Anti-Pseudomonas aeruginosa serotype O11 LPS immunoglobulin M monoclonal antibody panobacumab (KBPA101) confers protection in a murine model of acute lung infection

Mice immunized with Formalin-fixed mucoid Pseudomonas aeruginosa cells developed an immune response directed, in part, towards the P. aeruginosa glycocalyx. The polyclonal mouse sera produced good immunofluorescent staining of the P. aeruginosa glycocalyx and cell surface. A library of 250 hybridoma cell lines which produced monoclonal antibodies directed against P. aeruginosa was established. Twelve clones (4.8%) produced antibody which reacted with alginate in an enzyme-linked immunosorbent assay (ELISA). Clone Ps 53 was chosen for further study, cloned, and an ascites tumor established. Clone Ps 53 was chosen for further study because the antibody produced demonstrated a specificity similar to that of a recently isolated heparin – rat-lung lectin which recognizes alginates of the Homma nontypable P. aeruginosa strains. The Ps 53 clone produced an immunoglobulin M which reacted with P. aeruginosa alginate and produced good immunofluorescent staining of the P. aeruginosa glycocalyx. The Ps 53 monoclonal antibody has an apparent specificity for L-guluronic residues in ELISA. Competitive binding studies with various alginates and monosaccharides suggest that the C6 carboxyl group of uronic acids are recognized by the antibody and that the antigen-binding site is fairly large and may recognize a particular sequence or epitope of alginic acid which is rich in L-guluronic acid. The Ps 53 monoclonal antibody did not react uniformily with all P. aeruginosa alginates but did react with all of the alginates of the Homma nontypable strains tested, suggesting that acetylation or various modifications found in P. aeruginosa alginates may interfere with antibody binding and define specific epitopes. The Ps 53 antibody also reacted with purified outer membrane, indicating that some alginate or L-guluronic acid is intimately associated with outer membrane.

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Antibacterial efficacy of R-type pyocins against Pseudomonas aeruginosa on biofilms and in a murine model of acute lung infection

Journal of Antimicrobial Chemotherapy ◽

10.1093/jac/dkaa121 ◽

2020 ◽

Cited By ~ 1

Author(s):

Mar Redero ◽

Javier Aznar ◽

Ana I Prieto

Keyword(s):

Pseudomonas Aeruginosa ◽

High Risk ◽

Murine Model ◽

Biofilm Formation ◽

Bacterial Load ◽

Preventive Treatment ◽

Lung Infection ◽

Before And After ◽

Treatment Alternatives ◽

Potential Use

Abstract Background The appearance of MDR strains and the development of biofilms make Pseudomonas aeruginosa infections a therapeutic challenge. To overcome this scenario, bacteriocins have been proposed as a potential adjuvant or alternative to antibiotic treatment. Objectives To study the activity of R-pyocins on biofilms and in a murine model of pneumonia using a high-risk clone of P. aeruginosa. Methods The activity of R-pyocins on P. aeruginosa biofilms was tested on bacteria attached to a silicone surface, before and after biofilm formation. The effectiveness of R1-pyocin was studied in a murine model of pneumonia using ST175, a high-risk clone of P. aeruginosa. Results R-pyocins attacked adherent bacteria, preventing biofilm formation, and penetrated into the biofilm, killing P. aeruginosa within it, resulting in a dramatic reduction in bacterial load. R1-pyocin was active in a murine model of P. aeruginosa lung infection, administered before infection as a preventive treatment, and in acute pneumonia, with efficiency higher than standard colistin treatment. In addition, this work is the first to describe histopathological lung changes after administration of R-pyocins, contributing to the resolution of P. aeruginosa pneumonia in a murine model. Conclusions This work highlights the potential use of the R-pyocins as therapeutic agents, alone or as adjuvants, due to its effectiveness on biofilms and in a murine model of pneumonia using ST175, a high-risk clone of P. aeruginosa. It may thus be feasible to consider R-pyocins as a possible therapeutic alternative in XDR infections, where treatment alternatives are limited.

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Pharmacokinetics and Safety Profile of the Human Anti-Pseudomonas aeruginosa Serotype O11 Immunoglobulin M Monoclonal Antibody KBPA-101 in Healthy Volunteers

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01699-08 ◽

2009 ◽

Vol 53 (8) ◽

pp. 3442-3446 ◽

Cited By ~ 28

Author(s):

Hedvika Lazar ◽

Michael P. Horn ◽

Adrian W. Zuercher ◽

Martin A. Imboden ◽

Peter Durrer ◽

...

Keyword(s):

Monoclonal Antibody ◽

Pseudomonas Aeruginosa ◽

Body Weight ◽

Healthy Volunteers ◽

Human Monoclonal Antibody ◽

Dose Range ◽

Plasma Concentrations ◽

Immunoglobulin M ◽

Serious Adverse Events ◽

The Mean

ABSTRACT KBPA-101 is a human monoclonal antibody of the immunoglobulin M isotype, which is directed against the O-polysaccharide moiety of Pseudomonas aeruginosa serotype O11. This double-blind, dose escalation study evaluated the safety and pharmacokinetics of KBPA-101 in 32 healthy volunteers aged 19 to 46 years. Each subject received a single intravenous infusion of KBPA-101 at a dose of 0.1, 0.4, 1.2, or 4 mg/kg of body weight or placebo infused over 2 h. Plasma samples for pharmacokinetic assessments were taken before infusion as well as 0.25, 0.5, 1, 2, 2.5, 4, 6, 8, 12, 24, 36, and 48 h and 4, 7, 10, and 14 days after start of dosing. Plasma concentrations of KBPA-101 were detected with mean maximum concentrations of drug in plasma of 1,877, 7,571, 24,923, and 83,197 ng/ml following doses of 0.1, 0.4, 1.2, and 4.0 mg/kg body weight, respectively. The mean elimination half-life was between 70 and 95 h. The mean volume of distribution was between 4.76 and 5.47 liters. Clearance ranged between 0.039 and 0.120 liters/h. At the highest dose of 4.0 mg/kg, plasma KBPA-101 levels were greater than 5,000 ng/ml for 14 days. KBPA-101 exhibited linear kinetics across all doses. No anti-KBPA-101 antibodies were detected after dosing in any subject. Overall, the human monoclonal antibody KBPA-101 was well tolerated over the entire dose range in healthy volunteers, and no serious adverse events have been reported.

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Efficacy of species-specific protein antibiotics in a murine model of acute Pseudomonas aeruginosa lung infection

Scientific Reports ◽

10.1038/srep30201 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 30

Author(s):

Laura C. McCaughey ◽

Neil. D. Ritchie ◽

Gillian R. Douce ◽

Thomas J. Evans ◽

Daniel Walker

Keyword(s):

Pseudomonas Aeruginosa ◽

Murine Model ◽

Lung Infection ◽

Specific Protein ◽

Species Specific

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Therapeutic effects of a human antiflagella monoclonal antibody in a neutropenic murine model of Pseudomonas aeruginosa pneumonia.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.37.2.164 ◽

1993 ◽

Vol 37 (2) ◽

pp. 164-170 ◽

Cited By ~ 21

Author(s):

K Oishi ◽

F Sonoda ◽

A Iwagaki ◽

P Ponglertnapagorn ◽

K Watanabe ◽

...

Keyword(s):

Monoclonal Antibody ◽

Pseudomonas Aeruginosa ◽

Murine Model ◽

Therapeutic Effects

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Anti-Inflammatory Effects of RTD-1 in a Murine Model of Chronic Pseudomonas aeruginosa Lung Infection: Inhibition of NF-κB, Inflammasome Gene Expression, and Pro-IL-1β Biosynthesis

Antibiotics ◽

10.3390/antibiotics10091043 ◽

2021 ◽

Vol 10 (9) ◽

pp. 1043

Author(s):

Mansour A. Dughbaj ◽

Jordanna G. Jayne ◽

A Young J. Park ◽

Timothy J. Bensman ◽

Marquerita Algorri ◽

...

Keyword(s):

Gene Expression ◽

Pseudomonas Aeruginosa ◽

Microarray Analysis ◽

Lung Tissue ◽

Murine Model ◽

Lung Infection ◽

Cell Counts ◽

Tissue Homogenates ◽

Anti Inflammatory

Vicious cycles of chronic airway obstruction, lung infections with Pseudomonas aeruginosa, and neutrophil-dominated inflammation contribute to morbidity and mortality in cystic fibrosis (CF) patients. Rhesus theta defensin-1 (RTD-1) is an antimicrobial macrocyclic peptide with immunomodulatory properties. Our objective was to investigate the anti-inflammatory effect of RTD-1 in a murine model of chronic P. aeruginosa lung infection. Mice received nebulized RTD-1 daily for 6 days. Bacterial burden, leukocyte counts, and cytokine concentrations were evaluated. Microarray analysis was performed on bronchoalveolar lavage fluid (BALF) cells and lung tissue homogenates. In vitro effects of RTD-1 in THP-1 cells were assessed using quantitative reverse transcription PCR, enzyme-linked immunosorbent assays, immunoblots, confocal microscopy, enzymatic activity assays, and NF-κB-reporter assays. RTD-1 significantly reduced lung white blood cell counts on days 3 (−54.95%; p = 0.0003) and 7 (−31.71%; p = 0.0097). Microarray analysis of lung tissue homogenates and BALF cells revealed that RTD-1 significantly reduced proinflammatory gene expression, particularly inflammasome-related genes (nod-like receptor protein 3, Mediterranean fever gene, interleukin (IL)-1α, and IL-1β) relative to the control. In vitro studies demonstrated NF–κB activation was reduced two-fold (p ≤ 0.0001) by RTD-1 treatment. Immunoblots revealed that RTD-1 treatment inhibited proIL-1β biosynthesis. Additionally, RTD-1 treatment was associated with a reduction in caspase-1 activation (FC = −1.79; p = 0.0052). RTD-1 exhibited potent anti-inflammatory activity in chronically infected mice. Importantly, RTD-1 inhibits inflammasome activity, which is possibly a downstream effect of NF-κB modulation. These findings support that this immunomodulatory peptide may be a promising therapeutic for CF-associated lung disease.

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Influence of antibiotic and E5 monoclonal immunoglobulin M interactions on endotoxin release from Escherichia coli and Pseudomonas aeruginosa.

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.40.1.247 ◽

1996 ◽

Vol 40 (1) ◽

pp. 247-252 ◽

Cited By ~ 17

Author(s):

K C Lamp ◽

M J Rybak ◽

B J McGrath ◽

K K Summers

Keyword(s):

Escherichia Coli ◽

Monoclonal Antibody ◽

Pseudomonas Aeruginosa ◽

Bactericidal Activity ◽

Immunoglobulin M ◽

Monoclonal Immunoglobulin ◽

Bacterial Killing ◽

E Coli ◽

Factors Associated ◽

Medium Range

Recent controversy surrounding the activity of monoclonal antibodies against endotoxin highlights the necessity of identifying all factors associated with increased mortality, one of which is endotoxin concentrations. Antibiotics may induce different patterns of endotoxin release. We compared the release of free endotoxin (in endotoxin units per milliliter) over 6 h and changes in numbers of CFU of exponentially growing Escherichia coli and Pseudomonas aeruginosa (10(6) to 10(7) CFU/ml) cultured in chemically defined endotoxin-free broth combined with pooled human serum and/or 10 micrograms of E5 immunoglobulin M monoclonal antibody per ml. MICs and MBCs were tested in each medium at the same inoculum. The inoculum was exposed to antibiotics at a single fixed multiple of the MIC for each medium (range, two to eight times the MIC). E5 antibody had no effect on MICs, MBCs, bactericidal activity, or endotoxin release. In the presence of 50% serum, amikacin, ceftazidime, imipenem, and ofloxacin each killed equivalent amounts of E. coli over 6 h; however, ceftazidime induced the highest release of endotoxin. Amikacin and ofloxacin produced the most favorable ratio of endotoxin release to amount of bacterial killing. In the presence of 50% serum, ceftazidime and imipenem reduced the P. aeruginosa inoculum to the greatest extent over 6 h. Although its bactericidal activity was diminished, ofloxacin caused the lowest release of free endotoxin. Imipenem and ofloxacin showed similar low ratios of endotoxin release to bacterial killing. In summary, antibiotic class, presence of serum, and type of organism influenced bactericidal activity and endotoxin release.

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