Analysis of a predominant immunoglobulin population in the cerebrospinal fluid of a multiple sclerosis patient by means of an anti-idiotypic hybridoma antibody.

Abstract

We have produced hybridoma antibodies directed against immunoglobulins present in the cerebrospinal fluid ofa patient with multiple sclerosis (MS). One hybridoma antibody recognized an idiotypic determinant of an immunoglobulin population [an idiotype (Id)] which constituted approximately 1% of the immunoglobulin present in the cerebrospinal fluid. The Id focused in the pH range 8. 0-8.4. It was present at roughly 10-to 15-fold higher relative (compared to total immunoglobulin) concentration in cerebrospinal fluid than in serum of the homologous MS patient. The Id could not be detected in three cerebrospinal fluid samples and 28 serum samples of heterologous MS patients or in the serum of 43 optic neuritis patients. The Id persisted in the homologous MS patient at increased concentration over the entire (6 years) observation period. The Id could be shown to react with a Theiler murine encephalomyelitis virus strain WW which was isolated from mice after inoculation with periplaque white matter from brain in a histologically confirmed case of MS.The cerebrospinal fluid (CSF) ofpatients with multiple sclerosis (MS) frequently contains restricted (oligoclonal) populations of Igs that are increased in concentration relative to other CSF Ig populations (1)(2)(3)(4)(5) and are thought to be produced by intrathecally localized B cells (6-9). Many attempts have been made to identify the specificity ofthese predominant CSF Ig populations in the hope offinding some clues to the nature of the antigen(s) involved in the pathogenesis of MS. Those studies have revealed predominant CSF Ig populations reactive with various viruses, most notably measles (reviewed in ref. 10) and, in one case, reactive with smooth muscle antigens (11). These antibodies seem to account, however, only for a minor fraction of the oligoclonal . Thus, the specificity(ies) of most of the predominant Ig populations in the CSF of MS patients remains unknown.Recent studies by Baird et al. (15) and Nagelkerken et al. (16) have demonstrated the feasibility of producing anti-idiotypic antibodies by immunization of mice and guinea pigs, respectively, with CSF Ig of MS patients. Such anti-idiotypic antibodies may provide ideal reagents for investigation of the specificity ofpredominant CSF 1g. The present study describes the production of one such anti-idiotypic hybridoma antibody and its application in specificity analyses ofa predominant CSF Ig population. As controls, serum samples from 14 healthy Scandinavians were obtained, and a normal Ig pool was prepared from serum samples of 70 healthy North Americans. The pooled serum was precipitated with ammonium sulfate at 42% saturation. The precipitate was then redissolved in and dialyzed against 0.1 M K phosphate buffer at pH 7.4 and subsequently purified on DE-32 as described for CSF Ig. MATERIALSProduction of Murine Anti-CSF-Ig Hybridomas. A/J mice were immunized by four intraperitoneal inoculations with 100 ,ug of Ig (1:9 in complete Freund's adjuvant) partially purified from pooled (1974)(1975)(19...

Phylogenetic Analysis of the SpeciesTheilovirus: Emerging Murine and Human Pathogens

“…S2 and M2 are laboratory derivatives of BeAn 8386 and are therefore expected to be nearly identical; however, laboratory contamination has been suggested as the explanation for the close relationship between WW and TO4 (35). Around the time WW virus was isolated in 1977, other investigators were using TO4 virus in experiments at the Wistar Institute (15). TO4 virus was also included in the serological characterization of DA virus that was isolated 15 years after TO4 (8).…”

Molecular and immunogenic features of myelin lipids: incitants or modulators of multiple sclerosis?

“…Carefully monitoring conditions including influences of CSF IgG concentration, markers of oligodendrocytes, and contamination with other cells that was eliminated by judicious double staining, and controlling for species effects, they showed conclusively that oligodendrocytes are not the targets for CSF Igs [170]. Similarly, among reports of antibody specificity in MS, an early analysis of the predominant Igs in MS CSF and serum employing anti-idiotypic hybridomas not only anticipated the production in recent years of recombinant monoclonal antibodies from MS CSF B cells but also revealed a heterogeneity of antibody response between patients [171].…”

Multiple Sclerosis

“…The levels of these antibodies have been demonstrated to be 8-15 times higher in CSF than in serum. Gerhard et al 158 further demonstrated that the antiidiotypic antibodies have specificity towards Theiler murine encephalomyelitis virus. Only more extensive studies can evaluate the implications of these findings.…”

“…The titers of these antibodies correlated with the malignancy and disability of the disease. Recently, Gerhard et al 158 and Nagelkerken et al 159 elucidated the oligoclonal bands in CSF from MS patients by means of antiidiotypic antibodies. The levels of these antibodies have been demonstrated to be 8-15 times higher in CSF than in serum.…”