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Use of an automated apparatus to quantitate mumps virus- and poliovirus-neutralizing antibody is described. The automated titration equipment affords savings in effort, time, and reagents in conducting large-scale surveys for the determination of mumps- and poliovirus-neutralizing antibodies. This technique has been found to be reproducible and gives results comparable to other antibody assay methods.

By employing the techniques of immunofluorescence and radioimmunodiffusion using (32)P-labeled poliovirus as the antigen, the immunoglobulin response to poliovirus in serum, nasopharynx, spinal fluid, and in different segments of the central nervous system (CNS) was studied after intramuscular, oral, intranasal, and intrathalamic administration of inactivated (Salk), live attenuated (Sabin), or live virulent (Mahoney) type I poliovirus. Spinal fluid gammaG antibody was detected after immunization with Sabin or Mahoney virus and intramuscular administration of Salk vaccine. The response in the CNS was characterized by the appearance of gammaG antibody after oral or intrathalamic administration of Mahoney virus and rarely after intrathalamic inoculation of Sabin vaccine. The antibody activity in CNS was limited to the areas of poliovirus replication. Intrathalamic immunization with Mahoney virus resulted in local gammaG antibody production in the CNS in the absence of any detectable response in serum. Discrete foci of gammaG-containing cells were observed in those areas of CNS which contained poliovirus antibody. No immunoglobulin-containing cells or poliovirus antibody was seen in the CNS of monkeys immunized with intramuscularly or orally administered Sabin or Salk vaccine and in sham-immunized control monkeys. It is suggested that the CNS, when stimulated locally with a potent replicating viral antigen, may manifest a specific local antibody response, which is independent of the response in serum.

A simple, economical radioassay system employing disposable polypropylene microcentrifuge tubes was developed. Plastic adapters permitted automatic operation in liquid scintillation spectrometers. Counting efficiencies of (3)H, (14)C, (32)P, and (125)I in liquid scintillation cocktails and of (32)P by Cerenkov radiation (at lower efficiency in absence of added scintillator) were comparable to those in standard vials. Multipurpose use of the microtubes made the system versatile and expedient, e.g., collection of precipitates and radioassay in the same container. Collection of radioimmune precipitates was aided by a carrier inorganic precipitate, Mg(2)P(2)O(7).

The occurrence of a local production in the central nervous system (CNS) of antibodies against different selected viruses was analyzed by comparison of titers in serum and cerebrospinal fluid samples from groups of 50 patients with multiple sclerosis from Finland, Norway, and Sweden. Measles antibodies were determined in hemagglutination inhibition, hemolysis inhibition, and nucleocapsid complement fixation tests; mumps, parainfluenza virus type 1, and rubella virus antibodies were determined in hemagglutination inhibition tests; and herpes simplex virus type 1 antibodies were determined in passive hemagglutination tests. For reference purposes tests were also made for adenovirus antibodies in penton hemagglutination enhancement tests and poliovirus antibodies in neutralization enhancement tests. Among the 150 multiple sclerosis patients, a local production of antibodies against measles virus was found in the CNS in 57%, against rubella virus in 19%, mumps virus in 15%, herpes simplex virus type 1 in 11%, and parainfluenza virus type 1 (Sendai) in 3%. A local production in the CNS of antibodies against any of the viruses studied was found in 71% of multiple sclerosis patients. These included 48, 16, and 7% that produced antibodies to one, two, and three or more viruses, respectively.

Rabbits were immunized with the attenuated LSc, 2a(1)b or the virulent Mahoney strain of type 1 poliovirus, or with a partially purified preparation of the latter. It was found that the animals responded in a heterogeneous manner to antigens with a homogeneous virion population, such as LSc,2a(1)b and, to a lesser extent, the partially purified Mahoney antigen. This heterogeneous immune response permits the preparation of specific antisera for intratypic serodifferentiation tests. However, the unpurified Mahoney strain, which has a heterogeneous virion population, induced a homogeneous immune response; all the antisera had a wide spectrum of antibodies and could not distinguish the wild from the attenuated strain. The differentiation capacity of the specific antisera was found to be linked to either the IgM or the IgG. The possible implications of these findings for human vaccination against poliomyelitis are discussed.