Intrathecal Synthesis of Anti‐Borrelia burgdorferi Antibodies in Neuroborreliosis: a Study with Special Emphasis on Oligoclonal IgM Antibody Bands

Pre- and post-treatment (up to 3–26 months after antibiotic therapy) humoral immune responses were investigated in five neuroborreliosis patients. Anti-Borrelia (B.) burgdorferi IgG and IgM antibodies in CSF and serum were quantitated by capture ELISA. Agarose gel isoelectric focusing (AIF) and protein blotting were used to detect oligoclonal IgG and IgM bands as well as oligoclonal anti-B. burgdorferi IgG and IgM antibodies. These latter components were visualized by transfer to antigen-coated membranes (immunoblot) and immunoenzymatic staining. By ELISA, intrathecal anti-B. burgdorferi IgG and IgM antibody synthesis was detected in all initial specimens and continued 3–26 months after antibiotic therapy in four and three cases, respectively. AIF with protein blotting showed oligoclonal bands of total IgG as well as total IgM in the initial CSF specimens of all patients and persistence of such components occurred in four and five cases, respectively. By AIF and immunoblot, oligoclonal anti-B. burgdorferi IgG and IgM antibody bands could be detected in the CSF of every patient. IgG antibody bands were present in all initial CSF samples. The first specimen of one patient was negative for IgM antibody bands but such components appeared 3 weeks later. Oligoclonal CSF anti-B. burgdorferi IgG antibody components persisted over the entire follow-up periods in all but one case where they disappeared 6 weeks after treatment. The oligoclonal IgM antibodies in CSF vanished in two cases (after being present up to 4 and 11 months after antibiotic therapy) while they persisted over the entire (3–6 months after antibiotic therapy) follow-up periods in three cases. The specificity of the IgM antibody immunoblot technique was corroborated by control experiments, including antibody absorption studies and use of 41 kDa flagellar antigen.