Specific ELISA Test Research Articles

The engagement of the aryl hydrocarbon receptor by tryptophan derivatives can prevent the development of anti-FVIII antibodies in an experimental model of hemophilia A

Abstract Patients affected by haemophilia A require treatment with factor VIII (FVIII) protein. The most relevant complication is the development of neutralizing FVIII-specific antibodies or “inihibitors”. We reported that the inhibitor-positive status was associated with reduced activity of the immune-regulatory enzyme indoleamine 2,3-dioxygenase 1 (IDO1), that promotes regulatory effects via the production of tryptophan catabolites, known as kynurenines. Some of those tryptophan derivatives are endogenous ligands for the Aryl hydrocarbon receptor (AhR). In this study we tested the potential of tryptophan-related AhR ligands for inhibiting the development of anti-FVIII antibodies in hemophilic (F8 KO) mice. To this aim, F8 KO mice were treated with recombinant human FVIII (rhFVIII) alone or in combination with selected AhR ligands once weekly for four weeks. Antibody titers were tested by specific ELISA and Bethesda test. All mice treated with rhFVIII developed high-titer anti-FVIII antibodies after 4 weeks of treatment. Administration of a specific tryptophan metabolite prevented the generation of anti-FVIII antibodies in almost 80% of F8 KO mice. The protective effect of these AhR ligands was negated by co-administration of the AhR antagonist CH-223191 or in AhR KO mice. Similar results were obtained by administration of engineered gold nanoparticles loaded with the same tryptophan metabolite and rhFVIII. These results suggest that the engagement of AhR, by specific tryptophan derivatives, may be a possible new strategy to control the immune response to rhFVIII. Our findings might lead to the development of novel immunomodulatory interventions for preventing or eradicating inhibitors in hemophilia A patients.

Articular and subcutaneous adipose tissues of rheumatoid arthritis patients represent equal sources of immunoregulatory mesenchymal stem cells

Adipose-derived mesenchymal stem cells (ASCs) have immunoregulatory properties, but their activity is dependent on signals provided by the local microenvironment. It is likely that highly inflammatory milieu of rheumatoid joint affects ASCs activity. To test this hypothesis, the function of rheumatoid ASCs derived from articular adipose tissue (AT-ASCs) and ASCs derived from subcutaneous adipose tissue (Sc-ASCs) has been analysed. Articular adipose tissue (infrapatellar fat pad) and subcutaneous adipose tissue (from the site of skin closure with sutures) were obtained from rheumatoid arthritis (RA) patients undergoing total knee joint replacement surgery. ASCs were isolated accordingly to the routinely applied procedure, expanded and treated or not with IFNγ and TNF (10 ng/ml). To evaluate immunomodulatory properties of AT- and Sc-ASCs, co-cultures with peripheral blood mononuclear cells (PBMCs) from healthy donors have been set. Proliferation of activated PBMCs (3H-thymidine incorporation method), secretion of IL-10 and IL-17A in co-culture supernatants (specific ELISA tests) and T regulatory FoxP3+ cells (Tregs) percentage have been evaluated (flow cytometry). Performed experiments demonstrated that ASCs from both sources have comparable properties. They suppress proliferation of activated PBMCs to the similar extent, induce IL-10 secretion by resting PBMCs and moderately induce generation of FoxP3+ Treg cells. Interestingly, both AT-ASCs and Sc-ASCs cause increase of IL-17A secretion by activated PBMCs as well as induce up-regulation of IL-6 concentration in co-culture supernatants. We demonstrated that AT-ASCs and Sc-ASCs obtained from RA patients possess similar immunomodulatory properties despite different localization and distinct cytokine milieu of tissue of origin. Our results indicate that ASCs derived from rheumatoid adipose tissues are not strongly immunosuppressive in vitro and that they may contribute to the pathogenesis of RA due to IL-17A secretion enhancement.

A study of altered cytokine rhythms associated with successful implantation in cows

Implantation is a critical process wherein the dam accepts the semi-allogenic embryo, characterized by certain vital immunological changes involving certain factors and cytokines. The current study was undertaken in Sahiwal cows to analyze the changes in certain selective immunological components of the body during peri-implantation period. Post-insemination the study was performed for a period till day 40 and the plasma was used to estimate the levels of progesterone, interleukin 1β (IL-1β), IL-2, IL-4, IL-6, and leukemia inhibiting factor (LIF) using Bovine specific ELISA Test Kits. The phagocytic activity and mRNA expression of progesterone-induced blocking factor (PIBF) were also analyzed. It was observed that in pregnant cows, the expression of PIBF increased from day 16th onward and remained high throughout the study period whereas the phagocytic activity was higher in non-pregnant cows all through the study period. The IL-1β, IL-2, and IL-6 were significantly higher in non-pregnant cows from 16th to 18th day post-AI as compared to P cows. On the other hand, IL-4 and LIF showed a significantly higher concentration in pregnant cows during peri-implantation period. Hence, it can be concluded that early embryonic survival is dependent on interplay of certain critical immunological factors in the body.

Role of Serum Midkine Level as a Diagnostic Biomarker for Very Early and Early Hepatocellular Carcinoma

Background and study aim: AFP is the most commonly utilized biomarker for HCC although it has a low sensitivity and specificity for the disease. New biomarkers with better sensitivity and specificity are to be studied. The aim of this work is to evaluate the role of midkine as a biomarker for early detection of HCC and for detection of HCC in patients negative for AFP. Patients and Methods: This study included 46 HCC patients on top of Child Pugh class A cirrhosis (group A), 46 patients with Child Pugh class A cirrhosis without liver focal lesion(s) (group B) and 46 apparently healthy controls (group C). Demographics, clinical data, radiologic findings, biochemical profile including serum AFP and midkine assessment using specific ELISA tests of the study participants were entered in the study. Results: Serum midkine had a statistically significant better sensitivity, specificity, positive predictive value, negative predictive value and accuracy over serum AFP for diagnosing very early and early HCC from liver cirrhosis and for diagnosing AFP negative HCC from liver cirrhosis. Conclusion: Serum midkine is to be considered for diagnosis of very early and early HCC especially in AFP negative cases.

Clinical profile of dengue in a hitherto non-endemic region—A hospital based study from northeast India

Abstract Background Global burden of dengue is substantial, with an annual incidence of 50 million cases. India has witnessed gradual rise in its incidence, mainly from urban regions. North-eastern India, hitherto non-endemic for dengue, has reported sporadic cases of dengue lately. Objective To study the clinical profile of dengue in north-east India. Method and material Clinical profile of 218 dengue patients (≥15 years) was studied in a hospital-based observational study. Dengue was diagnosed by WHO criteria and confirmed by NS1 antigen and/or dengue specific IgM ELISA tests. Statistical analyses were done using SPSS and STATA Data analysis and statistical software. A ‘p value’ Results There was male preponderance (58.7%), affecting mainly younger (15–34 years) subjects (73.85%). Majority (96.33%) were without travel history, none had dengue previously. Serological confirmation was present in 77.06%. On admission, mean haemoglobin and platelet counts were low with normal leucocyte count and haematocrit. Platelet and leucocyte counts declined during hospital stay; 8.25% developed severe thrombocytopenia ( 3 ) necessitating platelet transfusion, 17.43% developed leukopenia. Systemic hypotension was found in 13.3%. Mortality was nil and average hospital stay was 5.36 days. Conclusion First attacks of dengue without travel history, in a hitherto non-endemic region suggest recent endemicity.

Epidemiological and Clinicopathological Studies of Sheep Naturally Infected with Foot and Mouth Disease Virus (SAT2) in Egypt

This study was done to evaluate the epidemiological pattern and cardiac markers of foot and mouth disease virus (FMDV, SAT2) in naturally infected sheep, in Dakahlia Governorate, Egypt. The specific antigen ELISA test confirmed positive FMDV, SAT2 in sheep with oral lesions (14.7%), lameness (22.9%) and in dead sheep (3.97%). Death was highest (p

130 ANTRAL FOLLICLE COUNTS, VULVA WIDTH, AND SERUM ANTI-MULLERIAN HORMONE CONCENTRATIONS IN BOVINE FEMALES OF THE TABAPUÃ BREED

Bovine genetic selection for fertility is difficult in part due to the low heritability of fertility traits and to the extension of the calving interval and the productive life length inherent to this species, which makes it harder to identify superior individuals from their records. In this regard, it is feasible to try to identify characteristics that are easy to detect and may be linked to superior reproductive efficiency as early as possible in females. The objective was to evaluate the possible relationships between vulva width, antral follicle counts (AFC), and the serum concentrations of anti-Mullerian hormone (AMH). Tabapuã (a Brazilian Zebu breed) heifers (n = 155) and cows (n = 422) from 4 herds were used in the study in which all antral follicles ≥3 mm were counted directly with an ultrasound unit. Follicle counts were classified into 2 classes, based on the methodology proposed by Burns et al. (2005) in dairy cattle, such that the low and intermediate AFC classes were merged into one class (low; n = 245) and the high and very high into another (high; n = 332). The vulva width was determined by measuring the distance between the lateral borders of the vulva with a digital caliper placed at a 90-degree angle from the half point of the rima vulvae. Single blood samples were collected from a subgroup of animals (n = 220) and the serum frozen (–20°C) after centrifugation for the AMH analysis, which was performed by specific bovine ELISA (Ansh Laboratories, Webster, TX, USA) test. All statistical analyses were performed with SAS (SAS Institute Inc., Cary, NC, USA). Data was submitted for analyses of variance using the PROCGENMOD procedure. Vulva width was larger (P = 0.02) in the class of animals of the high AFC (8.73 ± 0.11 cm) in comparison with those of the low AFC (8.39 ± 0.09 cm), considering the larger sample (n = 591). In the subsample (n = 220) the concentration of AMH was higher (P < 0.0001) in the high AFC class females (1.15 ± 0.07 ng mL–1; n = 95) in comparison with the low AFC class (0.50 ± 0.06 ng mL–1; n = 125). Vulva width, AFC, and AMH serum concentrations were interrelated and likely fertility indicative phenotypes under the present experimental conditions and should be further evaluated in larger samples in order to be considered in bovine genetic selection.

Detection of atypical Chlamydiaceae in roe deer (Capreolus capreolus)

Investigations on fecal samples, vaginal swabs and sera from roe deer (Capreolus capreolus) in south-western France led to the detection of a non-classified Chlamydiaceae strain. A total of 85 vaginal swabs were sampled from roe deer that had been captured in 2012 (n=42) and 2013 (n=43). Using a Chlamydiaceae family-specific real-time PCR, only one vaginal swab out of the 42 samples done in 2012 tested positive and was subsequently identified as Chlamydia (C.) psittaci. In contrast, 6/43 vaginal swab samples were positive in 2013. Four of these positive samples came from a single group of roe deer, captured in the Fabas plain. Fecal samples from this group of 9 females were subsequently analyzed, with 6 of them testing positive with the Chlamydiaceae-specific PCR. All positive samples collected in 2013 were negative when re-tested with C. abortus-, C. pecorum- and C. suis-specific real-time PCR assays. Sera from this group of 9 females were analyzed with two immunoassays (recomLine and ELISA). Whereas intense positive reactions with C. pneumoniae antigens were observed for all sera when tested with the recomLine test, none was positive with the C. abortus specific ELISA test. Comparative sequence analysis of the 16S, 23S rRNA and ompA gene sequences from 3 animals, as well as the MLST analysis from 2 animals, showed that this roe deer group likely harbored the same bacterium related to members of the family Chlamydiaceae. Notably, the roe deer strain formed a separate entity different from the currently recognized chlamydial species, with C. trachomatis, C. suis and C. muridarum appearing as its closest relatives.

Progesterone-induced stimulation of mammary tumorigenesis is due to the progesterone metabolite, 5α-dihydroprogesterone (5αP) and can be suppressed by the 5α-reductase inhibitor, finasteride

Progesterone has long been linked to breast cancer but its actual role as a cancer promoter has remained in dispute. Previous in vitro studies have shown that progesterone is converted to 5α-dihydroprogesterone (5αP) in breast tissue and human breast cell lines by the action of 5α-reductase, and that 5αP acts as a cancer-promoter hormone. Also studies with human breast cell lines in which the conversion of progesterone to 5αP is blocked by a 5α-reductase inhibitor, have shown that the in vitro stimulation in cell proliferation with progesterone treatments are not due to progesterone itself but to the metabolite 5αP. No similar in vivo study has been previously reported. The objective of the current studies was to determine in an in vivo mouse model if the presumptive progesterone-induced mammary tumorigenesis is due to the progesterone metabolite, 5αP. BALB/c mice were challenged with C4HD murine mammary cells, which have been shown to form tumors when treated with progesterone or the progestin, medroxyprogesterone acetate. Cells and mice were treated with various doses and combinations of progesterone, 5αP and/or the 5α-reductase inhibitor, finasteride, and the effects on cell proliferation and induction and growth of tumors were monitored. Hormone levels in serum and tumors were measured by specific RIA and ELISA tests. Proliferation of C4HD cells and induction and growth of tumors was stimulated by treatment with either progesterone or 5αP. The progesterone-induced stimulation was blocked by finasteride and reinstated by concomitant treatment with 5αP. The 5αP-induced tumors expressed high levels of ER, PR and ErbB-2. Hormone measurements showed significantly higher levels of 5αP in serum from mice with tumors than from mice without tumors, regardless of treatments, and 5αP levels were significantly higher (about 4-fold) in tumors than in respective sera, while progesterone levels did not differ between the compartments. The results indicate that the stimulation of C4HD tumor growth in BALB/c mice treated with progesterone is due to the progesterone metabolite 5αP formed at elevated levels in mammary cells as a result of the 5α-reductase action on progesterone. The results provide the first in vivo demonstration that stimulation of breast cell tumorigenesis and tumor growth accompanying progesterone treatment is due to the progesterone metabolite 5αP, and that breast tumorigenesis can be blocked with the 5α-reductase inhibitor, finasteride.

Changes in Hormones of Somatotropic Axis during Transition Period in Murrah Buffaloes (Bubalus bubalis) Supplemented with Vitamin E

The present study was aimed to assess the effect of vitamin E supplementation on GH-insulin–IGF-1 axis in Murrah buffaloes during transition period. The Murrah buffaloes (n = 12) were selected during their late gestation from National Dairy Research Institute (NDRI) livestock herd and divided randomly into two groups, comprising of 6 each. Buffaloes of group I were given only the control diet, while group II were supplemented with 2000 IU/day/head vitamin E along with control feed. Blood sample were collected from each buffalo at weekly interval from day -21 to day +21 relative to parturition by jugular vein- puncture. Growth hormone (GH) -IGF-1 and insulin were quantified by bovine specific ELISA test kit. The results revealed non significant difference in circulatory concentration of insulin and GH during transition period between the two groups. However, insulin and IGF-I magnitude of decline was significantly greater in Group I. It was concluded that decline in the levels of these hormones during transition period was a physiological phenomenon that was associated with negative energy balance. Dietary supplementation of Vitamin E had no direct influence on peripheral profiles of these hormones during transition period.

External Validation Of The HIT Expert Probability (HEP) Score, a Novel Pre-Test Probability Model For Heparin-Induced Thrombocytopenia Based On Broad Expert Opinion

IntroductionThe diagnosis of heparin-induced thrombocytopenia (HIT) is challenging due to the overlap of its clinical features with other entities, variable specificity of ELISA-based tests, poor availability of functional tests such as the serotonin release assay (SRA) and suboptimal positive predictive values of pre-test probability clinical scoring models, such as the 4T and Lillo-Le Louėt models. Improved diagnostic tools are desirable to guide early therapeutic decisions in patients with suspected HIT and to reduce inappropriate use of alternative anticoagulants. The HIT Expert Probability (HEP) Score, a pre-test probability clinical scoring model for HIT based on broad expert opinion, has recently been proposed to improve the diagnosis of HIT compared to the 4T score. We sought to externally and prospectively validate the HEP score. MethodsPre-test probability of HIT was prospectively assessed for 51 consecutive patients referred to the Cleveland Clinic Vascular Medicine Consultative Service for evaluation of possible HIT between August 1, 2012 and February 1, 2013. Electronic medical records were reviewed to obtain clinical, biochemical (including IgG specific HIT ELISA and SRA tests) and imaging data, and two Vascular Medicine fellows independently applied the 4T and HEP scores for each patient. Two independent HIT expert adjudicators rendered a diagnosis of HIT likely or unlikely. Both scorers and adjudicators were blinded to the Results of HIT laboratory testing, consultative service inputs and scores generated by other raters. ResultsThe median 4T and HEP scores were 4.5 [range 3.0, 6.0] and 5 [range 3.0, 8.5], respectively. Interobserver agreement was lower for the HEP Score compared to 4T score, intraclass correlation coefficient: 0.50 (95% CI 0.16–0.85) vs. 0.69 (0.54–0.86). There were no significant differences between area under receiver-operating characteristics curves of 4T and HEP score against HIT ELISA testing (0.62 vs. 0.58, P = 0.62), SRA (0.74 vs. 0.73, P = 0.97), expert panel diagnosis (0.71 vs. 0.79, P = 0.20), concordant HIT ELISA and SRA (0.76 vs. 0.72, P = 0.65), and positive SRA or positive HIT ELSA with HIT likely expert panel diagnosis (0.73 vs. 0.75, P = 0.80). The HEP score model was 100% sensitive and 17% specific for determining the presence of HIT as defined by positive SRA or positive HIT ELISA with HIT likely expert panel diagnosis compared to 4T score (sensitivity, 94%; specificity 38%) (Figure 1, Table 1). [Display omitted] ConclusionThe HEP and 4T scores are excellent screening pre-test probability models for HIT, however, in this prospective validation study, test characteristics for the diagnosis of HIT based on confirmatory laboratory testing and expert opinion are similar. Given the complexity of the HEP scoring model compared to that of the 4T model, further validation of the HEP score is warranted prior to widespread acceptance. Disclosures:Kim:Philips US: Consultancy; GE: Research Funding.

Seroepidemiological Survey of Crimean-Congo Hemorrhagic Fever Among Sheep in Mazandaran Province, Northern Iran

Crimean-Congo hemorrhagic fever (CCHF) is a disease prevalent among humans and animals and is endemic in Iran. Although CCHF has been reported in all of its neighboring provinces, in Mazandaran in northern Iran there have been no reports of any cases of human infection. This research has been carried out to clarify the epidemiological aspects of CCHF infection among sheep in various geographical regions of Mazandaran province. In this survey, 270 blood samples were collected from sheep in eastern, central, and western Mazandaran between 2010 and 2011, and the specific ELISA test for CCHF virus was carried out on the blood samples in the National Reference Laboratory in the Pasteur Institute, Tehran, Iran. The CCHF infection rate according to this study was 3.7%. A weak statistical relationship (p=0.063) was seen between the different geographical regions, with a gradual decrease in the infection rate noted, stretching from the eastern to the western portions of the province (eastern 6.8%, central 2.8%, and western 0%). Older sheep were 2.7 times more likely to be infected with the virus (OR 2.70; 95% CI 1.50,4.87; p<0.001). As the infection rate in Mazandaran is low among sheep, and as yet there have been no reports of human infection in this province, CCHF disease is not considered a serious health problem in Mazandaran. It is recommended that further research be carried out on other animals, high-risk human groups, and ticks, in order to more completely reveal the status of the disease in this province.

Dengue Infection and Miscarriage: A Prospective Case Control Study

BackgroundDengue is the most prevalent mosquito borne infection worldwide. Vertical transmissions after maternal dengue infection to the fetus and pregnancy losses in relation to dengue illness have been reported. The relationship of dengue to miscarriage is not known.MethodWe aimed to establish the relationship of recent dengue infection and miscarriage. Women who presented with miscarriage (up to 22 weeks gestation) to our hospital were approached to participate in the study. For each case of miscarriage, we recruited 3 controls with viable pregnancies at a similar gestation. A brief questionnaire on recent febrile illness and prior dengue infection was answered. Blood was drawn from participants, processed and the frozen serum was stored. Stored sera were thawed and then tested in batches with dengue specific IgM capture ELISA, dengue non-structural protein 1 (NS1) antigen and dengue specific IgG ELISA tests. Controls remained in the analysis if their pregnancies continued beyond 22 weeks gestation. Tests were run on 116 case and 341 control sera. One case (a misdiagnosed viable early pregnancy) plus 45 controls (39 lost to follow up and six subsequent late miscarriages) were excluded from analysis.FindingsDengue specific IgM or dengue NS1 antigen (indicating recent dengue infection) was positive in 6/115 (5·2%) cases and 5/296 (1·7%) controls RR 3·1 (95% CI 1·0–10) P = 0·047. Maternal age, gestational age, parity and ethnicity were dissimilar between cases and controls. After adjustments for these factors, recent dengue infection remained significantly more frequently detected in cases than controls (AOR 4·2 95% CI 1·2–14 P = 0·023).InterpretationRecent dengue infections were more frequently detected in women presenting with miscarriage than in controls whose pregnancies were viable. After adjustments for confounders, the positive association remained.

Serological Evaluation of Crimean-Congo Hemorrhagic Fever in Humans with High-Risk Professions Living in Enzootic Regions of Isfahan Province of Iran and Genetic Analysis of Circulating Strains

Crimean-Congo hemorrhagic fever (CCHF) is a zoonotic viral disease that is asymptomatic in infected livestock, but causes a serious threat to humans with a mortality rate up to 50%. Although the CCHF virus (CCHFV) is often transmitted by ticks, livestock-to-human and human-to-human transmission also occurs. In the current study, we focused on CCHF in the province of Isfahan, located in the center of Iran and deemed to be the second most infected province. Human and livestock sera and resident ticks in the livestock are collected from different regions of the province and analyzed with specific IgG ELISA and RT-PCR tests. Overall, 12% and 12.7% of studied human and livestock populations were IgG positive, respectively. The genome of CCHFV was detected in 9% of ticks resident in livestock involved in this survey. The CCHFV isolates from infected ticks were genetically examined. Nucleotide sequence of the S-segment revealed that the different isolates were closely related to each other, with nucleotide sequence identities higher than 98%. Phylogenetic analysis demonstrated that a variant isolate clustered with the Iraq strain. This high proportion of IgG-positive sera and nearly high proportion of infected ticks increases the risk of CCHF outbreaks in the province and probably posits a great danger to other provinces.

Cross-Reaction between the Crude Hydatid Cyst Fluid Antigens of Human and Animals Origin in Response to Human IgG Class and Subclasses

The current work aimed to evaluate the cross-reactivity of human immune sera against crude hydatid fluid antigens of sheep, human, mouse, cattle, as well as B fraction of cystic fluid antigen. 30 balb/c mice were infected with sheep hydatid cyct fluid antigen containing protoscolex after the viability of these protoscolices was assessed. ANOVA was used to test the difference of themean of optical density (OD) values among case and control groups. The highest human IgG class antibody was against antigen B (0.93) and the lowest against cattle HCF antigen (0.32). The differences between responses to these antigens were statistically significant (P < 0.001). The sensitivity and specificity of ELISA test used for evaluating the responses of human total IgG to different hydatid cyst fluid (HCF) antigens among the case and control groups were 100 and 95.8%, respectively. Cross-reaction of human IgG class and subclasses responses was found almost for all the antigens with the best reaction against human and cattle (HCF) antigens and antigen B using a ratio of mean OD value to each antigen divided by the cut-off point value for the same antigen. Human sera showed a considerable cross-reactivity against all antigens by using ELISA.

Assessing GastroPanel serum markers as a non-invasive method for the diagnosis of atrophic gastritis and &amp;lt;i&amp;gt;Helicobacter pylori&amp;lt;/i&amp;gt; infection

Gastric colonization by Helicobacter pylori increases the risk of gastric disorders, including atrophic gastritis which can be diagnosed based on levels of serum biomarkers like Gastrin and Pepsinogen. We therefore examined the efficacy of a serological-based method namely GastroPanel Blood kit, in diagnosing and scoring gastritis associated to Helicobacter pylori infection. Patients with dyspeptic symptoms were prospectively recruited on voluntary basis at the Yaounde Central Hospital and University Teaching Hospital, from March to July 2011. The degree of atrophy was classified according to levels in patient serum of pepsinogens I and II (PGI and PGII) and Gastrin 17 (G17) and compared with histological profiles as reference method. A specific ELISA test was used for the detection of H. pylori IgG antibodies. In total, 86 volunteers from 21 to 83 years old (mean = 46.4 ± 3.3) were enrolled, including 74.4% of women and 25.6% of men. The prevalence of gastritis was statistically similar between Gastro Blood Panel test and histology used as reference method (89.5% versus 83.7%: p > 0.20). Diagnosis based on serum makers showed high sensitivity (93.1%) in comparison with the reference method. However, the serological based method has diagnosed more atrophic gastritis than the reference (17.4% versus 7.0%: p 0.05). Furthermore, the prevalence of H. pylori infection did not differ significantly between serological method (84.9%) and reference method (81.4%). These results suggest that diagnosis of atrophic gastritis and H. pylori infection obtained with an optional serological method (GastroPanel) is in a strong agreement with the biopsy findings, and thus can be a useful non endoscopic assessment of stomach mucosal atrophy in patients with dyspepsia.

Sublingual immunization with an engineered Bacillus subtilis strain expressing tetanus toxin fragment C induces systemic and mucosal immune responses in piglets

Sublingual (SL) and intranasal (IN) administration of a Bacillus subtilis-based tetanus vaccine was tested in piglets, which more closely mimic the human immune system than mice. Piglets were immunized by the SL, IN or oral routes with vaccine expressing tetanus toxin fragment C, or commercial tetanus vaccine given by intramuscular injection as a control. Tetanus toxoid specific ELISA and passive neutralization tests were used to measure IgG and IgA levels in serum and mucosal secretions, and assess protective serum antibodies, respectively. The nature of the immune response was explored by MHC Class II, TGF-β1 expression, and ELISA assays for multiple cytokines. SL or IN immunization of piglets induced neutralizing tetanus toxoid specific serum antibody and local salivary and vaginal IgA responses. Standard tetanus vaccine resulted in systemic antibodies, whereas oral administration of the Bacillus-based vaccine was ineffective. Further analyses indicated a balanced Th1/Th2 response to SL or IN immunization. ConclusionThis study demonstrates for the first time that SL or IN administration is effective for inducing both systemic and mucosal responses in a piglet model, indicating that SL or IN delivery of a B. subtilis-based tetanus vaccine can be a simple, non-invasive, low cost strategy to induce immunity to tetanus.

Elevated Japanese Encephalitis Virus Activity Monitored by Domestic Sentinel Piglets in Thailand

Twenty-nine domestic piglets from pig farms located in three provinces of Thailand between 2003 and 2004 were used as sentinel animals for Japanese encephalitis virus (JEV) circulation. Piglets were used as sentinel to underline, on one hand, the role of domestic pigs as JEV amplifying host and, on another hand, to point out the interest of using sentinel animals for Japanese encephalitis surveillance. JEV activity was demonstrated through i/ antibody detection using a specific ELISA test for the identification of Immunoglobulins of class M and G, ii/ virus isolation on cell culture, after experimental mosquito inoculation for virus amplification. Almost 100% and 83% of the piglets, respectively, had specific IgG and IgM JEV antibodies and 35% yielded a virus isolate. Piglets of the growing farm industry act as virus amplifier increasing the risk of transmission for the human community. Conclusively, since piglets JEV infection appears early in life and is generally clinically unnoticed, it represents an exceptional sentinel model for human health threats, which has to be considered by health authorities.

Seroprevalence of Borrelia IgG antibodies among young Swedish children in relation to reported tick bites, symptoms and previous treatment for Lyme borreliosis: a population-based survey

BackgroundLyme borreliosis (LB) is the most common tickborne infection in Sweden and the seroprevalence of Borrelia immunoglobulin G (IgG) antibodies varies between 2% and 26%. The seroprevalence in young Swedish...

Mass spectrometry assay as an alternative to the enzyme-linked immunosorbent assay test for biomarker quantitation in ecotoxicology: Application to vitellogenin in Crustacea ( Gammarus fossarum)

Vitellogenin (Vg) is a widespread biomarker for measuring exposure to endocrine disruptors. Vg quantification is usually done by using the ELISA test (enzyme-linked immunosorbent assay). Since this test is specific to a target protein, it can rarely be used with other species due to low cross-reactivity across species. Therefore alternative analytical methods have to be considered as the development of a specific and sensitive ELISA test for each new target is time-consuming and may prove unsuccessful. This paper focuses on the development of a quantitative assay by liquid chromatography tandem mass spectrometry (LC-MS/MS) of vitelogenin in an invertebrate ( Gammarus fossarum) for which no ELISA kit is available. The linearity of the method was within the concentration range 2.5–25,000 pg/mL and the limit of detection was estimated at 0.75 pg/mL of Vg. This method has been demonstrated to be an alternative to existing immunological methods for quantifying Vg in invertebrates due to its greater sensitivity, specificity and reproducibility (intra- and inter-assay below 15%). This assay was applied for Vg determination in female G. fossarum following exposure to a known endocrine disruptor, methyl farnesoate, in crustaceans. The availability of a quantitative G. fossarum LC-MS/MS assay should open the way for further studies to evaluate xenoestrogen effects in aquatic male G. fossarum.