LipL32 Antigen Research Articles

Assessment of Risk Factors in Synanthropic and Wild Rodents Infected by Pathogenic Leptospira spp. Captured in Southern Chile.

Simple SummaryLeptospirosis is a zoonosis caused by pathogenic Leptospira, and synanthropic and wildlife species of rodents are an important source of infection; however, much of the information about the progression of the infection was obtained from lab murine models. The aim of this study was to assess infection status and risk factors by pathogenic Leptospira in synanthropic and wild rodent species and describe histopathological lesions in several organs from naturally infected animals. In this study, 121 rodents from three synanthropic species and two wild species were trapped within dairy farms in Southern Chile, where the bacteria were present. Liver, heart, kidney, and lungs from trapped animals were analyzed by different techniques to detect if the lesions present were produced by the bacteria. A large proportion of animals were identified as infected that were not detected by the microscopic agglutination test. There is a lower risk of infection in the fall compared to the rest of the seasons, and the synanthropic species has a lower risk of infection in comparison with wildlife species. Immunohistochemistry and quantitative real-time lipL32 polymerase chain reaction contributed to identifying the presence of pathogenic Leptospira in related histological lesions and 50% more infections than serology.Leptospirosis is caused by pathogenic Leptospira, and synanthropic and wildlife species of rodents are an important source of infection; however, much of the information about infection progression was obtained from murine models. The aim of this study was to assess infection status and risk factors associated with pathogenic Leptospira in synanthropic and wild rodent species and describe histopathological lesions in several organs from naturally infected animals. In a cross-sectional study, 121 rodents from three synanthropic species and two wild species were trapped in dairy farms in Southern Chile. Liver, heart, kidney, and lungs from trapped animals were fixed in formalin and stained with hematoxylin–eosin. Tissues with lesions consistent with Leptospira infection were tested by immunohistochemistry (IHC) and real-time polymerase chain reaction (qPCR) using the LipL32 antigen. Risk factors were assessed by a conditional mixed-logistic regression model. More than half (56.7%) of the negative reactors to the microscopic agglutination test were identified as infected either by IHC/qPCR. A lower risk of infection compared to the rest of the seasons was found in the fall, and the synanthropic species have a lower risk of infection in comparison with the wildlife species. IHC and qPCR contributed to the identification of pathogenic Leptospira in related histological lesions and 50% more infections than serology.

Leptospirosis: a diagnostic conundrum.

Leptospirosis is a serious public health concern worldwide. It is highly endemic in the Andaman Islands and its prevalence is increasing in other Indian states. Clinical features are non-specific and diagnosis relies on laboratory confirmation. The gold standard is microscopic agglutination testing, but this is not widely available. Real-time polymerase chain reaction testing of LipL32 antigen provides the earliest detection of pathogenic Leptospira in the body. We found it to be 100% specific, but it should be used in the first 10 days of illness for reliable results.

The use of halloysite clay and carboxyl-functionalised multi-walled carbon nanotubes for recombinant LipL32 antigen delivery enhanced the IgG response.

We studied the feasibility of using halloysite clay nanotubes (HNTs) and carboxyl-functionalised multi-walled carbon nanotubes (COOH-MWCNTs) as antigen carriers to improve immune responses against a recombinant LipL32 protein (rLipL32). Immunisation using the HNTs or COOH-MWCNTs significantly increased the rLipL32-specific IgG antibody titres (p < 0.05) of Golden Syrian hamsters. None of the vaccines tested conferred protection against a challenge using a virulent Leptospira interrogans strain. These results demonstrated that nanotubes can be used as antigen carriers for delivery in hosts and the induction of a humoral immune response against purified leptospiral antigens used in subunit vaccine preparations.

Production of leptospiral LipL32 antigen in Pichia pastoris and its use in an enzyme-linked immunosorbent assay

Production of leptospiral LipL32 antigen in Pichia pastoris and its use in an enzyme-linked immunosorbent assay

Rapid hybridization of chitosan-gold-antibodies via metal-free click in water-based systems: a model approach for naked-eye detectable antigen sensors.

A surface plasmon resonance (SPR) expression after hybridization of chitosan-gold nanoparticle-antibody (CS-AuNPs-Ab) based on: i) metal-free click chemistry, and, ii) in water system as an approach for a rapid antigen sensing, is proposed. The chitosan-hydroxybenzyl triazole complex enables us to carry out the conjugation of mPEG and trifluoromethylated oxanorbornadiene (OND) in water. CS-mPEG-OND further allows metal-free click to hybridize chitosan (CS) with azido-modified gold nanoparticles (azido-AuNPs) in aqueous solution at room temperature. The CS-mPEG-OND conjugated with LipL32 antibody (Ab) not only effectively binds with LipL32 antigen (Ag) but also performs the cycloaddition with azido-AuNPs to display a change in color within 2 min. The phenomenon leads to a simple and efficient naked-eye antigen detection technique.

Production of leptospiral LipL32 antigen in Pichia pastoris and its use in an enzyme-linked immunosorbent assay

The production of recombinant LipL32 protein using Escherichia coli has been used extensively for the development of vaccines and diagnostic tests for leptospirosis. However, E. coli has demonstrated limitations, including low yield and lack of post-translational modifications. In this study, rLipL32 was produced in eukaryotic expression system (Pichia pastoris) and evaluated the antigen by enzyme-linked immunosorbent assay (ELISA). The yield obtained from the culture supernatant reached 270 mg/L and ELISA showed an accuracy of 95.34%. In summary, the production of rLipL32 using P. pastoris did not impair the antigenic characteristics of this antigen and ensured its use for detecting the leptospiral antibodies in swine sera.

Serological Evidence of Leptospiral Infection in Pig Populations in Different Districts in Japan

Serum samples were collected from 938 pigs of 24 farms in Hokkaido, Kagoshima, and Okinawa prefectures in Japan in 2001-2005. Enzyme-linked immunosorbent assay (ELISA) was used for the detection of antibodies to LipL32 antigen which is common to Leptospira interrogans. Samples positive in ELISA were then investigated by microscopic agglutination test for the identification of causal leptospires. Antibodies specific to leptospires of serovars Copenhageni, Bratislava, Australis and Javanica were detected in serum samples of pigs from each of the three districts. In addition, antibodies to leptospires of serovars Autumnalis and Tarassovi were predominantly detected in those from Kagoshima. The present study, thus, revealed that leptospires belonging to different serovars prevail in the pig population in Japan. In addition, it is the first detection of antibodies to leptospires belonging to serovars Javanica and Tarassovi in pigs in Japan.

Evaluation of different ways of presenting LipL32 to the immune system with the aim of developing a recombinant vaccine against leptospirosis

Leptospirosis, caused by bacteria of the genus Leptospira, is a direct zoonosis with wide geographical distribution. The implications in terms of public health and the economical losses caused by leptospirosis justify the use of a vaccine against Leptospira in human or animal populations at risk. In this study, we used the external membrane protein LipL32 as a model antigen, as it is highly immunogenic. The LipL32 coding sequence was cloned into several expression vectors: (i) pTarget, to create a DNA vaccine; (ii) pUS973, pUS974, and pUS977 for expression in BCG (rBCG); and (iii) pAE, to express the recombinant protein in Escherichia coli, for a subunit vaccine. Mice were immunized with the various constructs, and the immune response was evaluated. The highest humoral immune response was elicited by the subunit vaccine (rLipL32). However, with rBCG, the titer was still rising at the end of the experiment. The serum of vaccinated animals was able to recognize LipL32 on the membrane of the Leptospira, detected by indirect immunofluorescence. A monoclonal antibody anti-LipL32 was shown to inhibit the growth of Leptospira in vitro, indicating potential protection induced by the LipL32 antigen.

Recombinant Antigen-based Latex Agglutination Test for Rapid Serodiagnosis of Leptospirosis

A rapid recombinant antigen-based latex agglutination test (LAT) has been developed to detect specific anti-leptospiral antibodies from human and dog sera. The recombinant LipL32 antigen developed and used for detecting the antibodies is specific in detection of the pathogenic serovars of Leptospira as the expression of the LipL32 antigen is restricted only to the pathogenic leptospires. The sensitized latex beads are stable and could be stored at 4 degrees C for more than three months without showing loss of activity for both weakly and strongly positive samples. The test is found to be sensitive, specific and accurate as compared to the standard microscopic agglutination test (MAT). Moreover, the recombinant antigen-coated latex beads could detect the specific anti-leptospiral antibodies in the acute phase of the illness. The test is simple and inexpensive, and is rapid in the management of large numbers of patients.