Back to Search
Start Over
Opsono-Adherence Assay to Evaluate Functional Antibodies in Vaccine Development Against Bacillus anthracis and Other Encapsulated Pathogens.
- Source :
-
Journal of visualized experiments : JoVE [J Vis Exp] 2020 May 19 (159). Date of Electronic Publication: 2020 May 19. - Publication Year :
- 2020
-
Abstract
- The opsono-adherence assay is a functional assay that enumerates the attachment of bacterial pathogens to professional phagocytes. Because adherence is requisite to phagocytosis and killing, the assay is an alternative method to opsono-phagocytic killing assays. An advantage of the opsono-adherence assay is the option of using inactivated pathogens and mammalian cell lines, which allows standardization across multiple experiments. The use of an inactivated pathogen in the assay also facilitates work with biosafety level 3 infectious agents and other virulent pathogens. In our work, the opsono-adherence assay was used to assess the functional ability of antibodies, from sera of animals immunized with an anthrax capsule-based vaccine, to induce adherence of fixed Bacillus anthracis to a mouse macrophage cell line, RAW 264.7. Automated fluorescence microscopy was used to capture images of bacilli adhering to macrophages. Increased adherence was correlated with the presence of anti-capsule antibodies in the serum. Non-human primates that exhibited high serum anti-capsule antibody concentrations were protected from anthrax challenge. Thus, the opsono-adherence assay can be used to elucidate the biological functions of antigen specific antibodies in sera, to evaluate the efficacy of vaccine candidates and other therapeutics, and to serve as a possible correlate of immunity.
- Subjects :
- Animals
Anthrax microbiology
Anthrax prevention & control
Antigens, Bacterial immunology
Fluorescein-5-isothiocyanate metabolism
Fluorescence
Humans
Macrophages immunology
Mice
Primates immunology
Primates microbiology
RAW 264.7 Cells
Anthrax immunology
Anthrax Vaccines immunology
Antibodies, Bacterial immunology
Bacillus anthracis immunology
Bacterial Adhesion
Opsonin Proteins immunology
Subjects
Details
- Language :
- English
- ISSN :
- 1940-087X
- Issue :
- 159
- Database :
- MEDLINE
- Journal :
- Journal of visualized experiments : JoVE
- Publication Type :
- Academic Journal
- Accession number :
- 32510489
- Full Text :
- https://doi.org/10.3791/60873