Specific Serum IgM Research Articles

Cellular immune response in convalescents from Ixodes tick-borne borreliosis

Ixodes tick-borne borreliosis is a natural focal transmissive infectious multi-system disease featured with complex pathogenesis, which multiple aspects remain unclarified. The persistence stage during this infection associated with prolonged Borrelia presence in metastatic foci and repeated multiple dissemination is most complicated for clinical practice. It is assumed that the chronic process can be caused by an inadequate immune response associated with activated autoimmune mechanisms leading to emergence of permanent irreversible (degenerative and atrophic) changes in affected organs. Patients who experienced tick-borne borreliosis need dynamic observation for assessing disease prognosis providing a maintenance therapy. The purpose of the study was to evaluate an opportunity of using cellular immunity indices for predicting disease transition to a chronic course.Materials and methods. The study was carried out at the Medical Association of the Far East Branch of the Russian Academy of Sciences. Case report form data, serum and blood plasma samples collected from 22 patients aged 29–83 years old were examined. Immunological examination data from patients with ixodes tick-borne borreliosis (12–18 months after the onset of acute period) were analyzed. Specific IgM and IgG against Borrelia burgdorferi were determined by using the OMNICS diagnostic test system (St. Petersburg). Lymphocyte immunophenotyping was performed on BD FACSCalibur flow cytometer (Becton Dickinson, USA) by using doublelabeled monoclonal antibodies (Beckman Coulter, France).Results. A variability of activated peripheral blood lymphocytes was found in patients with tick-borne borreliosis evidencing about individual immune response. An imbalanced cellular immune response recorded in seronegative convalescents from tick-borne borreliosis, may be an indirect finding of ongoing Borrelia infection. Finding of specific serum IgG and IgM antibodies in convalescents at late stage coupled to impaired immune system is a warning sign presuming a risk to developing autoimmune reactions. Detection of specific IgM antibodies at late timepoint combined with increased immune cytotoxic potential may be referred to a marker of possible disease transition to chronic course.

Protective immunity against CyHV-3 infection via different prime-boost vaccination regimens using CyHV-3 ORF131-based DNA/protein subunit vaccines in carp Cyprinus carpio var. Jian

Cyprinid Herpesvirus 3 (CyHV-3), also known as Koi Herpesvirus (KHV), causes Koi Herpesvirus Disease (KHVD) which leads to serious economic losses worldwide. To exploit DNA/subunit vaccine candidates, CyHV-3 ORF131 gene and cDNA was cloned and analyzed in the present study. Major B cell epitopes of deduced CyHV-3 pORF131 was also predicted. Then the complete CDS of CyHV-3 ORF131 was inserted into pEGFP-N1 vector and a modified pYD1/EBY100 system to construct the DNA and subunit vaccine, respectively. Subsequently, carp were immunized with homologous and heterologous prime-boost regimens relying on the constructed DNA and oral subunit vaccines. Then the protective immunity generated from different vaccines and regimens as well as the capacity of yeast (Saccharomyces cerevisiae) as an oral vaccine vehicle was evaluated. Our study confirmed that CyHV-3 ORF131 gene consisted of 2 introns and 3 exons encoding a 428 amino acids peptide. Further analysis indicated that four fragments of CyHV-3 pORF131 contained the major B cell epitopes (Cys20~Val140, Ser169~Tyr245, Thr258~Pro390, Phe414~Gln428), which could be linked and expressed in E. coli (BL21) as a truncated pORF131. The expression of full-length CyHV-3 pORF131 by pEGFP-N1 and yeast surface display was verified by In vitro assays before vaccination. Immunization of carp with CyHV-3 ORF131 DNA and subunit vaccines could evoke the activation of immune-related genes such as CXCa, CXCR1, IL-1β, TNF-α, INF-a1, Mx-1, IgM, IgT1 and production of specific serum IgM measured by ELISA. RPS (relative percent of survival) ranging from 53.33% to 66.67% was acquired post challenge test. Moreover, flow cytometry analysis illustrated the delivery of surface-displayed CyHV-3 pORF131 to midgut after oral gavage. Thus, our findings suggest that CyHV-3 ORF131 can serve as DNA/subunit vaccines candidate and the yeast as an ideal oral vaccine vehicle.

Epidemiological and clinical analysis of community-acquired Mycoplasma pneumonia in children from a Spanish population, 2010–2015

Introduction and objectivesMycoplasma pneumoniae (MP) is one of the most common etiological agents of community-acquired pneumonia (CAP) in children. We aimed to describe the clinical and epidemiological characteristics, treatment and outcome of children diagnosed with community-acquired MP pneumonia (CAMP) in a tertiary hospital in Valencia, Spain. Material and methodsMedical records of children <14 years with CAMP were retrospectively reviewed from January 2010 to December 2015. Patients with radiological evidence of pneumonia and microbiological confirmation of MP (PCR from nasopharyngeal swab and/or serum specific IgM) were considered CAMP. ResultsOne hundred and sixty two children were diagnosed with CAMP; median age 6 years (IQR: 4–9). The positive MP test rate among children with CAP progressively increased with age as did the empirical use of macrolides. There were two peaks of cases in 2011 and in 2015, being July, August, November and December the seasons with the higher number of cases. The most frequent radiological pattern was segmental infiltrate (62.3%) and 22 (13.6%) children had pleural effusion. It was noteworthy the mild symptomatology and low levels of inflammatory parameters that children with CAMP had. A macrolide was empirically initiated in 68.5% of cases. Hospital admission rate was inversely proportional to patient's age. ConclusionsAccording to this study, older, less symptomatic patients and with lower inflammatory parameters had the greatest rate of MP infection among children with CAP and thus they could benefit of empiric macrolide therapy. Therefore, knowing the epidemiology of a geographical area may be important for the management of CAP in children.

Epidemiological and clinical analysis of community-acquired Mycoplasma pneumonia in children from a Spanish population, 2010-2015

ResumenIntroducción y objetivosMycoplasma pneumoniae (MP) es uno de los agentes etiológicos más comunes de las neumonías adquiridas en la comunidad (NAC) en niños. Objetivo: describir las características clínicas y epidemiológicas, tratamiento y evolución de los pacientes con NAC por MP (NACM) en un hospital terciario de Valencia, España.Material y métodosSe revisaron retrospectivamente las historias clínicas de los niños < 14 años con NACM entre enero de 2010 y diciembre de 2015. Los pacientes con evidencia radiológica de neumonía y confirmación microbiológica de MP (PCR de exudado nasofaríngeo y/o anticuerpos IgM específicos frente a MP) se consideraron NACM.ResultadosUn total de 162 pacientes se diagnosticaron de NACM; mediana de edad de 6 años (rango intercuartílico: 4-9 años). La proporción de pruebas positivas para MP en pacientes con NAC, así como el uso empírico de macrólidos, aumentó progresivamente con la edad. Hubo un pico de casos en 2011 y en 2015, con un máximo de casos en julio, agosto, noviembre y diciembre. El patrón radiológico más frecuente fue el infiltrado segmentario (62,3%), mientras que 22 (13,6%) presentaron derrame pleural. Los niños con NACM desarrollaron una clínica leve, con poca elevación de parámetros inflamatorios. Se inició tratamiento empírico con un macrólido en el 68,5% de los casos. La necesidad de ingreso hospitalario fue inversamente proporcional a la edad del paciente.ConclusionesSegún este estudio, los niños con NAC de mayor edad tuvieron la mayor proporción de infección por MP, siendo poco sintomáticos y con escasa elevación de parámetros inflamatorios, pudiéndose beneficiar del tratamiento empírico con macrólidos. Por consiguiente, conocer la epidemiología de un área geográfica podría ser importante para el abordaje de las NAC en niños.

West-Nile virus encephalitis in an immunocompetent pediatric patient: successful recovery

BackgroundWest Nile virus (WNV) is a mosquito-borne RNA virus belonging to the Flaviviridae family. Symptomatic infection happens in only about 20% of the cases, while WNV neuroinvasive disease (WNND) is rare and accounts for less than 1%. There is insufficient information about natural history and clinical course in children, because underdiagnosis is common, and reports are scarce. On the other hand, Europe has seen a dramatic increase of WNV infections in the last decades, and the Po valley itself, in Northern Italy, has become an endemic region since 2013.Case presentationWe hereby report a case of West-Nile virus neuroinvasive disease in a 12-year-old boy. This is one of the very few cases diagnosed in the Italian pediatric population. The clinical presentation was compatible with acute encephalitis. Diagnosis was made by detection of specific IgM in both serum and cerebrospinal fluid. He finally was discharged with complete recovery, and no neurologic sequelae after a 12-months follow up period.ConclusionsGiven its non-specific clinical presentation, the diffusion of WNV constitutes a crucial and emerging concern. Even though rare, neuroinvasive WNV infection should always be suspected in pediatric patients, living or traveling in endemic areas, presenting with meningitis, encephalitis or acute flaccid paralysis during the WNV transmission season.

Recombinant phosphoglucomutase and CAMP factor as potential subunit vaccine antigens induced high protection against Streptococcus iniae infection in flounder (Paralichthys olivaceus).

The aim of this study was to screen vaccine candidates from virulence factors of Streptococcus iniae in flounder model. The immunogenicity of recombinant phosphoglucomutase (rPGM) and rCAMP factor was confirmed by Western blot. The percentage of surface membrane immunoglobulin-positive (sIg+ ) lymphocytes in peripheral blood leucocytes, the specific and total serum IgM and the activity of acid phosphatase (ACP) and peroxidase (POD) in flounder were determined with flow cytometry, ELISA and commercial enzyme activity kits, respectively, after intraperitoneal immunization with rPGM and rCAMP factor. The results showed that rPGM and rCAMP factor could induce significant rise in sIg+ lymphocytes, specific serum IgM and activities of ACP and POD. Additionally, the relative percent survival rate of the vaccinated flounder was 64 and 54% in challenge experiment using S. iniae, respectively. These results indicated that rPGM and rCAMP factor could evoke humoural and innate immune response in flounder and provide high-efficiency immunoprotection against S. iniae infection. Phosphoglucomutase (PGM) and CAMP factor were promising vaccine candidates against S. iniae in flounder. Phosphoglucomutase and CAMP factor have the potential to be vaccine candidates, which provide important information for us to develop the effective subunit vaccines, especially the multivaccine, against S .iniae in aquaculture.

Comparative study of the adjuvant potential of four Th0 cytokines of flounder (Paralichthys olivaceus) on an E. tarda subunit vaccine

Cytokines have the potential as adjuvants for the application of vaccines in mammals. However, the adjuvant potential of teleost cytokines was limited. In the present work, the adjuvant effects of four recombinant cytokines including IL-1β, IL-8, TNF-α and G-CSF on E. tarda subunit vaccine rOmpV were comparatively investigated in flounder (Paralichthys olivaceus). Compared with control, the levels of specific serum antibodies and IgM + B lymphocytes were significantly enhanced by rIL-1β, rIL-8 and rG-CSF, whereas rIL-1β and rIL-8 induced significantly higher levels than rG-CSF. All four cytokines enhanced the expression of genes involved in humoral and/or cellular immunities, whereas rIL-1β and rIL-8 induced highest levels of genes involved in humoral immunities and cellular immunities, respectively. Compared to the relative percent survivals (RPS) of control group (40%) and rOmpV plus rG-CSF group (54%), rOmpV plus rIL-1β or rIL-8 produced higher RPS of 75% and 68%, respectively. Our results indicated that rIL-1β and rIL-8 are promising adjuvants for subunit vaccines against E. tarda.

Diagnosing tick-borne encephalitis: a re-evaluation of notified cases

We set out to investigate the serological response of TBE virus (TBEV)-specific IgM and IgG antibodies in stored serum and cerebrospinal fluid (CSF) in notified TBE patients, in order to confirm or reject the diagnosis. We applied the ELISA methods used in clinical practice, Enzygnost and Immunozym, and assessed RT-PCR as a diagnostic tool. A total of 173 TBE cases were notified to the Public Health Agency. Samples from 129 patients were eligible for the study. Stored serum samples were found for 111 patients and CSF samples for 88 patients. All serum samples were analyzed with both Enzygnost and Immunozym, as well as an additional 140 control samples. CSF samples, including samples from ten controls, were analyzed with Immunozym. RT-PCR for TBEV was performed on 126 serum, two whole blood, 96 CSF, two feces and four nasopharynx samples. Only two of 111 notified patients lacked detectable TBEV IgM in serum, from whom one sample was RT-PCR positive. According to the ECDC definition, 117/129 (90.7%) of the reported TBE cases were confirmed. Positive RT-PCR results were obtained in eight patients, one from whole blood and eight from serum samples. Four out of eight of the RT-PCR positive patients were TBEV-IgM positive and none had detectable TBEV-specific IgG. All of the tested CSF, feces and nasopharynx samples were RT-PCR-negative. TBEV-specific IgG was detected in 88.4% and IgM in 31.6% of the CSF samples. RT-PCR on serum samples and CSF IgG antibodies can be used as complementary methods in TBE diagnostics, not least early in the disease course.

Serologically diagnosed acute human bocavirus 1 infection in childhood community-acquired pneumonia.

AimTo assess the role of human bocavirus 1 (HBoV1) as a causative agent of non‐severe community‐acquired pneumonia (CAP) in children.MethodsPatients aged 2‐59 months with non‐severe CAP (respiratory complaints and radiographic pulmonary infiltrate/consolidation) attending a University Hospital in Salvador, Brazil were enrolled in a prospective cohort. From 820 recruited children in a clinical trial (http://ClinicalTrials.gov NCT01200706), nasopharyngeal aspirate (NPA), and acute and convalescent serum samples were obtained from 759 (92.6%) patients. NPAs were tested for 16 respiratory viruses by PCR. Acute HBoV1 infection was confirmed by measuring specific IgM and IgG responses in paired serum samples.ResultsRespiratory viruses were detected in 693 (91.3%; 95%CI: 89.1‐93.2) CAP cases by PCR. HBoV1‐DNA was detected in 159 (20.9%; 95%CI: 18.2‐24.0) cases. Of these 159 PCR positive cases, acute HBoV1 infection was confirmed serologically in 38 cases (23.9%; 95%CI: 17.8‐31.0). Overall, acute HBoV1 infection was confirmed in 5.0% (38/759) of non‐severe CAP patients. HBoV1 was detected in 151 cases with at least one other virus making 31.7% of all multiple virus (n = 477) detections. Among all 759 cases, 216 had one respiratory virus detected, and sole HBoV1 was detected in only 8 (3.7%). Acute HBoV1 infection was serologically diagnosed in 34 (22.5%) HBoV1‐DNA‐positive cases with another virus, compared to 4 (50.0%) cases with sole virus detection (p = 0.09).ConclusionHBoV1 was detected by PCR in one fifth of the children with non‐severe CAP and acute HBoV1 infection was serologically confirmed in one quarter of these cases.

A second wave of monocyte expansion may contribute to the severe dengue in 2014 outbreak in China (HUM4P.272)

Abstract The 2014 dengue outbreak, the largest one in the last ten years in China, resulted in 13,800 hospitalized patients and more than 300 severe dengue cases in Guangzhou. Both of the viral strain and the host immune background may contribute to severe dengue. Here we report the monocyte response in dengue fever (DF) and severe dengue (SD) during the acute infection of seven dengue patients (DF n=4 and SD n=3) using multiparameter flow-cytometry with a panel of 9-surface markers. We also compared dengue specific IgG, IgM and NS1 in serum at the same time points for each patient. Six of the seven patients were diagnosed as secondary DENV1 infection, while one DF diagnosed as primary DENV1 infection. NS1 was nearly cleared in all patients when the serum IgG and IgM reached the high level at day 8-10 from the onset of fever. Obvious monocyte expansion was observed in both of DF and SD groups, frequently companied with the expansion of CD14++CD16+ subset. But the pattern of the response in the two groups was different at the recovery phase. The monocyte expansion in DF group maintained at a high level at day 7-10. Interestingly, the SD group underwent a second wave of monocyte expansion, in parallel with clinical course, starting from day 7 to day 10, especially the CD16+CD64+ monocyte subset. The characteristics of monocyte responses in DF and SD groups will advance our understanding of human dengue pathogenesis and may implicate the potential targets for early therapy.

Tick-borne encephalitis: A review of epidemiology, clinical characteristics, and management.

Tick-borne encephalitis is an infection of central nervous system caused by tick-borne encephalitis virus transmitted to humans predominantly by tick bites. During the last few decades the incidence of the disease has been increasing and poses a growing health problem in almost all endemic European and Asian countries. Most cases occur during the highest period of tick activity, in Central Europe mainly from April to November. Tick-borne encephalitis is more common in adults than in children. Clinical spectrum of the disease ranges from mild meningitis to severe meningoencephalitis with or without paralysis. Rare clinical manifestations are an abortive form of the disease and a chronic progressive form. A post-encephalitic syndrome, causing long-lasting morbidity that often affects the quality of life develops in up to 50% of patients after acute tick-borne encephalitis. Clinical course and outcome vary by subtype of tick-borne encephalitis virus (the disease caused by the European subtype has milder course and better outcome than the disease caused by Siberian and Far-Easter subtypes), age of patients (increasing age is associated with less favorable outcome), and host genetic factors. Since clinical features and laboratory results of blood and cerebrospinal fluid are nonspecific, the diagnosis must be confirmed by microbiologic findings. The routine laboratory confirmation of the tick-borne encephalitis virus infection is based mainly on the detection of specific IgM and IgG antibodies in serum (and cerebrospinal fluid), usually by enzyme-linked immunosorbent assay. There is no specific antiviral treatment for tick-borne encephalitis. Vaccination can effectively prevent the disease and is indicated for persons living in or visiting tick-borne encephalitis endemic areas.

The Seroprevalence of Some Pathogen Specific IgM in Children with Acute Respiratory Tract Infections in Guangzhou Region, 2011 - 2012.

Acute respiratory tract infections (ARTIs) are the leading cause of morbidity and death in children < 5 years worldwide. The aim of this study is to analyze the seroprevalence of nine pathogen specific IgMs in children with ARTIs with respect to gender, age, and seasonality in the Guangzhou region. Serum samples were collected from 20160 children with ARTIs admitted to the Guangzhou Women and Children's Medical Center between 2011 and 2012. Serum-specific IgM antibodies to nine respiratory pathogens, Mycoplasma pneumonia (MP), Legionella pneumophila (LP), Coxiella burnetii (C. burnetii), Chlamydophila pneumonia (CP), adenovirus (ADV), respiratory syncytial virus (RSV), type A and type B influenza virus (IVA and IVB), and parainfluenza virus (PIV), were detected using immunofluorescence assay. The male-to-female ratio of all patients was 1.9:1. The median age was 3 years and 8 months with a significant difference in seropositivity to respiratory tract pathogens between children from different age groups. Seropositivity was detected in 43.53% of the children with the top three pathogens being MP (33.15%), RSV (10.27%), and ADV (6.63%), followed by IVB (2.63%), LP (2.25%), IVA (1.59%), PIV (1.57%), CP (0.27%), and C. burnetii (0.13%). The prevalence of single, double, and triple seropositivity was 70.20% (6160/8775), 25.22% (2213/8775), and 4.57% (401/8775), respectively. The total IgM seropositivity for any kind of pathogen in the nine kinds of pathogens peaked in winter (46.53%), while the nadir was observed in summer (41.97%). The top three seroprevalence of nine kinds of pathogen specific IgM was MP, followed by RSV and ADV. The epidemic pathogen specific IgM had a season-specific seropositivity distribution. Seroprevalence of the pathogen should be a focus of attention.

Immunological and biochemical changes in young children with recurrent course of respiratory infections and cytomegaloviral infection

Cytomegalovirus (CMV) infection in practically healthy children of different ages ranges from 50% to 64%. Both acute course of infection and prolonged persistance of pathogen is accompanied by immune deficiency formation that may become one of factors in charge of high level of infection morbidity in future as recurrent respiratory infections (RRI). At the same time significant spread of deficiency states in infants - deficiency of iron ion or vitamin D - is also accompanied by changes in immunological reactivity, especially when combined with intracellular infections. Purpose of the work was to study some immune and biochemical changes in young children (n=37) with recurrent respiratory diseases and cytomegaloviral infection. Materials and methods. In 51 children the CMV infection was verificated by simultaneous detection of specific anti-CMV IgG and anti-CMV IgM in blood serum and viral DNA finding by polymerase chain reaction. The observation group consisted of 37 (72.5%) children infected with CMV as well as suffering from recurrent respiratory infections. Comparison group was represented by the remaining 14 (27.5%) patients with episodic respiratory infections (ERI). In RRI group the patients with the frequent respiratory diseases with bacterial implications four and more times during the second year of life were included. The levels of serum iron and 25-hydroxy-cholecalciferol (vitamine 25(ОН)D) were detected in blood serum of all children by ELECSYS-test. In the early convalescence period after respiratory infections comparative immunograms assessment was conducted by three main components: phagocytic activity of neutrophils, the system of cellular immunity and quality of humoral immunity. In the vast majority (72.5%) of the children age up to 1yo with verified CMV infection respiratory infections of viral and bacterial etiology gained a recurrent course (p <0.05). In the early convalescence period after respiratory infection process in children with CMV and RRI the immune response is characterized by a decreasing of the functional and metabolic activity of neutrophile granulocytes (р<0,05) and imbalance of humoral immunity consisting in decreased antibody generation (IgА (р<0,05), IgМ (р<0,05), tendency to IgG decreasing). These changes are compensated by the increasing of the total number of B-lymphocytes (р<0,05). We founded a significantly lower average level of 25-hydroxy-cholecalciferol (32,37|23,41-39,95|ng/ml) in children of the observation group. In the comparison group this rate was 44,21|35,38-52,82|ng/ml, р<0,05. We also detected a correlation dependence between concentration of vitamin 25(OH)D and antiCMV IgG (r = 0,35, p <0.05), decreased concentration of each of them is clinically manifested in pathogen persistence and recurrence course of respiratory infections in children. In the patients from observation group serum iron concentration was significantly lower than the corresponding data of those children who suffered from the respiratory diseases occasionally: 4,05|3,74-5,05| mcmol/l and 8,6|4,92-11,72| mcmol/l (р<0,05) respectively. There was a slight correlation dependence between serum iron level and blood anti-CMV IgG-antibodies level (r=0,28, р<0,05). So, the fact of the important role of iron ions in the mechanisms which regulate the functional activity of CD4+-cells was confirmed (r=0,49, р<0,05).

Recombinant TNFα as oral vaccine adjuvant protects European sea bass against vibriosis: Insights into the role of the CCL25/CCR9 axis

Vibrio anguillarum is the main causative agent of vibriosis in cultured sea bass. Unfortunately, available vaccines against this disease do not achieve the desired protection. In this study, to accomplish uptake, processing, and presentation of luminal antigens, a commercial sea bass oral vaccine against V. anguillarum was improved with the addition of recombinant fish-self tumor necrosis factor α (rTNFα), as adjuvant. To explore mechanisms, systemic and local responses were analyzed through serum specific IgM titers, gene expression, lymphocytes spatial distribution in the gut, and in vitro functional assays. We found along the trial, over expressed transcripts of genes encoding cytokines and antimicrobial molecules at the gut of rTNFα supplied group. Orally immunized fish with vaccine alone confer protection against V. anguillarum challenge throughout a short time period. In contrast, adjuvant-treated group significantly extended the response. In both cases, achieved protection was independent of serum IgM. Yet, IgT transcripts were found to increase in the gut of rTNFα-treated fish. More importantly, fish treated with rTNFα showed a dramatic change of their T lymphocytes distribution and localization in gut mucosal tissue, suggesting specific antigen recognition and further intraepithelial T lymphocytes (IEL) activation. To determine the mechanism behind IEL infiltration, we characterized the constitutive and activated pattern of chemokines in sea bass hematopoietic tissues, identifying for the first time in fish gut, an intimate relation between the chemokine ligand/receptor CCL25/CCR9. Ex-vivo, chemotaxis analyses confirmed these findings. Together, our results demonstrate that improved oral vaccines targeting key cytokines may provide a means to selectively modulate fish immune defence.

SAT0043 Impaired B cell immunity in IL-27R knock-out mice in collagen induced arthritis

BackgroundCollagen induced arthritis (CIA), a mouse model for rheumatoid arthritis, is both B cell and T helper (Th) 17 cell mediated. Interleukin 27 receptor (IL27R) signaling upregulates IL12 receptor expression...

Situación de la tos ferina en la Comunidad Valenciana: ¿asistimos a una reactivación epidémica?

IntroductionPertussis is a highly contagious vaccine-preventable disease. An increasing incidence has been reported in several countries around the world in the last few years. The aim of this study is to analyze the situation of pertussis in the Valencian Community (Spain), with the purpose of verifying the increased incidence of the illness. MethodsA descriptive analysis of probable and confirmed cases detected during 2011 was conducted. Methods used for Bordetella pertussis detection from clinical samples were: culture isolation, polymerase chain reaction (PCR)-based detection of bacterial DNA, and/or detection of specific IgM antibodies. The disease incidence and other epidemiological variables were estimated. These results were compared to data collected in previous years (2008-2010). The Epidemiological Surveillance Analysis and Microbiological Surveillance Network integrated systems of the regional Department of Health were used as sources of information. ResultsIn 2011, 249 cases of pertussis were detected (incidence rate of 4.89×105 inhabitants). This rate is statistically significantly higher than those reported in 2008 (0.73×105 inhabitants), 2009 (0.53×105 inhabitants), and 2010 (0.53×105 inhabitants). The highest incidence rate was observed in cases younger than one year old (252.97×105 inhabitants), with marked differences compared to the rest of age groups. More than two-thirds (69%) of reported cases were confirmed by a laboratory test. Detection of specific serum IgM antibodies was positive in 10% of cases, culture isolation was positive in 17%, and PCR-based detection of bacterial DNA in 35% of cases. ConclusionsThe results of this study show a clear increase of pertussis incidence in the Valencian Community during 2011.

Innovative vaccination protocol against vibriosis in Dicentrarchus labrax (L.) juveniles: Improvement of immune parameters and protection to challenge

The effect of vaccination on immune parameters of European sea bass, Dicentrarchus labrax, is not fully established, as well as surveyed throughout rearing till the commercial size. Furthermore, available information on the possible role of booster treatments is scarce.Sea bass juveniles were vaccinated against Listonella anguillarum using a commercial bivalent formulation administered by immersion (priming: 95dph; booster: 165dph) or by immersion (priming: 95dph; booster: 165dph) and subsequent i.p. injection (booster: 233dph).Serum specific IgM and numbers of IgM+ cells in head kidney and spleen evidenced B-cell responses mainly after the immersion booster, accompanied by increased TcR-β transcripts and leucocyte respiratory burst.Immune enhancement was confirmed by the protection towards i.p. challenges with a virulent strain. RPS accounted for >70% in fish immersion-boosted and near 100% in fish further boosted i.p.Differently from usual farm practices, this innovative vaccination protocol proved to be highly effective. Booster treatments are therefore strongly recommended.

Clinical and radiological spectrum of Japanese encephalitis

IntroductionJapanese encephalitis (JE) is mosquito-borne flaviviral encephalitis that remains to be a major health problem in India — it still continues to cause havoc in many parts of the country. We undertook the study to analyze the clinical & radiological spectrum of JE in adults and children. MethodThis prospective study consists of 148 patients with JE. The diagnosis of JE was based on clinical, epidemiological, radiological features and demonstration of JE virus specific IgM in CSF and serum by JE virus immunoglobulin M capture enzyme-linked immunosorbent assay (MAC ELISA). All patients underwent a detailed neurological examination, CSF study & neuroimaging of brain (either CT or MRI or both). All patients were followed-up at regular interval. ResultSeizures were present in adults (52.88%) and in children (43.18%). Dystonia was more common in children 19 (43.18%) compared to adults 19 (18.2%), and Parkinsonian features were observed in both groups 47 (45.19%) of the adults and 20 (45.45%) of the children. JE-specific IgM antibody was detected in both CSF and serum in 81.7%. In neuroimaging, apart from classical involvement of thalami, basal ganglia & midbrain, prominent involvement of hippocampus and other areas of the cortex was also found in 27 (45.6%) patients. Presence of thalamic lesion in CT/MRI showed significant relationship to the development of dystonia. However, no correlation was found between the neuroimaging features and poor clinical outcome. Twenty three patients (15.5%) died during acute phase of illness. On multivariate logistic regression analysis age, prolonged fever, Glasgow coma scale, recurrent seizures and reflex changes were found to be the predictors of outcome at the time of discharge. ConclusionA trend of severe and frequent involvement in younger patients with dystonia and other movement disorders was observed. It should be emphasized that presence of atypical cranial CT/MRI features in JE was not unknown and they need to be differentiated from herpes simplex encephalitis in appropriate clinical setting.

Evaluation of serum specific IgM detection in diagnosis of respiratory viral infections in children

The present study was designed to explore the practical application of the rapid etiological diagnosis by detecting specific IgM antibody against common respiratory viruses in children with acute lower respiratory infections (ALRI). Clinical specimens including nasopharyngeal aspirates and serum of acute phase from hospitalized children were collected from 207 infants and children with acute lower respiratory infections from March 2009 to September 2010. Seven common respiratory virus antigens were identified from the collected nasopharyngeal aspirates by direct immunofluorescence assay (DFA). ELISA was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB and PIV, while indirect immunofluorescence assay (IFA) was used to detect specific IgM antibody against RSV, ADV, IFVA, IFVB, PIV1, PIV2 and PIV3 in collected acute phase serum. The overall positive rates to detect viral antigen by using DFA, ELISA and IFA was 67.6%, 57.5% and 39.6%, respectively. The consistent rate of ELISA and IFA versus accepted DFA were 21.7% and 31.4%, respectively. The average days from onset of the symptoms to blood sample collection for those with the consistent results by ELISA and DFA were 12.0 d for ADV, 9.6 d for PIV2, 9.5 d for IFV, and 5.3 d for RSV, respectively, and by IFA and DFA were 15.0 d for PIV3, 9.2 d for ADV, and 7.4 d for RSV, respectively. Among all age groups, the consistent rate of serum viral IgM and antigen detections was highest in children younger than 3 years old. Although there were differences between serum IgM antibody and viral antigen detections, specific IgM antibody detection was of value in early and rapid etiological diagnosis of pediatric ALRI, especially for young children. It could provide serologic evidence of respiratory virus infection. The diagnostic rate of pathogen could be improved if it was used in combination with viral antigen diagnostic methods.

Augmentation of immunity to influenza A virus by manipulating a host counter inflammatory pathway (49.5)

Abstract Reactions to pathogens are usually tuned to effect immunity and limit tissue damage.With chronic infections, where immunity is unsuccessful, tissue damage would be more severe were it not for the operation of numerous host counter inflammatory events. One such event is the inhibitory effects of some galectins binding to their receptors on effector T cells. A potential downside of counter inflammatory events is that they may constrain the effectiveness of immunity to acute infections. We present evidence for this situation in mice acutely infected with Influenza A virus (IAV). We show that virus specific CD8+T cells in both bronchoalveolar lavage (BAL) and spleen up regulate Tim-3 following IAV infection. In an ex-vivo apoptosis assay, addition of Galectin-9 (Gal-9) to the splenocytes (d10pi) caused apoptosis of both Tim-3+and DbNPtet+ CD8+T cells. Furthermore, Galectin-9 knockout (G9KO) mice mounted more robust acute phase virus-specific CD8+T cell response and cleared virus more rapidly than WT mice. G9KO mice also generated higher virus specific serum IgM, IgG and a more rapid IgA response than WT. When IAV immune mice were challenged with a heterologous IAV, G9KO mice mounted a 4 fold stronger IAV specific CD8+T cell response than WT mice. Tim-3 fusion protein infusion into infected WT mice enhanced CD8+T cell responses to IAV. Our results could indicate that manipulating galectin signals may represent a convenient approach to improve immune responses to some vaccines.