The use of plaque-forming techniques in animal virology has markedly increased flae sensitivity and precision of measuring neutralizing antibody, and this together with other developments such as the ready availability of large quantities of relatively pure viral antigens and new techniques for characterizing antibody have prompted a reexamination of several aspects of the formation and properties of viral neutralizing antibody. In the work being reported a poliovirus-rabbit antibody system has been extensively studied with particular emphasis on experiments concerned with the length of the induction period, changes in properties of antibody produced at different times after an antigenic stimulus, differences in !9S and 7S antibody formation and on the ability of animals to display "immunological memory." The combination of high sensitivity and high accuracy in measuring antibody to poliovirus (PV) makes an excellent tool permitting for example the study of the very early appearance of circulating antibody.In this work, to be reported in a series of papers, we have sought to (a) analyze 19S and 7S type antibody formation in terms of antigen dose requirement for induction and maintained synthesis, kinetics, duration of immunological memory, and sensitivity to whole body x-irradiation; (b) examine the progressive changes in physicochemical and biological properties of antibody with time after antigenic stimulus; and (c) to investigate the properties of antibody formed in vitro by in ~/vo primed cells. In addition the nature of "normal" antibody and its significance in relation to the immune response has been examined and will be reported.Early in this study it was found (1-3) that the induction period was of the order of several hours or less and that macroglobulin antibody was demonstrable 8 to 12 hours after antigen injection when sufficiently sensitive methods * This investigation was supported in part by research grant E-4350 from the National Institute of Allergy and Infectious Diseases, United States Pubfic Health Service. were employed for a n t i b o d y detection. I t was further reported (2) t h a t either a transitory or an enduring response was elicited depending upon the dose of antigen used, and t h a t a correlation existed between the t y p e of response and the molecular forms (19S, 7S) of a n t i b o d y synthesized. T h e present report extends these earlier observations on the length of the induction period, the antigen dose requirements for induction of 19S and 7S a n t i b o d y and on the kinetics of their formation.
Materials and MethodsVirus.--Methods for growth of HeLa cell monolayers and techniques of preparation and assay of PV have been described (4), The Brunhilde strain (type 1) of poliovirus and Coxsackie B-4 virus 1 were used. The viruses were cloned and propagated on HeLa cell monolayers. Prior to storage at --70 ° or --15°C stock virus was partly purified and concentrated by differential centrifugatinn. Such preparations contained about 1 #g total protein per 10 ~'4 plaque-formin...