionic strength-dependent hemolytic activity, previously not detected. Repeated intravenous administration of uninfected control CBA/J male mice with peptide HP (2-20), however, caused neither morbidity nor mortality. These findings strongly evidence the therapeutic efficacy and safety values of peptide HP (2-20) as a lead drug for the treatment of acquired candidiasis.

Treatment efficacy of the lead RNAIII-inhibiting peptide YSPWTNF-NH2 in acquired Staphylococcus aureus sepsis: a histopathological assessment.

The quorum-sensing interfering RNAIII-inhibiting peptide (RIP) YSPXTNF and its synthetic analogues YSPWTNF and YSPITNF have been shown to prevent and suppress diseases caused by Staphylococcus aureus at different body sites in different animal models. This study was designed to investigate histopathologically the therapeutic efficacy of lead peptide RIP YSPWTNF-NH(2) in the subcutaneous air sac murine model of acquired S. aureus sepsis. Two experimental protocols were evaluated: an infection/therapy protocol, for which twenty BALB/c mice per group were infected with a subcutaneous inoculum of S. aureus strain ATCC 25923 ( [Formula: see text] colony forming units) that were either pretreated or not with 150microg of peptide RIP, and a safety protocol, for which three uninfected mice per group received treatment with either 150microg of peptide RIP or saline. Therapeutic efficacy was assessed by clinical examination for a period of 20 days and histopathology at 12, 24, 36, 48, 96 and 168h after inoculation. Treatment safety was assessed histopathologically at 24, 48 and 264h after inoculation. Subcutaneous administration of uninfected control mice with a single dose of peptide RIP YSPWTNF caused no significant histopathology in most organs examined, except for slight to moderate lung and liver congestions. In contrast to the situation with the untreated infected control group mice that presented with histopathological alterations consistent with the diagnosis of rapidly progressive and highly erosive disease (100% mortality by day 3), treatment of infected animals with peptide RIP YSPWTNF had a profound therapeutic effect on survival rate (67% by day 20) and on disease progression. The histopathological examination confirmed the clinical findings showing that extensive tissue damage at the site of the infection and in organs were greatly suppressed in the peptide RIP-treated animals.