Further Studies of Viral Antibodies in the Cerebrospinal Fluid of Patients with Multiple Sclerosis: Vaccinia and Parainfluenza Type 1

In an earlier paper (1) we reported elevated measles antibody levels in cerebrospinal fluid (CSF) from a large series of patients with multiple sclerosis (MS) as compared to patients with other neurological diseases. Since our report, ter Meulen et al. (2) have isolated parainfluenza virus type 1 from brain tissue from 2 patients with MS, and Kempe (3) has found antibody to vaccinia virus in a high proportion of CSF specimens from MS patients, but not controls. Support of the parainfluenza virus isolations by antibody data, or confirmation of the vaccinia antibody study would considerably strengthen the implication that either virus could in at least some patients be causally related to MS. We have, therefore, gone back to our original series of CSF specimens, and report here the results of tests for neutralizing antibody to these viruses.

Materials and Methods. Specimens. CSF from 127 patients with MS and 78 patients with other neurological diseases were tested for antibodies to vaccinia virus. This is substantially the same series, with fewer controls, on which measles antibody titers were earlier reported, and is fully described there (1). CSF specimens from approximately one half of this series (62 MS and 36 control patients) were tested for antibody to parainfluenza virus type 1.

Antibody assays. Vaccinia virus, originating from the IHD strain (4), and grown in HeLa cells, was obtained from Dr. Samuel Baron, NIAID, NIH. Equal volumes of the undiluted CSF specimens were mixed with 600 pfu of sonicated vaccinia virus in Eagles # 2 medium, incubated with occasional agitation for 2 hr at 37°, and then further diluted 1:10 before 0.2 ml volumes were added to triplicate vertical 25 mm flat-bottom glass vials (5) containing sheets of confluent L-cells.