RESUMEN
Extremely drug-resistant (XDR) Acinetobacter baumannii is a notorious and frequently encountered pathogen demanding novel therapeutic interventions. An initial monoclonal antibody (MAb), C8, raised against A. baumannii capsule, proved a highly effective treatment against a minority of clinical isolates. To overcome this limitation, we broadened coverage by developing a second antibody for use in a combination regimen. We sought to develop an additional anti-A. baumannii MAb through hybridoma technology by immunizing mice with sublethal inocula of virulent, XDR clinical isolates not bound by MAb C8. We identified a new antibacterial MAb, 65, which bound to strains in a pattern distinct from and complementary to that of MAb C8. MAb 65 enhanced macrophage opsonophagocytosis of targeted strains and markedly improved survival in lethal bacteremic sepsis and aspiration pneumonia murine models of A. baumannii infection. MAb 65 was also synergistic with colistin, substantially enhancing protection compared to monotherapy. Treatment with MAb 65 significantly reduced blood bacterial density, ameliorated cytokine production (interleukin-1ß [IL-1ß], IL-6, IL-10, and tumor necrosis factor), and sepsis biomarkers. We describe a novel MAb targeting A. baumannii that broadens immunotherapeutic strain coverage, is highly potent and effective, and synergistically improves outcomes in combination with antibiotics.
Asunto(s)
Infecciones por Acinetobacter/inmunología , Acinetobacter baumannii/inmunología , Anticuerpos Monoclonales/inmunología , Infecciones por Acinetobacter/sangre , Infecciones por Acinetobacter/microbiología , Animales , Antibacterianos/inmunología , Anticuerpos Antibacterianos/inmunología , Biomarcadores/sangre , Colistina/inmunología , Citocinas/sangre , Citocinas/inmunología , Farmacorresistencia Bacteriana Múltiple/inmunología , Ratones , Pruebas de Sensibilidad Microbiana/métodos , Sepsis/sangre , Sepsis/inmunología , Sepsis/microbiologíaAsunto(s)
Antibacterianos/efectos adversos , Colistina/efectos adversos , Desensibilización Inmunológica , Hipersensibilidad a las Drogas/terapia , Antibacterianos/inmunología , Colistina/inmunología , Fibrosis Quística/tratamiento farmacológico , Fibrosis Quística/inmunología , Hipersensibilidad a las Drogas/inmunología , Humanos , Masculino , Persona de Mediana Edad , Pruebas CutáneasRESUMEN
UNLABELLED: The purpose of this study was to take advantage of the nematode Caenorhabditis elegans to perform a whole-animal chemical screen to identify potential immune activators that may confer protection against bacterial infections. We identified 45 marketed drugs, out of 1,120 studied compounds, that are capable of activating a conserved p38/PMK-1 mitogen-activated protein kinase pathway required for innate immunity. One of these drugs, the last-resort antibiotic colistin, protected against infections by the Gram-negative pathogens Yersinia pestis and Pseudomonas aeruginosa but not by the Gram-positive pathogens Enterococcus faecalis and Staphylococcus aureus. Protection was independent of the antibacterial activity of colistin, since the drug was administered prophylactically prior to the infections and it was also effective against antibiotic-resistant bacteria. Immune activation by colistin is mediated not only by the p38/PMK-1 pathway but also by the conserved FOXO transcription factor DAF-16 and the transcription factor SKN-1. Furthermore, p38/PMK-1 was found to be required in the intestine for immune activation by colistin. Enhanced p38/PMK-1-mediated immune responses by colistin did not reduce the bacterial burden, indicating that the pathway plays a role in the development of host tolerance to infections by Gram-negative bacteria. IMPORTANCE: The innate immune system represents the front line of our defenses against invading microorganisms. Given the ever-increasing resistance to antibiotics developed by bacterial pathogens, the possibility of boosting immune defenses represents an interesting, complementary approach to conventional antibiotic treatments. Here we report that the antibiotic colistin can protect against infections by a mechanism that is independent of its microbicidal activity. Prophylactic treatment with colistin activates a conserved p38/PMK-1 pathway in the intestine that helps the host better tolerate a bacterial infection. Since p38/PMK-1-mediated immune responses appear to be conserved from plants to mammals, colistin may also activate immunity in higher organisms, including humans. Antibiotics with immunomodulatory properties have the potential of improving the long-term outcome of patients with chronic infectious diseases.
Asunto(s)
Caenorhabditis elegans/inmunología , Caenorhabditis elegans/microbiología , Colistina/inmunología , Colistina/farmacología , Evaluación Preclínica de Medicamentos/métodos , Inmunidad Innata , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Animales , Infecciones Bacterianas/inmunología , Infecciones Bacterianas/prevención & control , Caenorhabditis elegans/efectos de los fármacos , Caenorhabditis elegans/metabolismo , Enterococcus faecalis/efectos de los fármacos , Enterococcus faecalis/inmunología , Inmunomodulación , Análisis por Micromatrices , Modelos Animales , Pseudomonas aeruginosa/efectos de los fármacos , Pseudomonas aeruginosa/inmunología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus aureus/inmunología , Yersinia pestis/efectos de los fármacos , Yersinia pestis/inmunología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Acinetobacter baumannii is a leading cause of multidrug-resistant infections worldwide. This organism poses a particular challenge due to its ability to acquire resistance to new antibiotics through adaptation or mutation. This study was undertaken to determine the mechanisms governing the adaptability of A. baumannii to the antibiotic colistin. Screening of a transposon mutant library identified over 30 genes involved in inducible colistin resistance in A. baumannii. One of the genes identified was lpsB, which encodes a glycosyltransferase involved in lipopolysaccharide (LPS) synthesis. We demonstrate that loss of LpsB function results in increased sensitivity to both colistin and cationic antimicrobial peptides of the innate immune system. Moreover, LpsB is critical for pathogenesis in a pulmonary model of infection. Taken together, these data define bacterial processes required for intrinsic colistin tolerance in A. baumannii and underscore the importance of outer membrane structure in both antibiotic resistance and the pathogenesis of A. baumannii.