Development of a monoclonal antibody specific to burbot (Lota lota) IgM and optimization of an ELISA to measure anti-Aeromonas sp. antibody titers following pathogen challenge

• Published in: Fish & shellfish immunology 2023-06, Vol.137, p.108775-108775, Article 108775
• Main Authors: Oliver, Luke P., Bruce, Timothy J., Ma, Jie, Jones, Evan M., Cain, Kenneth D.
• Format: Article
• Language: English
• Subjects: Aeromonas; Aeromonas sp; Animals; Antibodies, Monoclonal; Antibody; Burbot; Enzyme-Linked Immunosorbent Assay - veterinary; Fishes; Gadiformes; Monoclonal
• ISSN: 1050-4648; 1095-9947
• DOI: 10.1016/j.fsi.2023.108775

Burbot (Lota lota) are an ideal candidate for cool or cold-water aquaculture and are gaining interest because of their high economic value, low temperature requirements, and fast growth rate. Limited information exists on the innate and adaptive immune systems of this species. This is partly due to the lack of species-specific tools to determine antibody responses following disease or vaccination or to characterize the immune response in general. An anti-IgM monoclonal antibody (mAb 27C) was developed and characterized via enzyme-linked immunosorbent assay (ELISA) and Western blot for species specificity, affinity to the heavy chain of burbot IgM, and cross-reactivity to other reagents used in the analysis. The 27C monoclonal antibody was further utilized to develop an ELISA protocol to measure the specific antibody response of burbot following exposure to two pathogenic strains of Aeromonas sp. (A141 and IR004). This ELISA confirmed that vaccinated burbot that survived the challenge with either strain developed statistically higher titers of anti-Aeromonas antibodies specific for the relative strain when compared to fish that were not vaccinated or challenged. Western blot analysis further demonstrated that burbot surviving challenge had serum IgM that recognized distinct antigens specific to the strain they were challenged with, A141 bound to antigens in the 50-250Kda range and IR004 bound to a distinct 150Kda antigen. Western blots further indicated that each strain shared antigenic regions regardless of experimental Aeromonas strain exposure. Finally, immunofluorescent staining confirmed that mAb 27C binds to membrane-bound IgM (presumably B cells) on burbot head kidney cells. Taken together, results from this study demonstrate that mAb 27C specifically recognized burbot IgM and will be an important tool to further characterize the adaptive and cellular immune responses of this fish species. [Display omitted] •Production of a monoclonal antibody (mAb27C), specific to the heavy chain of burbot IgM.•mAb 27C detects differences in Aeromonas sp. (strains A141 and IR004) antibody titers after exposure or vaccination.•mAb 27C detects immunogenic regions of Aeromonas sp. (strains A141 and IR004) via western blot.•mAb 27C detects IgM + cells via immunocytochemistry.