Development and evaluation of time-resolved rapid immunofluorescence test for detection of TSOL18 specific antibody in porcine cysticercosis infections.

BACKGROUND: Porcine cysticercosis, a serious zoonotic parasitic disease, is caused by the larvae of Taenia solium and has been acknowledged by the World Organization for Animal Health. The current detection methods of Cysticercus cellulosae cannot meet the needs of large-scale and rapid detection in the field. We hypothesized that the immunofluorescence chromatography test strip (ICS) for detecting Cysticercus cellulosae, according to optimization of a series of reaction systems was conducted, and sensitivity, specificity, and stability testing, and was finally compared with ELISA. This method utilizes Eu3+-labeled time-resolved fluorescent microspheres (TRFM) coupled with TSOL18 antigen to detect TSOL18 antibodies in infected pig sera. RESULTS: ICS and autopsy have highly consistent diagnostic results (n = 133), as determined by Cohen's κ analysis (κ = 0.925). And the results showed that the proposed ICS are high sensitivity (0.9459) with specificity (0.9792). The ICS was unable to detect positive samples of other parasites. It can be stored for at least six months at 4℃. CONCLUSIONS: In summary, we established a TRFM-ICS method with higher sensitivity and specificity than indirect ELISA. Results obtained from serum samples can be read within 10 min, indicating a rapid, user-friendly test suitable for large-scale field detection.

The effects of thioredoxin peroxidase from Cysticercus cellulosae excretory-secretory antigens on TGF-ß signaling pathway and Th17 cells differentiation in Jurkat cells by transcriptomics.

Thioredoxin peroxidase (TPx) protein from the excretory-secretory antigens (ESAs) of Cysticercus cellulosae (C. cellulosae) has been shown to regulate the differentiation of host Treg and Th17 cells, resulting in an immunosuppressive response dominated by Treg cells. However, the molecular mechanism by which TPx protein from the ESAs of C. cellulosae regulates the imbalance of host Treg/Th17 cell differentiation has not been reported. TPx protein from porcine C. cellulosae ESAs was used to stimulate Jurkat cells activated with PMA and ionomycin at 0, 24, 48, and 72 h. Transcriptomic analysis was performed to investigate the signaling pathways associated with Jurkat cells differentiation regulated by TPx protein from C. cellulosae ESAs. Gene Set Enrichment Analysis (GSEA) revealed that TPx protein from porcine C. cellulosae ESAs could induce upregulation of the TGF-ß signaling pathway and downregulation of Th17 cell differentiation in Jurkat cells. TPx protein from porcine C. cellulosae ESAs can activate the TGF-ß signaling pathway in Jurkat cells, thereby regulating the differentiation of Treg/Th17 cells and leading to an immunosuppressive response dominated by Treg cells, enabling evasion of the host immune attack. This study provides a foundation for further validation of these pathways and further elucidates the molecular mechanisms underlying immune evasion caused by porcine C. cellulosae.

Regulation of DNA methylation on key parasitism genes of Cysticercus cellulosae revealed by integrative epigenomic-transcriptomic analyses.

BACKGROUND: The life cycle of Taenia solium is characterized by different stages of development, requiring various kinds of hosts that can appropriately harbor the eggs (proglottids), the oncospheres, the larvae and the adults. Similar to other metazoan pathogens, T. solium undergoes transcriptional and developmental regulation via epigenetics during its complex lifecycle and host interactions. RESULT: In the present study, we integrated whole-genome bisulfite sequencing and RNA-seq technologies to characterize the genome-wide DNA methylation and its effect on transcription of Cysticercus cellulosae of T. solium. We confirm that the T. solium genome in the cysticercus stage is epigenetically modified by DNA methylation in a pattern similar to that of other invertebrate genomes, i.e., sparsely or moderately methylated. We also observed an enrichment of non-CpG methylation in defined genetic elements of the T. solium genome. Furthermore, an integrative analysis of both the transcriptome and the DNA methylome indicated a strong correlation between these two datasets, suggesting that gene expression might be tightly regulated by DNA methylation. Importantly, our data suggested that DNA methylation might play an important role in repressing key parasitism-related genes, including genes encoding excretion-secretion proteins, thereby raising the possibility of targeting DNA methylation processes as a useful strategy in therapeutics of cysticercosis.

Swine cysticercosis and associated risk factors in non-technified pig breeding in semi-arid region of Sergipe state, Brazil.

The aim of this study was to determine the frequency of Taenia solium anti-metacestode antibodies in slaughtered pigs in a semi-arid region of the "Alto Sertão" of Sergipe state, Brazil, and verify the risk factors associated with swine cysticercosis. For this, 230 samples of swine blood from two slaughterhouses were collected and analyzed by indirect ELISA. The pigs came from five non-technical properties in the semi-arid region of the Alto Sertão of Sergipe state. Searches for cysts in the skeletal muscles of the pigs were performed during slaughter. In addition, an epidemiological questionnaire was applied to the pigs' original properties to determine risk factors. Besides that, the official health services database was evaluated for confirmed cases of neurocysticercosis and taeniasis in humans in the last 5 years, living in the studied region. Seropositivity in pigs was 12.6%, with no significant difference between males and females. No cysts were found in the carcasses of the slaughtered pigs. A positive association was found for properties that discharge domestic sewage into the environment, in river or streams, increasing the risk of positivity by 5.72 times. When analyzing the database of official agencies, there were no records of cases of neurocysticercosis or taeniasis in the resident population in the last 5 years. However, there were frequent cases of idiopathic epilepsy. The results demonstrate that study area is endemic for swine cysticercosis and serves as a warning of the possibility of the occurrence of the taeniasis-cysticercosis complex.

Regulation of piglet T-cell immune responses by thioredoxin peroxidase from Cysticercus cellulosae excretory-secretory antigens.

Taenia solium (T. solium) cysticercosis is a serious threat to human health and animal husbandry. During parasitization, Cysticercus cellulosae (C. cellulosae) can excrete and secrete antigens that modulate the host's T-cell immune responses. However, the composition of C. cellulosae excretory-secretory antigens (ESAs) is complex. This study sought to identify the key molecules in C. cellulosae ESAs involved in regulating T-cell immune responses. Thus, we screened for thioredoxin peroxidase (TPx), with the highest differential expression, as the key target by label-free quantification proteomics of C. cellulosae and its ESAs. In addition, we verified whether TPx protein mainly exists in C. cellulosae ESAs. The TPx recombinant protein was prepared by eukaryotic expression, and ESAs were used as the experimental group to further investigate the effect of TPx protein on the immune response of piglet T cells in vitro. TPx protein induced an increase in CD4+ T cells in piglet peripheral blood mononuclear cells (PBMCs), while CD8+ T cells did not change significantly. This resulted in an imbalance in the CD4+/CD8+ T-cell ratio and an increase in CD4+CD25+Foxp3+ Treg cells in the PBMCs. In addition, TPx protein initiated T helper 2 (Th2)-type immune responses by secreting IL-4 and IL-10 and suppressed Th1/Th17-type immune responses. The results showed that ESAs were involved in regulating piglet T-cell immune responses cells. This suggests that TPx protein found in ESAs plays an essential role to help the parasite evade host immune attack. Moreover, this lays a foundation for the subsequent exploration of the mechanism through which TPx protein regulates signaling molecules to influence T-cell differentiation.

Rapid Detection of Cysticercus cellulosae by an Up-Converting Phosphor Technology-Based Lateral-Flow Assay.

Cysticercosis is a neglected tropical disease caused by the larvae of Taenia solium in pigs and humans. The current diagnosis of porcine cysticercosis is difficult, and traditional pathological tests cannot meet the needs of detection. This study established a UPT-LF assay for the detection of Cysticercus cellulosae. UCP particles were bound to two antigens, TSOL18 and GP50; samples were captured, and the signal from the UCP particles was converted into a detectable signal for analysis using a biosensor. Compared to ELISA, UPT-LF has higher sensitivity and specificity, with a sensitivity of 93.59% and 97.44%, respectively, in the case of TSOL18 and GP50 antigens and a specificity of 100% for both. Given its rapidness, small volume, high sensitivity and specificity, and good stability and reproducibility, this method could be used in the diagnosis of cysticercosis.

Cysticercus cellulosae Regulates T-Cell Responses and Interacts With the Host Immune System by Excreting and Secreting Antigens.

Cysticercus cellulosae (C. cellulosae) excretes and secretes antigens during the parasitic process to regulate the host immune response; however, resulting immune response and cytokine production in the host during infection still remains unclear. We used C. cellulosae crude antigens (CAs) as controls to explore the effect of excretory secretory antigens (ESAs) on T-cell immune responses in piglets. C. cellulosae ESAs induced imbalanced CD4+/CD8+ T-cell proportions, increased the CD4+Foxp3+ and CD8+Foxp3+ T-cell frequencies, and induced lymphocytes to produce interleukin-10, which was mainly attributed to CD4+ and CD4-CD8- T cells. The ESAs also induced Th2-type immune responses. The results showed that the ability of C. cellulosae to escape the host immune attacks and establish a persistent infection may be related to host immune response regulation by the ESAs.

Neurocysticercosis in Child Bearing Women: An Overlooked Condition in Mozambique and a Potentially Missed Diagnosis in Women Presenting with Eclampsia.

Several studies suggest that neurocysticercosis might be one of the main reasons for onset of seizures in adolescents and young adults in Mozambique, however, no studies have evaluated the burden and impact of the disease in women of childbearing age. This is also true for other parasitic diseases such as toxoplasmosis, toxocariasis and onchocerciasis, although the latter has been reported in some regions in central and northern Mozambique. We describe herein an 18 year old female who was admitted to Hospital Central de Quelimane, Zambézia Province in Mozambique, 8 days after delivery with a misdiagnosis of eclampsia that turned out to be neurocysticercosis. We discuss challenges in the diagnosis of neurocysticercosis in women of childbearing age and the potential for missed diagnosis in those who present with eclampsia and other conditions associated with seizures.

Morphological characterization of Cysticercus cellulosae in naturally infected pigs in Punjab (India).

Porcine cysticercosis is an important medical and veterinary concern in the developing world. The present study was carried out to determine the morphological characteristics of Cysticercus cellulosae, so as to differentiate the C. viscerotropica (larval form of T. asiatica) which is having only rudimentary hooks on their rostellum. Morphological analysis was conducted on 22 Cysticercus positive samples. Measurements for number of hooks (large and small), the total length and blade length of large and small hooks per rostellum were carried out as per previous studies. Microscopic examination of all the cysts showed typical characteristic of T. solium i.e. presence of hooks in all the cysts. The results indicated absence of T. asiatica from naturally infected pigs in Punjab (India).

Orofacial cysticercosis: Report of a rare case with review of literature.

Cysticercosis is common in developing countries in which the combination of rural society, crowding, and poor sanitation facilities allows greater contact between humans and pigs and thus more opportunities for fecal contamination of food and water occurs. They are rarely located in oral and perioral tissues, particularly in the muscles of mastication, muscle of the facial expression, suprahyoid muscles, and postcervical musculature and also as in the tongue, buccal mucosa, and lip. Cysticercosis is a potentially fatal parasitic disease that rarely found in the maxillofacial region in humans. This paper reports the case of a young female patient presented with isolated lesion of cysticercosis involving buccinator muscle. In conclusion, we suggest that cysticercosis should be considered in the differential diagnosis of intraoral solitary nodules within the oral and maxillofacial region, especially in endemic areas. High-resolution ultrasonography is an excellent noninvasive and cost-effective modality for the diagnosis and also suggests that localized parasitic infections such as Cysticercus cellulosae can be successfully treated with conservative management using oral antiparasitic (antihelminthic) medication.

Rising trends of neurocysticercosis: A serological report from tertiary-care hospital in South India.

INTRODUCTION: Taenia solium is a common two-host parasitic cestode, residing in both humans (definitive) and pigs (intermediate). Invasion of this parasitic cyst into central nervous system leads to a condition known as neurocysticercosis (NCC). The World Health Organization (WHO) considers NCC as one of the "most neglected" tropical zoonotic diseases. The disease is presented with pleomorphic clinical manifestations, of which epilepsy is the most common. Diagnosis of NCC is carried out by serological tests and imaging methods. Only a few studies from Andhra Pradesh, Kerala, Tamil Nadu, and Pondicherry are available regarding the seropositive levels of NCC in South India. MATERIALS AND METHODS: A descriptive analysis was carried out on NCC suspected patients attending outpatient or inpatient department of different clinics majorly from neurology, medicine, pediatrics, ophthalmology, and skin at Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, a tertiary care hospital in South India. A total of 391 patient samples (either serum or cerebrospinal fluid or urine) for 5 years from January 2011 to December 2015 were taken into the study. Serological investigations such as enzyme-linked immunosorbent assay and enzyme-linked immunoelectro transfer blot were performed for assessing the seropositivity levels of NCC. RESULTS: The overall seropositive cases of NCC in the study population were found to be 32.5% of which positive male cases (59.1%) exceeding females (40.9%). The frequency of adult positive cases (77.2%) was more than that of pediatrics cases (22.8%) with an average of 30.9 years of age. CONCLUSIONS: NCC seropositive levels show an increasing trend with the study period. This necessitates a proper attention to the unnoticed spread of the parasitic disease, which affects the quality of life in the community. Quality screening and diagnostic strategy should be implied along with proper awareness for preventive measure practices have to be set up to reduce the impact of morbidity caused by NCC.

Imaging and Serological-Evidence of Neurocysticercosis Among Patients with Seizures in Odisha, an Unexplored Eastern Coastal Province in India.

INTRODUCTION: Neurocysticercosis being a potential to human transmitted disease, is the major cause of seizures and a public health problem in tropical countries. Though India is known to be highly endemic, there are many provinces where reports are still unavailable thereby underestimating its actual burden. MATERIALS AND METHODS: Anti-Cysticercus IgG antibodies in sera from cases presenting with seizures were screened by ELISA in a preliminary study in Odisha state which is a province in Eastern coastal India that was never explored before. Patients presenting with recent onset of seizures within age group 5 to 50 years, either local residents of Odisha or inhabitants from other parts of the country living for at least one year period in the study area were included. RESULTS: The present study showed 43.75% cases with seizures to be confirmed neurocysticercosis (NCC) based on serology and brain imaging. However, statistically no association was established between anti-Cysticercus antibody detection and radio imaging characteristics (location, number of lesions, and stage). CONCLUSION: This is the first study in Odisha presenting a series of cases with serological evidence of exposure to the parasite along with imaging characteristics which was consistent with NCC. It is recommended that NCC must be considered for a differential diagnosis in each active epilepsy case irrespective of prior prevalence information in all unexplored provinces in India and other endemic regions; also a compulsory reporting is warranted in order to aid in quantifying its actual burden.

Oral Cysticercosis- A Diagnostic Dilemma.

Cysticercosis, a helminthic disease commonly seen in India, Latin America, Eastern Europe and Southern Africa, results from extraintestinal encystation of the larval form of Taenia solium. It is a condition in which man acts as intermediate host instead of definitive host. The most frequent sites of cysticercosis are subcutaneous layers, brain, muscles, heart, liver, lungs, and peritoneum. Oral cysticercosis is considered rare and cause cystic swellings or nodules in the mouth and a precise clinical diagnosis is not usually established. Here, we report a case of oral cysticercosis in a 32-year-old female occurring in the mentalis muscle presenting as asymptomatic nodule.

Oral cysticercosis: a diagnostic difficulty.

Cysticercosis is a rare disease caused by the ingestion of the parasite Cysticercus cellulosae, a larval stage of Taenia solium. The definitive host is human who harbors the adult worm and may accidentally or incidentally become the host. The larval form of cyst is commonly seen in the brain, meninges and eyes. Cases in the maxillofacial region including oral cavity and cheek muscles are rarely reported. Cysticercosis is not commonly considered in the diagnosis of swellings of the head and neck and a diagnostic and therapeutic dilemma for clinicians. Hence, they are of utmost interest to the practitioner and have to be studied. We present an unusual case of cysticercosis presenting as a solitary cystic nodule in the upper left vestibule of the oral cavity in an 18 year male and the diagnosis was made on histopathological examination.

Diagnóstico laboratorial da neurocisticercose: revisão e perspectivas / Laboratory diagnosis of neurocysticercosis: review and perspectives

A neurocisticercose é causada por Cysticercus cellulose, a forma larval de Taenia solium, quando este se aloja no sistema nervoso central. O seu diagnóstico é realizado com base em dados clínicos, epidemiológicos, demonstração do agente etiológico pelas técnicas de imagem e testes laboratoriais. No presente estudo, apresentamos uma revisão do diagnóstico laboratorial, com ênfase no desempenho dos testes para pesquisa de anticorpos específicos e detecção de antígenos circulantes, utilização de antígeno homólogo ou heterólogo, nativo e recombinante, bem como a aplicação de métodos moleculares.

Neurocysticercosis is caused by Cysticercus cellulosae, the larval form of Taenia solium, when it lodges in the central nervous system. The diagnosis of neurocysticercosis is based on clinical and epidemiological data, neuroimaging findings of etiological agent and serologic test results. Herein we present a review of clinical diagnosis, emphasizing test performance for specific antibody and antigen detection, the use of homologous or heterologous antigen, native and recombinant antigens as well as the application of molecular methods.

Comparação de técnicas imunológicas in-house empregadas na rotina diagnóstica de cisticercose humana / Comparative analysis on in-house immunologic assays used for performing the routine laboratorial diagnosis of human cysticercosis

Neste estudo foram analisados os resultados obtidos do diagnóstico de cisticercose no Centro de Imunologia do Instituto Adolfo Lutz (IAL), no período de março/2007 a julho/2010. A detecção de anticorpos específicos em 522 amostras de soro e líquido cefalorraquidiano (LCR)foi realizada pelas técnicas de imunofluorescência indireta (IFI) e hemaglutinação indireta (HAI). A frequência de amostras reagentes foi de 11,0% no LCR e 8,2% no soro. Em 50% das amostras não houve informações sobre suspeita clínica de neurocisticercose dos pacientes, sendo disponíveis nos18,3% e 16,4%, em amostras, respectivamente, de LCR e soro. Nas amostras de paciente com suspeita de neurocisticercose, a positividade foi de 22,6% (LCR) e de 18,4% (soro). Houve associação entre a suspeita clínica e a positividade dos testes (p>0.05). A maioria das amostras testadas foi proveniente do Estado de São Paulo, e 16,9% de amostras de LCR e 35,9% de amostras séricas foram enviados de outros Estados do país. Os ensaios de IFI e HAI apresentaram teste de concordância Kappa de 86%. Pela indisponibilidade de kits de reagentes diagnósticos de cisticercose em amostras de LCR no mercado, os testes in-house produzidos no IAL têm sido de grande relevância para os serviços de saúde pública.

Epidemiologic pattern of patients with neurocysticercosis diagnosed by computed tomography in Curitiba, Brazil / Perfil epidemiológico de pacientes com neurocisticercose diagnosticados por tomografia computadorizada em Curitiba, Brazil

The aim of this study was to drawn an epidemiological pattern of neurocystisticercosis (NCC) patients diagnosed by computed tomography at the major private diagnostic center in Curitiba, Brazil. A total of 1,009 medical files of consecutive patients presenting neurological indications were diagnosed by computed tomography from July 2007 to April 2008. Patient data included sex, age, municipality and tomography findings were analysed by Epi-info version 6.0.1. software. Most patients (81.10 percent) were living in Curitiba. A total of 91/1,009 cases (9.02 percent) were considered positive to NCC; 88 (96.7 percent) patients had inactive form of NCC and only 3 (3.2 percent) patients had cysts in granulomatous process. No patients had both forms. The prevalence of NCC cases in studied group was greater in patients between 51 to 60 years old, however, difference between sex was not significant. This epidemiological pattern of NCC was similar to the first NCC study in Curitiba, performed in 1995-1996 with 9.24 percent of positive cases.

Determinou-se o perfil epidemiológico da neurocisticercose (NCC) em pacientes diagnosticados por tomografia computadorizada (TC) no maior centro privado de diagnósticos de Curitiba, Brasil. Foram analisados 1009 registros médicos de pacientes consecutivos com indicações neurológicas submetidos a TC entre julho de 2007 a abril de 2008. Os dados dos pacientes que incluíram sexo, idade, município de residência e achados tomográficos foram analisados pelo software Epi-info versão 6.01. A maioria dos pacientes (80,10 por cento) era procedente de Curitiba; 91/1.009 casos (9,02 por cento) foram positivos para NCC; 88 (96,7 por cento) apresentaram a forma inativa e apenas em 3 (3,2 por cento) cistos em processo granulomatoso; não houve formas mistas. A prevalência de casos de NCC foi maior entre 51 e 60 anos. Não houve diferença significativa para o sexo entre os casos. O perfil dos pacientes diagnosticados para NCC por TC neste estudo é semelhante ao estudo anterior realizado em Curitiba entre 1995-1996, com 9,24 por cento de casos de NCC.

Evaluation of an enzyme immunoassay for clinical diagnosis of neurocysticercosis in symptomatic patients / Avaliação de um teste imunoenzimático para o diagnóstico clínico de neurocisticercose em pacientes sintomáticos

INTRODUCTION: Neurocysticercosis is an infection of the human central nervous system caused by the metacestode larvae of Taenia solium. Neurocysticercosis is the most common parasitic disease in developing countries. Epilepsy is the most common clinical manifestation. Difficulties in confirming the diagnosis motivated the evaluation of the enzyme-linked immunosorbent assay on cerebral spinal fluid (CSF). METHODS: Twenty-two patients with NCC and 44 control patients were studied. CSF was analyzed using a commercial ELISA kit developed for NCC. Sensitivity and specificity were measured and a multivariate logistic regression was performed. RESULTS: Sensitivity and specificity of ELISA were 31.8 percent and 100 percent, respectively, with accuracy of 77.3 percent. Only the size of the lesions proved to be important for performance of the test. CONCLUSIONS: The results showed that ELISA contributes to the diagnosis of neurocysticercosis if the result is negative or if the patient has a lesion of 2 cm or more.

INTRODUÇÃO: Neurocisticercose é a infecção do sistema nervoso central causada pela larva metacestódea da Taenia solium. Neurocisticercose é a parasitose mais comum nos países em desenvolvimento. Epilepsia é a sua manifestação clínica mais comum. A dificuldade para confirmar o diagnóstico motivou a avaliação do ensaio imunoenzimático ligado à enzima no líquido cérebro-espinhal. MÉTODOS: Vinte e dois pacientes com NCC e 44 pacientes controles foram estudados. Líquido cérebro-espinhal foi analisado por um kit ELISA comercial desenvolvido para NCC. A sensibilidade e especificidade foram medidas e uma análise multivariada de regressão logística foi realizada. RESULTADOS: A sensibilidade e a especificidade de ELISA foram, respectivamente, 31,8 por cento e 100 por cento, com acurácia de 77,3 por cento. Apenas o tamanho das lesões mostrou-se importante para o desempenho do teste. CONCLUSÕES: Este estudo concluiu que ELISA contribui para o diagnóstico de NCC, caso o teste seja negativo ou caso o paciente seja portador de lesão cuja dimensão seja igual ou maior que dois centímetros.

Frequência de anticorpos anti-Cysticercus cellulosae em população de risco para cisticercose e em segmento de população considerado supostamente normal, em regiões do Estado de São Paulo, Brasil / Frequency of anti-cysticercus cellulosae antibodies in high risk population for cysticercosis and in a segment considered as normal population, in areas of Säo Paulo state, Brazil

Examinaram-se, através de reação de fixação de complemento, os soros de 234 pacientes internados em hospital psiquiátrico localizado no município de Presidente Prudente, considerados de risco para infecção cisticercótica, além de 454 soros de gestantes procedentes da Região Administrativa de Santos e 397 soros de indivíduos considerados supostamente normais, procedentes da Região Administrativa de Presidente Prudente. O antígeno utilizado na reação de fixação de complemento foi obtido através de extração metílica, à temperatura ambiente, dos císticercos tratados com acetona. Consideraram-se positivas as reações em que ocorreu fixação de complemento a partir da diluição 1 :2. Dos 1.085 soros testados, 27 apresentaram atividade anticomplementar e 17 (1,6%) mostraram-se reagentes. Todavia, quando se consideraram, separadamente, os grupos procedentes de Santos, Presidente Prudente e os doentes mentais, percebe-se diferença significativa nos resultados: assim, os índices de freqüência foram, respectivamente, 0,88% e 1,00% para os indivíduos procedentes de Santos e Presidente Prudente e considerados supostamente normais e 3,8% para os doentes mentais. Os resultados indicam que não é desprezível a ocorrência de anticorpos anti-Cysticercus cellulosae em nosso meio, especialmente entre pacientes de hospitais psiquiátricos (AU).