Leptospira Antigens Research Articles

Coinfection with Orthohantavirus and Leptospira spp. in Rats Collected from Markets in Indonesia.

Background: Rats are an important reservoir animal for several zoonotic pathogens worldwide, including hantaviruses and Leptospira spp., which are the causative agents of hemorrhagic fever with renal syndrome, hantavirus cardiopulmonary syndrome, and leptospirosis. Although a previous study indicated a high frequency of antihantaviral antibodies in patients with acute fever in Indonesia, circulating hantaviruses and their reservoir animals in the country remain limited. Materials and Methods: The presence of hantavirus in rats captured in the urban area of Bogor, Indonesia, from which Leptospira spp. were isolated using PCR, followed by DNA sequencing. Immunohistochemical analyses were performed to detect hantaviral and leptospiral antigens in rat kidney tissues. Results: Seoul of Orthohantavirus seoulense (SEOV) RNA was detected from 24 of 80 Rattus norvegicus (30%). SEOV and Leptospira coinfection was detected in 10 of 80 rats (12.5%). Immunohistochemistry revealed that hantavirus antigens were positively stained in the interstitial capillaries and cells, whereas Leptospira antigens were stained in the luminal side of the renal tubules. Conclusion: This study revealed a high prevalence of SEOV and SEOV and Leptospira coinfection among rats in the urban areas of Bogor, Indonesia, indicating a potential risk of rat-borne zoonotic diseases in the area.

Enhancement of clinical signs in C3H/HeJ mice vaccinated with a highly immunogenic Leptospira methyl-accepting chemotaxis protein following challenge.

Leptospirosis is the most widespread zoonosis and a life-threatening disease in humans and animals. Licensed killed whole-cell vaccines are available for animals; however, they do not offer heterologous protection, do not induce long-term protection, or prevent renal colonization. In this study, we characterized an immunogenic Leptospira methyl-accepting chemotaxis protein (MCP) identified through a reverse vaccinology approach, predicted its structure, and tested the protective efficacy of a recombinant MCP fragment in the C3H/HeJ mice model. The predicted structure of the full-length MCP revealed an architecture typical for topology class I MCPs. A single dose of MCP vaccine elicited a significant IgG antibody response in immunized mice compared to controls (P < 0.0001), especially the IgG1 and IgG2a subclasses. The vaccination with MCP, despite eliciting a robust immune response, did not protect mice from disease and renal colonization. However, survival curves significantly differed between groups, and the MCP-vaccinated group developed clinical signs faster than the control group. There were differences in gross and histopathological changes between the MCP-vaccinated and control groups. The factors leading to enhanced disease process in vaccinated animals need further investigation. We speculate that anti-MCP antibodies may block the MCP signaling cascade and may limit chemotaxis, preventing Leptospira from reaching its destination, but facilitating its maintenance and replication in the blood stream. Such a phenomenon may exist in endemic areas where humans are highly exposed to Leptospira antigens, and the presence of antibodies might lead to disease enhancement. The role of this protein in Leptospira pathogenesis should be further evaluated to comprehend the lack of protection and potential exacerbation of the disease process. The absence of immune correlates of protection from Leptospira infection is still a major limitation of this field and efforts to gather this knowledge are needed.

Potentially fatal cerebral venous sinus thrombosis followed by a hemorrhagic stroke in a vaccinated patient: Do not rule out COVID-19 as a reason

Stroke is amongst the leading causes of morbidity and mortality globally. Majority of strokes are ischemic and up to 1/3rd ischemic strokes have infective etiology. In this post-pandemic era, SARS-CoV-2 is an established infectious agent which can precipitate it by various mechanisms. Though post-vaccination incidence of COVID-19 has dropped drastically, still new cases get reported with typical or atypical presentations. Irrespective of causation, malignant cerebral edema in stroke necessitates salvage neurosurgery. Hereby authors present an interesting diagnostic chase in a young female who presented in altered sensorium, underwent emergency decompressive craniectomy and neurologically recovered over subsequent days. Despite being double vaccinated, she developed potentially fatal COVID-encephalitis, cerebral venous sinus thrombosis (CVST), ischemic infarcts and hemorrhagic transformation. Suprisingly, there were cross reacting antibodies against antigens of herpes simplex virus (HSV), leptospira and cytomegalovirus (CMV) yielding false positivity on serology screening test, which spoofed us while carrying out infectious disease work up of stroke.

Humoral immune response and changes in peritoneal cell populations in rats immunized against two Leptospira serovars; serovar patoc and serovar pyrogenes

BackgroundLeptospirosis is a zoonotic disease caused by Leptospira species. Variations in lipopolysaccharide (LPS) structure in Leptospira are known to be associated with the serovar diversity and antigenicity. Development of immunodiagnostics for early detection of leptospirosis based on immune responses against different pathogenic antigens as well as development of vaccines are important. Hence, this study has assessed the immune response generated against leptospiral LPS and whole antigen preparations of pathogenic and saprophytic Leptospira and specific changes in peritoneal cells was also studied to elucidate the cellular responses associated with immune response of Wistar rats.MethodsDuring the study, immune response induced by two types of Leptospira antigen preparations of two selected serovars was compared. Changes in the specific peritoneal cell subpopulations following immunizations of rats were analyzed using flow cytometry.ResultsOf the two antigen preparations tested, the LPS extract induced a higher IgM immune response as opposed to the sonicated antigen preparation. Of the two serovars tested, L. interrogans serovar Pyrogenes had induced a higher IgM response compared to that by L. biflexa serovar Patoc. Considering the IgG titers, equivalent responses were observed with all four antigen preparations. Significant increases in lymphocytes were observed following immunization with LPS of both serovars. Interestingly, the B2 cell percentages increased significantly during the immunization period. Further, significant correlations were observed with both IgM and IgG responses and percentage of B2 cells in the peritoneal cavity (PC).ConclusionLPS extract of L. interrogans serovar Pyrogenes induced higher IgM response while the IgG response was equivalent among the four antigen preparations tested. Significant increase of B2 cell percentage in the peritoneal cavity during the immunization reflects the accumulation of B2 cells in the PC which may play considerable role in generating humoral response against Leptospira antigens.

Genetic Analysis of LigA, LipL32, Loa22, and OmpL1 Antigens from Leptospira spp. Sejroe Serogroup: How Close We Are To a Vaccine Against Bovine Leptospirosis?

Bovine leptospirosis has as main causative agents Leptospira spp. from Sejroe serogroup. Vaccination is a crucial step to control this infection. The use of conserved proteins among Leptospira spp. is of great importance for a protective immune response. The aim of the present study is to genetically analyze antigens of Leptospira spp. from Sejroe serogroup strains isolated from cattle for a preliminary evaluation of vaccine candidates. Genes associated with antigenicity-LigA, LipL32, Loa22, and OmpL1-were analyzed through bioinformatic and immunoinformatic tools. Despite high diversity observed in strains, on an amino acid level, highly conserved regions were observed (> 90%), particularly in LipL32 gene. Moreover, highly conserved amino acid regions (> 30 aa) were observed in all genes, regardless of species, geographical origin or biological source of isolation. Superposed structures of protein fragments including all the predicted MHC-II and B-Cell epitopes were demonstrated. Results presented herein are preliminary, but a fundamental step towards the development of an efficient vaccine against bovine leptospirosis, a silent but enormously concerning disease.

The Clinical and Laboratory Profiles of Immunocompetent Patients With Short-Duration Fever With Neutropenia in a Tertiary Care Hospital in Pune, India.

Background Management of a febrile patient is based on understanding the pathophysiology of an abnormal temperature and temperature regulation, impacts of fever, and its treatment. In the current study, we aimed to characterize and compare the epidemiological, etiologic, microbiological, serological, clinical, and outcome traits of febrile patientswith acute neutropenia admitted to a tertiary care center in Western Maharashtra. Methods Adult patientswith a history of fever of less than two weeks' duration and without any immunosuppressive state were screened with predefined inclusion and exclusion criteria. General anddemographic information (age andgender), and clinical examinations (type and duration of fever) were recorded. Biochemical, hematologic(total and differential cell counts), and immunologicmeasurements (rapid malaria, dengue, Leptospira, and viral hepatitis antigen antibodies) were performed. Data were analyzed using an appropriate statistical package. Results A total of 403 (214 males) young adults (aged: 29±11 years) with clinical presentation of fever were studied. The majority (n=361, 89.6%) had low-grade continuous fever with an average duration of 3±1 (mean±standard deviation (SD)) days. Headache and myalgia were the common symptoms present, and patients had an average hospital stay of 4±1 days. Dengue (55%) was the most common cause of febrile neutropenia, and all patients recovered well without antibiotics and granulocyte colony-stimulating factor. The mean C-reactive protein (CRP) level was 61.4±4.4 mg/L. CRP and procalcitonin (PCT) were directly correlated with the degree of neutropenia and inverselycorrelated with total leucocyte count (TLC). Conclusions It was highlighted from this study that antibiotics are not necessary for viral infections that have been diagnosed to stop the development of secondary bacterial infections. A clinician should be aware of "when not to use antibiotics,"or the world will soon have to deal with superbugs.

Next generation sequencing of transcribed genes in ruminant γδ T cell populations

Bovine γδ T cells are distinguished by expression of WC1, hybrid pattern recognition receptors and co-receptors to the T cell receptor (TCR), or their absence. WC1 molecules bind pathogens and the ability of γδ T cells to respond to pathogens largely correlates with their expression of particular WC1 genes. Following activation, the TCR and WC1 molecules co-localize and knocking down WC1 abrogates the ability of WC1-expressing γδ T cells to respond to antigen. It is known that these two major populations, WC1+ and WC1-, differ in their TCR gene expression and previous studies showed other differences using semi-quantitative RT-PCR and serial analysis of gene expression. Differences in genes expressed would influence the functional outcome when WC1+ vs. WC1- γδ T cells respond to pathogens. To identify unique aspects of their transcriptome, here we performed RNA-Seq of flow cytometrically sorted bovine WC1+ and WC1- γδ T cells and compared them to all mononuclear cells in blood. The greatest differences in gene expression were found between γδ T cells and other mononuclear cells and included those involved in lymphocyte activation and effector processes. Only minor differences occurred between ex vivo WC1+ vs. WC1- γδ T cells with those gene products being involved in cell adhesion and chemotaxis. After culturing cells from primed animals with Leptospira antigens major difference in the transcriptome was evident, with over 600 genes significantly differentially expressed including those focused on cytokine signaling. Unexpectedly, antigen-responding and non-responding populations of WC1+ γδ T cells had few differences in their transcriptomes outside of cytotoxic factors although they had more WC1–1, WC1–2 and WC1–13 transcripts. Through differential gene expression we were able to define properties of ex vivo and stimulated WC1+ cells which will be useful in understanding their functional biology.

The effect of natural infection with different Leptospira interrogans serovars on oxidative stress biomarkers and acute-phase responses in horses and cattle.

Leptospirosis is one of the important zoonotic bacterial diseases with a worldwide distribution that is often subclinical. We aimed to investigate the oxidant/antioxidant balance and acute-phase response in naturally infected horses and cattle with Leptospira interrogans. A total of 600 serum samples from horses and cattle were examined for L. interrogans antibodies using the microscopic agglutination test to determine anti-Leptospira IgG antibodies against a panel of eight important Leptospira antigens in Iran. Then, serum total antioxidant capacity, catalase, glutathione peroxidase, and malondialdehyde activities, and nitric oxide, total protein, serum amyloid A, haptoglobin, and albumin concentrations were measured in seropositive and seronegative samples. Serum catalase activities and malondialdehyde, serum amyloid A, and haptoglobin ‌concentrations in seropositive cattle and horses were significantly higher (P<.05) than in those that were seronegative. Antibody titers increased from 1:100 to 1:200 in cattle with L. interrogans infection, resulting in a decrease in the serum total antioxidant capacity (P<.05), an increase in serum glutathione peroxidase (P<.01) activity and nitric oxide (P<.05) , serum amyloid A (P<.01), and haptoglobin (P<.05) concentrations. Following the increase in the agglutinating antibody titers in horses infected with L. interrogans, the serum total antioxidant capacity (P<.01) decreased, and serum nitric oxide (P<.05), malondialdehyde (P<.05), and serum amyloid A (P<.01) concentrations were increased. In this study, horses and cattle had extensive changes in oxidant/antioxidant equilibrium and acute-phase protein concentrations when infected with L. interrogans. We also demonstrated a direct link between antibody titers and the type of leptospiral serovar using serum oxidative and inflammatory markers.

Absence of serological or molecular evidence of Leptospira infection in farmed swine in the Hong Kong Special Administrative Region

Leptospirosis is an important zoonotic disease with several maintenance host species including swine. A cross sectional survey was undertaken between January to October 2020 to investigate the prevalence of leptospirosis in farmed swine in the Hong Kong Special Administrative Region (HKSAR) of China. Serum samples were collected from swine on seven farms (15 swine per farm; ten multiparous sows and five twelve-week-old weaners), while kidney samples were collected from 64 swine submitted for routine post-mortem (26 farms; average 2.4 swine per farm, range 1–6). Microscopic agglutination tests (MAT) to a panel of 24 Leptospira antigens did not reveal any evidence of seroconversion at a titre of 1:100. Polymerase chain reaction (PCR) testing of the kidney samples for Leptospira DNA did not detect any evidence of infection. Bayesian methods were used to compute the probability that the leptospirosis prevalence in farmed swine in the HKSAR was <3%, given none of the 105 swine sampled were positive on the MAT. The results of this study demonstrate no serological or molecular evidence of leptospirosis in farmed swine in the HKSAR. Subsequent statistical analysis supports the conclusion that the prevalence of leptospirosis in farmed swine in the HKSAR is negligible at present.

Development of in-house ELISAs as an alternative method for the serodiagnosis of leptospirosis

BackgroundLeptospirosis is most often diagnosed clinically, and a laboratory test with high diagnostic accuracy is required. MethodsIgM and IgG ELISAs using Leptospira antigens were established and evaluated in relation to the microscopic agglutination test (MAT). Antigen preparation consisted of saprophytic Leptospira biflexa to detect genus-specific antibodies (genus-specific ELISA) and a pool of the five most prevalent Leptospira interrogans serovars in Sri Lanka to detect serovar-specific antibodies (serovar-specific ELISA). IgM and IgG immune responses were studied in severe and mild leptospirosis patients (n = 100 in each group). ResultsThe ELISAs showed high repeatability and reproducibility. The serovar-specific IgM-ELISA showed a sensitivity of 80.2% and specificity of 89%; the genus-specific IgM-ELISA showed a sensitivity of 83.3% and specificity of 91%. The serovar- and genus-specific IgG-ELISAs showed sensitivities of 73.3% and 81.7%, respectively, and specificities of 83.3% and 83.3%, respectively. The commercial IgM-ELISA showed a sensitivity of 79.2% and specificity of 93%. The commercial IgG-ELISA showed a sensitivity of 50% and specificity of 96.7%. IgM levels observed in mild and severe leptospirosis patients were significantly higher than in the healthy control group, with mean absorbance values of 0.770, 0.778, and 0.163, respectively. Severe leptospirosis patients had significantly higher mean anti-leptospiral IgG levels compared to both mild leptospirosis patients and healthy control group subjects (0.643, 0.358, and 0.116, respectively; ANOVA, p < 0.001). The presence of anti-leptospiral IgG above an optical density of 0.643 at 1:100 could predict a high risk of severe disease. ConclusionThe serovar-specific in-house ELISA could be used for the laboratory diagnosis of leptospirosis in endemic settings. The high levels of anti-leptospiral IgG observed suggest its value as a predictor of disease severity.

Goat γδ T cell subpopulations defined by WC1 expression, responses to pathogens and cytokine production

The major functions of γδ T cells in mammals overlap with those of αβ T cells but differ in that γδ T cells are rapid responders and see different types of antigens. While γδ T cells have been shown to be a major population of circulating lymphocytes in artiodactyl species such as cattle, sheep, and pigs, less is known about these cells in goats, an important agricultural species. We have recently shown that WC1, a γδ T cell-specific family of hybrid pattern recognition receptors/co-receptors, is a multigenic family in goats expanded beyond what occurs in cattle. This study was conducted to address some of the limitations of previous studies in determining the proportions of γδ T cells, WC1+ γδ T cells as well as the WC1.1+ and WC1.2+ subpopulations in blood and to evaluate their responses to various pathogens. Previously, the proportion of caprine γδ T cells was determined using a monoclonal antibody (mAb) 86D that we show here does not react with all γδ T cells thereby underestimating their contribution to the lymphocyte population. Using a mAb reactive with the TCRδ constant region we found the proportion of γδ T cells in blood was not significantly less than that of either CD4 or CD8 T cells and did not decrease with age after 6 months. γδ T cells that expressed WC1 ranged from ~20 to 85% of the total γδ T cells. Less than half of those were classified as WC1.1+ or WC1.2+ by mAb staining thus indicating a third major WC1+ population. We found that naïve γδ T cells proliferated in cultures of PBMC stimulated with antigens of Leptospira or Mycobacterium avium paratuberculosis (MAP) more than they did in control medium cultures or in those stimulated with M. bovis BCG antigens and that the responding γδ T cells included both WC1+ and WC1- cells. In ex vivo PMA/ionomycin-stimulated cultures of WC1- γδ T cells but not WC1+ cells produced both IL-17 and IFNγ. In longterm cultures with Leptospira or MAP both WC1- and WC1+ cells proliferated but only WC1- γδ T cells produced IL-17. In conclusion, goats have a substantial number of WC1- and WC1+ γδ T cells in PBMC that do not decrease with animal age after 6 months; both populations respond to bacterial antigens as naïve cells but in these cultures only the WC1- γδ cells produc IL-17 and IFNγ .

Cloning, expression and purification of Leptospira LigB antigen in Escherichia coli

Leptospira is one of the most common zoonotic diseases in the tropics and subtropics. Humans are infected by exposure to Leptospira contained water or food sources. Leptospirosis usually breaks out after the flood and causes several consequences for people and economy. Leptospirosis disease, if not rapidly detected and treated promptly, it causes serious consequences such as acute hepatitis-kidneys, meningitis and bleeding, heart and nerve complications, and severe illness can lead to death. Therefore, quick and accurate detection of Leptospira pathogen plays a very important role in Leptospirosis disease treatment. Among antigens of Leptospira, a conserved domain of LigB antigen (Leptospiral immunoglobulin-like protein) was reported that is present in the most of pathogenic serovars of Leptospira, but not in the non-pathogenic Leptospira biflexa, thus this conserved domain was used for production of Leptospirosis detection kits as well as vaccine for Leptospirosis. In order to create a kit for Leptospirosis diagonostic, especially detect anti-Leptospira antibodies in Leptospira infected serum and plasma samples, about 1kb gene fragment encoding for conserved domain of LigB (about 36 kb in molecular weight) was used as the material for producing of LigB protein by DNA recombinant technology. In this study, we present the results for cloning, expressing a conserved domain of LigB antigen in E. coli cells and purifying protein by affinity chromatography collumn. The result indicates that recombinant LigB protein was successfully expressed in E. coli Rosetta 1 and purified by Hitrap chealating collumn. The LigB protein concentration after purification reached 60 mg/L medium with 98% purity. This purified protein will be used as the materials for creating Leptospirosis kit.

Detection and modeling of anti-Leptospira IgG prevalence in cats from Lisbon area and its correlation to retroviral infections, lifestyle, clinical and hematologic changes

Leptospirosis is a zoonosis of global importance caused by Leptospira species. Rodents are the main reservoirs, known to shed the bacteria in urine, thus contaminating water and soil and infecting other animals and people. Leptospirosis has been re-emerging in both developing and developed countries including Europe. It has been hypothesized that cats could be asymptomatic carriers of Leptospira. This study aims to evaluate cats’ exposure to Leptospira in Lisbon, Portugal, by measuring IgG titres and correlating them with possible factors that may increase the risk of exposure in urban cats. Two hundred and forty-three samples were collected from the biobank. An ELISA test followed by a seroprevalence analysis using a finite mixture model was performed to detect and measure anti-Leptospira IgG antibodies titres. In parallel, a survey was conducted to identify possible risk factors for seropositivity.According to the ELISA test protocol, only twenty-three cats (9.5%; 95% CI =(6.1%;13.9%)) could be considered as seropositive to Leptospira antigens. However, when the same data were analysed by the best different mixture models, one hundred and forty-four cats (59.3%; 95%CI = (52.8%; 65.5%)) could be classified as intermediate and high antibody responders to Leptospira antigens. Seropositivity to Feline Immunodeficiency Virus infection (FIV) was found to be the only significant risk factor associated with anti-Leptospira IgG antibodies. In conclusion, the present studies raises the possibility of a higher exposure of cats to Leptospira than previously thought due to the identification of a subpopulation of cats with intermediate antibody levels.

Prevalence of Leptospira serogroups in buffaloes from the Brazilian Amazon.

Although Brazil has one of the largest buffalo populations in the Americas, buffalo leptospirosis is still poorly explored when compared to that in bovines; thus, the aim of this research was to carry out a large serological study for leptospirosis in this species in the Brazilian Amazon. For this, we collected 1,405 serum samples from buffaloes raised in the Amazon delta region, which is considered a major area of buffalo production in Brazil. The test used was a microscopic agglutination test (MAT) adopting 34 Leptospira antigens, some of which have never been tested for buffaloes in Brazil, including autochthonous strains; in total, 20 serogroups were evaluated. From the total of 1,405 serum samples, 894 (63.6%) reacted in the MAT to at least one of the 20 serogroups, and 511 (36.4%) did not react. The serogroups Sejroe, Autumnalis and Pomona were the most prevalent, with titres ranging from 100 to 12,800, and the autochthonous strains used were not significant in relation to the reference serovars. Leptospirosis in buffaloes seems to have a serological profile similar to leptospirosis in cattle, mainly due to the prevalence of the Sejroe serogroup; however, the results of this study suggested that in the Brazilian Amazon, Leptospira strains that are serologically distinct from the autochthonous strains isolated in the southeastern region of Brazil may be circulating in these animals. Other serovars could also be inserted into the panel of antigens used in MAT for serological studies on buffaloes.

Bovine immune response to leptospira antigen in different novel adjuvants and vaccine delivery platforms

Leptospirosis is a global zoonosis causing significant economic losses for cattle production. Current cattle vaccines against leptospirosis need improvement to provide efficacy against multiple serovars, reduce shedding in urine, and to induce earlier and more robust immune responses. In this study, Leptospira borgpetersenii serovar Hardjo strain 203 antigen was combined with novel adjuvants (a biodegradable polyanhydride compressed rod implant (VPEAR), poly(diaminosulfide) microparticles, a water-oil-water emulsion adjuvant, and aluminum hydroxide) to develop novel vaccines. Cattle were immunized twice, at a 4 week interval, with inoculums containing adjuvants alone or leptospira antigens and immune responses were compared to responses of cattle receiving a commercial monovalent leptospirosis vaccine (Spirovac). All animals were inoculated with a single dose of Spirovac at 20 weeks to assess antigen recall responses. Serum antibody responses were increased (P > 0.05) at 8 and 20 weeks after vaccination in cattle receiving inoculums containing leptospira antigens combined with water-oil-emulsion, poly(diaminosulfide) microparticles (PNSN-MP), or aluminum hydroxide and in cattle vaccinated with Spirovac. Humoral responses were predominantly IgG1 isotypes. Antigen-specific proliferative responses were detected after initial vaccination in cattle vaccinated with Spirovac, PNSN-MP and water-oil-water treatments. Most proliferative responses occurring within CD4+ and gamma delta T cell populations expressing CD45RO and CD25 markers, a response consistent with an effector memory phenotype. Antigen-specific immune responses were not detected in cattle vaccinated with VPEAR after initial inoculation, but were detected in the antigen recall responses. PBMCs from cattle vaccinated with Spirovac, oil-water-oil, or PNSN-MP treatments had increased (P < 0.05) IL-17A release after in vitro stimulation with leptospirosis antigens, whereas all groups produced IFN-γ and IL-17A after in vitro stimulation during the antigen recall response. Our data demonstrates that combining leptospirosis antigens with these adjuvants enhances immunogenicity in cattle.Interpretative Summary: Vaccination of livestock is a key mechanism for minimizing transmission of leptospirosis, a zoonotic disease. Leptospirosis vaccines for cattle need to be improved to provide greater levels of protection from kidney colonization, better immune responses, and protection against multiple serovars. This could be accomplished using new vaccine adjuvants. In this study, several novel adjuvants were evaluated for their ability to induce effective immune responses in cattle to leptospira antigens as compared to currently available vaccines. Data suggested that vaccines containing biodegradable polymer microparticles and oil-emulsion adjuvants induced similar or greater immune responses as compared to a commercial vaccine. Our data suggest these new vaccine formulations warrant further investigation as new vaccine formulations for cattle and other livestock.

Molecular Mimicry between Betaine Aldehyde Dehydrogenase of Leptospira and Retinal Dehydrogenase 1 of Human Lens: A Potential Trigger for Cataract Formation in Leptospiral Uveitis Patients

ABSTRACT Purpose: Rapidly progressing cataract is one of the ocular manifestations in leptospiral uveitis patients. We examined whether molecular mimicry between the leptospira antigens and lens proteins exists that could result in cataract in these patients. Methods: Immunoblot analysis using patient sera was done with proteins from normal lens and cataract lens from leptospiral uveitis patients and the cross-reacting lens proteins were identified by mass spectrometry analysis. Results: Retinal dehydrogenase 1 and crystallins (α-B, α-A2, β-B2), were recognized by the antibodies in the serum of leptospiral uveitis patients. And, retinal dehydrogenase 1 is homologous to the leptospiral protein, betaine aldehyde dehydrogenase. Conclusions: Leptospiral uveitis patient serum contains antibodies that cross-react with multiple lens proteins that have a role in maintaining lens transparency. And, these antibodies could act as a potential trigger for cataractogenesis.

Evaluation of Postvaccinal Antibody Responses to Three Brands of Combined Canine Vaccines Containing Canine Parvovirus Type 2 Following Single Administration in Nigerian Local Dogs (Canis lupus familiaris)

The current level of morbidity and mortality among dogs due to canine parvovirus infection despite vaccination raises concerns about the effectiveness of commercially available polyvalent modified live vaccine containing Distemper, Adenovirus, Leptospira, Parvovirus and Parainfluenza antigens (DHLPPi) in Nigeria. This study was conducted to evaluate canine parvovirus type 2 (CPV-2) antibody responses to three different brands of DHLPPi vaccines.Fifteen Nigerian local dogs, age 8- 14 weeks; were randomly selected. The dogs were divided into 3 groups (A, B and C) each containing 5 dogs. The dogs were vaccinated with respective polyvalent modified live vaccines containing Distemper, Adenovirus, Leptospira, Parvovirus and Parainfluenza antigens (DHLPPi) tagged A, B and C. Blood samples were collected before vaccination weekly for four weeks and on day 90 post vaccination. The sera from the collected blood samples were subjected to Indirect Enzyme Linked Immunosorbent Assay (ELISA). The mean titre values were analysed using ANOVA test and student t-test. Group A had a statistical significant (P<0.05) higher antibody titre when compared with that of group B on day 21. A statistical significant (P<0.05) increase in antibody response was found in group Bwhen compared to group A on days 14 and 28 and group C on day 14. Group C had significantly higher antibody titre when compared to group A on days 7 and 28 and group B on day 21. The group C antibody titres were found to be the consistently higher than those of groups A except on days 14 and 21 and and group B on every other days except on days 14 and 90. In conclusion, the three vaccines were found to be immunogenic, but vaccines B and C gave adequate and consistently high immune response whilevaccine A gave the least response. The occurrence CPV enteritis in dogs despite vaccination may therefore, be attributed to vaccine failure as a result of poor immunogenicity of some vaccines used in routine vaccination of dogs and other factors that may affect the quality of vaccine or ability of the host to produce adequate antibody to administered vaccines.

Severe Leptospirosis Features in the Spleen Indicate Cellular Immunosuppression Similar to That Found in Septic Shock.

Objectives: To compare microscopic and immunologic features in the spleens of patients who died of pulmonary hemorrhage and shock caused by leptospirosis (11 cases) or Gram-positive/-negative bacterial septic shock (10 cases) to those from control spleens (12 cases from splenectomy).Methodology: Histological features in the red pulp and white pulp were analyzed using archived samples by a semi quantitative score. Immunohistochemistry was used for the recognition of immune cell markers, cytokines, caspase-3 and Leptospira antigens.Results: The control group differed significantly from the leptospirosis and septic shock patients which demonstrate strong similarities: diffuse congestion in the red pulp with a moderate to intense infiltration of plasma cells and polymorphonuclear cells; follicles with marked atrophy; high density of CD20+ cells; low density of NK, TCD4+ and active caspase-3 positive cells and strong expression of IL-10; leptospirosis patients had higher S100 and TNF-α positive cells in the spleen than the other groups.Conclusion: The results suggest that an immunosuppressive state develops at the terminal stage of severe leptospirosis with pulmonary hemorrhage and shock similar to that of patients with septic shock, with diffuse endothelial activation in the spleen, splenitis, and signs of disturbance in the innate and adaptive immunity in the spleen. The presence of leptospiral antigens in 73% of the spleens of the leptospirosis patients suggests the etiological agent contributes directly to the pathogenesis of the lesions. Our results support therapeutic approaches involving antibiotic and immunomodulatory treatments for leptospirosis patients and suggest that leptospirosis patients, which are usually young men with no co-morbidities, form a good group for studying sepsis and septic shock.

Leptospirosis complicated with Guillain Barre syndrome, papillitis and thrombotic thrombocytopenic Purpura; a case report

BackgroundLeptospirosis is a zoonosis commonly prevalent in tropical countries. Clinical course of leptospirosis varies from mild to severe disease. Here we present a case of leptospirosis complicated with Guillain-Barre Syndrome (GBS), papillitis, and Thrombotic Thrombocytopenic Purpura(TTP).Case presentationA 21-year-old Asian male presented with fever, myalgia, oliguria and dyspnoea where he was managed as for leptospirosis complicated with pulmonary haemorrhages and acute renal failure. Leptospirosis was confirmed by Microscopic Agglutination Test(MAT) with a fourfold rise in antibody titre between acute and convalescent serum. The highest antibody titre was against Leptospira antigen serogroup Semaranga (strain Patoc) (1:1280) followed by serogroup Australis (strain Australis) (1:640) and serogroup Autumnalis (strain Bankgkinang) (1:320). Two weeks later he developed blindness, ascending weakness of lower limbs with global areflexia and an acute inflammatory demyelinating polyradiculopathy(AIDP) variant GBS was confirmed with nerve conduction studies. TTP complicated the picture several days later. He was initiated on plasmapheresis where clinical improvement was seen after 14 cycles. He had an incomplete neurological recovery with permanent vision loss but completely recovered from TTP. He also had permanent renal impairment.ConclusionLeptospirosis should be suspected and treated empirically in the relevant clinical settings where it can present with an atypical clinical picture as in our case with an acute febrile illness followed by GBS as well as TTP.

Apparently high Leptospira antibody seropositivity in donkeys for slaughter in three municipalities in Durango, Mexico.

We determined the seropositive percentage and the determinants of Leptospira exposure in domestic donkeys presented for slaughter in the State of Durango, Mexico. We examined 194 donkeys in 4 gathering premises. Sera of donkeys were analyzed for anti- Leptospira IgG antibodies using a microscopic agglutination assay with a panel of 7 Leptospira antigens. The association between Leptospira seropositivity and general characteristics of donkeys was assessed by bivariate and multivariate analyses. Antibodies against Leptospira (for at least 1 of the 7 Leptospira serovars examined) were found in 151 of 194 (77.8%) donkeys. Seropositive donkeys were found in all regions, municipalities, and premises examined. The predominant serovar was L. interrogans Icterohaemorrhagiae ( n = 117; 60.3%), followed by L. interrogans Sejroe ( n = 96; 49.5%), L. interrogans Canicola ( n = 10; 5.2%), L. kirschneri Grippotyphosa ( n = 6; 3.1%), L. borgpetersenii Ballum ( n = 5; 2.6 %), L. biflexa Semaranga ( n = 3; 1.5%), and L. noguchii Panama ( n = 2; 1%). Leptospira seropositivity was associated with gathering premises (OR = 2.64; 95% CI: 1.27-5.46; p = 0.009) and municipalities (OR = 0.11; 95% CI: 0.01-0.78; p = 0.02). Our results demonstrate an apparently high seropositive percentage of Leptospira infection in the donkeys studied in Mexico.