Development of Opsonic Mouse Monoclonal Antibodies against Multidrug-Resistant Enterococci

Autor: Ermioni Kalfopoulou, Suzana Malić, Roberto Adamo, Karmela Miklić, Tihana Lenac Rovis, Johannes Huebner, Stipan Jonjić, Diana Laverde, Felipe Romero-Saavedra, Filippo Carboni
Jazyk: angličtina
Rok vydání: 2019
Předmět:
0301 basic medicine
medicine.drug_class
Enterococcus faecium
030106 microbiology
Immunology
SagA
Biology
Monoclonal antibody
Microbiology
Enterococcus faecalis
opsonic
Mice
03 medical and health sciences
Antigen
Polysaccharides
medicine
Animals
Bacterial Capsules
opsonophagocytic assay
Antigens
Bacterial
hybridoma technology
BIOMEDICINE AND HEALTHCARE. Basic Medical Sciences
Antibodies
Monoclonal
Drug Resistance
Microbial
Opsonin Proteins
biology.organism_classification
capsular polysaccharide
3. Good health
Multiple drug resistance
DHG
diheteroglycan
monoclonal antibodies
030104 developmental biology
Infectious Diseases
Microbial Immunity and Vaccines
biology.protein
Hybridoma technology
Parasitology
Immunotherapy
Bacterial antigen
Antibody
BIOMEDICINA I ZDRAVSTVO. Temeljne medicinske znanosti
Zdroj: Infection and Immunity
Volume 87
Issue 9
ISSN: 0019-9567
1098-5522
DOI: 10.1128/iai.00276-19
Popis: Multidrug-resistant enterococci are major causes of hospital-acquired infections. Immunotherapy with monoclonal antibodies (MAbs) targeting bacterial antigens would be a valuable treatment option in this setting. Here, we describe the development of two MAbs through hybridoma technology that target antigens from the most clinically relevant enterococcal species.
Multidrug-resistant enterococci are major causes of hospital-acquired infections. Immunotherapy with monoclonal antibodies (MAbs) targeting bacterial antigens would be a valuable treatment option in this setting. Here, we describe the development of two MAbs through hybridoma technology that target antigens from the most clinically relevant enterococcal species. Diheteroglycan (DHG), a well-characterized capsular polysaccharide of Enterococcus faecalis, and the secreted antigen A (SagA), an immunogenic protein from Enterococcus faecium, are both immunogens that have been proven to raise opsonic and cross-reactive antibodies against enterococcal strains. For this purpose, a conjugated form of the native DHG with SagA was used to raise the antibodies in mice, while enzyme-linked immunosorbent assay and opsonophagocytic assay were combined in the selection process of hybridoma cells producing immunoreactive and opsonic antibodies targeting the selected antigens. From this process, two highly specific IgG1(κ) MAbs were obtained, one against the polysaccharide (DHG.01) and one against the protein (SagA.01). Both MAbs exhibited good opsonic killing against the target bacterial strains: DHG.01 showed 90% killing against E. faecalis type 2, and SagA.01 showed 40% killing against E. faecium 11231/6. In addition, both MAbs showed cross-reactivity toward other E. faecalis and E. faecium strains. The sequences from the variable regions of the heavy and light chains were reconstructed in expression vectors, and the activity of the MAbs upon expression in eukaryotic cells was confirmed with the same immunological assays. In summary, we identified two opsonic MAbs against enterococci which could be used for therapeutic or prophylactic approaches against enterococcal infections.
Databáze: OpenAIRE
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