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Towards a second generation of inactivated vaccines
Views mettre, P. and Paoletti, E.A. Recombinant fowlpox virus induces protective immunity in non-avian species. Vaccine 1988, 6, 497 5 Tripathy, D.N. and Hanson, L.E. Immunity to fowlpox. Am. J. Vet. Res. 1975, 36, 541 6 Beveridge, W.I.B. and Hart, L. Pox diseases. In: Animal Health in Australia. Australian Government Publishing Service, Canberra, 1985 pp. 57-61 7 Nelson, J.B. The behaviour of poxviruses in the respiratory tract IV. The nasal instillation of fowlpox virus in chickens and mice. J. Exp. Med. 1941, 74, 203 8 Mayr, A. and Mahnel, H. Charakterisierung
eines vom Rhinozeros isolierten Huhnerpockenvirus. Arch. Ges. Virusforsch 1970, 31, 51 9 Da Massa, A.J. The role of Culex tarsalis in the transmission of fowlpox virus. Avian Dis. 1966, 10, 57 10 Taylor, J., Weinberg, R., Languet, B., Desmettre, P. and Paoletti, E. Fowlpox virus based recombinant vaccines. In: Technological Advances in Vaccine Development. UCLA Symposium, 1988, in press 11 Boyle, D.B. and Coupar, B.E.H. Construction of recombinant fowlpox viruses as vectors for poultry vaccines. Virus Res. 1988, 10, 343
Results of potency studies The results of potency studies are shown in Table 2. The hydrosoluble fraction gave the highest protection 2, higher even than with whole virus. T ho duration of immunity was stu J with this antigen. SPF animals were given two doses 2 weeks apart and challenged 180 days after primary immunization. A protection of 74% was obtained. In addition, the presence of a cell-mediated response was confirmed by the macrophage inhibition test. These two basic experiments on safety as well as on efficacy focused attention on the herpes hydrosoluble fraction which was chosen as the antigen for our vaccine. Further electrophoretic analysis showed its glycoprotein nature 4. Agar gel precipitation techniques enabled the standardization of the glycoprotein content in each dose of vaccine. The cat served as a model for the development of this type of subunit vaccine. Other applications have been developed: pseudorabies in swine, rhinopneumonitis in horse and infectious bovine rhinotracheitis in cattle.
Towards a second generation of inactivated vaccines Use of herpesvirus (feline rhinotracheitis virus) subunits for the prevention of cat flu Prevention o f feline upper respiratory disease, commonly known as "cat flu' is traditionally achieved by two types of vaccination: live vaccines administered by intranasal or by parenteral inoculation or parenteral inactivated vaccines with an adjuvant. Both of these have been associated with side effects. The objective of the development o f a new generation o f cat flu vaccines was to market a vaccine which was perfectly safe in all epidemiological situations and was as potent as the current products available on the market.
Efforts were targeted towards the herpes virus which was the most likely candidate for explaining the above-mentioned postvaccinal reactions. The herpes virus was first split into four different antigenic fractions, according to the method described by Fargeaud et al. 4 The properties of each fraction were studied with regard to safety in carrier cats and potency in specific-pathogen-free animals. S a f e t y in c a r r i e r cats Field reports of reactions following the use of inactivated vaccines were recorded in cats which had previously been exposed to feline herpesvirus and which had become carriers. To reproduce such phenomena experimentally, cats were artificially infected. The different antigenic fractions and two control antigens were inoculated into these cats by the intradermat route and the skin reactions were measured. The ability of the complete virus to induce hypersensitivity reactions was thus confirmed and the capsidic fraction was found to induce similar reactions. In contrast, cats inoculated 0264-410X/88/060468-02 $03.00 © 1988 Butterworth & Co. (Publishers) Ltd.
468
Vaccine, Vol. 6, December 1988
with the hydrosoluble fraction were free from such reactions, as were control cats (not carrying the herpes virus) (Table 1).
A.L. Saint-G~rand RhOne M&ieux, Lyon, France
Table 1 Demonstration of the cutaneous hypersensitivity state in recovering cats. Results are given as degree of intensity (in mm) of the reactions after 48 h Cat No.
Seroneutralization titre (Iog~0)
CV
PV
LF
HF
CF
CC
MC
Control 1 2 3 4 5
0 1.7 1.9 0.7 1.4 1.0
1 5 2 5 8 7
0 8 1 5 7 8
0 1 0 1 2 5
0 0 0 0 1 0
0 5 0 5 8 5
0 0 0 0 0 0
0 0 0 0 0 0
Reaction intensity (mm)
CV, Crude virus; PV, purified virus; LF, lipidic fraction; MC, culture medium; HF, hydrosoluble fraction; CF, capsidic fraction; CC, control cells
Table 2 Activity of the different fractions of feline herpes virus No.
Protection at challenge
Seroneutralization titre at day
Vaccine
of cats
0
14
28
49
(%)
CV PV LF HF HF (P) CF Controls
6 6 6 6 4 6 4
~<0.24 ~<0.24 ~<0.24 ~<0.24 ~<0.24 ~<0.24
0.54 0.40 0.30 0.28 40.24 ~<0.34
1.80 1.70 0.94 0.86 1.62 1.22 ~<0.24
2.30 2.36 2.14 2.47 2.91 2.40 1.59
64 65 46 76 82 52 0
Seroneutralization titre is expressed as the mean median protective dose (PD~o). CV, Crude virus; PV, purified virus; LF, lipidic fraction; HF, hydrosoluble fraction; HF (P), polymerized hydrosoluble fraction; CF, capsidic fraction
Letters
References 1 Benoit-Jeannin, C. Rhinotracheite f61ine. Virus et immunite. PhD thesis, Universit~ C. Bernard, Lyon, 1983 2 Code of Federal Regulations. Animals and Animal Products: Feline Calcivirus Vaccine Publication No. 113.149, USA, 1983, p. 447 3 Chappuis, G., Benoit-Jeannin, C. and Far-
geaud, D. Rhinotrach@ite f61ine: vaccin inactiv@ purifi@ et mod@le experimental. Dev. Biol. Standard. 1981, 52, 485 4 Fargeaud, D., Benoit-Jeannin, C., Kato, F. and Chappuis, G. Biochemical study of the feline herpesvirus. 1. Identification of glycoproteins by affinity. Arch. Virol. 1984, 80, 69 5 Gaskell, R.M. and Wardley, R.C. Feline viral
respiratory disease: a review with particular reference to its epizootology and control. J. Small Anita. Practice 1977, 19, 1 Gaskell, C. and Gaskell, R. Respiratory diseases of cats. In Practice, November 1980, 2,6 Kahn, D.E. Report on intranasal feline rhinotracheitis calicivirus vaccine criticized. VM/SAC, January 1977, 8
Letters Intramuscular versus subcutaneous injection for hepatitis B vaccine The site of injection is an important parameter of antibody responses against vaccines 1. Protection against hepatitis B (HBV) is achieved when anti-HBs antibodies are >10 IU 1-1 (Ref. 2). However, using the vaccination schedule proposed by the manufacturer, the antiHBs antibody level remains below this threshold in 68-93% of the subjects 1, depending on the route of injection, the origin of the vaccine and eventually, the age and sex of the patients. During a mass vaccination campaign of health care workers, 21% remained at sub-threshold antibody titres after three subcutaneous injections of Hevac vaccine (lot L5045, Pasteur Institute, Paris, France) in the suprascapular region. A n t i b o d y titres remained <10 IU 1-1 in 38% (54/142) of male and 17% (103/602) of female subjects (;(2 30, p<10-4). A random selection of 85 subjects from the group whose A b titre remained <10 IU 1-~ was made for an intramuscular (i.m.) injection in the deltoid or a subcutaneous challenge in the suprascapular region. After the fourth booster injection, anti-HBs antibody remained undetectable in 41% of the subjects, regardless of the site of injection (18/42, 17/43 for i.m. and subcutaneous, respectively). The magnitude of the antibody response in the other subjects was, however, clearly higher when the challenge injection was given intramuscularly (mean 62 IU 1-1, versus 222 IU 1-1, p < 5 x l 0 -3, Wilcoxon text). Two other studies have examined the role of the site of injection in low responders after anti-HBV vaccination. Horowitz compared deltoid versus intradermal boosters in subjects with low antibody titres 3 years after intradeltoid vaccination 3. Pead randomized gluteal versus deltoid boosters after intragluteal vaccination failure4. 0264-410X/88/060469-01 $03.00 ~) 1988 Butterworth & Co. (Publishers) Ltd.
In both studies, and in our findings, the site of booster injection had no effect on the proportion of responders. On the other hand, deltoid injection resulted in a larger antibody response than gluteal injection (usually into subcutaneous fat). Intradermal injection gave the same magnitude of antibody response as deltoid injection, but using less antigen (2 Ixg instead of 20 p.g). From these previous studies and our results, we suggest that poor immunizations to HBV vaccine are heterogeneous, resulting from either real refractory subjects, who do not respond to any kind of booster, or low responders who could be better vaccinated using a more immunogenic site of injection, such as the deltoid muscle. Unfortunately, unpleasant local reactions sometimes occur in normal subjects when vaccinated in the arm 5 or intradermally (42% in Ref. 3). Considering the high incidence of local reaction after intradermal injection, we propose, from a practical point of view, that the HBV vaccination should always be initiated by i.m. deltoid injection and subsequent
injections should be subcutaneous if any local reaction occurs.
Christian Fessard, Olivier Riche and Jacques H.M. Cohen Immunology Laboratory, R. Debrd Hospital, 51092 Reims, France References 1 Centers for Disease Control. Suboptimal response to hepatitis B vaccine given by injection into the buttock. Morbid. Mortal Week. Rep. 1985, 34, 105 2 Goudeau, A., Coursaget, P., Barin, F., Dubois, F., Chiron, J.P., Denis, F. and Diop Mar, I. Prevention of hepatitis B by active and passive-active immunization. In: 1981 International Symposium on Viral Hepatitis (Eds Szmuness, W., Alter, H.J. and Maynard, J.E.) Franklin Institute Press, Philadelphia, 1982, pp. 509-525 3 Horowitz, M.M, Ershler, W.B., McKinney, W.P. and Battiola, R.J. Duration of immunity after hepatitis B vaccination. Efficacy of lowdose booster vaccine. Ann. Intern. Med. 1988, 108, 185 4 Pead, P.J. Immune responses to hepatitis B vaccination in hospital staff. Biomed. Pharmacother. 1986, 40, 251 5 Dienstag, J.L, Werner, B.G., Polk, B.F. et a/. Hepatitis B vaccine in health care personnel: safety, immunogenicity and indicators of efficacy. Ann. Intern. Med. 1984, 101, 34
Blood group A substances are not present in currently available pneumococcal vaccine The short paper by Nurmi et al.1 appears to duplicate previously published data: and states that 'After pneumococcal vaccination, a long-lasting antibody response to blood group A substance was found in 10 of 11 subjects with O or B blood group but not in any of the nine
vaccinated subjects of A or AB." They base their observations on studies of 14-valent Pneumovax lot HY4689 (Fi) NC 0532. Lot HY4689 (Fi) NC 0532 tested by Nurmi et al. dates from 1978 and would have expired long ago: we therefore assume that the authors must Vaccine, Vol. 6, D e c e m b e r 1988
469
eines vom Rhinozeros isolierten Huhnerpockenvirus. Arch. Ges. Virusforsch 1970, 31, 51 9 Da Massa, A.J. The role of Culex tarsalis in the transmission of fowlpox virus. Avian Dis. 1966, 10, 57 10 Taylor, J., Weinberg, R., Languet, B., Desmettre, P. and Paoletti, E. Fowlpox virus based recombinant vaccines. In: Technological Advances in Vaccine Development. UCLA Symposium, 1988, in press 11 Boyle, D.B. and Coupar, B.E.H. Construction of recombinant fowlpox viruses as vectors for poultry vaccines. Virus Res. 1988, 10, 343
Results of potency studies The results of potency studies are shown in Table 2. The hydrosoluble fraction gave the highest protection 2, higher even than with whole virus. T ho duration of immunity was stu J with this antigen. SPF animals were given two doses 2 weeks apart and challenged 180 days after primary immunization. A protection of 74% was obtained. In addition, the presence of a cell-mediated response was confirmed by the macrophage inhibition test. These two basic experiments on safety as well as on efficacy focused attention on the herpes hydrosoluble fraction which was chosen as the antigen for our vaccine. Further electrophoretic analysis showed its glycoprotein nature 4. Agar gel precipitation techniques enabled the standardization of the glycoprotein content in each dose of vaccine. The cat served as a model for the development of this type of subunit vaccine. Other applications have been developed: pseudorabies in swine, rhinopneumonitis in horse and infectious bovine rhinotracheitis in cattle.
Towards a second generation of inactivated vaccines Use of herpesvirus (feline rhinotracheitis virus) subunits for the prevention of cat flu Prevention o f feline upper respiratory disease, commonly known as "cat flu' is traditionally achieved by two types of vaccination: live vaccines administered by intranasal or by parenteral inoculation or parenteral inactivated vaccines with an adjuvant. Both of these have been associated with side effects. The objective of the development o f a new generation o f cat flu vaccines was to market a vaccine which was perfectly safe in all epidemiological situations and was as potent as the current products available on the market.
Efforts were targeted towards the herpes virus which was the most likely candidate for explaining the above-mentioned postvaccinal reactions. The herpes virus was first split into four different antigenic fractions, according to the method described by Fargeaud et al. 4 The properties of each fraction were studied with regard to safety in carrier cats and potency in specific-pathogen-free animals. S a f e t y in c a r r i e r cats Field reports of reactions following the use of inactivated vaccines were recorded in cats which had previously been exposed to feline herpesvirus and which had become carriers. To reproduce such phenomena experimentally, cats were artificially infected. The different antigenic fractions and two control antigens were inoculated into these cats by the intradermat route and the skin reactions were measured. The ability of the complete virus to induce hypersensitivity reactions was thus confirmed and the capsidic fraction was found to induce similar reactions. In contrast, cats inoculated 0264-410X/88/060468-02 $03.00 © 1988 Butterworth & Co. (Publishers) Ltd.
468
Vaccine, Vol. 6, December 1988
with the hydrosoluble fraction were free from such reactions, as were control cats (not carrying the herpes virus) (Table 1).
A.L. Saint-G~rand RhOne M&ieux, Lyon, France
Table 1 Demonstration of the cutaneous hypersensitivity state in recovering cats. Results are given as degree of intensity (in mm) of the reactions after 48 h Cat No.
Seroneutralization titre (Iog~0)
CV
PV
LF
HF
CF
CC
MC
Control 1 2 3 4 5
0 1.7 1.9 0.7 1.4 1.0
1 5 2 5 8 7
0 8 1 5 7 8
0 1 0 1 2 5
0 0 0 0 1 0
0 5 0 5 8 5
0 0 0 0 0 0
0 0 0 0 0 0
Reaction intensity (mm)
CV, Crude virus; PV, purified virus; LF, lipidic fraction; MC, culture medium; HF, hydrosoluble fraction; CF, capsidic fraction; CC, control cells
Table 2 Activity of the different fractions of feline herpes virus No.
Protection at challenge
Seroneutralization titre at day
Vaccine
of cats
0
14
28
49
(%)
CV PV LF HF HF (P) CF Controls
6 6 6 6 4 6 4
~<0.24 ~<0.24 ~<0.24 ~<0.24 ~<0.24 ~<0.24
0.54 0.40 0.30 0.28 40.24 ~<0.34
1.80 1.70 0.94 0.86 1.62 1.22 ~<0.24
2.30 2.36 2.14 2.47 2.91 2.40 1.59
64 65 46 76 82 52 0
Seroneutralization titre is expressed as the mean median protective dose (PD~o). CV, Crude virus; PV, purified virus; LF, lipidic fraction; HF, hydrosoluble fraction; HF (P), polymerized hydrosoluble fraction; CF, capsidic fraction
Letters
References 1 Benoit-Jeannin, C. Rhinotracheite f61ine. Virus et immunite. PhD thesis, Universit~ C. Bernard, Lyon, 1983 2 Code of Federal Regulations. Animals and Animal Products: Feline Calcivirus Vaccine Publication No. 113.149, USA, 1983, p. 447 3 Chappuis, G., Benoit-Jeannin, C. and Far-
geaud, D. Rhinotrach@ite f61ine: vaccin inactiv@ purifi@ et mod@le experimental. Dev. Biol. Standard. 1981, 52, 485 4 Fargeaud, D., Benoit-Jeannin, C., Kato, F. and Chappuis, G. Biochemical study of the feline herpesvirus. 1. Identification of glycoproteins by affinity. Arch. Virol. 1984, 80, 69 5 Gaskell, R.M. and Wardley, R.C. Feline viral
respiratory disease: a review with particular reference to its epizootology and control. J. Small Anita. Practice 1977, 19, 1 Gaskell, C. and Gaskell, R. Respiratory diseases of cats. In Practice, November 1980, 2,6 Kahn, D.E. Report on intranasal feline rhinotracheitis calicivirus vaccine criticized. VM/SAC, January 1977, 8
Letters Intramuscular versus subcutaneous injection for hepatitis B vaccine The site of injection is an important parameter of antibody responses against vaccines 1. Protection against hepatitis B (HBV) is achieved when anti-HBs antibodies are >10 IU 1-1 (Ref. 2). However, using the vaccination schedule proposed by the manufacturer, the antiHBs antibody level remains below this threshold in 68-93% of the subjects 1, depending on the route of injection, the origin of the vaccine and eventually, the age and sex of the patients. During a mass vaccination campaign of health care workers, 21% remained at sub-threshold antibody titres after three subcutaneous injections of Hevac vaccine (lot L5045, Pasteur Institute, Paris, France) in the suprascapular region. A n t i b o d y titres remained <10 IU 1-1 in 38% (54/142) of male and 17% (103/602) of female subjects (;(2 30, p<10-4). A random selection of 85 subjects from the group whose A b titre remained <10 IU 1-~ was made for an intramuscular (i.m.) injection in the deltoid or a subcutaneous challenge in the suprascapular region. After the fourth booster injection, anti-HBs antibody remained undetectable in 41% of the subjects, regardless of the site of injection (18/42, 17/43 for i.m. and subcutaneous, respectively). The magnitude of the antibody response in the other subjects was, however, clearly higher when the challenge injection was given intramuscularly (mean 62 IU 1-1, versus 222 IU 1-1, p < 5 x l 0 -3, Wilcoxon text). Two other studies have examined the role of the site of injection in low responders after anti-HBV vaccination. Horowitz compared deltoid versus intradermal boosters in subjects with low antibody titres 3 years after intradeltoid vaccination 3. Pead randomized gluteal versus deltoid boosters after intragluteal vaccination failure4. 0264-410X/88/060469-01 $03.00 ~) 1988 Butterworth & Co. (Publishers) Ltd.
In both studies, and in our findings, the site of booster injection had no effect on the proportion of responders. On the other hand, deltoid injection resulted in a larger antibody response than gluteal injection (usually into subcutaneous fat). Intradermal injection gave the same magnitude of antibody response as deltoid injection, but using less antigen (2 Ixg instead of 20 p.g). From these previous studies and our results, we suggest that poor immunizations to HBV vaccine are heterogeneous, resulting from either real refractory subjects, who do not respond to any kind of booster, or low responders who could be better vaccinated using a more immunogenic site of injection, such as the deltoid muscle. Unfortunately, unpleasant local reactions sometimes occur in normal subjects when vaccinated in the arm 5 or intradermally (42% in Ref. 3). Considering the high incidence of local reaction after intradermal injection, we propose, from a practical point of view, that the HBV vaccination should always be initiated by i.m. deltoid injection and subsequent
injections should be subcutaneous if any local reaction occurs.
Christian Fessard, Olivier Riche and Jacques H.M. Cohen Immunology Laboratory, R. Debrd Hospital, 51092 Reims, France References 1 Centers for Disease Control. Suboptimal response to hepatitis B vaccine given by injection into the buttock. Morbid. Mortal Week. Rep. 1985, 34, 105 2 Goudeau, A., Coursaget, P., Barin, F., Dubois, F., Chiron, J.P., Denis, F. and Diop Mar, I. Prevention of hepatitis B by active and passive-active immunization. In: 1981 International Symposium on Viral Hepatitis (Eds Szmuness, W., Alter, H.J. and Maynard, J.E.) Franklin Institute Press, Philadelphia, 1982, pp. 509-525 3 Horowitz, M.M, Ershler, W.B., McKinney, W.P. and Battiola, R.J. Duration of immunity after hepatitis B vaccination. Efficacy of lowdose booster vaccine. Ann. Intern. Med. 1988, 108, 185 4 Pead, P.J. Immune responses to hepatitis B vaccination in hospital staff. Biomed. Pharmacother. 1986, 40, 251 5 Dienstag, J.L, Werner, B.G., Polk, B.F. et a/. Hepatitis B vaccine in health care personnel: safety, immunogenicity and indicators of efficacy. Ann. Intern. Med. 1984, 101, 34
Blood group A substances are not present in currently available pneumococcal vaccine The short paper by Nurmi et al.1 appears to duplicate previously published data: and states that 'After pneumococcal vaccination, a long-lasting antibody response to blood group A substance was found in 10 of 11 subjects with O or B blood group but not in any of the nine
vaccinated subjects of A or AB." They base their observations on studies of 14-valent Pneumovax lot HY4689 (Fi) NC 0532. Lot HY4689 (Fi) NC 0532 tested by Nurmi et al. dates from 1978 and would have expired long ago: we therefore assume that the authors must Vaccine, Vol. 6, D e c e m b e r 1988
469