Hydatid Patients Research Articles

Diagnostic Efficiency of Heat Shocked Protoscoleces Extract Antigens for Human Cystic Echinococcosis by ELISA

Diagnosis of cystic echinococcosis is complex and has to be confirmed by the combination of immunological tests and imaging techniques. In this study heat shock proteins were induced and their immunoreactivity was assessed by ELISA. Sera were collected from 34 hydatid patients who were admitted to the Rizgary Teaching Hospital through October 2013 to July 2017, in addition to 29 healthy donors and 18 non-hydatid cases. For heat shock response, two batches of 25000 protoscoleces (Ps) were incubated separately at 42°C and 45 °C for 4 hours. Heat treated and normal Ps were disrupted and the extracts were divided into two parts. One part was directly used as source of antigens (PE, PE42 and PE45) and the other one was partially purified on Sephadex G150. The immunoreactivity of these antigens, as well as hydatid fluidwas assessed by ELISA. The cutoff value to differentiate positive from negative sera was established by receiver operating characteristic (ROC) analysis. Two peaks of PE42 and three peaks of PE45 resulted by Sephadex G150. Extracts of 42ºC treated Ps resulted in two protein peaks and were used as PE42P1 and PE42P2 antigens. For 45°C treated Ps, the chromatography patterns resulted in three protein peaks and were used as PE45P1, PE45P2 and PE45P3 antigens. Highest rates of sensitivity, specificity and diagnostic accuracy were detected with PE42P2 (91.2%) and PE45P2 (91.2%). Sensitivity of ELISA was consistent for liver cysts with all applied antigens. Hydatid antigens extracted from heat treated Ps markedly raised the sensitivity of ELISA to detect anti-hydatid IgG.

A PIONEER USAGE OF GOLD NANOPARTICLES IN SANDWICH ELISA BASED ON CIRCULATING HYDATID ANTIGEN AS A TOOL FOR IMPROVEMENT OF SERODIAGNOSTIC TESTING

Human cystic echinococcosis is a zoonosis occurring due to dogs handling and exposure toEchinococcus granulosus ova in their stools being an accidental intermediate host. Serologyremains the only sure and important tool to diagnose this disease. We aimed to improve thedetection of circulating antigen by sandwich ELISA through its binding to gold nanoparticleand proving the role of minute nanoparticles in serodiagnosis. Gold nanoparticles (AuNPs)were employed for the capture of the antigens of protoscolices (pAg) in the patient’s serum bysandwich ELISA. Cross-reactivity for antigens from Hymenolepis nana, Entrobius vermicularis,and Fasciola gigantica was ruled out by using anti-protoscolices polyclonal IgG antibodies(ppAb). Antigen of sonicated protoscolices which were removed from camel lungcysts, was purified and injected to a New-zealand white rabbit giving ppAb which then loadedon AuNPs being used as a diagnostic indicator for circulating pAg by both sandwich ELISAand nanogold sandwich ELISA techniques. Nanogold sandwich ELISA was able to give positiveresults with 96.3% of hydatid patients and 5% of non-hydatid patients while sandwichELISA showed 81.4% and 20% positive cases of the same groups respectively. The sensitivityand specificity of nanogold sandwich ELISA were 96.3% & 95% against 81.5% & 80% forthose of sandwich ELISA respectively. The conjugation of AuNPs to anti-Echinococcus IgGantibodies seems to be beneficial to increase PPV, NPV, and efficiency and reduce the overallcost of the assay as less antibody was needed.

Evaluation of Oxidative Stress and Antioxidants in Iraqi Patients with Hydatid Disease

Antioxidants and oxidative stress status are clinically important in the detection of many diseases. In order to estimate the role of oxidative stress in hydatid disease pathogenesis, the antioxidant levels and oxidative stress status were examined in the patients of hydatid cysts. Thirty patients having active hydatid cyst of Echinococcus granulosus, previously diagnosed by X-ray, were adopted as patients’ group. Additionally, 25 healthy individuals who did not have hydatid infection or any other parasitic infection constituted the control group. Antioxidant status and oxidative stress levels were determined using biochemical tests including plasma Malondialdehyde (MDA) level, and erythrocyte Catalase (CAT), Glutathione Peroxidase (GSH Px), Superoxide Dismutase (SOD), Glutathione-S-Transferase (GST) activity, in addition to erythrocyte Glutathione concentration GSH-conc. The statistical evaluation of plasma MDA levels showed significantly higher levels in hydatid patients than in healthy controls, while erythrocyte SOD, GSH-Px, CAT, GST and GSH levels were significantly declined in hydatid patients compared to the control which can be used as diagnostic markers in the laboratory diagnosis of the disease. To conclude, hydatid patients show elevated oxidative stress status, and therefore, the antioxidant therapy should be considered in addition to the routine medicines in this group of patients.

A 53 KDa Glycan Antigen of Hydatid Cyst Wall May Involve in Evasion from Host Immune System

Background:Recent studies have shown that similar host glycan antigens are expressed by helminths such as Echinococcus granulosus hydatid cysts to evade from host immune system. In this work to investigate these antigens further, immunological cross-reactivity between human sera and hydatid cyst wall antigens has been investigated.Materials and Methods:Hydatid cyst wall antigens were used in enzyme-linked immunosorbent assay and Western immunoblotting and probed with pooled sera of hydatidosis patients and healthy controls. Sodium metaperiodate treatment was used to investigate glycan antigens.Results:A band with molecular weight about 53 KDa reacted with both hydatid patients' sera and also normal human sera. It has been shown that this band was a glycan antigen.Conclusions:A 53 KDa glycan antigen of hydatid cyst wall that reacted with all human sera may have an important role for evasion from host immune system.

Common Spinal Parasites.

To report the four most common spinal parasites for providing insight into definitive therapy. Twelve patients with spinal parasites were diagnosed between 2009 and 2016. A definite diagnosis was established in the form of histopathology (n=9) and response to drug therapy (n=3). The minimum follow-up was 0.9 years and ranged to a maximum of 8 years, with a median of 3 years. Twelve patients aged between 8 and 69 years were reviewed, including nine hydatidosis and three non-hydatid cases. Occupational exposure to the endemic area and unclean food were the main routes of infestation. Spinal parasites can present symptoms that correlated with the level of the lesion. There was complete paraplegia in four patients and paraparesis in six patients before surgery. Seven hydatid patients underwent posterior decompression and pedicle screw fixation with posterolateral fusion. Two non-hydatid patients experienced laminoplasty after posterior decompression. Six hydatid patients had reoccurrences and two patients with intramedullary hydatidosis died of severe complications one year after surgery. Spinal hydatidosis significantly differs from the three other non-hydatid parasites in diagnosis and treatment. The final prognosis of spinal hydatidosis remains poor, especially for intramedullary hydatidosis.

Role of Imaging in Diagnosis, Predicting Biological Activity and in Treatment Plan of Hydatid Disease

Cystic Echinococcosis (CE) is a zoonotic parasitic disease caused by the larval stage of the cestode Echinococcus granulosus. Hydatid cyst is a significant universal medical problem. In human cestode involves any organ and develops to hydatid cyst. Liver is the first and most frequently involved organ as the liver is the primary filter station of portal circulation. Though hydatid disease is benign disease, traditionally surgery is the treatment of choice for complete elimination of infective focus, which is associated with considerable mortality, morbidity. Systemic chemotherapy and percutaneous drainage have evolved in the recent past as an alternative to conventional surgery. The safety and success rate of these methods is influenced by the size, stage, location of the cysts and associated complications. Benzimidazoles can be considered in multiple, small hydatid cysts, but large cysts (cysts > 6 cm), bone marrow depression, pregnancy are contraindications of pharmacotherapy. Percutaneous drainage combined with ALB is a safe and effective treatment in liver hydatid patients with a surgical contraindication and younger cysts, but the presence of certain radiological signs (pericystic ducts and exophytic components) is a contraindication to non-surgical management because of the danger of biliary obstruction. However, the surgical treatment technique also cannot be standardized, should be tailored according to the cyst relation to the Broncho, biliary, vascular structures and associated complications if any. This review will focus on role of imaging in establishing the diagnosis, in determining cyst location, size, stage of the cyst and in identifying any associated complications, so that enable the clinician to perform recommended therapeutic procedures for the different cyst types and tailor the procedure performed based on cyst relation to broncho, biliary, vascular structures and associated complications.

Epidemiological, Clinical and Paraclinical Study of Hydatid Cysts in Three Educational Medical Centers in 10 Years

Background: Echinococcosis or hydatidosis, caused by the larval stage of Echinococcus granulosus (E. granulosus), is an important public health problem in many areas of the world and Iran is a country of endemic situation for hydatidosis In the present study, we evaluated epidemiological, complications and clinical characteristics of hydatidosis at three University Medical Centers in Tehran over a 10-year period. M aterials and Methods: This is a descriptive cross-sectional study performed in patients with hydatid cysts. Information about age, gender, number of cysts, organ involvement, morbidity and mortality and relapse were collected from medical records of hydatid patients. Paraclinic information such as CT Scan, MRI, ultrasound, complete blood count, pathological diagnosis and complication of disease were collected. R es ults: Overall, 81 patients, 35 (43.2%) male and 46 (56.8%) female, who were diagnosed as having hydatid cyst by clinical and radiological findings, with pathologic documentation were studied in three university medical center registries over a 10-year period (2003- 2012) in Tehran. Fourteen patients (17% of cases) had complications resulting from this disease. Patients' age ranged from 5 to 86 years, and the peak prevalence of the disease was between 20 and 40 (34% of cases). C onclusion: Iran is a country of endemic situation for hydatidosis. Prevalence rate of hydatidosis in Iran was reported to be 0.61-2 in 100000 populations . The highest rate of infection and complications were in patients of 20-40 years age. Clinical examination revealed that abdominal pain was the most common complaint and was present in 51.7% of the cases. Other most common complain were cough, abdominal mass, dyspnea, icterus, chest pain, dyspepsia, back pain and seizure; and it was result of occupying effect of cysts in organs. This is similar with previous studies in Iran

Antibodies to Echinococcus granulosus Egg Antigens in Hydatid Patients: Characterization by Immunoblot and Diagnostic Value

Cystic echinococcosis/hydatidosis, caused by Echinococcus granulosus, is a chronic zoonotic larval cestode infection in humans. Human hydatidosis is a public health parasitic disease that is cosmopolitan in its distribution. Several years may last between contamination with E. granulosus eggs and development of symptomatic cysts. Antigens expressed by embryos at early differentiation steps between the embryo and the young cyst may be interesting as markers of early infection or reinfection. This study investigates antibody response (IgG) to E. granulosus egg antigens by Immunoblot (IB) and ELISA in 20 hydatid patients surgically confirmed. We have also tested sera using protoscoleces and hydatid cyst fluid (HCF) from E. granulosus. The results show that hydatid patients reacted to eggs as well as protoscoleces and HCF. However, eggs share antigens in common with other life-cycle stages, but also support the notion that they may possess some unique stage-specific antigens determinants. The possible functional significance and the potential use of combined assessment of these antigens in IB with Ig G response in ELISA are discussed.

P016 Impairment of TH1/TH17 host protective responses by macrophages alternative activation as a mechanism for helminthes survival

Human hydatidosis is caused by the larval stage of the metacestode Echinococcus granulosus (E.g.). It is one of the world’s major zoonotic infections. It usually manifests as unilocular cyst(s) mainly located in the liver. The prolonged coexistence of the parasite and its host involve the laminated layer (LL). This acellular layer represents an important barrier against immunological and physiological reactions of the host.We have previously highlighted an evident role of IFN γ in parasite killing by NOS2 (Nitric oxide Synthase2) pathway 1 . Moreover, IL-10/Arginase pathways seem to be implicated in parasite escape to host defense 2 . Moreover, LL impairs Th1/Th17 protective responses and allows the parasite to survive. This role may be mediated by Treg up-regulation by LL. In the present work, we aim to identify the early effect of LL in parasite establishment. In this sense, the effect of LL on macrophages phenotype and expression of some M1/M2 markers was investigated. NOS2 and Arginase activities were evaluated in cultures performed with PBMC of hydatid patients and healthy donors. Moreover, implication of mannose receptor (MR) and TGF β on Arginase activity was evaluated using mannose (MR antagonist) and Anti-TGF β . Activity of NADPH oxydase (ROS production) was also evaluated by chemiluminescence in cultures performed with human monocytes primed with NADPH oxydase agonists (like opsonized zymosan). Finally, TLR2, TLR4, CD14 and CD23 expression by the same cells was evaluated by flow cytometric immunoassay. We have found that NOS2 and NADPH activities are inhibited by LL. This effect is associated with Arginase activity stimulation. Interestingly, MR and TGF β seem to be implicated in the Arginase induction by LL. Moreover, LL increases TLR2 and CD14 expression and decreases TLR4 and CD23 expression by monocytes. Collectively, ours finding suggest that e.g. laminated layer induced M2 phenotype. These macrophages induce Treg cells and impair M1/Th1/Th17 protective responses allowing parasite survival. Inhibition of these mechanisms seems to be important issue to address during the design of anti-hydatic treatment.

Interleukin-17A correlates with interleukin-6 production in human cystic echinococcosis: a possible involvement of IL-17A in immunoprotection against Echinococcus granulosus infection

Hydatidosis is a parasitic disease caused by the development, in humans and other mammals, of the larval form of Taenia, Echinococcus granulosus. It is one of the world's major zoonotic infections. This study aimed to examine interleukin-6 (IL-6), interferon-γ (IFN-γ) and interleukin-17A (IL-17A) production in patients with cystic echinococcosis (CE), and the role of IL-17A in the modulation of the immune response against the extracellular parasite, E. granulosus. A relationship between IL-6, IL-17A production and C reactive Protein (CRP) levels was also assessed. IL-6, IFN-γ, IL-17A and CRP production were determined in serum from Algerian hydatid patients. Cytokine production was also measured in supernatants from cultures of peripheral blood mononuclear cells (PBMCs) from hydatid patients stimulated by a major parasitic antigen (antigen-5). The increased activity of IL-6, IFN-γ and IL-17A were observed in most serum samples from patients. In contrast, healthy controls showed only minor levels. Similarly, high levels of CRP were detected. Our in vitro results indicate a positive correlation between IL-6, IFN-γ and IL-17A production in PBMC culture supernatants. However, IL-6, IFN-γ and IL-17A activity was low in serum and supernatants of PBMC cultures from relapsing patients, and there was no evidence of an immune response against parasitic antigen. Collectively, our results show that IL-17A was produced during human cystic echinococcosis, and was involved in the host defense mechanisms against the extracellular parasite E. granulosus. Our data suggest that IL-17A plays an immunoprotective role in this parasitic, helminth infection.

Involvement of IL-10 and IL-4 in evasion strategies of Echinococcus granulosus to host immune response

Human echinococcosis is one of the world's major zoonotic infections. In this study, our aim is to clarify one of the strategies used by the parasite for evasion and prolonged infestation in the host. We wished to provide further immunological evidence for the involvement of IL-10/IL-4 in these mechanisms. In this regard, we investigated the effects of IL-4 and IL-10 on protoscoleces (PSC: the larval form of the parasite), co-cultured with patient PBMC. Furthermore, we used IL-4 and IL-10 antibodies to confirm this effect. Our results showed that IL-4 and IL-10 reduced PSC killing. This reduction correlates with an decrease in NO production by PBMC. In conclusion, the results reported here suggest that IL-4/IL-10 impairs the Th1 protective response and allows the parasite to survive in hydatid patients.

Étude du profil cytokinique de patients atteints d’hydatidose : une possible application en matière d’immunosurveillance

Hydatidosis is a severe parasitic disease caused by infection with metacestode of the tapeworms Echinococcus granulosus. In human, the larval forms develop into large cysts especially in the liver, lung and brain. Our aim in this study was to investigate Th1 and Th2 cytokines production in hydatid disease in order to evaluate implication of Th1/Th2 ratio in the evolution of pathology according to the cystic localization, clinical stage and clinical evolution. Interferon-gamma (IFN-gamma), interleukine-12 (Il-12), interleukine-16 (Il-16), interleukine-18 (Il-18), interleukine-4 (Il-4), interleukine-5 (Il-5), interleukine-10 (Il-10) and interleukine-13 (Il-13) production is determined in sera from hydatid Algerian patients (n = 177) with liver, lung, liver and lung associated, spleen, kidney, osseo, heart and multiples hydatid cyst and in sera from patients with clinical complications (calcified liver cysts; infected lung cysts; vomique lung cysts and patients who relapsed) and according clinical stage (before surgical extirpation of the cyst and after surgical extirpation of the cyst). Cytokines are evaluated by enzyme-linked immunosorbent Kits (ELISA Immunotech). The coexistence of elevated activities of IFN-gamma, Il-12, Il-16, Il-18, Il-4, Il-5, Il-10, and Il-13 is observed in most of sera from hydatid patients. In contrast, healthy controls showed minor levels. These results support Th1 and Th2 cell subsets activation in human hydatidosis. The comparison of Th1/Th2 production shows that the induction of these mediators is related to the cystic localization, the clinical stage and clinical evolution. Collectively, our data indicates that Th1 cytokines are related to the protective immunity, in contrast Th2 cytokines are responsible to the susceptibility to disease and associated with chronicle stage, clinical complications and secondary locations.

In vitro production of cytokines by peripheral blood mononuclear cells from hydatid patients.

The role of cytokines in human hydatidosis (Echinococcus granulosus infection) was evaluated in immunoassays determining production of IL-4, IL-10 and interferon-gamma (IFN-gamma) in peripheral blood mononuclear cell (PBMC) cultures from 30 hydatid patients and 14 uninfected controls. In cell cultures from hydatid patients parasite and non-parasite antigen stimulation significantly increased IL-4 production (P < or 0.005). Spontaneous and mitogen-driven IL-4 production was similar in patients and controls. IL-10 and IFN-gamma production did not differ statistically in the two groups, even though some hydatid patients produced these cytokines in large amounts. Notably, antigen-driven IFN-gamma concentrations were invariably higher in patients than in uninfected controls. Data analysis showed a relationship between IgE and IgG4 responses and parasite-driven cytokine production. High IgE and IgG4 responders produced high IL-4 and IL-10 concentrations. High IgE responders showed decreased IFN-gamma production, but high IgG4 responders had IFN-gamma levels slightly higher than those of low responders. Cytokine response patterns did not relate to the clinical stage of disease. The significantly increased IL-4 and the high IL-10 concentrations found in PBMC from many hydatid patients in this study are consistent with Th2 cell activation in human hydatidosis. The presence of antigen-driven IFN-gamma production in patients with E. granulosus infection implies concurrent intervention of the Th1 or Th0 cell subset.

Immunological markers indicating the effectiveness of pharmacological treatment in human hydatid disease.

The relation of interferon-gamma (IFN-gamma), IL-4, IL-10 production and specific IgE, total IgG, IgG subclass expression to the effectiveness of pharmacological treatment in human hydatid disease (Echinococcus granulosus infection) was evaluated in 27 hydatid patients divided into four clinical groups according to their response to albendazole/mebendazole therapy (full, partial, low and non-responders). After parasite antigen stimulation, peripheral blood mononuclear cells (PBMC) from full responders produced significantly more IFN-gamma (P = 0.038), significantly less IL-4 (P = 0.001) and less IL-10 than PBMC from non-responders. PBMC from partial and low responders produced intermediate cytokine concentrations. ELISA determining immunoglobulin production showed that sera from all non-responders had IgE and IgG4 antibodies, both regulated by IL-4. In contrast to IgG4, IgE decreased rapidly in full responders. Full responders also showed the highest percentage of IgG3 reactions. Qualitative analysis of total IgG responses in hydatid patients' sera determined by immunoblotting showed that binding profiles to hydatid cyst fluid antigen differed in the four groups of treated patients. Non-responders had the highest percentage of reactions to all subunits of antigens 5 and B, and full responders had the highest percentage of reactions to antigen 5 alone. The high IFN-gamma production associated with a lack of IL-4 and low IL-10 production in the full responders, and vice versa the high IL-4 and IL-10 production associated with lack of or low IFN-gamma production in the non-responders implies Th1 cell activation in protective immunity and Th2 cell activation in susceptibility to hydatid disease. IgE may be a useful marker of therapeutic success in hydatid patients with pretreatment specific IgE antibodies. IgG subclass responses and differential immunoglobulin subclass binding pattern to hydatid antigens may also be useful in the immunosurveillance of hydatid disease.

In VitroAntihydatic Action of IFN-γIs Dependent on the Nitric Oxide Pathway

Hydatidosis is a widely endemic helminthic disease vectored in human by the larval stage of the metacestode Echinococcus granulosus. It is characterized by the long-term coexistence of chronic infection with detectable humoral and cellular responses against the macroparasite. Previous studies demonstrated interferon-gamma (IFN-gamma) and nitric oxide (NO) production (in vivo and in vitro) during hydatidosis. In this study, we tested the hypothesis that NO production after IFN-gamma induction may constitute a host defense against E. granulosus. We also investigated the IFN-gamma effect on protoscolices (larval form of the parasite) viability in coculture with hydatid patients' peripheral blood mononuclear cells (PBMC). PBMCs from hydatic patients incubated with IFN-gamma (100 U/mL) alone are effective in the killing of protoscolices. This scolicidal activity is concomitant with elevation of nitrite levels. NO release and cytotoxic activity are inhibited by N-monomethyl-L-arginine (L-NMMA), a specific inhibitor of the NO pathway and increased by L-arginine, an NO precursor, and tetrahydrobiopterin (BH4), a nitric oxide synthase (NOS) cofactor. Our results indicate that IFN-gamma mediated iNOS induction as one of host defense mechanism against human E. granulosus infection.

Antibody responses in human cystic hydatid disease to the laminated layer of Echinococcus granulosus

The laminated layer of hydatid cysts of Echinococcus granulosus represents a considerable amount of parasite material. Its antigenic role, however, is unclear. Extracts of laminated layer taken from sheep cysts were analysed in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS/PAGE) and were found to contain bands at 55 and 25-29 kDa, which reacted with an anti-sheep IgG antibody probe, indicating that these were likely to be host-contaminating components within the layer. However, the same bands were also recognised by a significant proportion of human hydatid patients, particularly by IgG4 antibodies, and not by negative control individuals. These individuals did not recognise immunoglobulin heavy and light chains in a sheep serum extract in the same manner. It seems likely that there are either host or parasite antigenic components at similar molecular weights or that certain parasite antigens may share epitopes with sheep immunoglobulins. The antigens at 25-29 kDa were found to be glycoproteins by lectin blot analysis and may be important markers of disease status.

Review of surgical management of cystic hydatid disease in a resource limited setting: Turkana, Kenya.

Cystic hydatid disease is a zoonotic infection of humans caused by infection with the larval stage of Echinococcus granulosus. The prevalence rate (5-10%) in Turkana, Northern Kenya is among the highest worldwide. With an increase in foreign travel and migration of peoples, practitioners need to be aware of diseases common in these countries and their rarer manifestations. The objective of this study was to review the management of this disease in a poorly resourced high-prevalence area. The surgical records of Kakuma Mission Hospital, Turkana from July 1981 to May 2002 were reviewed. A total of 710 (female : male, 3 : 2) surgical procedures for hydatid disease were recorded, the average age being 27 years (range, 3-65 years). There were 663 patients, and therefore 47 patients had repeat procedures. A total of 52.8% (n = 375) had hepatic cysts (248 right sided, 30 left sided, 97 site undefined), and 10.1% (n = 72) had multiple abdominal cysts. Other intra-abdominal sites included 8.16% mesenteric (n = 58), 2.9% retroperitoneal (n = 21), 3.5% spleen (n = 25), the abdominal wall (n = 4). There was no information on the site of disease in 5.9% of cases (n = 42). More unusual sites were retro-orbital, gluteal, zygomatic, brachial, parotid, uterine, tibial and foot. The largest volume of cyst fluid was 7 l. There was one intraoperative death and one postoperative death recorded. Endocystectomy +/- a scolicidal was the commonest procedure performed. However, 165 percutaneous aspiration injection of a scolicidal agent and reaspiration procedures were performed, six being on pregnant women. This is one of the largest series of surgical hydatid patients from Africa and demonstrates the wide diversity in cyst location.

Immune response and in vivo production of cytokines in patients with liver hydatidosis.

Cytokines play an important role in the human immunological response, but the exact role of cytokines in the human immune response against parasites, especially against Echinococcus granulosus, remains unclear. IL-1, IL-2, IL-4 and tumour necrosis factor (TNF) levels in peripheral blood of 21 patients with liver hydatidosis were evaluated before surgical treatment, and the levels of IgA, IgM, IgG, IgE, specific IgE against E. granulosus, C3, C4 and DF complement fractions and CD20, CD3, CD4, CD8 and CD16 cell percentages were also determined, as was the relationship between these variables and cytokine levels. Data from hydatid patients were compared with data obtained from 21 healthy volunteers. Hydatid patients showed increases of IgG, IgE, IgEs and IL-2 (P < 0.01), and decreases of IL-1 and TNF levels (P < 0.001), but these variables (respectively) increased in patients showing cysts in the central area of the liver or with a wide opening of cysts in the biliary tract. The increase of IL-1, IL-2 and IL-4 showed a close relationship with the number, characteristics and above all the location of cysts within the liver itself. IgG and IL-4 levels and also IgG and IgE levels showed a significant correlation (P < 0.05).

Production of IL-5 and IL-6 by peripheral blood mononuclear cells (PBMC) from patients with Echinococcus granulosus infection

The role of Th2 cytokines in human hydatidosis was evaluated in ELISA determining IL-5 and IL-6 production in PBMC cultures from 27 pharmacologically treated hydatid patients and from 13 uninfected controls. PBMC from patients produced large amounts of parasite antigen-driven IL-5, whereas PBMC from uninfected individuals produced none. In contrast, PBMC from patients and from uninfected controls produced large amounts of parasite antigen-driven IL-6. Immunoglobulin isotype analysis revealed that IL-5 production correlated significantly with IgE and IgG4 expression (IL-5/IgE r = 0.5, P < 0.05; IL-5/IgG4 r = 0.6, P < 0.05). The high IL-5 levels in supernatants from patients' PBMC did not correspond to an increase in eosinophils. Neither IL-5 nor IL-6 production showed an association with the outcome of therapy. Overall, these findings confirm that the lymphocytes of individuals with Echinococcus granulosus infection contain Th2-like subpopulations.

Double aldehyde stabilisation of erythrocytes in the indirect hemagglutination for echinococcosis

Sheep erythrocytes were stabilised with glutaraldehyde tanned and fixed with formalin in the indirect hemagglutination test (IHA-GF) and sensitised with hydatid antigen for the diagnosis of human cystic echinococcosis (CE). The sensitivity of this method was compared to that prepared with fresh tanned cells (IHA-TA) in 278 sera from hydatid patients. The sensitivity of IHA-GF (87.8%) was higher than that of IHA-TA (85.6%), the difference being insignificant. Higher geometric mean titres were obtained by IHA-GF (1:13 300) than by IHA-TA (1:11 600). The use of two sorts of aldehydes proved to be a satisfactory method, showing high sensitivity, a very good specificity and some advantages. The sensitised cells retained their diagnostic effectiveness for at least 15–18 months when stored at 4 °C. The technique is inexpensive and rapid, allowing the testing of a large number of sera. The method reduces the variation of the results and improves the reproducibility of the test. When the minimal diagnostic titre—1:400 is used the specificity of IHA-GF might increase by 2.9% while the sensitivity might decline by only 1.4%. The IHA-GF demonstrated better immunodiagnostic characteristics than enzyme linked immunosorbent assay (ELISA) and Latex agglutination test (LAT). The IHA-GF should be considered as an useful method in the range of classical diagnosis for the serology of CE. The clinical diagnostic potential should be increased by a combination of at least two tests: IHA-GF and ELISA or LAT.