Veterinary Immunology and Immunopathology: Sciencedirect

Download as pdf or txt
Download as pdf or txt
You are on page 1of 3

Veterinary Immunology and Immunopathology 219 (2020) 109985

Contents lists available at ScienceDirect

Veterinary Immunology and Immunopathology


journal homepage: www.elsevier.com/locate/vetimm

A canine vaccine against Leptospira serovars Icterohaemorrhagiae, Canicola T


and Grippotyphosa provides cross protection against Leptospira serovar
Copenhageni
J. Bouveta,*, L. Lemaitrea, C. Carioub, M. Scottoa, C. Blainb, F. Oberlia, L. Cupillardb, P.M. Guigala
a
Boehringer Ingelheim Animal Health, Centre de Recherche de Saint-Vulbas, Parc Industriel de la Plaine de l’Ain, 805 Allée des Cyprès, 01150 Saint-Vulbas, France
b
Boehringer Ingelheim Animal Health, Lyon Porte des Alpes, rue de l’aviation, 69800 Saint-Priest, France

ARTICLE INFO ABSTRACT

Keywords: Efficacy of the Leptospira components of multivalent vaccine DAPPi-L was previously demonstrated against
Leptospirosis virulent challenge with three serovars of Leptospira interrogans (Canicola, Icterohaemorrhagiae and
Dog Grippotyphosa) carried out 14 days after primary vaccination.
Vaccine In this study we demonstrate that this vaccine provides, two weeks after vaccination, an additional protection
Cross protection
(prevention of mortality, clinical signs, renal infection, bacterial excretion, renal carriage and renal lesions)
Copenhageni
against fatal leptospirosis due to Leptospira interrogans serovar Copenhageni (serovar of major medical im-
portance).

1. Introduction 2. Materials and methods

Leptospirosis, one of the major zoonotic diseases, is due to a bac- Fourteen Beagle puppies2 aged between 8 and 9 weeks were pro-
terial infection involving multiple serovars from pathogenic Leptospira vided by a commercial supplier (kennel regularly controlled for ser-
spp affecting a broad range of mammals (Levett, 2001). In humans ology against pathogenic Leptospira) and allocated randomly into two
severe leptospirosis is most frequently associated to serogroup Icter- groups of 7 animals each according to age and litter. These animals
ohaemorrhagiae, in particular serovar Copenhageni (Bharti et al., were seronegative for serovars Icterohaemorrhagiae, Canicola, Grip-
2003). potyphosa, Pomona, Sejroe, Hebdomonadis, Australis, Autumnalis and
In dogs, epidemiological investigation based on serological data in Hardjo (MAT analysis performed by National Reference Centre for
Europe shows that serovars Icterohaemorrhagiae and Copenhageni are Leptospira – CNRL, Pasteur Institute, Paris, France). On days 0 and 28,
highly prevalent, with up to 20 % of positive dogs in some countries vaccinated puppies received one subcutaneous injection of EUR-
(Ellis, 2010; Delaude et al., 2017). ICAN®DAPPi-Lmulti, a freeze-dried vaccine containing four live atte-
We have previously demonstrated that EURICAN®DAPPi-Lmulti1 nuated organisms (canine distemper virus, canine adenovirus type 2,
provides a full protection against fatal leptospirosis caused by Leptospira canine parvovirus type 2 and canine parainfluenza virus) rehydrated
interrogans serogroup Icterohaemorrhagiae serovar Icterohaemor- with a liquid vaccine containing three inactivated strains of Leptospira
rhagiae with protection against mortality, clinical signs of disease, in- interrogans (serovars Canicola, Icterohaemorrhagiae and Grippoty-
fection, bacterial excretion, renal carriage and renal lesions (Bouvet phosa) whereas control puppies received one subcutaneous injection of
et al., 2016). In this paper we describe the level of protection brought only EURICAN®DAPPi. Two weeks after the second vaccine injection,
by the same vaccine against a severe leptospirosis challenge with Lep- each animal was intraperitoneally challenged with 1.9 × 109 Leptospira
tospira interrogans serogroup Icterohaemorrhagiae serovar Copenha- interrogans serogroup Icterohaemorrhagiae serovar Copenhageni strain
geni. (isolated from diseased dog and provided by KIT Royal Tropical


Corresponding author.
E-mail address: [email protected] (J. Bouvet).
1
Eurican is registered trademark of Boehringer Ingelheim in France and elsewhere
2
all animal experiments conducted for this study have been reviewed by the Boehringer Ingelheim Ethical Committee (registered under n°13 at the French Ministère de
l’Education Nationale, de l’Enseignement Supérieur et de la Recherche) before conducting the study. Approval confirms that all experiments conform to the relevant regulatory
standards defined by the European and French Laws (directive EU2010/63 and Decret n° 2013-118)

https://doi.org/10.1016/j.vetimm.2019.109985
Received 17 December 2018; Received in revised form 13 November 2019; Accepted 14 November 2019
0165-2427/ © 2019 Elsevier B.V. All rights reserved.
J. Bouvet, et al. Veterinary Immunology and Immunopathology 219 (2020) 109985

Table 1 period) and GBS (Global Biochemical Score, defined for each animal as
Clinical signs following challenge in the 7 control dogs (percentage and the sum of all daily scores derived from biochemical and haematolo-
number of affected dogs, and maximum duration of clinical sign). gical results during post-challenge monitoring period) of vaccinated
Euthanasia 57 % (4) and control animals (Table 2).
Apathy/prostration 43 % (3) - 2 days ICS = 50 All blood and urine samples were negative before challenge. After
Dehydration 71 % (4) - 4 days ICS = 48 challenge, controls showed two to six positive blood samples, one to six
Vomiting 29 % (2) - 2 days ICS = 20
positive urine samples, all had positive kidney samples and moderate to
Diarrhoea 29 % (2) - 1 day ICS = 13
Icterus/Petechiae 57 % (4) - 4 days ICS = 92 severe kidney lesions. On the contrary, all vaccinated puppies remained
GCS 223 negative for blood, urine and kidney, without any kidney lesions (tables
2 and 3).
ICS = Individual Clinical Score. Protection against leptospirosis through inactivated vaccines relies
GCS = Global Clinical Score. mainly on humoral immunity (Adler and Pena Moctezuma, 2010), di-
rected towards Leptospira lipopolysaccharide (LPS) molecules as evi-
Institute, Amsterdam, The Netherlands; identity confirmed by CNRL, denced by the transfer of passive immunity through anti-LPS antibodies
using molecular analysis), previously cultured in Ellinghausen-McCul- (Jost et al., 1986). As a consequence, protection elicited by these vac-
lough-Johnson-Harris medium. Non-blinded clinical examination, la- cines is restricted to infection by serovars with related LPS antigen
boratory analyses (biological parameters) and statistical analysis of the (Adler, 2015).
results were performed as described in Bouvet et al. (2016). Detection Although it is generally accepted that strains entering in the com-
of leptospires in blood, urine and kidney was carried out using a position of bacterin vaccines can provide protection against other ser-
quantitative real-time PCR assay targeting LipL32 gene (Bourhy et al., ovars within the same serogroup, only few supportive experimental
2011; Blanchard et al., 2018). Each sample was tested in duplicate and evidence is available, mainly in rodent models (Sonrier et al., 2001;
result was expressed as negative (no detection of LipL32 in both assay Rosario et al., 2012). Proof of efficacy in the target species against all
samples) or positive (at least one assay sample with detection of six clinical criteria is necessary to comply with European regulations
LipL32). and claim such intra-serogoup protection of commercial vaccines.
Our study demonstrates that a primary course of two doses of
3. Results and discussion EURICAN® DAPPi-Lmulti, provided an onset of immunity of 2 weeks
post-vaccination with full protection against mortality, clinical signs,
No adverse events were recorded following the vaccination and all renal infection, bacterial excretion, renal carriage and renal lesions
animals remained healthy until challenge was performed. caused by L. interrogans serogroup Icterohaemorrhagiae serovar
After challenge, all control puppies had at least two altered biolo- Copenhageni. This vaccine can thus prevent infection of dogs against a
gical parameters and 5 animals out of 7 (71 %) were clinically diseased serovar of major medical importance in the context of a zoonotic dis-
with alteration of general condition, dehydration, digestive signs (vo- ease
miting, diarrhoea) and cutaneo-mucosal signs (icterus, petechiae) di-
versely associated (Table 1). In vaccinated puppies, no change was
recorded for all biological parameters. All animals remained in good Declaration of Competing Interest
health and did not show any clinical signs. A significant difference was
found between GCS (Global Clinical Score, defined for each animal as Authors of this paper are employees of Boehringer Ingelheim
the sum of all daily clinical scores during post-challenge monitoring Animal Health, the manufacturer of EURICAN®DAPPi-Lmulti.

Table 2
Percentage and number of affected animals - Statistical results - Vaccine claims for Copenhageni serovar.
Parameter V C p value* Claim

euthanasia 0 % (0/7) 57 % (4/7) – • prevent mortality


clinical signs
GBS
0
0
%
%
(0/7)
(0/7)
71 % (5/7)
100 % (7/7)
0.011
< 0.001
• prevent clinical signs
leptospiraemia 0 % (0/7) 100 % (7/7) < 0.001 • prevent renal infection
leptospiruria 0 % (0/7) 100 % (7/7) < 0.001 • prevent bacterial excretion
positive kidney 0 % (0/7) 100 % (7/7) < 0.001 • prevent renal carriage
kidney lesions 0 % (0/7) 100 % (7/7) < 0.001 • prevent renal lesions
V: vaccinated dogs; C: control dogs.
* in bold, significant difference between groups.
GBS: Global Biochemical and hematological Score.

Table 3
Percentage and number of positive dogs for blood, urine and kidney and histopathology (kidney).
Days after challenge

Blood Urine Kidney

Group 2 3 4 5 8 11 3 5 8 11 14 21 28 detection histology


V 0% 0% 0% 0% 0% 0% 0% 0% 0 % (0/7) 0 % (0/7) 0% 0% 0% 0 % (0/7) 0 % (0/7)
(0/7) (0/7) (0/7) (0/7) (0/7) (0/7) (0/7) (0/7) (0/7) (0/7) (0/7)
C 57 % 86 % 71 % 57 % 43 % 20 % 0% 33 % 100 % 100 % 100 % 100 % 100 % 100 % (7/7) 100 % (7/7)
(4/7) (6/7) (5/7) (4/7) (3/7) (4/5*) (0/7) (2/6**) (5/5**) (5/5*) (3/3) (3/3) (3/3)

V= vaccinated dogs; C= control dogs (*one dog euthanised on day 6, on dog euthanised on day 8, one dog euthanised on day 9 and one dog euthanised on day 11 -
last sample accounted at the next timepoint; ** no urine sample for one animal on T5 and 2 animals on T8).

2
J. Bouvet, et al. Veterinary Immunology and Immunopathology 219 (2020) 109985

Acknowledgement Bourhy, P., Bremont, S., Zinini, F., Giry, C., Picardeau, M., 2011. Comparison of real-time
PCR assays for detection of pathogenic Leptospira spp. In blood and identification of
variations in target sequences. J. Clin. Microbiol. 49, 2154–2160.
The authors wish to acknowledge technicians from Boehringer Bouvet, J., Cariou, C., Valfort, W., Villard, S., Hilaire, F., Oberli, F., Cupillard, L., Guigal,
Ingelheim Animal Health R&D Animal Science and Welfare department, P.M., 2016. Efficacy of a multivalent DAPPi-Lmulti canine vaccine against mortality,
Research Laboratory and Clinical Analysis Laboratory for their con- clinical signs, infection, bacterial excretion, renal carriage and renal lesions caused by
Leptospira experimental challenges. Vaccine Rep 6, 23–28.
tribution to this work. Delaude, A., Rodriguez-Campos, S., Dreyfus, A., Counotte, M.J., Francey, T.,
Schweighauser, A., Lettry, S., Schuller, S., 2017. Canine leptospirosis in Switzerland -
References a prospective cross-sectional study examining seroprevalence, risk factors and ur-
inary shedding of pathogenic leptospires. Prev. Vet. Med. 141, 48–60.
Ellis, W.A., 2010. Control of canine leptospirosis in Europe: time for a change? Vet. Rec.
Adler, B., 2015. Vaccines against leptospirosis. Curr. Top. Microbiol. Immunol. 387, 167, 602–605.
251–272. Jost, B.H., Adler, B., Vinh, T., Faine, S., 1986. A monoclonal antibody reacting with a
Adler, B., Pena Moctezuma, A., 2010. Leptospira and leptospirosis. Vet. Microbiol. 140, determinant on leptospiral lipopolysaccharide protects guinea pigs against leptos-
287–296. pirosis. J. Med. Microbiol. 22, 269–275.
Bharti, A.R., Nally, J.E., Ricaldi, J.N., Matthias, M.A., Diaz, M.M., Lovett, M.A., Levett, Levett, P.N., 2001. Leptospirosis. Clin. Microbiol. Rev. 14, 296–326.
P.N., Gilman, R.H., Willig, M.R., Gotuzzo, E., Vinetz, J.M., 2003. Peru-United States Rosario, L.A., Arencibia, D.F., Suarez, Y.E., Infante, J.F., Valdes, B.Y., Batistat, N., 2012.
Leptospirosis Consortium, Leptospirosis: a zoonotic disease of global importance. Cross-protection among unrelated Leptospira pathogens serovars: an unfinished
Lancet. 12, 757–771. story. Adv. Clin. Exp. Med. 21, 581–589.
Blanchard, S., Cariou, C., Bouvet, J., Valfort, W., Cupillard, L., Poulet, H., Oberli, F., De Sonrier, C., Branger, C., Michel, V., Ruvoen-Clouet, N., Ganiere, J.P., Andre-Fontaine, G.,
Saint-Vis, B., 2018. Quantitative real-time Leptospira PCR assays: a quicker alter- 2001. Evidence of cross-protection within Leptospira interrogans in an experimental
native to the culture method for the detection of pathogenic Leptospira species in dog model. Vaccine 19, 86–94.
urine, blood and kidney for clinical follow-up. In: 3Rd European Leptospirosis Society
Meeting. Alghero.

You might also like