Analysis of serum and lymphocyte surface IgM of healthy and immunodeficient horses with monoclonal antibodies.

The research is about the creation and utilization of nine monoclonal antibodies that solely bond with equine (horse) immunoglobulin M (IgM), and their further testing on various horse breeds and other animal species for their efficiency in detecting and quantifying IgM molecules.

Preparation of Monoclonal Antibodies

• The researchers prepared nine monoclonal antibodies that reacted with equine (horse) immunoglobulin M (IgM) and not other equine immunoglobulins or serum proteins.
• These antibodies were produced by cells (C 1.9) that could precipitate with IgM and bind to staphylococcal protein A.
• These cells were then triple-cloned (C 1.9/3.2) for the further characterization of the antibodies.

Testing of Monoclonal Antibodies

• A specific monoclonal antibody, termed as C 1.9/3.2, was tested on serum IgM from horses of several breeds.
• The researchers employed studies with iodine-125 labeled IgM to ascertain this monoclonal antibody’s reactivity to an IgM determinant (an antigen-specific region), which was found on all IgM molecules.
• They used this monoclonal antibody to determine the quantity of IgM in the serum of adult horses and presuckling foals.

Application of Monoclonal Antibodies on B Lymphocytes

• Immuno-fluorescent testing demonstrated that the C 1.9/3.2 monoclonal antibody recognized mu-chain determinant (a segment of the heavy chain of IgM) on live B lymphocytes.
• Moreover, cells containing IgM could be distinguished in acetone-fixed frozen sections of lymphoid tissue with the help of this antibody.

Testing on Multiple Animal Species

• Lastly, the team investigated whether the C 1.9/3.2 antibody could also identify IgM molecules in several other animal species.
• The positive results suggested that this monoclonal antibody could be a useful tool in studies of IgM in not only horses but also other animals.