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Journal of Hygiene THE AMERICAN JOURNAL OF HYGIENE VOL. 74 JULY, 1961 NO. 1 AVIAN EMBEYO RABIES IMMUNIZATION II. A COMPARISON OF THE ANTIGENICITY OF. HIGH EGG-PASSAGE AND DUCK-EMBRYO VACCINES ADMINISTERED INTRADERMALLY Downloaded from https://academic.oup.com/aje/article/74/1/1/136363 by guest on 30 September 2021 IN MAN BY PAUL R. SCHNURRENBERGER, GEORGE R. ANDERSON, JACK H. RUSSELL AND FREDERICK H. WENTWORTH* (Received for publication November 22, 1961) Several earlier studies have demon- been possible to make valid conclusions strated the feasibility of avian embryo concerning their relative efficiency. pre-exposure rabies vaccination in the It was the purpose of the study re- human population (1-4). The chick ported here to make such a direct com- embryo vaccine (HEP), which contains parison in a selected pre-exposure hu- high egg-passage attenuated rabies vi- man group. It was also our intent to rus, had been administered primarily in compare the results of 3 participating a series of 3 intradermal inoculations of laboratories performing parallel serum 0.2 ml each at 5-day intervals, with the neutralization studies in an effort to recommendation that a booster inocula- gain information relative to the sensi- tion of 0.2 ml be given subsequently. tivity and reproducibility of this test. Such a procedure has two advantages Freshman and sophomore veterinary over those presently employed for rabies students at the Ohio State University vaccination: it reduces the risk of Veterinary College were selected for test anaphylactic reactions and provides subjects.2 The vaccines used were desig- minimal inconvenience to the recipient. nated A and B.3 The students were In a recent report describing experi- assigned these letters at random and ence with the duck-embryo rabies vac- received the corresponding vaccine cine (DEV) (5), administered in the throughout the study. The subjects, same fashion as described for the HEP inoculators and laboratories were not vaccine, it was suggested that further informed of the true identity of the investigation of this material in high 2 risk groups might be profitable. Studies Our sincere appreciation is extended to Dr. John H. Helwig and the other faculty members reported to date were not designed to of the College of Veterinary Medicine, The compare the antigenicity of the two Ohio State University, who made this study vaccines directly and it has thus not possible. 3 We are grateful to Dr. J. M. Ruegsegger i Division of Communicable Diseases and Di- of Lederle Laboratories and Dr. F. B. Peck, Jr., vision of Laboratories, Ohio Department of of Eli Lilly & Company for furnishing the Health, Columbus, Ohio. vaccines used in this study. AM.J.HYG. 1961, VOL. 74: 1-6 SCHNURRENBEKGEB, ANDERSON, RUSSELL AND WENTWORTH vaccines until after the laboratory re- The sera were inactivated at 56 C sults were recorded. for 30 minutes. Four-fold dilutions of Two persons did the inoculating. This serum were mixed with an equal volume necessitated a study design which would of CVS fixed rabies virus and the mix- differentiate the effect of the vaccine ture was incubated in a water bath at and inoculator technique. This was ac- 37 C for 90 minutes. The challenge complished by alternating inoculators virus was diluted in 10 per cent inacti- so that no subject was given 3 inocula- vated guinea-pig serum in saline. After tions by the same person. incubation the sera were placed in ice Vaccines and vaccination procedures and 5 or 6 Swiss white mice weighing Downloaded from https://academic.oup.com/aje/article/74/1/1/136363 by guest on 30 September 2021 were the same as those described in approximately 10 grams each were in- previous communications (4, 5). The oculated intracerebrally with 0.03 ml of high egg-passage vaccine is a live prod- the serum virus mixture. The mice were uct, Flury strain, attenuated by 180 to observed for 15 days and 50 per cent 210 passages on chick embryos. Duck- mortality end points were calculated ac- embryo vaccine, however, is the CVS cording to the method of Eeed and strain of fixed virus, grown on duck Muench (6). Appropriate controls were embryos and inactivated with beta included with each test group. propiolactone. Both vaccines were ad- Laboratory EL used approximately 100 ministered in a series of 4 intradermal LD50 (5) of challenge virus; Laboratory inoculations of 0.2 ml on the medial as- L used 100 to 300 LD50 (7) ; and Lab- pect of the forearm. The right and left oratory O used approximately 300 LD50 forearms were alternated on successive of virus. inoculations. The primary series of 3 The antibody titer of the serum was inoculations was administered at 5-day expressed as the reciprocal of the final intervals while the fourth or booster serum dilution protecting 50 per cent dose was given 171 days after the of the mice. original inoculation. Blood samples were obtained prior to vaccination, 31 RESULTS days after the first inoculation, prior to the booster, and 7 and 30 days after the Pre-immunization sera from 119 sub- booster. jects with no history of prior antirabic treatment were examined. Three of Each serum specimen was divided into these had demonstrable neutralizing 3 aliquots and placed into screw-capped antibody. tubes. Aliquots obtained from each A total of 111 subjects with negative specimen were sent to the two pharma- pre-immunization sera received the pri- ceutical houses supplying the vaccine mary series of inoculations at the pre- while the remaining portions were stored scribed intervals and their postim- at — 20 C by our own laboratory until munization sera were tested. In each tested. of the three laboratories, antibody was The serum neutralization test (SN) demonstrated in a higher percentage of used by the participating laboratories 4 the subjects who had received the duek- was performed as follows: embryo vaccine than of those who had * We would like to acknowledge our thanks been given the high egg-passage vaccine to H. M. Powell of Eli Lilly and Company and (table 1). On the basis of the end- Jack Black of Lederle Laboratories who so graciously performed the serum neutralization point titers, this difference was statisti- studies. cally significant (P < 01) for the re- AVIAN EMBRYO RABIES IMMUNIZATION TABLE 1 Serum neutralizing liters follomng 3 intradermal inoculations of rabies vaccine, as determined by 3 laboratories on serum, aliquots Lab EL LabL LabO Vaccine HEP DEV HEP DEV HEP DEV Number of subjects 5C 55 56 52 21 24 Per cent Per cent Per cent Per cent Per cent Per cent Downloaded from https://academic.oup.com/aje/article/74/1/1/136363 by guest on 30 September 2021 Titer Negative 33.9 11.1 41.1 23.1 37.5 23.8 (reciprocal) 1-3 17.9 10.7 15.9 30.8 37.5 33.4 4-15 35.6 60.0 35.6 26.9 8.3 28.5 16-128 12.6 18.2 7.4 19.2 16.7 14.3 Total responding 66.1 88.9 58.9 76.9 62.5 76.2 suits obtained by Laboratory EL, but sults of Laboratory EL, which showed was not significant at this level for those the greatest apparent advantage for obtained by the other two laboratories. HEP, the statistical difference was ques- This apparent advantage did not per- tionable (P = .02). sist, for the results obtained by all 3 As may be seen in figure 1, there was laboratories on the pre-booster sera no essential difference between titers ob- failed to demonstrate a significant dif- tained on the 77 individuals with sera ference between the two groups. available 7 and 30 days after the fourth According to the results from all 3 inoculations. In view of this, it is not laboratories with reference to sera ob- surprising that no significant difference tained 7 days after the booster inocula- in response could be detected between tion, HEP vaccine appeared to be the two vaccines 30 days after the slightly more antigenic than DEV vac- booster (table 2). cine. However, judging from the re- Analysis of the results in terms of TABLE 2 Serum neutralizing titers SO days after a fourth intradermal injection of rabies vaccine, as determined by 8 laboratories on serum aliquots Lab EL Lab L LabO Vaccine HEP DEV HEP DEV HEP DEV Number of subjects 51 47 51 51 10 15 Per cent Per cent Per cent Per cent Per cent Per cent Titer Negative 7.7 14.9 11.8 21.6 10.0 20.0 (reciprocal) 1-3 2.0 4.3 9.8 7.8 20.0 26.6 4-15 17.5 19.1 15.7 27.5 20.0 26.7 16 or greater 72.8 61.7 62.7 43.1 50.0 26.7 Total responding 92.3 85.1 88.2 78.4 90.0 80.0 SCHNURRENBEHGER, ANDERSON, RUSSELL AND WENTWORTH • significant titer (1 in 5, Laboratory EL) / failed to exhibit any neutralizing ca- rf> pacity when tested in another labora- / / • tory. The subject from whom this was (> / ?8 o° obtained showed no antibody response to ( / o • / £• / °o° / 3 injections of the HBP vaccine. ( o ' o Q The relative sensitivity of the tech- o / niques as employed by the different 1 / V > O / laboratories is demonstrated by the 8 8 < / geometric mean antibody titers reported Downloaded from https://academic.oup.com/aje/article/74/1/1/136363 by guest on 30 September 2021 • / ft ••' / by the 3 laboratories on the post-pri- / / / mary sera. For Laboratory EL it was S 1 in 5.2; for L, 1 in 3.6; and 0, 1 in 3.1.
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