Comp. lmmun. Microbiol. infect. Dis., Vol. 1, pp. 1 9 3 - 1 9 6 . © Pergamon Press Ltd., 1979. Printed in Great Britain
0147-9571/79/0101-0193502.00/0
C O M B I N E D V A C C I N A T I O N OF CATS A G A I N S T P A N L E U C O P E N I A A N D RABIES A. BRUN, G. CHAPPUIS, P. PRECAUSTA, J. P. SOULEBOT a n d J. TERRt~ Iffa-Merieux, 254, rue Marcel M6rieux, 69007 Lyon, France
Abstract--In a previous paper [3], we demonstrated that it was possible to vaccinate cats with a combined vaccine against Panleucopenia and Rabies. The purpose of this paper is to sum up the tests for safety and potency, carried out in cats vaccinated with a new combined vaccine against Panleucopenia and Rabies. This vaccine is different from that mentioned in the previous paper. The new Rabies vaccine is a vaccine adjuvanted by aluminium hydroxide. First of all, we shall examine the means and methods implemented and then, present the comparative results obtained with the two types of Rabies vaccine (whether adjuvanted or not) combined with the Panleucopenia live vaccine.
Key words: Panleucopenia, Rabies, combined vaccine
VACCINATION DE CHATS A L'AIDE D'UN VACCIN ASSOCIE, PANLEUCOPENIE-RAGE Resume--Dans un precedent article [3], nous avions d6montre la possibilit+ de vacciner le chat l'aide d'un vaccin associ6 de la Panleucop~nie et de la Rage. Le present travail a pour but de faire le bilan des essais d'innocuit6 et d'activit~ r+alis+s sur des chats vaccin6s ~, l'aide d'un nouveau vaccin associ+, Panleucop+nie-Rage, diff6rent du pr6c6dent. Le nouveau vaccin antirabique est un vaccin adjuv+ par un gel d'hydroxyde d'alumine. Nous envisagerons tout d'abord les moyens et les m6thodes mis en oeuvre, et pr6senterons les rdsultats comparatifs obtenus avec les deux types de vaccin antirabique (adjuv6 ou non) associ+ au vaccin vivant de la Panleucop+nie.
Mots clefs: Panleucopenie, Rage, vaccin associ+
MATERIALS AND METHOD
1.1. The following vaccines are used 1.1.1. Panleucopenia monovalent vaccine. T h e m o d i f i e d virus u s e d for t h e p r e p a r a t i o n o f t h e P a n l e u c o p e n i a m o n o v a l e n t v a c c i n e is c u l t u r e d o n c a t k i d n e y cells. T h e v a c c i n e is i s s u e d u n d e r t h e f r e e z e - d r i e d f o r m a n d its titre is a b o v e 10 3 D C P E s 0 p e r dose. 1.1.2. Rabies monovalent vaccine. T h e r a b i e s v i r u s is m u l t i p l i e d o n cells c o m i n g f r o m n e w b o r n h a m s t e r s . It is c o m p l e t e l y i n a c t i v a t e d w i t h b e t a p r o p i o l a c t o n e [ 1 ]. T h e v a c c i n e is i s s u e d u n d e r t h e liquid f o r m . I n t h e first case, the v a c c i n e c o n t a i n s n o a d j u v a n t . I n t h e s e c o n d case, it c o n t a i n s a n a d j u v a n t : a l u m i n i u m h y d r o x i d e . T h e R a b i e s liquid v a c c i n e is u s e d to dilute t h e P a n l e u c o p e n i a f r e e z e - d r i e d v a c c i n e . T h e v o l u m e o f a d o s e o f v a c c i n e is 1 ml. 193
194
A. BRUN, G. CHAPPUIS,P. PRECAUSTA,J. P. SOULEBOTand J. TERRI~
1.2. Animals The 39 cats of European breed of 3 to 5 months of age come from our breeding farm in protected atmosphere [2] and are sensitive to Panleucopenia and Rabies. 1.3. Experiments in cats All the vaccines described have met the requirements of the safety and potency tests classically carried out in our Institute: --specific and non specific safety in mice, guinea-pigs and cats; - - p o t e n c y : by titration on cell culture for the Panleucopenia vaccine and NIH test on mice for the Rabies vaccine. We shall describe here only the tests carried out in cats. The animals are vaccinated by the subcutaneous or intramuscular route on Day 0 with one dose of vaccine. The clinical observation of animals and regular leucocyte counts make it possible to check the safety of the vaccines used. Some cats are challenged against Panleucopenia on Day 28 by the intraperitoneal route with the challenge strain PLI CU4. Leucocyte counts are carried out within the days following the challenge. The animals are sacrificed 14 days after the challenge. Blood samples are taken on Days 0 and 28. They are used for the potency test carried out by means of the antibodies titration.
1.4. Antibodies titration The titration of Panleucopenia seroneutralizing antibodies is carried out by the seroneutralization technique on kitten kidney cells [3]. The titration of Rabies seroneutralizing antibodies is carried out by the seroneutralization technique in mice [3]. All the titres are expressed in log of basis 10 of the inverse of the neutralizing dilution 50% (before mixing with the virus).
2. RESULTS
2.1. Safety and potency tests of the Panleucopenia valence All the cats vaccinated with the combined vaccines remain in good health and the number of white blood cells never vary significantly. It is possible to follow on Fig. 1. the evolution of the average number of white blood cells after challenge in cats vaccinated with: - - t h e non adjuvanted combined vaccine against Panleucopenia and Rabies; - - t h e adjuvanted combined vaccine against Panleucopenia and Rabies and in control cats. The eleven control cats show a decrease by 75% in the number of white blood cells and 4 out of them die from Panleucopenia. But no significant variation in the number of white blood cells nor clinical manifestation of Panleucopenia were noted in the two groups of vaccinated animals.
Combined vaccination of cats against panleucopenia and rabies o o
o
E
~z
+,o
E
ioooo .....S Iooo J
Panleucopenia+ rabies/ adjuvanted Vaccinated .... Panleucopenio÷rabies non adjuvanted Controls
.c
I
% d z
II dO
5
4
1
I
I
I
6
8
14
Time, days Fig. 1. Potency of the Panleucopenia after the virulent challenge.
Table 1. Rabies antibodies titre Without adjuvant
With adjuvant
1.35 1.88 2.10 2.35 1.30 1.24 1.95 1.24 1.60 1.66 1.30 1.24 2.10
1.95 1.86 2.22 2.55 2.70 2.44 2.70 2.70 2.70 2.70 2.70 2.70 2.70 2.55 1.02
m:
1.64 _+0.24
m:
2.4 + 0.26
195
196
A. BRUN, G. CHAPPUIS,P. PRECAUSTA,J. P. SOULEBOTand J. TERRE
The average antibodies titre on D a y 28 before challenge is: 3. l for the animals vaccinated with the non adjuvanted vaccine against Panleucopenia and Rabies. 3.3 for the animals vaccinated with the adjuvanted vaccine against Panleucopenia and Rabies. 2.2. Safety and potency tests o f the Rabies valence The vaccinated cats remained in good health. Table 1 shows the antibodies titre on D a y 28. The average antibodies titre is: 1.64 for the 13 cats vaccinated with the non adjuvanted vaccine against Panleucopenia and Rabies. 2.4 for the 15 cats vaccinated with the adjuvanted vaccine against Panleucopenia and Rabies. These significantly different results were obtained after a single vaccination. Table 2. Titre of Rabies neutralizing antibodies after the first vaccination Rabies vaccine without adjuvant
Rabies vaccine with adjuvant
1.64_+ 0.24 n=13
2.4 _+0.26 n=15
3. CONCLUSION Two combined vaccines against Panleucopenia and Rabies are compared. These vaccines differ in the presence or absence of an adjuvant, aluminium hydroxide, in the Rabies valence. These vaccines are perfectly safe and of equivalent potency for the Panleucopenia valence. The adjuvanted vaccine causes the a p p e a r a n c e o f neutralizing antibodies at a higher level for the Rabies valence, about 6 times higher after the first vaccination.
REFERENCES 1. Precausta, P., Vaccin antirabique inactiv~ ~.usage vdt6rinaire pr6par6 f partir de culture cellulaire, lmmunobiol. Stand. lnter. Symp. rabies, II, Lyon, 9-11 decembre 1972, p. 162 Karger, Basel (1974). 2. Brun, A. and Chappuis, G., L'elevage de chats et de chiens virus free. Rev. Mdd. Vdt. 127, 889-899 (1976). 3. Brun, A., Chappuis, G., Precausta, P. and Terr~, J., lmmunisation des chats contre la Panleucopenie et la Rage. Rev. Mdd. Vdt. 127, 1575-1580(1976).
0147-9571/79/0101-0193502.00/0
C O M B I N E D V A C C I N A T I O N OF CATS A G A I N S T P A N L E U C O P E N I A A N D RABIES A. BRUN, G. CHAPPUIS, P. PRECAUSTA, J. P. SOULEBOT a n d J. TERRt~ Iffa-Merieux, 254, rue Marcel M6rieux, 69007 Lyon, France
Abstract--In a previous paper [3], we demonstrated that it was possible to vaccinate cats with a combined vaccine against Panleucopenia and Rabies. The purpose of this paper is to sum up the tests for safety and potency, carried out in cats vaccinated with a new combined vaccine against Panleucopenia and Rabies. This vaccine is different from that mentioned in the previous paper. The new Rabies vaccine is a vaccine adjuvanted by aluminium hydroxide. First of all, we shall examine the means and methods implemented and then, present the comparative results obtained with the two types of Rabies vaccine (whether adjuvanted or not) combined with the Panleucopenia live vaccine.
Key words: Panleucopenia, Rabies, combined vaccine
VACCINATION DE CHATS A L'AIDE D'UN VACCIN ASSOCIE, PANLEUCOPENIE-RAGE Resume--Dans un precedent article [3], nous avions d6montre la possibilit+ de vacciner le chat l'aide d'un vaccin associ6 de la Panleucop~nie et de la Rage. Le present travail a pour but de faire le bilan des essais d'innocuit6 et d'activit~ r+alis+s sur des chats vaccin6s ~, l'aide d'un nouveau vaccin associ+, Panleucop+nie-Rage, diff6rent du pr6c6dent. Le nouveau vaccin antirabique est un vaccin adjuv+ par un gel d'hydroxyde d'alumine. Nous envisagerons tout d'abord les moyens et les m6thodes mis en oeuvre, et pr6senterons les rdsultats comparatifs obtenus avec les deux types de vaccin antirabique (adjuv6 ou non) associ+ au vaccin vivant de la Panleucop+nie.
Mots clefs: Panleucopenie, Rage, vaccin associ+
MATERIALS AND METHOD
1.1. The following vaccines are used 1.1.1. Panleucopenia monovalent vaccine. T h e m o d i f i e d virus u s e d for t h e p r e p a r a t i o n o f t h e P a n l e u c o p e n i a m o n o v a l e n t v a c c i n e is c u l t u r e d o n c a t k i d n e y cells. T h e v a c c i n e is i s s u e d u n d e r t h e f r e e z e - d r i e d f o r m a n d its titre is a b o v e 10 3 D C P E s 0 p e r dose. 1.1.2. Rabies monovalent vaccine. T h e r a b i e s v i r u s is m u l t i p l i e d o n cells c o m i n g f r o m n e w b o r n h a m s t e r s . It is c o m p l e t e l y i n a c t i v a t e d w i t h b e t a p r o p i o l a c t o n e [ 1 ]. T h e v a c c i n e is i s s u e d u n d e r t h e liquid f o r m . I n t h e first case, the v a c c i n e c o n t a i n s n o a d j u v a n t . I n t h e s e c o n d case, it c o n t a i n s a n a d j u v a n t : a l u m i n i u m h y d r o x i d e . T h e R a b i e s liquid v a c c i n e is u s e d to dilute t h e P a n l e u c o p e n i a f r e e z e - d r i e d v a c c i n e . T h e v o l u m e o f a d o s e o f v a c c i n e is 1 ml. 193
194
A. BRUN, G. CHAPPUIS,P. PRECAUSTA,J. P. SOULEBOTand J. TERRI~
1.2. Animals The 39 cats of European breed of 3 to 5 months of age come from our breeding farm in protected atmosphere [2] and are sensitive to Panleucopenia and Rabies. 1.3. Experiments in cats All the vaccines described have met the requirements of the safety and potency tests classically carried out in our Institute: --specific and non specific safety in mice, guinea-pigs and cats; - - p o t e n c y : by titration on cell culture for the Panleucopenia vaccine and NIH test on mice for the Rabies vaccine. We shall describe here only the tests carried out in cats. The animals are vaccinated by the subcutaneous or intramuscular route on Day 0 with one dose of vaccine. The clinical observation of animals and regular leucocyte counts make it possible to check the safety of the vaccines used. Some cats are challenged against Panleucopenia on Day 28 by the intraperitoneal route with the challenge strain PLI CU4. Leucocyte counts are carried out within the days following the challenge. The animals are sacrificed 14 days after the challenge. Blood samples are taken on Days 0 and 28. They are used for the potency test carried out by means of the antibodies titration.
1.4. Antibodies titration The titration of Panleucopenia seroneutralizing antibodies is carried out by the seroneutralization technique on kitten kidney cells [3]. The titration of Rabies seroneutralizing antibodies is carried out by the seroneutralization technique in mice [3]. All the titres are expressed in log of basis 10 of the inverse of the neutralizing dilution 50% (before mixing with the virus).
2. RESULTS
2.1. Safety and potency tests of the Panleucopenia valence All the cats vaccinated with the combined vaccines remain in good health and the number of white blood cells never vary significantly. It is possible to follow on Fig. 1. the evolution of the average number of white blood cells after challenge in cats vaccinated with: - - t h e non adjuvanted combined vaccine against Panleucopenia and Rabies; - - t h e adjuvanted combined vaccine against Panleucopenia and Rabies and in control cats. The eleven control cats show a decrease by 75% in the number of white blood cells and 4 out of them die from Panleucopenia. But no significant variation in the number of white blood cells nor clinical manifestation of Panleucopenia were noted in the two groups of vaccinated animals.
Combined vaccination of cats against panleucopenia and rabies o o
o
E
~z
+,o
E
ioooo .....S Iooo J
Panleucopenia+ rabies/ adjuvanted Vaccinated .... Panleucopenio÷rabies non adjuvanted Controls
.c
I
% d z
II dO
5
4
1
I
I
I
6
8
14
Time, days Fig. 1. Potency of the Panleucopenia after the virulent challenge.
Table 1. Rabies antibodies titre Without adjuvant
With adjuvant
1.35 1.88 2.10 2.35 1.30 1.24 1.95 1.24 1.60 1.66 1.30 1.24 2.10
1.95 1.86 2.22 2.55 2.70 2.44 2.70 2.70 2.70 2.70 2.70 2.70 2.70 2.55 1.02
m:
1.64 _+0.24
m:
2.4 + 0.26
195
196
A. BRUN, G. CHAPPUIS,P. PRECAUSTA,J. P. SOULEBOTand J. TERRE
The average antibodies titre on D a y 28 before challenge is: 3. l for the animals vaccinated with the non adjuvanted vaccine against Panleucopenia and Rabies. 3.3 for the animals vaccinated with the adjuvanted vaccine against Panleucopenia and Rabies. 2.2. Safety and potency tests o f the Rabies valence The vaccinated cats remained in good health. Table 1 shows the antibodies titre on D a y 28. The average antibodies titre is: 1.64 for the 13 cats vaccinated with the non adjuvanted vaccine against Panleucopenia and Rabies. 2.4 for the 15 cats vaccinated with the adjuvanted vaccine against Panleucopenia and Rabies. These significantly different results were obtained after a single vaccination. Table 2. Titre of Rabies neutralizing antibodies after the first vaccination Rabies vaccine without adjuvant
Rabies vaccine with adjuvant
1.64_+ 0.24 n=13
2.4 _+0.26 n=15
3. CONCLUSION Two combined vaccines against Panleucopenia and Rabies are compared. These vaccines differ in the presence or absence of an adjuvant, aluminium hydroxide, in the Rabies valence. These vaccines are perfectly safe and of equivalent potency for the Panleucopenia valence. The adjuvanted vaccine causes the a p p e a r a n c e o f neutralizing antibodies at a higher level for the Rabies valence, about 6 times higher after the first vaccination.
REFERENCES 1. Precausta, P., Vaccin antirabique inactiv~ ~.usage vdt6rinaire pr6par6 f partir de culture cellulaire, lmmunobiol. Stand. lnter. Symp. rabies, II, Lyon, 9-11 decembre 1972, p. 162 Karger, Basel (1974). 2. Brun, A. and Chappuis, G., L'elevage de chats et de chiens virus free. Rev. Mdd. Vdt. 127, 889-899 (1976). 3. Brun, A., Chappuis, G., Precausta, P. and Terr~, J., lmmunisation des chats contre la Panleucopenie et la Rage. Rev. Mdd. Vdt. 127, 1575-1580(1976).
