Canine Vaccine Research Articles

An Oil-Based Adjuvant Improves Immune Responses Induced by Canine Adenovirus-Vectored Vaccine in Mice.

There is a significant need for highly effective vaccines against emerging and common veterinary infectious diseases. Canine adenovirus type 2 (CAV2) vectors allow rapid development of multiple vaccines and have demonstrated their potential in animal models. In this study, we compared the immunogenicity of a non-replicating CAV2 vector encoding the rabies virus glycoprotein with and without MontanideTM ISA 201 VG, an oil-based adjuvant. All vaccinated mice rapidly achieved rabies seroconversion, which was associated with complete vaccine protection. The adjuvant increased rabies antibody titers without any significant effect on the anti-CAV2 serological responses. An RT2 Profiler™ PCR array was conducted to identify host antiviral genes modulated in the blood samples 24 h after vaccination. Functional analysis of differentially expressed genes revealed the up-regulation of the RIG-I, TLRs, NLRs, and IFNs signaling pathways. These results demonstrate that a water-in-oil-in-water adjuvant can shape the immune responses to an antigen encoded by an adenovirus, thereby enhancing the protection conferred by live recombinant vaccines. The characterization of early vaccine responses provides a better understanding of the mechanisms underlying the efficacy of CAV2-vectored vaccines.

The association between rLiHyp1 protein plus adjuvant and amphotericin B is an effective immunotherapy against visceral leishmaniasis in mice

Treatment of visceral leishmaniasis (VL) is compromised by drug toxicity, high cost and/or the emergence of resistant strains. Though canine vaccines are available, there are no licensed prophylactic human vaccines. One strategy to improve clinical outcome for infected patients is immunotherapy, which associates a chemotherapy that acts directly to reduce parasitism and the administration of an immunogen-adjuvant that activates the host protective Th1-type immune response. In this study, we evaluated an immunotherapy protocol in a murine model by combining recombinant (r)LiHyp1 (a hypothetical amastigote-specific Leishmania protein protective against Leishmania infantum infection), with monophosphoryl-lipid A (MPLA) as adjuvant and amphotericin B (AmpB) as reference antileishmanial drug. We used this protocol to treat L. infantum infected-BALB/c mice, and parasitological, immunological and toxicological evaluations were performed at 1 and 30 days after treatment. Results showed that mice treated with rLiHyp1/MPLA/AmpB presented the lowest parasite burden in all organs evaluated, when both a limiting dilution technique and qPCR were used. In addition, these animals produced higher levels of IFN-γ and IL-12 cytokines and IgG2a isotype antibody, which were associated with lower production of IL-4 and IL-10 and IgG1 isotype. Furthermore, low levels of renal and hepatic damage markers were found in animals treated with rLiHyp1/MPLA/AmpB possibly reflecting the lower parasite load, as compared to the other groups. We conclude that the rLiHyp1/MPLA/AmpB combination could be considered in future studies as an immunotherapy protocol to treat against VL.

Influence of age and vaccination interval on canine parvovirus, distemper virus, and adenovirus serum antibody titers

Canine core vaccine titer screenings are becoming increasingly popular in veterinary practice as a tool to guide vaccination decisions, despite a lack of supportive, peer-reviewed evidence-based literature. Additionally, it has been suggested that the canine core vaccine duration of host protective immunity can persist past the currently recommended vaccination interval. Thus, this study evaluated serum antibody titers against three core antigens in dogs with known vaccination histories and lifestyles, analyzing the effect of life stage, exposure risk, and time since last vaccination (TSLV). Clinically healthy dogs (n = 188) presenting to the primary care services of three colleges of veterinary medicine were selected to represent a variety of ages, breeds, and vaccination history. Serum antibody titers for canine parvovirus (CPV), canine distemper virus (CDV), and canine adenovirus-2 (CAV2) were measured via virus neutralization and hemagglutination inhibition. CAV2 and CPV titers decreased, while CDV titers had a decreasing trend with increasing time since last vaccination or vaccination interval. When assessing circulating antibody levels historially associated with protective immunity across various vaccination intervals, 62% (95%CI 36–82%; 8/13) of dogs had positive titers for CDV 5 years post last vaccination, while 92% (95%CI 67–99%; 12/13) of dogs were positive for CAV2 and CPV. Both advanced age and life stage were associated with lower titers and thus, identify a canine population cohort likely at higher disease risk. The results of this study revealed that patient duration of core vaccine-mediated immunity changes with a number of variables, with animal aging and time since vaccination influencing host humoral immunity. This provides further support for the performance of canine core antibody titers to assess whether a vaccine booster and/or specific type of booster is warranted.

Re-emergence of canine influenza in the United States of America; a call for One Health approach; a letter to the editor

Re-emergence of canine influenza in the United States of America; a call for One Health approach; a letter to the editor

Development of a monoclonal antibody recognizing novel linear neutralizing epitope on H protein of canine distemper virus vaccine strains (America-1 genotype)

Canine distemper virus (CDV) is an economically important virus responsible for canine distemper (CD), a highly contagious disease that afflicts various animal species worldwide. The hemagglutinin (H) protein is the major neutralizing target of virus. Therefore, it is often considered as immunogen to prepare neutralizing antibodies. The accurate identification of neutralizing epitope will provide important antigenic information and extend the knowledge of mechanisms of virus neutralization. In this study, we generated a neutralizing monoclonal antibody (mAb) 4C6 against CDV H protein, and defined the minimal linear epitope 238DIEREFDT245, which was highly conserved in America-1 genotype of CDV strains (vaccines). The mAb 4C6 could not react with a CDV strain that had two substitutions of D238Y and R241G in the epitope, which appeared in most CDV strains of the other genotypes. Besides, a few different amino acid mutations in the epitope were also included. Collectively, the epitope 238DIEREFDT245 was variable in the other genotypes of CDV strains. The epitope 238DIEREFDT245 was exposed to the surface of CDV H protein, showing good antigenicity. These data will provide insights into structure, function and antigenicity of H protein and lay the foundation for the development of diagnostic technologies and vaccine design for CDV.

Mucosal Vaccination with Live Attenuated Bordetella bronchiseptica Protects against Challenge in Wistar Rats.

Bordetella bronchiseptica (Bb) is a Gram-negative bacterium responsible for canine infectious respiratory disease complex (CIRDC). Several vaccines targeting this pathogen are currently licensed for use in dogs, but their mechanism of action and the correlates of protection are not fully understood. To investigate this, we used a rat model to examine the immune responses induced and the protection conferred by a canine mucosal vaccine after challenge. Wistar rats were vaccinated orally or intranasally on D0 and D21 with a live attenuated Bb vaccine strain. At D35, the rats of all groups were inoculated with 103 CFU of a pathogenic strain of B. bronchiseptica. Animals vaccinated via either the intranasal or the oral route had Bb-specific IgG and IgM in their serum and Bb-specific IgA in nasal lavages. Bacterial load in the trachea, lung, and nasal lavages was lower in vaccinated animals than in non-vaccinated control animals. Interestingly, coughing improved in the group vaccinated intranasally, but not in the orally vaccinated or control group. These results suggest that mucosal vaccination can induce mucosal immune responses and provide protection against a Bb challenge. This study also highlights the advantages of a rat model as a tool for studying candidate vaccines and routes of administration for dogs.

Proteomic analysis of canine vaccines

To use proteomic analysis to identify qualitatively and quantitatively mammalian protein components of commercial veterinary vaccines against canine distemper, leptospirosis, borreliosis, and rabies. 25 licensed veterinary vaccines (from 4 different manufacturers) against canine distemper and leptospirosis, borreliosis, and rabies (3-year and 1-year durations of immunity). Duplicate samples from a single-lot vial of each vaccine were prepared by acetone precipitation and proteolysis with trypsin and Lys-C protease mix. Peptides mixtures (1 μg) were analyzed by liquid chromatography-tandem mass spectrometry using an Orbitrap Fusion Lumos mass spectrometer. Liquid chromatography-tandem mass spectroscopy data were searched against a Bos taurus protein database using MaxQuant to identify and quantify mammalian proteins in the vaccines. Identified proteins were classified by function and network analysis to visualize interactions. The largest number of mammalian proteins was identified in 3-year rabies vaccines (median, 243 proteins; range, 184 to 339 proteins) and 1-year rabies vaccines (median, 193 proteins; range, 169 to 350 proteins). Borrelia and leptospirosis-distemper (L&D) vaccines had the lowest number of proteins. Rabies vaccines had the highest number of identified proteins in common (n = 316); 33 were unique to 1-year products and 44 were found in 3-year products. Borrelia and L&D vaccines had 16 and 22 uniquely identified proteins, respectively. The protein classifications were primarily modulators of protein-binding activity, enzymes, transfer-carrier proteins, cytoskeletal proteins, defense-immunity proteins, calcium-binding proteins, and extracellular matrix proteins. This study demonstrates proteomics application to evaluate quality differences among different vaccines, identifying potential stimulants of desirable and undesirable immune responses.

A commercial monovalent canine parvovirus vaccine performs better than a commercial combination vaccine in puppies

Thirty puppies were randomly divided in to 3 groups for comparative evaluation of two commercial CPV vaccines. Each group was further subdivided in to < 6 months and < 3 months -old puppies and either vaccinated with a monovalent vaccine: Primodog, a combination vaccine: Duramune or maintained as a non-vaccinated control. Humoral immune response was determined by Hemagglutination Inhibition (HAI) on 21 and 35 -days after vaccination. The geometric mean titer (GMT) induced by Duramune, 21 and 35 -days post-vaccination was GMT 73.3 and 137.2, respectively. Comparatively, Primodog demonstrated higher GMT on 21 and 35 -days after vaccination: 97.0 and 168.9, respectively. The older puppies (< 6 months old) demonstrated higher seroconversion to both vaccines.

MELANOCYTIC NEOPLASIA IN PANTHERA SPECIES: CLINICAL PRESENTATIONS, PATHOLOGIC FINDINGS AND RESPONSES TO TREATMENT.

Neoplasia is a common cause of morbidity and mortality in captive nondomestic felids. Seven tigers (Panthera tigris), two African lions (Panthera leo), and two snow leopards (Panthera uncia) were diagnosed with melanocytic neoplasia (10 malignant melanomas, two benign melanocytomas) over a 20-yr period. Animals were 10-19 yr old and 5/7 tigers were phenotypically white. Malignant melanoma tumor location included skin (n = 4), oral mucosa (n = 2), nasal planum (n = 1), iris/uvea (n = 2), and lip margin (n = 1); melanocytomas were found in skin (n = 2). Metastasis to regional lymph nodes was seen at diagnosis in 3/7 melanoma cases. Thoracic radiography (n = 6) and/or computed tomography (n = 2) did not detect pulmonary metastasis at diagnosis but were useful for detection later in the disease course. Median survival time (MST) for all cases ranged from 1 mon - 40 mon. Seven cases with malignant melanoma underwent treatment, which included surgery, radiation therapy, and administration of the canine melanoma vaccine (Oncept®) or a combination of these treatments; MST was 5-40 mon for these cases. While multimodal therapy may provide an improved survival time, the majority of animals with malignant melanoma invariably died from neoplastic disease. Necropsy confirmed metastasis of malignant melanoma in 7/9 animals; sites included lung, liver, lymph node, kidney, mesentery, pleural cavity, heart, stomach, spleen, and adrenal gland. This case series describes the clinical and histologic findings of melanocytic neoplasia in nondomestic felids as well as multimodal treatment strategies incorporating the canine melanoma vaccine.

Dietary yeast beta 1,3/1,6 glucan supplemented to adult Labrador Retrievers alters peripheral blood immune cell responses to vaccination challenge without affecting protective immunity.

Yeast-derived 1,3/1,6 beta-glucans may alter host immunity to produce robust and quickly resolved responses that align with companion animal health goals. In adult dogs, immunomodulation by yeast 1,3/1,6 beta-glucans in extruded kibble diet have not been well-documented. The study objective was to evaluate systemic immune responses in dogs fed kibble diets with two yeast 1,3/1,6 beta-glucans doses before and after vaccine challenge. Twenty-four adult Labrador Retrievers were assigned to 3 dietary treatments consisting of a basal diet (control) supplemented with 0.012 or 0.023% (0.5 or 1X, respectively) yeast 1,3/1,6 beta-glucan with equal sex representation within each treatment (8 dogs/diet). Animals were fed experimental diets for a 29d acclimation period, after which baseline blood samples were collected before administration of a combination canine distemper virus, parvovirus, and adenovirus-2 vaccine. Blood samples were collected weekly for 21d following vaccination with whole blood for CBC analysis, serum for titer and cytokine assays, and peripheral blood mononuclear cells (PBMC) isolated for flow cytometric immune cell profiling. Data were analyzed using the MIXED procedure with diet and timepoint fixed effects. Serum titer was analyzed by Kruskal-Wallis test (SAS 9.4; P≤0.05). Prior to vaccination, beta-glucan diets did not affect serum cytokines, antibody titer, or immune cell populations. In the first 7 days post-vaccination (dpv), PBMC CD21 low B cells increased in 36.5-58.1% in all groups but the magnitude of change was lesser in the 0.5X beta-glucan diet resulting in 25.6% lower CD21 low populations compared to control-fed dogs (P=0.007). By 21dpv, B cell populations recovered to baseline levels in dogs fed 1X beta-glucan, but CD21 high cells remained elevated 50.5% in dogs fed 0.5X beta-glucan diets compared to baseline (P<0.0001). While no differences in serum titer or cytokines were observed, feeding both beta-glucan diets maintained stable blood monocytes whereas a 53.0% decrease between baseline and 14dpv was observed in control-fed dogs (P=0.01). Collectively, these outcomes suggest that a 1X dose of 1,3/1,6 yeast beta-glucan in extruded kibble diets altered monocytes associated with trained immunity, did not reduce PBMC CD21 low B cell responsiveness, and simultaneously contributed to B cell population resolution by 21dpv in adult dogs. Additional research to assess the functionality of these changes is needed.

Cellular and humoral immune responsiveness to inactivated Leptospira interrogans in dogs vaccinated with a tetravalent Leptospira vaccine.

Vaccination is commonly used to protect dogs against leptospirosis, however, memory immune responses induced by canine Leptospira vaccines have not been studied. In the present study, antibody and T cell mediated responses were assessed in dogs before and 2weeks after annual revaccination with a commercial tetravalent Leptospira vaccine containing serogroups Canicola and Australis. Vaccination significantly increased average log2 IgG titers from 6.50 to 8.41 in year 1, from 5.99 to 7.32 in year 2, from 5.32 to 8.32 in year 3 and from 5.32 to 7.82 in year 4. The CXCL-10 levels, induced by in vitro stimulation of PBMC with Canicola and Australis, respectively, significantly increased from 1039.05pg/ml and 1037.38pg/ml before vaccination to 2547.73pg/ml and 2730.38pg/ml after vaccination. IFN-γ levels increased from 85.60pg/ml and 178.13pg/ml before vaccination to 538.62pg/ml and 210.97pg/ml after vaccination. The percentage of proliferating CD4+ T cells in response to respective Leptospira strains significantly increased from 1.43% and 1.25% before vaccination to 24.11% and 14.64% after vaccination. Similar responses were also found in the CD8+ T cell subset. Vaccination also significantly enhanced the percentages of central memory CD4+ T cells from 12% to 26.97% and 27.65%, central memory CD8+ T cells from 3% to 9.47% and 7.55%, and effector CD8+ T cells from 3% to 7.6% and 6.42%, as defined by the expression of CD45RA and CD62L, following stimulation with Canicola and Australis, respectively. Lastly, enhanced expression of the activation marker CD25 on T cells after vaccination was found. Together, our results show that next to IgG responses, also T cell responses are induced in dogs upon annual revaccination with a tetravalent Leptospira vaccine, potentially contributing to protection.

An Assessment and Comparison of the Efficacy of Two Licensed Tetravalent Leptospira Vaccines for Dogs Using an Improved Challenge Model.

An objective of the present studies was to compare the protective effects of two European licensed canine tetravalent leptospirosis vaccines, Nobivac L4 and Versican Plus L4. Four similar challenge studies in beagle dogs were performed. In each study the dogs were assigned to one of three treatment groups as follows: group 1, Nobivac L4; group 2, Versican Plus L4; group 3, non-vaccinated control group. Two vaccinations were followed by a Leptospira challenge. Strains of the following serogroups were used for challenge: study 1, Grippotyphosa; study 2, Icterohaemorrhagiae; study 3, Canicola; study 4, Australis. Parameters of efficacy were antibody titres; body temperature; clinical signs; cultures of Leptospira bacteria from the blood, urine, kidney and liver; rapid urinalysis; macroscopic and histopathological examination at necropsy. It was concluded that compared to vaccination with Versican Plus L4, vaccination with Nobivac L4 resulted in generally better control of leptospirosis disease parameters after the challenge including a complete prevention of the clinical signs following a Grippotyphosa and Icterohaemorrhagiae challenge. In contrast, vaccination with Versican Plus L4 only prevented infection by Australis and shedding by Grippotyphosa and Australis but it did not lead to any statistically significant reduction of either infection or shedding following an Icterohaemorrhagiae challenge.

2022 AAHA Canine Vaccination Guidelines

ABSTRACTThese guidelines are an update and extension of previous AAHA peer-reviewed canine vaccination guidelines published in 2017. Vaccination is a cornerstone of canine preventive healthcare and one of the most cost-effective ways of maintaining a dog’s health, longevity, and quality of life. Canine vaccination also serves a public health function by forming a barrier against several zoonotic diseases affecting dogs and humans. Canine vaccines are broadly categorized as containing core and noncore immunizing antigens, with administration recommendations based on assessment of individual patient risk factors. The guidelines include a comprehensive table listing canine core and noncore vaccines and a recommended vaccination and revaccination schedule for each vaccine. The guidelines explain the relevance of different vaccine formulations, including those containing modified-live virus, inactivated, and recombinant immunizing agents. Factors that potentially affect vaccine efficacy are addressed, including the patient’s prevaccination immune status and vaccine duration of immunity. Because animal shelters are one of the most challenging environments for prevention and control of infectious diseases, the guidelines also provide recommendations for vaccination of dogs presented at or housed in animal shelters, including the appropriate response to an infectious disease outbreak in the shelter setting. The guidelines explain how practitioners can interpret a patient’s serological status, including maternally derived antibody titers, as indicators of immune status and suitability for vaccination. Other topics covered include factors associated with postvaccination adverse events, vaccine storage and handling to preserve product efficacy, interpreting product labeling to ensure proper vaccine use, and using client education and healthcare team training to raise awareness of the importance of vaccinations.

Transcriptome and proteome analysis of innate immune responses to inactivated Leptospira and bivalent Leptospira vaccines in canine 030-D cells

Mandatory potency testing of Leptospira vaccine batches relies partially on in vivo procedures, requiring large numbers of laboratory animals. Cell-based assays could replace in vivo tests for vaccine quality control if biomarkers indicative of Leptospira vaccine potency are identified. We investigated innate immune responsiveness induced by inactivated L. interrogans serogroups Canicola and Icterohaemorrhagiae, and two bivalent, non-adjuvanted canine Leptospira vaccines containing the same serogroups. First, the transcriptome and proteome analysis of a canine monocyte/macrophage 030-D cell line stimulated with Leptospira strains, and vaccine B revealed more than 900 DEGs and 23 DEPs in common to these three stimuli. Second, comparison of responses induced by vaccine B and vaccine D revealed a large overlap in DEGs and DEPs as well, suggesting potential to identify biomarkers indicative of Leptospira vaccine quality. Because not many common DEPs were identified, we selected seven molecules from the identified DEGs, associated with pathways related to innate immunity, of which CXCL-10, IL-1β, SAA, and complement C3 showed increased secretion upon stimulation with both Leptospira vaccines. These molecules could be interesting targets for development of biomarker-based assays for Leptospira vaccine quality control in the future. Additionally, this study contributes to the understanding of the mechanisms by which Leptospira vaccines induce innate immune responses in the dog.

Vaccination with human tyrosinase DNA as a therapy for equine intraocular melanoma—4 cases: 2016–2021

SummaryThis report describes four cases of intraocular melanoma which were treated with a course of human tyrosine DNA canine melanoma vaccine. Two cases continued to show progression of the tumour. One case presented with bilateral intraocular melanomas and demonstrated clinical progression in one eye within 5 months, and apparent quiescence of disease in the other, though subtle deterioration was noted at a 2‐year follow‐up. The final case presented with recurrent ulceration due to an intraocular melanoma and demonstrated a good clinical response with a reduction in the frequency and severity of the episodes of ulceration. The small number of cases in this report precludes firm conclusions, but these reported outcomes suggest limited justification for the routine recommendation of melanoma vaccines to horses with intraocular melanomas.

Multi-institutional retrospective study of canine foot pad malignant melanomas: 20 cases.

Melanomas arising from the foot pad are a rare clinical entity in dogs. The biologic behaviour of foot pad malignant melanoma is not well understood, and these tumours are infrequently described. The objective of this study was to evaluate the clinical characteristics of primary canine foot pad melanoma in a larger cohort of patients. Eligible cases were solicited from the American College of Veterinary Internal Medicine (ACVIM) Oncology listserv for retrospective review. Included dogs had a cytologic and/or histologic diagnosis of foot pad melanoma evaluated by a board-certified clinical or anatomic pathologist. Dogs with cutaneous, oral, digital, subungual or interdigital melanomas were excluded. A total of 20 cases were included. Eleven dogs received various adjuvant therapies including chemotherapy, radiation therapy, and/or the ONCEPT canine melanoma vaccine following surgery. At diagnosis, regional lymph node metastasis was observed in four dogs (20%). Seven dogs developed subsequent regional and/or distant metastasis for an overall metastatic rate of 55%. The progression-free interval (PFI) was 101 days (range, 20-960 days). The median survival time (MST) was 240 days (range, 25-479 days). For dogs receiving adjuvant therapy, the MST was 159 days (range, 25-387 days). Canine foot pad melanoma is a rare neoplasm that can exhibit an aggressive behaviour.

Biological and Genetic Characterization of Canine Distemper Virus Vaccine Candidate Named as CD1901-100

Biological and Genetic Characterization of Canine Distemper Virus Vaccine Candidate Named as CD1901-100

Canine Adenovirus 1 Isolation Bioinformatics Analysis of the Fiber.

Canine adenovirus type 1 (CAdV-1) is a double-stranded DNA virus, which is the causative agent of fox encephalitis. The Fiber protein is one of the structural proteins in CAdV-1, which mediates virion binding to the coxsackievirus and adenovirus receptor on host cells. The suspected virus was cultured in the MDCK cells, and it was determined through the cytopathic effects, sequencing and electron microscopy. The informatics analysis of the Fiber was done using online bioinformatics servers. The CAdV-1-JL2021 strain was isolated successfully, and were most similar to the CAdV-1 strain circulating in Italy. The occurrence of negative selection and recombination were found in the CAdV-1-JL2021 and CAdV-2-AC_000020.1. Host cell membrane was its subcellular localization. The CAdV-1-JL2021 Fiber (ON164651) had 6 glycosylation sites and 107 phosphorylation sites, exerted adhesion receptor-mediated virion attachment to host cell, which was the same as CAdV-2-AC_000020.1 Fiber. The Fiber tertiary structure of the CAdV-1-JL2021 and CAdV-2-AC_000020.1 was different, but they had the same coxsackievirus and adenovirus receptor. “VATTSPTLTFAYPLIKNNNH” were predicted to be the potential CAdV-1 B cell linear epitope. The MHC-I binding peptide “KLGVKPTTY” were both presented in the CAdV-1-JL2021 and CAdV-2-AC_000020.1 Fiber and it is useful to design the canine adenovirus vaccine.

Compatibility between a rabies vaccine and two canine combined vaccines against canine distemper, adenovirosis, parvovirosis, parainfluenza virus disease and leptospirosis, with or without canine coronavirus

In many countries, vaccination programs still require dogs to be vaccinated against rabies in addition to Canine distemper virus (CDV), adenovirus (CAV), parvovirus (CPV), parainfluenza virus (CPiV), Leptospira (L) or Canine coronavirus (CCV= Cv). Few vaccines containing all these antigens are commercially available and, unless compatibility between the vaccines was demonstrated, concurrent administration of a DAPPi-L(Cv) vaccine and a vaccine against rabies should not be recommended. This may be of concern for practitioners who wish to vaccinate dogs with all components on the same day. This study aimed at evaluating immunological compatibility between a monovalent rabies vaccine (Rabisin™) and two large combination vaccines against CDV, CAV, CPV, CPiV with 2 leptospira components +Cv (Recombitek® C6/Cv) or with 4 Leptospira components (Recombitek® C8), when injected concomitantly at two separate injection sites.Fourteen days after administration of the rabies vaccine, with or without concomitant administration of combo vaccines, all dogs had seroconverted against rabies and maintained protective titers over the duration of the study. In addition, 100% of the puppies vaccinated with one or the other combo vaccines seroconverted against CDV, CAV, CPV, CPiV (CCV) and Leptospira, whatever the vaccination group. Lack of immunological interference between Rabisin™ and all components of the Recombitek® C6/Cv or Recombitek® C8 Combo vaccines was demonstrated by non-inferiority analysis, except for CDV in the Recombitek®C8+ Rabisin™ group. Based on these results, a concomitant administration of Rabisin™ with Recombitek® C6/Cv or Recombitek® C8 can be recommended in daily practice, which can be essential for facilitating vaccination compliance.

Activation of Canine, Mouse and Human TLR2 and TLR4 by Inactivated Leptospira Vaccine Strains.

Canine Leptospira vaccines contain inactivated strains of pathogenic Leptospira, the causative agents of leptospirosis. For an effective response to vaccination, activation of the innate immune system via pattern recognition receptors such as TLRs is crucial. However, it is not known which TLRs are activated by Leptospira in dogs. To investigate the involvement of canine TLR2, TLR4, and TLR5 in the recognition of Leptospira, we stimulated canine moDC and reporter cells expressing canine TLR2 with either whole-inactivated bacteria or purified LPS of Leptospira strains, representing the serogroups generally used in canine leptospirosis vaccines. Using the endotoxin neutralizing reagent polymyxin B and TLR4 antagonist RS-LPS, we demonstrate that Leptospira LPS and canine TLR4 are involved in IL-1β production as well as in the uptake of inactivated Leptospira in canine moDC. Furthermore, polymyxin B only partially inhibited IL-1β production induced by inactivated Leptospira, suggesting that next to TLR4, also other TLRs may be involved. The observed activation of canine TLR2-expressing reporter cells by inactivated Leptospira strains indicates that TLR2 could be one of these TLRs. Next, we analyzed TLR2 and TLR4 activating capabilities by the same Leptospira strains using human and mouse TLR-expressing reporter cells. Inactivated Leptospira and leptospiral LPS activated not only mouse, but also human TLR4 and this activation was shown to be LPS dependent in both cases. Additionally, inactivated Leptospira activated mouse and human TLR2-expressing reporter cell lines. In our study, we could not identify significant species differences in the recognition of Leptospira by TLR2 and TLR4 between dog, human and mouse. Lastly, we show that these inactivated Leptospira strains are recognized by both mouse and human TLR5 reporter cells only after exposure to additional heat-treatment. Unfortunately, we were not able to confirm this in the canine system. Our data show that TLR2 and TLR4 are involved in the recognition of Leptospira strains used in the production of canine Leptospira vaccines. This study contributes to the understanding of Leptospira-induced innate immune responses in dogs, humans, and mice. Future studies are needed to further explore the role of canine TLR2, TLR4 and TLR5 in the induction of vaccine-mediated immunity against Leptospira.