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1.
Vaccine ; 33(32): 3963-9, 2015 Jul 31.
Article in English | MEDLINE | ID: mdl-26100922

ABSTRACT

The safety and protective efficacy of a new octavalent combination vaccine containing inactivated Erysipelothrix rhusiopathiae, Parvovirus, and Leptospira interrogans (sensu lato) serogroups Canicola, Icterohaemorrhagiae, Australis (Bratislava), Grippotyphosa, Pomona and Tarassovi - Porcilis(®) Ery+Parvo+Lepto - was evaluated in laboratory studies and under field conditions. The safety (2× overdose and repeated dose) was tested in 26 gilts. In this study, neither vaccine related temperature increase nor other systemic reactions were observed after intramuscular vaccination. No local reactions were observed except for one animal that had a small local reaction (2cm diameter) that lasted for 5 days after the third vaccination. Efficacy was tested in 40 gilts. A group of 20 gilts was vaccinated at 20 and 24 weeks of age with Porcilis(®) Ery+Parvo+Lepto and a group of 20 age- and source-matched animals served as the control group. The gilts were inseminated at 41 weeks or 66 weeks of age and were challenged with serovar Pomona 10 weeks after insemination, corresponding to 6 months (n=2×10) and 12 months (n=2×10) after the last vaccination. After both the 6- and 12-month challenges the control animals developed clinical signs (fever, lethargy and anorexia) and leptospiraemia as determined by positive blood culture. In addition, both the 6- and 12-month challenges resulted in death of 21% and 27% of the total number of foetuses in the control groups, respectively. Clinical signs and leptospiraemia were statistically significantly lower in vaccinated gilts after both the 6- and 12-month challenges. In addition, foetal death was statistically significantly lower (3% and 2%, respectively) in vaccinated gilts after both the 6- and 12 month challenges. The vaccine was tested further under field conditions on a Portuguese farm with a history of an increasing abortion rate associated with a Leptospira serovar Pomona infection (confirmed by PCR and serology). This study was designed as an observational-longitudinal field study. At the start of the study, all breeding sows and replacement gilts on the farm were vaccinated twice with Porcilis(®) Ery+Parvo+Lepto at an interval of 4 weeks. Starting six months after the primary vaccination schedule, the animals were re-vaccinated during the second week of every subsequent lactation. New replacement gilts were vaccinated using the same schedule. After vaccination, the abortion rate reduced rapidly from 12.6% in winter months of 2012 (December 2011 to March 2012) to 0.5% in winter months of 2013, a statistical significant decrease of 96%. The total number of abortions on the farm decreased from 55 in 2012 to 6 in 2013. Thereafter, the abortion rate remained stable and in the period December 2013 to April 2014 was still low (0.6%). In conclusion, the present studies demonstrate that the octavalent Porcilis(®) Ery+Parvo+Lepto vaccine can be safely used in gilts and sows and induces significant protection, for the duration of at least one year, against serovar Pomona induced clinical signs, leptospiraemia and foetal death. Protection against Pomona associated reproductive failure was confirmed under field conditions where a significant reduction in abortion rate was observed.


Subject(s)
Bacterial Vaccines/immunology , Erysipelothrix Infections/prevention & control , Leptospira interrogans serovar pomona/immunology , Leptospirosis/veterinary , Parvoviridae Infections/veterinary , Swine Diseases/prevention & control , Viral Vaccines/immunology , Abortion, Induced , Animals , Bacteremia/prevention & control , Bacterial Vaccines/administration & dosage , Bacterial Vaccines/adverse effects , Drug-Related Side Effects and Adverse Reactions/pathology , Fetal Death , Fever/prevention & control , Injections, Intramuscular , Leptospirosis/prevention & control , Longitudinal Studies , Parvoviridae Infections/prevention & control , Portugal , Survival Analysis , Swine , Vaccines, Combined/administration & dosage , Vaccines, Combined/adverse effects , Vaccines, Combined/immunology , Vaccines, Inactivated/administration & dosage , Vaccines, Inactivated/adverse effects , Vaccines, Inactivated/immunology , Viral Vaccines/administration & dosage , Viral Vaccines/adverse effects
2.
Rev Argent Microbiol ; 45(1): 34-8, 2013.
Article in English | MEDLINE | ID: mdl-23560786

ABSTRACT

Leptospira reactivity in stray and household dogs in Campeche as well as associated risk factors to the seropositivity in household dogs have been herein determined. The survey included 323 dogs, 142 of which were stray dogs and 181 household dogs. Nine Leptospira interrogans serovars were tested by the microagglutination test. Reactivity was 21.3 % (69/323), 17.2 % corresponded to household dogs and 26.7 % to stray dogs. Leptospira Canicola (29 %), Leptospira Hardjo (22.58 %), and Leptospira Icterohaemorrhagiae (16.12 %) were the most common serovars reacting against the serum of household animals, while Leptospira Canicola (15.78 %), Leptospira icterohaemorrhagiae (13.15 %), and Leptospira Pomona (7.89 %) were those reacting in stray dogs. Results showed that all dogs have been in contact with different Leptospira serovars and outdoor exposure is the main infection risk factor.


Subject(s)
Antibodies, Bacterial/blood , Dog Diseases/epidemiology , Leptospira/immunology , Leptospirosis/veterinary , Animals , Animals, Wild , Disease Reservoirs , Dog Diseases/diagnosis , Dogs , Environmental Exposure , Leptospira interrogans serovar canicola/immunology , Leptospira interrogans serovar icterohaemorrhagiae/immunology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/epidemiology , Mexico/epidemiology , Pets , Risk Factors , Seroepidemiologic Studies , Urban Health , Zoonoses
3.
Rev. argent. microbiol ; 45(1): 34-8, mar. 2013.
Article in Spanish | LILACS, BINACIS | ID: biblio-1171771

ABSTRACT

Leptospira reactivity in stray and household dogs in Campeche as well as associated risk factors to the seropositivity in household dogs have been herein determined. The survey included 323 dogs, 142 of which were stray dogs and 181 household dogs. Nine Leptospira interrogans serovars were tested by the microagglutination test. Reactivity was 21.3


corresponded to household dogs and 26.7


), and Leptospira Icterohaemorrhagiae (16.12


) were the most common serovars reacting against the serum of household animals, while Leptospira Canicola (15.78


), and Leptospira Pomona (7.89


) were those reacting in stray dogs. Results showed that all dogs have been in contact with different Leptospira serovars and outdoor exposure is the main infection risk factor.


Subject(s)
Antibodies, Bacterial/blood , Dog Diseases/epidemiology , Leptospira/immunology , Leptospirosis/veterinary , Animals , Animals, Wild , Pets , Dogs , Dog Diseases/diagnosis , Seroepidemiologic Studies , Environmental Exposure , Risk Factors , Leptospira interrogans serovar canicola/immunology , Leptospira interrogans serovar icterohaemorrhagiae/immunology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/epidemiology , Mexico/epidemiology , Disease Reservoirs , Urban Health , Zoonoses
4.
Rev. Argent. Microbiol. ; 45(1): 34-8, 2013 Jan-Mar.
Article in Spanish | BINACIS | ID: bin-133181

ABSTRACT

Leptospira reactivity in stray and household dogs in Campeche as well as associated risk factors to the seropositivity in household dogs have been herein determined. The survey included 323 dogs, 142 of which were stray dogs and 181 household dogs. Nine Leptospira interrogans serovars were tested by the microagglutination test. Reactivity was 21.3


(69/323), 17.2


corresponded to household dogs and 26.7


to stray dogs. Leptospira Canicola (29


), Leptospira Hardjo (22.58


), and Leptospira Icterohaemorrhagiae (16.12


) were the most common serovars reacting against the serum of household animals, while Leptospira Canicola (15.78


), Leptospira icterohaemorrhagiae (13.15


), and Leptospira Pomona (7.89


) were those reacting in stray dogs. Results showed that all dogs have been in contact with different Leptospira serovars and outdoor exposure is the main infection risk factor.


Subject(s)
Antibodies, Bacterial/blood , Dog Diseases/epidemiology , Leptospira/immunology , Leptospirosis/veterinary , Animals , Animals, Wild , Disease Reservoirs , Dog Diseases/diagnosis , Dogs , Environmental Exposure , Leptospira interrogans serovar canicola/immunology , Leptospira interrogans serovar icterohaemorrhagiae/immunology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/epidemiology , Mexico/epidemiology , Pets , Risk Factors , Seroepidemiologic Studies , Urban Health , Zoonoses
5.
Vet Immunol Immunopathol ; 145(1-2): 546-50, 2012 Jan 15.
Article in English | MEDLINE | ID: mdl-22227076

ABSTRACT

Tertiary lymphoid organs (TLOs) are structures that are morphologically and functionally similar to secondary lymphoid organs. TLOs usually arise in a background of chronic inflammation. Several histological patterns of interstitial nephritis have been documented in porcine leptospirosis. Among them the lympho-follicular pattern is characterized by infiltrates of mononuclear cells organized in lymphoid follicle-like structures. Immunohistological analysis of 5 cases of porcine lympho-follicular nephritis associated with Leptospira Pomona infection demonstrated the presence of inflammatory cell populations, including B cells, T cells, macrophages and follicular dendritic cells (FDCs), which were compartmentalized as in TLOs. Immunohistochemistry for Leptospira Pomona revealed an intimate association between leptospiral antigen and FDCs. Overexpression of MHCII in different populations of both professional and non-professional antigen presenting cells was also demonstrated. FDCs play role during TLOs induction for their ability to retain non-self antigens in the form of immune complexes, thus causing persistent T cell activation, generation of a complex cytokine network and stimulation of humoral immunity. Sustained bacterial antigen presentation in the context of chronic leptospiral nephritis, may also lead to autoimmune mechanisms involved in the generation of TLOs. Whether lymphoid neogenesis and TLOs play a protective role in porcine leptospiral nephritis is still unclear.


Subject(s)
Kidney/microbiology , Leptospira interrogans serovar pomona , Leptospirosis/veterinary , Lymphoid Tissue/microbiology , Nephritis, Interstitial/veterinary , Swine Diseases/microbiology , Animals , Chronic Disease , Kidney/immunology , Kidney/pathology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/immunology , Leptospirosis/pathology , Lymphoid Tissue/immunology , Lymphoid Tissue/pathology , Nephritis, Interstitial/immunology , Nephritis, Interstitial/microbiology , Nephritis, Interstitial/pathology , Swine , Swine Diseases/immunology , Swine Diseases/pathology
6.
Mem Inst Oswaldo Cruz ; 106(6): 763-8, 2011 Sep.
Article in English | MEDLINE | ID: mdl-22012234

ABSTRACT

This work reports a survey of Leptospira spp in pampas deer (Ozotoceros bezoarticus) in the Pantanal wetlands of the state of Mato Grosso do Sul, Brazil by serology and polymerase chain reaction (PCR). Seventy pampas deer were captured in the dry season and surveyed using PCR, microscopic agglutination test (MAT) (n = 51) and by both techniques (n = 47). PCR detected infections in two pampas deer and MAT detected infections in three. Through sequencing and phylogenetic analyses, the PCR-amplified fragment detected in deer was identified as Leptospira interrogans. Serovars Pomona and Butembo were detected using MAT and the highest titre was 200 for serovar Pomona. Epidemiological aspects of the findings are discussed.


Subject(s)
Antibodies, Bacterial/blood , Deer/microbiology , Leptospira interrogans/isolation & purification , Leptospirosis/veterinary , Agglutination Tests/veterinary , Animals , Brazil/epidemiology , Female , Leptospira interrogans/immunology , Leptospira interrogans serovar pomona/immunology , Leptospira interrogans serovar pomona/isolation & purification , Leptospirosis/diagnosis , Leptospirosis/epidemiology , Male , Phylogeny , Polymerase Chain Reaction/veterinary , Seasons , Wetlands
7.
Frontiers in Immunology ; 2: 1-4, Oct. 2011.
Article in English | Sec. Est. Saúde SP, SESSP-IBPROD, Sec. Est. Saúde SP, SESSP-IBACERVO | ID: biblio-1062905

ABSTRACT

Leptospirosis isazoonosis caused by pathogenic bacteria from the genus Leptospira.Thediseaserepresents a seriouspublic health problem inunderdeveloped tropical countries.Leptospire sinfecthosts throughsmallabrasions in the skin or mucousmembranesand they rapidly disseminateto target organs.Thecapacityof some pathogenic leptospira lstrains toacquire thenegative complementregulators factorH(FH)andC4bbindingpro teincorrelateswith their abilityto survivein humans e rum.Inthis study weasses sed the functional consequences oft heagemaculardegeneration-associatedpolymorphismFHHis402orFHTyr402onFH Leptospira interactions.Inbinding assaysusing sub-satura tingamounts ofFH, theFHTyr402 variant interacted with all the strainstested more stronglythanthe FHHis402variant.Athigher concentrations, differences tendedto disappear. We then compared co factor activities displayed by FH. His402and FH Tyr402 bound tothe surface ofL.interrogans. Bothvariantsex ibit similar activity as cofactors for FactorI mediated cleavage of C3b,thusindicating that they do not differin their capacity toregulate the complement cascade.


Subject(s)
Complement Factor H/analysis , Complement Factor H/immunology , Leptospira interrogans serovar pomona/genetics , Leptospira interrogans serovar pomona/immunology , Leptospira/immunology , Complement Factor H/isolation & purification , Polymorphism, Genetic/immunology
8.
Mem. Inst. Oswaldo Cruz ; 106(6): 763-768, Sept. 2011. tab
Article in English | LILACS | ID: lil-602063

ABSTRACT

This work reports a survey of Leptospira spp in pampas deer (Ozotoceros bezoarticus) in the Pantanal wetlands of the state of Mato Grosso do Sul, Brazil by serology and polymerase chain reaction (PCR). Seventy pampas deer were captured in the dry season and surveyed using PCR, microscopic agglutination test (MAT) (n = 51) and by both techniques (n = 47). PCR detected infections in two pampas deer and MAT detected infections in three. Through sequencing and phylogenetic analyses, the PCR-amplified fragment detected in deer was identified as Leptospira interrogans. Serovars Pomona and Butembo were detected using MAT and the highest titre was 200 for serovar Pomona. Epidemiological aspects of the findings are discussed.


Subject(s)
Animals , Female , Male , Antibodies, Bacterial/blood , Deer/microbiology , Leptospira interrogans/isolation & purification , Leptospirosis/veterinary , Agglutination Tests/veterinary , Brazil/epidemiology , Leptospira interrogans serovar pomona/immunology , Leptospira interrogans serovar pomona/isolation & purification , Leptospira interrogans/immunology , Leptospirosis/diagnosis , Leptospirosis/epidemiology , Phylogeny , Polymerase Chain Reaction/veterinary , Seasons , Wetlands
9.
Rev Argent Microbiol ; 43(1): 42-4, 2011.
Article in Spanish | MEDLINE | ID: mdl-21491066

ABSTRACT

Leptospirosis is an infectious disease resulting in significant economic losses in livestock production. This disease causes abortion, embryo death, death of calves within the first few days of life and mastitis. We report a leptospirosis outbreak in calf growing and fattening. Histopathological and hemoparasite studies, immunofluorescence, and bacterial cultures were performed. A strain of Leptospira interrogans serovar Pomona was isolated from samples collected from dead calves.


Subject(s)
Cattle Diseases/epidemiology , Disease Outbreaks/veterinary , Leptospira interrogans serovar pomona/isolation & purification , Leptospirosis/veterinary , Agglutination Tests , Animals , Argentina/epidemiology , Bacterial Vaccines/economics , Brain/microbiology , Brain/pathology , Cattle , Cattle Diseases/diagnosis , Cattle Diseases/economics , Cattle Diseases/microbiology , Cattle Diseases/pathology , Disease Outbreaks/economics , Guinea Pigs , Leptospira interrogans serovar pomona/immunology , Leptospirosis/diagnosis , Leptospirosis/economics , Leptospirosis/epidemiology , Leptospirosis/microbiology , Leptospirosis/pathology , Vaccination/economics , Vaccination/veterinary , Viscera/microbiology , Viscera/pathology
10.
Vaccine ; 27(44): 6129-36, 2009 Oct 19.
Article in English | MEDLINE | ID: mdl-19715780

ABSTRACT

Leptospirosis is an important zoonotic disease worldwide. Subunit vaccines are an attractive intervention strategy against this disease, but potent, non-toxic adjuvants are necessary components to any effective vaccine. Among various adjuvant candidates, liposomes have garnered recent attention for their capacity as carriers of vaccines. In the present study we prepared novel liposomes using total polar lipids from the nonpathogenic bacterium, Mycobacterium smegmatis (designated smegmosomes). The potential for smegmosomes as a vaccine delivery/adjuvant system was evaluated with novel leptospira protective antigens (Lp0607, Lp1118, Lp1454) and compared with conventional aluminum hydroxide adjuvant (alum) in a hamster model of leptospirosis. Four-week-old hamsters were immunized subcutaneously twice at three weeks intervals and either bled at various time points to evaluate antibody responses, sacrificed to isolate splenocytes for lymphocyte proliferation and cytokine profiles in response to recall antigen, or challenged intraperitoneally with a modified lethal dose (10X MLD(50)) of virulent Leptospira interrogans serovar Pomona. Our results demonstrate that smegmosomes carrying antigens are better adjuvants than alum as revealed by enhanced and long term antibody response, lymphocyte proliferation and significant enhancement in both Th1 (IFN-gamma) and Th2 (IL-4, IL-10) cytokine production. Additionally, smegmosomes were found to induce memory responses that are significantly higher than those of alum. Above all, smegmosomes were observed to impart a significantly higher level of protection than alum as revealed by enhanced survival, reduced histopathological lesions and bacterial load in vital organs. Taken together, the data of the present study suggests that smegmosomes will serve well as a promising delivery vehicle/adjuvant system that can induce both Th1 and Th2 type immune responses and provide a novel tool in development of improved vaccines for leptospirosis and other infectious diseases.


Subject(s)
Bacterial Vaccines/immunology , Leptospirosis/immunology , Leptospirosis/prevention & control , Liposomes/immunology , Adjuvants, Immunologic , Aluminum Hydroxide/immunology , Animals , Antibodies, Bacterial/blood , Antigens, Bacterial/immunology , Cell Proliferation , Cricetinae , Disease Models, Animal , Female , Interferon-gamma/blood , Interleukin-10/blood , Interleukin-4/blood , Leptospira interrogans serovar pomona/immunology , Mycobacterium smegmatis/immunology , Recombinant Proteins/immunology
11.
Vaccine ; 27(47): 6537-45, 2009 Nov 05.
Article in English | MEDLINE | ID: mdl-19729088

ABSTRACT

We prepared novel liposomes from total polar lipids of non-pathogenic Leptospira biflexa serovar Potac (designated leptosomes) and evaluated their vaccine delivery/adjuvant potential with novel protective antigens (Lp0607, Lp1118 and Lp1454) of L. interrogans serovar Pomona in a hamster model. The immune response induced by three individual antigens and protective efficacy were evaluated and compared to those induced by same antigens entrapped with PC-liposomes and E. coli lipid liposomes (escheriosomes). Four-week-old hamsters were immunized subcutaneously twice at a 3-week interval, bled at various time points to evaluate antibody response and sacrificed to isolate splenocytes for lymphocyte proliferation and cytokine profiles in response to recall antigen. For the challenge test, 10x MLD(50) (modified lethal dose 50%) of virulent L. interrogans serovar Pomona were administered intraperitoneally. Our results demonstrate that leptosome are better adjuvant than PC-liposomes as revealed by enhanced long term antibody response, lymphocyte proliferation and significant enhancement of both Th1 (IFN-gamma) and Th2 (IL-4 and IL-10) cytokines. Additionally, leptosomes and escheriosomes induced significantly higher level of memory responses than PC-liposome did. Moreover, the novel leptosomal vaccine induced significantly higher levels of protection than those prepared with PC-liposomes as revealed by enhanced survival, reduced histopathological lesions in vital organs and reduced leptospiral load in kidneys. Taken together, the results of the present study clearly reveal that both leptosomes and escheriosomes have emerged as promising delivery vehicles/adjuvants that can be widely exploited with newly discovered antigens in future leptospira vaccines.


Subject(s)
Antigens, Bacterial/immunology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/prevention & control , Liposomes/immunology , Adjuvants, Immunologic/pharmacology , Animals , Antibodies, Bacterial/blood , Cell Proliferation , Cricetinae , Cytokines/immunology , Female , Immunity, Humoral , Immunologic Memory , Kidney/microbiology , Kidney/pathology , Leptospirosis/immunology , Lymphocyte Activation , Phospholipids/immunology , Recombinant Proteins/immunology
12.
J S Afr Vet Assoc ; 80(1): 45-9, 2009 Mar.
Article in English | MEDLINE | ID: mdl-19653519

ABSTRACT

A serological survey of leptospirosis in cattle originating from rural communities of the province of KwaZulu-Natal (KZN) in South Africa was carried out between March 2001 and December 2003. The survey was designed as a 2-stage survey, using the local diptank as the primary sampling point. In total, 2021 animals from 379 diptanks in 33 magisterial districts were sampled and tested with the microscopic agglutination test (MAT). The apparent prevalence at district level was adjusted for clustering and diagnostic test sensitivity and specificity and displayed in maps. The prevalence of leptospirosis in cattle originating from communal grazing areas of KZN was found to be 19.4% with a 95% confidence interval of 14.8-24.1%. At district level the prevalence of leptospirosis varied from 0 to 63% of cattle. Bovine leptospirosis was found to occur in communal grazing areas throughout the province with the exception of 2 districts. The southeastern regions showed a higher prevalence than other areas of the province; while in some of the northern and western districts a lower prevalence was noted. Several serovars were detected by the MAT and although Leptospira interrogans serovar pomona occurred most frequently, serovars tarrasovi, bratislava, hardjo, canicola and icterohaemorrhagica were also frequently identified. The findings of the survey are discussed.


Subject(s)
Antibodies, Bacterial/blood , Cattle Diseases/epidemiology , Leptospira/immunology , Leptospirosis/veterinary , Animals , Cattle , Cattle Diseases/transmission , Cluster Analysis , Female , Hemagglutination Tests/veterinary , Humans , Leptospira/isolation & purification , Leptospira interrogans serovar pomona/immunology , Leptospira interrogans serovar pomona/isolation & purification , Leptospirosis/epidemiology , Leptospirosis/transmission , Male , Rural Population , Seroepidemiologic Studies , South Africa/epidemiology , Zoonoses
13.
Vet Pathol ; 46(5): 800-9, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19179617

ABSTRACT

Class II major histocompatibility complex (MHCII) is required for the presentation of antigens to CD4 helper T cells. During nephritis, not only primary antigen presenting cells such as histiocytes and lymphocytes, but also cytokine-stimulated tubular epithelial cells express MHCII. Leptospirosis in fattening pigs is characterized by several degrees of nephritis, from absence of lesions to severe multifocal tubulo-interstitial inflammation. Renal tissue from 20 8-month-old pigs with spontaneous nephritis and 6 control pigs without renal lesions were investigated for leptospirosis by indirect immunohistochemistry (IHC) and polymerase chain reaction (PCR). IHC for MHCII also was performed on renal samples. Serum samples were tested for different serovars of Leptospira interrogans. Control pigs were free of interstitial nephritis and negative for leptospirosis by all tests. In pigs with nephritis, serology was positive for serovar Pomona in 19/20 pigs. In 16 of these 19 pigs, leptospiral renal infection was confirmed by PCR and/or indirect IHC. Nephritic lesions were classified histologically into perivascular lymphocytic (4 pigs), lymphofollicular (6 pigs), lymphohistiocytic (8 pigs), and neutrophilic (2 pigs) pattern. MHCII expression by histiocytes and lymphocytes was observed in all lesions. Prominent MHCII expression in regenerating tubular epithelium was observed in lymphofollicular and lymphohistiocytic nephritis. No tubular colocalization between leptospiral and MHCII antigen was observed. Results suggest that during leptospiral nephritis, MHCII contributes to the intensity of the inflammatory response. Furthermore de novo MHCII expression in regenerating tubules may play a role in the defence mechanism against leptospiral tubular colonization.


Subject(s)
Histocompatibility Antigens Class II/immunology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/veterinary , Nephritis, Interstitial/veterinary , Swine Diseases/microbiology , Animals , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Histocompatibility Antigens Class II/analysis , Immunohistochemistry/veterinary , Leptospira interrogans serovar pomona/genetics , Leptospirosis/immunology , Leptospirosis/microbiology , Nephritis, Interstitial/immunology , Nephritis, Interstitial/microbiology , Polymerase Chain Reaction/veterinary , Statistics, Nonparametric , Swine , Swine Diseases/immunology
14.
Vaccine ; 27(3): 378-87, 2009 Jan 14.
Article in English | MEDLINE | ID: mdl-19022317

ABSTRACT

Subunit vaccines are attractive as an intervention strategy against leptospirosis, an important zoonotic disease afflicting both humans and livestock. However, the success of subunit vaccines has been hampered by weak or short-term immunity and unavailability of nontoxic, potent adjuvants. In the present study, the variable region of recombinant Leptospira immunoglobulin like protein A (LigAvar) incorporated into conventional liposomes and PLGA microspheres produced robust immune responses that induced significant protection against virulent Leptospira interrogans serovar Pomona challenge in hamsters. Four-week-old hamsters were immunized subcutaneously with LigAvar incorporated into conventional liposomes or adsorbed on aluminum hydroxide (alum) and subsequently boosted after 3 weeks. Additionally, LigAvar incorporated into PLGA microspheres was evaluated as a single dose vaccine. All animals were challenged intraperitoneally 3 weeks after booster with a lethal dose (10 x MLD50) of virulent L. interrogans serovar Pomona. Animals were bled at various time points to evaluate antibody response, then sacrificed. Splenocytes were isolated and assayed for lymphocyte proliferation and cytokine profiles in response to recall antigen. Our results indicate that both liposomes and microspheres prove to be better adjuvants compared to conventional alum as revealed by enhanced antibody titers, lymphocyte proliferation and significant enhancement in both Th1(IL-12, IFN-gamma) and Th2 (IL-4, IL-10) cytokines. Moreover, LigAvar associated with liposomes and microspheres is able to provide better protection than LigAvar with alum as revealed by enhanced survival and reduced histopathological lesions in vital organs. Taken together, the data of the present study suggests that both liposomes and PLGA microspheres are promising adjuvants for use with future subunit vaccines for prevention of leptospirosis.


Subject(s)
Adjuvants, Immunologic/pharmacology , Antigens, Bacterial/immunology , Bacterial Vaccines/immunology , Lactic Acid/pharmacology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/prevention & control , Liposomes/pharmacology , Polyglycolic Acid/pharmacology , Animals , Antibodies, Viral/blood , Cell Proliferation , Cricetinae , Cytokines/biosynthesis , Female , Immunization, Secondary , Kidney/microbiology , Kidney/pathology , Leptospirosis/immunology , Leukocytes, Mononuclear/immunology , Liver/microbiology , Lung/microbiology , Mesocricetus , Microspheres , Polylactic Acid-Polyglycolic Acid Copolymer , Spleen/immunology , Survival Analysis , Vaccines, Subunit/immunology
15.
Microbes Infect ; 11(2): 230-7, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19070678

ABSTRACT

Leptospiral immunoglobulin-like protein (LigB) was truncated into conserved (LigBcon) and variable (varB1, varB2) fragments and expressed as GST/His-tag fusion proteins. Four-week-old hamsters were immunized with equal amounts of each fragment individually or combined in alum adjuvant at days 0 and 21 and subsequently challenged three weeks after the booster with 2.5 LD(50) live virulent Leptospira interrogans serovar Pomona. Our results demonstrate that immunization with LigB produced strong humoral immune responses as revealed by high titers against each fragment and significant enhancement in Th2 cytokines (IL-4, IL-10). A significant activation of CMI is revealed by enhanced proliferation of lymphocytes and up regulation of Th1 cytokines (IL-12p40, IFN-gamma) was also noted. Of the peptides studied, rLigBcon was able to impart maximum protection (71%), followed by rVarB1 (54%), whereas rVarB2 was not able to impart a significant level of protection (33%) against lethal infection as revealed by enhanced survival and reduced severity of histopathological lesions in vital organs (viz. kidney, liver, spleen) of the immunized animals. Moreover, concurrent administration of all three fragments significantly enhanced the protective efficacy of the vaccine (83%). Overall, our results clearly demonstrate that LigB has emerged as novel protective antigen that can be used in future subunit vaccines against leptospirosis.


Subject(s)
Antigens, Bacterial/immunology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/prevention & control , Adjuvants, Immunologic/administration & dosage , Adjuvants, Immunologic/pharmacology , Alum Compounds/administration & dosage , Alum Compounds/pharmacology , Animals , Antibodies, Bacterial/blood , Antigens, Bacterial/genetics , Cell Proliferation , Cricetinae , Immunization, Secondary , Interleukin-10/metabolism , Interleukin-4/metabolism , Kidney/pathology , Leptospira interrogans serovar pomona/genetics , Liver/pathology , Lymphocytes/immunology , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Spleen/pathology , Survival Analysis , Vaccines, Subunit/genetics , Vaccines, Subunit/immunology
16.
Asian Pac J Allergy Immunol ; 26(4): 229-36, 2008 Dec.
Article in English | MEDLINE | ID: mdl-19317342

ABSTRACT

Leptospira interrogans, the causative agent of leptospirosis, is an important zoonotic bacterium. The mechanisms and roles of cytokine induction in both humans and animals remain unclear. Therefore, the IFN-gamma, IL-6, IL-10 and IL-12 levels were measured by enzyme-linked immunosorbent assay (ELISA) in human THP-1 and mouse RAW 264.7 monocyte cell lines following stimulation with heat-killed Leptospira interrogans serogroup Pomona serovar Pomona, L. biflexa, E. coli or Salmonella group B. The production of IL-6 and IL-12 were higher and rose more rapidly in the RAW 264.7 cells with all bacteria. The IL-10 was not detected in the RAW 264.7 cells when induced by leptospires. The IFN-gamma level in human peripheral blood mononuclear cells (PBMCs) induced by leptospires was also significantly lower than with other bacteria. When IL-10 and IL-12 mRNA expressions were detected in hamster's spleen, their patterns were similar to what was observed in THP-1 in that IL-12 was only slightly increased while IL-10 expression was high. Moreover, the IFN-gamma expression could not be detected in hamsters. The more potent cytokine responses in the RAW 264.7 cells may indirectly reflect the disease outcome in mice which render them to be a good reservoir of leptospirosis. Whether these cytokines have contributed to immunoprotection during the L. interrogans infection remains to be further investigated.


Subject(s)
Cytokines/biosynthesis , Leptospira interrogans serovar pomona/immunology , Leptospirosis/immunology , Leukocytes, Mononuclear/immunology , Animals , Cell Line , Humans , Interferon-gamma/biosynthesis , Interleukin-10/biosynthesis , Interleukin-12/biosynthesis , Interleukin-6/biosynthesis , Leukocytes, Mononuclear/microbiology , Mice
17.
Vaccine ; 26(2): 277-87, 2008 Jan 10.
Article in English | MEDLINE | ID: mdl-18055070

ABSTRACT

We demonstrated earlier that immunization with recombinant Leptospira immunoglobulin like protein A (LigA) induced significant protection against virulent Leptospira interrogans serovar Pomona challenge in hamsters. However, the protective immune mechanism remains unclear. In the present study we demonstrated the protective efficacy of a LigA DNA vaccine and evaluated the immune mechanism underlying the protection against leptospirosis in hamsters. The LigA DNA vaccine was constructed in two truncated forms as the conserved portion (LigAcon) and a variable portion (LigAvar). Four-week-old hamsters were immunized three times at two-week intervals with vector alone or an equal amount of a recombinant construct containing either LigAcon or LigAvar. All animals were challenged intraperitoneally 2 weeks after the last immunization with a dose (LD50=10(8)) of virulent L. interrogans serovar Pomona. Prior to challenge, four animals were sacrificed, the spleen was removed aseptically, and splenocytes were assayed for lymphocyte proliferation and cytokine profiles in response to recall antigen. The protective efficacy was evaluated on the basis of survival and histopathological lesions in the kidney. The immuno-protective mechanism was assessed on the basis of Th1/Th2 profile of cytokines in immunized animals. Our results indicate that immunization with LigA DNA vaccine provides significant protection against leptospirosis. We suggest that immuno-protection is conferred by both humoral and cellular immunity as revealed by an increase in antibody titers during subsequent boosters, significant proliferation of lymphocytes and enhancement of both Th1 and Th2 cytokines. Taken together, the present study suggests that a LigA DNA vaccine is a promising candidate for prevention of leptospirosis.


Subject(s)
Antigens, Bacterial/immunology , Leptospira interrogans serovar pomona/immunology , Leptospirosis/prevention & control , Vaccines, DNA/immunology , Animals , Antibodies, Bacterial/blood , Antigens, Bacterial/genetics , Cell Proliferation , Cricetinae , Cytokines/biosynthesis , Cytokines/immunology , Female , Immunization, Secondary , Kidney/pathology , Leptospirosis/immunology , Lymphocytes/immunology , Spleen/immunology , Survival Analysis , Vaccines, DNA/genetics
18.
Rev. Soc. Bras. Med. Trop ; 40(6): 648-652, nov.-dez. 2007. ilus, graf, tab
Article in Portuguese | LILACS | ID: lil-471345

ABSTRACT

O presente trabalho teve por objetivo identificar a presença da Leptospira interrogans sorovar pomona em camundongos geneticamente selecionados para a alta e baixa resposta a anticorpos. Todos os animais foram submetidos ao isolamento bacteriano, imunohistoquímica (imunoperoxidase) em cortes de tecido renal e coloração através da hematoxilina-eosina. A técnica de imunoperoxidase apresentou-se pouco mais sensível em relação ao cultivo, entretanto, ambas foram bons parâmetros de identificação do agente. Presença de lesões renais mais intensas ocorreram em períodos em que houve maior número de bactérias isoladas em meio de cultivo. Camundongos da linhagem HIV-A conseguiram eliminar as leptospiras com maior eficiência e rapidez em relação as linhagem LIV-A, entretanto o estudo demonstrou que ambas linhagens da seleção IV-A foram eficientes em controlar o processo infeccioso.


The present work had the objective of identifying the presence of Leptospira interrogans serovar pomona in mice that had been genetically selected for high and low response to antibodies. All the animals were subjected to bacterial isolation, immunohistochemical analysis (immunoperoxidase) in renal tissue sections and hematoxylin-eosin staining. The immunoperoxidase technique was little more sensitive than culturing, but both were good parameters for agent identification. More severe renal lesions were present at times when there were greater numbers of bacteria isolated in culture medium. Mice of the lineage HIV-A were able to eliminate the Leptospira more efficiently and faster than the lineage LIV-A could. However, the study demonstrated that both lineages of the IV-A selection were efficient in controlling the infectious process.


Subject(s)
Animals , Male , Mice , Culture Media , Immunoenzyme Techniques , Kidney/microbiology , Leptospira interrogans serovar pomona/isolation & purification , Eosine Yellowish-(YS) , Hematoxylin , Host-Pathogen Interactions , Kidney/pathology , Leptospira interrogans serovar pomona/immunology , Mice, Mutant Strains , Sensitivity and Specificity , Staining and Labeling , Time Factors
19.
N Z Vet J ; 55(3): 102-8, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17534410

ABSTRACT

Current knowledge of leptospirosis in farmed deer in New Zealand is reviewed. Over the past 25 years, leptospirosis has been reported to occur in individual cases as well as in herd outbreaks in farmed deer and in human cases linked to farmed deer. Serological studies and evidence from bacterial culture suggest infection is widespread. Mixing of young stock from several sources appears to be a significant risk factor for outbreaks. The culture of Leptospira interrogans serovars Hardjobovis, Pomona and Copenhageni has been reported. Infection with serovar Hardjobovis had the highest prevalence, either individually or mixed with serovar Pomona. Infection with serovar Copenhageni appears uncommon and its pathogenicity in deer is unproven. Titres to serovars Australis, Ballum, Balcanica and Tarassovi have been reported. Deer appear to be maintenance hosts for serovar Hardjobovis, incidental or accidental hosts and probably a maintenance population for serovar Pomona, since some infections persist for several months, and accidental hosts for serovar Copenhageni. Serovar Pomona appears to produce clinical and probably subclinical disease, whereas serovar Hardjobovis appears to cause only subclinical disease, although the relative risk of disease causation has not been determined. Clinical disease is usually manifested by haemolysis, jaundice, renal lesions, haemoglobinuria and often by sudden death. Renal lesions are commonly observed at slaughter and many are associated with leptospiral infections. Occupationally, slaughterhouse workers appear to be at greatest risk of contracting the disease from deer. Vaccination produces serological responses, but its effectiveness in protecting against disease, and prevention or reduction of shedding in urine, has not yet been confirmed in deer. More robust knowledge of the epidemiology of leptospiral infections in deer, and the effectiveness of vaccines and vaccination regimes, is needed to assist the deer industry to develop a strategy to manage this disease.


Subject(s)
Deer/microbiology , Leptospira interrogans/isolation & purification , Leptospirosis/veterinary , Vaccination/veterinary , Animals , Animals, Domestic , Disease Outbreaks/veterinary , Female , Humans , Leptospira interrogans/immunology , Leptospira interrogans/pathogenicity , Leptospira interrogans serovar hebdomadis/immunology , Leptospira interrogans serovar hebdomadis/isolation & purification , Leptospira interrogans serovar hebdomadis/pathogenicity , Leptospira interrogans serovar pomona/immunology , Leptospira interrogans serovar pomona/isolation & purification , Leptospira interrogans serovar pomona/pathogenicity , Leptospirosis/epidemiology , Leptospirosis/pathology , Leptospirosis/prevention & control , Male , New Zealand/epidemiology , Risk Factors , Seroepidemiologic Studies , Species Specificity , Zoonoses
20.
Rev Soc Bras Med Trop ; 40(6): 648-52, 2007.
Article in Portuguese | MEDLINE | ID: mdl-18200418

ABSTRACT

The present work had the objective of identifying the presence of Leptospira interrogans serovar pomona in mice that had been genetically selected for high and low response to antibodies. All the animals were subjected to bacterial isolation, immunohistochemical analysis (immunoperoxidase) in renal tissue sections and hematoxylin-eosin staining. The immunoperoxidase technique was little more sensitive than culturing, but both were good parameters for agent identification. More severe renal lesions were present at times when there were greater numbers of bacteria isolated in culture medium. Mice of the lineage HIV-A were able to eliminate the Leptospira more efficiently and faster than the lineage LIV-A could. However, the study demonstrated that both lineages of the IV-A selection were efficient in controlling the infectious process.


Subject(s)
Culture Media , Immunoenzyme Techniques , Kidney/microbiology , Leptospira interrogans serovar pomona/isolation & purification , Animals , Eosine Yellowish-(YS) , Hematoxylin , Host-Pathogen Interactions , Kidney/pathology , Leptospira interrogans serovar pomona/immunology , Male , Mice , Mice, Mutant Strains , Sensitivity and Specificity , Staining and Labeling , Time Factors
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