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Description of additional terms and methods used in the Study of Healthcare Personnel with Influenza and other Respiratory Viruses in Israel. (DOCX 77 kb)

Introduction Influenza vaccine is recommended for the entire population in Israel. We assessed influenza vaccine effectiveness (VE) for the 2014/15 and 2015/16 seasons in Israel, for the first time. Methods: Combined nose and throat swab specimens were collected from patients with influenza-like illness (ILI) presenting to sentinel primary care clinics and tested for influenza virus by RT-PCR. VE of the trivalent inactivated vaccine (TIV) was assessed using test-negative case–control design. Results: During the 2014/15 season 1,142 samples were collected; 327 (28.6%) were positive for influenza, 83.8% A(H3N2), 5.8% A(H1N1)pdm09, 9.2% B and 1.2% A un-subtyped. Adjusted VE against all influenza viruses for this influenza season was −4.8% (95% confidence interval (CI): −54.8 to 29.0) and against influenza A(H3N2), it was −15.8% (95% CI: −72.8 to 22.4). For the 2015/16 season, 1,919 samples were collected; 853 (44.4%) were positive for influenza, 43.5% A(H1N1)pdm09, 57% B, 0.7% A(H3N2) and 11 samples positive for both A(H1N1)pdm09 and B. Adjusted VE against all influenza viruses for this influenza season was 8.8% (95% CI: −25.1 to 33.5), against influenza A(H1N1)pdm09, it was 32.3% (95% CI: (−4.3 to 56.1) and against influenza B, it was −2.2% (95% CI: (−47.0 to 29.0). Conclusions: Using samples from patients with ILI visiting sentinel clinics in Israel, we demonstrated the feasibility of influenza VE estimation in Israel.

West Nile Virus (WNV) is endemic in Israel and has been the cause of several outbreaks in recent years. In 2000, a countrywide mosquito survey was established to monitor WNV activity and characterize viral genotypes in Israel. We analyzed data from 7135 pools containing 277 186 mosquitoes collected over the past 15 years and, here, report partial sequences of WNV genomes obtained from 102 of the 336 positive mosquito pools. Phylogenetic analysis demonstrated that cluster 4 and the Mediterranean and Eastern European subtypes of cluster 2 within WNV lineage 1 circulated in Israel, as did WNV lineage 2, highlighting a high genetic diversity of WNV genotypes in our region. As a major crossroads for bird migration between Africa and Eurasia and with a long history of human infection, Israel serves as a resource hub for WNV in Africa and Eurasia and provides valuable information on WNV circulation in these regions.

Summary Objectives To enhance timely surveillance of bacterial enteric pathogens, space-time cluster analysis was introduced in Israel in May 2013. Methods Stool isolation data of Salmonella , Shigella , and Campylobacter from patients of a large Health Maintenance Organization were analyzed weekly by ArcGIS and SaTScan, and cluster results were sent promptly to local departments of health (LDOHs). Results During eighteen months, we identified 52 Shigella sonnei clusters, two Salmonella clusters, and no Campylobacter clusters. S. sonnei clusters lasted from one to 33 days and included three to 30 individuals. Thirty-one (60%) of the S. sonnei clusters were known to LDOHs prior to cluster analysis. Clusters not previously known by the LDOHs prompted epidemiologic investigations. In 31 of the 37 (84%) confirmed clusters, educational institutes (nursery schools, kindergartens, and a primary school) were involved. Conclusions Cluster analysis demonstrated capability to complement enteric disease surveillance. Scaling up the system can further enhance timely detection and control of outbreaks.

Bacteria naturally release membrane vesicles (MVs) under a variety of growth environments. Their production is associated with virulence due to their capacity to concentrate toxins and immunomodulatory molecules. In this report, we show that the 2 medically important species of mycobacteria, Mycobacterium tuberculosis and Mycobacterium bovis bacille Calmette-Guérin, release MVs when growing in both liquid culture and within murine phagocytic cells in vitro and in vivo. We documented MV production in a variety of virulent and nonvirulent mycobacterial species, indicating that release of MVs is a property conserved among mycobacterial species. Extensive proteomic analysis revealed that only MVs from the virulent strains contained TLR2 lipoprotein agonists. The interaction of MVs with macrophages isolated from mice stimulated the release of cytokines and chemokines in a TLR2-dependent fashion, and infusion of MVs into mouse lungs elicited a florid inflammatory response in WT but not TLR2-deficient mice. When MVs were administered to mice before M. tuberculosis pulmonary infection, an accelerated local inflammatory response with increased bacterial replication was seen in the lungs and spleens. Our results provide strong evidence that actively released mycobacterial vesicles are a delivery mechanism for immunologically active molecules that contribute to mycobacterial virulence. These findings may open up new horizons for understanding the pathogenesis of tuberculosis and developing vaccines.

An attenuated Mycobacterium tuberculosis RD1 knockout and pantothenate auxotroph (mc(2)6030) vaccine administered at 2 weeks of age failed to protect calves from low dose, aerosol M. bovis challenge at 2.5 months of age. In contrast, M. bovis bacille Calmette Guerin (BCG)-vaccinates had reduced tuberculosis-associated pathology as compared to non- and mc(2)6030-vaccinates. Mycobacterial colonization was not impacted by vaccination. Positive prognostic indicators associated with reduced pathology in the BCG-vaccinated group were decreased antigen induced IFN-gamma, iNOS, IL-4, and MIP1-alpha responses, increased antigen induced FoxP3 expression, and a diminished activation phenotype (i.e., downward arrow CD25+ and CD44+ cells and upward arrow CD62L+ cells) in mycobacterial-stimulated mononuclear cell cultures. The calf sensitization and challenge model provides an informative screen for candidate tuberculosis vaccines before their evaluation in costly non-human, primates.

The emergence of multidrug-resistant tuberculosis strains and the coinfection with HIV, together with advances in immunology, have led to renewed interest regarding ways to exploit the immune responses against Mycobacterium tuberculosis therapeutically. Here we review the fundamentals of tuberculosis therapy in view of the epidemiological and clinical challenges, and explore the experience with immune-based therapies for the treatment of active tuberculosis. These immune-based therapies are discussed here with the aim of assessing their potential use as adjuncts to chemotherapy.

The seasonal influenza vaccine is currently the most effective preventive modality against influenza infection. Nasopharyngeal samples of vaccinated and non-vaccinated patients presenting with Influenza-like-illness (ILI) were collected from over 20 outpatient clinics located in different geographic parts of Israel and were tested for the presence of influenza viruses (influenza A and influenza B). Here we show, that in the 2014-2015 season, the vaccine that included the A/Texas/50/2012 H3N2 virus was ineffective. Significant numbers of individuals vaccinated with the 2014-2015 vaccine, of all ages, were infected with influenza A (H3N2), manifesting similar symptoms as the non-vaccinated group. We further demonstrate that the Israeli circulating influenza A(H3N2) virus was different than that included in the 2014-2015 northern hemisphere vaccine, and that antibodies elicited by this vaccine were significantly less efficient in neutralizing influenza A(H3N2) infection.

BACKGROUND Increased upper respiratory infection (URI) among children at the beginning of school year is well known to parents and pediatricians. However, this phenomenon is not well documented or characterized. METHODS Computerized datasets from a large health maintenance organization in Israel were used to calculate the weekly rates of URI among children 3-14 years old for the years 2007-2012. In addition, nasopharyngeal swabs were collected in 2010-2012 from children with URI symptoms and controls during school opening time. Swabs were tested by real-time polymerase chain reaction for the presence of respiratory viruses. RESULTS Time-series analysis demonstrated a peak of URI in September each year. The peaks reached their height 2 weeks after school opening and returned to baseline within 4-7 weeks. The main 3 viruses detected both in URI patients and in healthy controls during the first weeks of school opening were rhinovirus, adenovirus and enterovirus. The detection rate of any respiratory virus, and of rhinovirus in particular, was significantly higher among cases than among controls (54% vs. 16%, P < 0.001 for any virus, and 35% vs. 6.0%, P < 0.01 for rhinovirus). When adjusting for age and sex cases had 5.8 times more viral detection when compared with controls. Upper respiratory symptoms were significantly more prevalent among the virus-positive cases when compared with negative ones. CONCLUSIONS Back-to-school illness consisting of URI has a distinct epidemiological pattern demonstrating a rapid rise peaking within 2 weeks of school opening and is associated predominantly with rhinovirus.

For many decades, the dominant view in the field of mycobacterial immunology has been that host defense against Mycobacterium tuberculosis relies exclusively on cell-mediated immunity. In search of new solutions to the overwhelming problem of tuberculosis, investigators set out several years ago to evaluate the help that can be offered by antibody-mediated immunity in host defense against M. tuberculosis, with the possibility that it may lead to the development of a novel and effective vaccine strategy. The literature on studies of antibody-mediated immunity against M. tuberculosis can be divided into several general categories: serological studies, passive antibody studies, animal studies, in vitro studies, and human studies. Monoclonal antibody (MAb) technology, described for the first time in the 1970s, allowed the selection of individual antibodies with particular antigen specificities. Vaccines presently used in humans belong to one of three main categories: inactivated, live attenuated, and subunit vaccines. Adhesion of microbes to host tissues is a significant step in the colonization of the host and the establishment of infection. The majority of these vaccines are thought to provide protection by eliciting protective antibody responses. The progress made in recent years is encouraging and should stimulate interest in evaluating the mechanisms by which antibodies may contribute to host defense against M. tuberculosis. The future challenges are to systematically dissect the conditions required for optimal antibody-mediated immunity against M. tuberculosis and to develop vaccine candidates that will work by eliciting protective antibody responses.

Arabinomannan (AM) is a polysaccharide of the mycobacterial capsule. The capsular polysaccharides of various microorganisms are diverse, and this diversity is important for classification of organisms into serotypes and vaccine development. In the present study we examined the prevalence and diversity of AM amongMycobacterium tuberculosisstrains using four AM-binding monoclonal antibodies (MAbs). One of these MAbs, MAb 9d8, is known to bind to AM specifically. By whole-cell enzyme-linked immunosorbent assay (ELISA), the AM recognized by MAb 9d8 was detected on the surfaces of 9 of 11 strains, while 2 strains showed no reactivity with MAb 9d8. However, the AM recognized by MAb 9d8 was found in the culture supernatants of all 11M. tuberculosisstrains tested, as demonstrated by capture ELISA. Other AM-binding MAbs reacted both with the surfaces and with the culture supernatants of all 11 strains. Mice immunized with an experimental AM-recombinantPseudomonas aeruginosaexoprotein A (rEPA) conjugate vaccine had an increased antibody response to AM and a moderate reduction in the numbers of CFU in their organs 7 days after challenge. Our results indicate that AM was detected in allM. tuberculosisstrains tested, with differences in epitope distributions of certain strains. In addition, our results suggest that an experimental AM-rEPA vaccine has a moderate effect on the numbers of CFU in organs early after infection.

The outermost layer ofMycobacterium tuberculosisis composed primarily of two polysaccharides, glucan (GC) and arabinomannan. To analyze the surface polysaccharide composition ofM. tuberculosis, we generated a monoclonal antibody (MAb) that bindsM. tuberculosisGC and is known as MAb 24c5. Immunofluorescence and whole-mount immunoelectron microscopy indicated that GC is on the outermost portion of the bacteria.M. tuberculosisstrains Erdman and CDC 1551 were analyzed for their ability to bind MAb 24c5 after in vitro growth in media with and without the detergent Tween 80. MAb 24c5 bound to Erdman and CDC 1551 at all culture times with only slightly greater apparent affinity after extended culture in the absence of Tween 80, indicating that a stable amount of GC polysaccharide antigen is associated with the cell surface ofM. tuberculosis. An enzyme-linked immunosorbent assay indicated that GC is antigenically similar to glycogen, and the amount of GC antigen increased in the media ofM. tuberculosiscultures grown either with or without the detergent Tween 80. Other nontuberculosis mycobacteria have antigenically similar GCs on their surfaces after in vitro growth. Inoculation of mice with live bacilli but not inoculation with dead bacilli elicited a strong antibody response to GC consistent with production of this antigen in vivo. Our results provide a more comprehensive picture of theM. tuberculosiscell envelope and the conditions that allow expression ofM. tuberculosisGC.

Cryptococcus neoformans is present in areas contaminated with pigeon droppings. Unrecognized infections are hypothesized to occur commonly among immunocompetent individuals. We used serology to estimate prevalence of cryptococcal infection in immunocompetent children from 3 regions. Our results indicate unrecognized cryptococcal infections are extremely common in Bronx children, but uncommon in children from Dutchess County, NY and the Philippines.

Murine mAbs reactive with the surface ofMycobacterium tuberculosiswere assayed for their ability to affect the course of infection in mice challenged with virulent organisms. An IgG3 mAb (9d8) specific for arabinomannan and reactive with purified antigen from a clinical isolate ofM. tuberculosisconferred partial protection on mice after respiratory challenge (30–60% survival >75 days;P≤ 0.05). Control mice pretreated with an irrelevant mAb of the same isotype succumbed to tuberculosis within 30 days. Mice with gene disruptions in interferon γ and major histocompatibility complex Class II also were partially protected from challenge. The protective mAb was neither bactericidal nor inhibitory of infection or bacterial replication. Nevertheless, it profoundly altered the nature of the granulomas in the infected lungs. Mice treated with mAb 9d8 and challenged withM. tuberculosislocalized the pathogen within granuloma centers, suggesting that the mAb conferred protection by enhancing a cellular immune response.

SUMMARY Fifty years after the introduction of the first effective antimicrobial agents against Mycobacterium tuberculosis, this pathogen continues to be a tremendous public health problem. The rise in the number of resistant strains and the difficulties involved in the therapy of tuberculosis in immunocompromised AIDS patients have renewed the interest in the development of effective vaccines. To evaluate whether a potential vaccine against tuberculosis could prevent infection by eliciting a protective antibody response, we reviewed the history of antibody-mediated immunity against tuberculosis. Review of the literature of the past 100 years demonstrates that there is sufficient evidence to conclude that antibody-mediated immunity can modify the course of infection in certain situations. Based on our findings and on what is known in other systems, we propose that the role of antibody-mediated immunity to M. tuberculosis be reexamined, using advanced technology.

Bacille Calmette Guerin (BCG) is effective to prevent miliary and meningeal TB in infants [1]. The reports about the efficacy of this vaccine for the prevention of adults pulmonary TB are contradictory and the consensus is that the protection conferred by BCG against this form of TB is questionable [1]. The wide use of BCG vaccination has been unable to prevent nearly two million deaths associated with TB that are produced every year. Currently the World Health Organization no longer recommends BCG vaccination in children born from HIVpositive mothers which complicate the implementation of BCG vaccination programs [2]. The imple‐ mentation of standard drug treatment for TB is difficult in the areas of the highest incidence of the disease. Treatment is further complicated by the limited effectiveness of the current therapeutic schemes against drug resistant strains of TB [3-5].

While influenza A viruses comprise a heterogeneous group of clinically relevant influenza viruses, influenza B viruses form a more homogeneous cluster, divided mainly into two lineages: Victoria and Yamagata. This divergence has complicated seasonal influenza vaccine design, which traditionally contained two seasonal influenza A virus strains and one influenza B virus strain. We examined the distribution of the two influenza B virus lineages in Israel, between 2011-2014, in hospitalized and in non-hospitalized (community) influenza B virus-infected patients. We showed that influenza B virus infections can lead to hospitalization and demonstrated that during some winter seasons, both influenza B virus lineages circulated simultaneously in Israel. We further show that the influenza B virus Yamagata lineage was dominant, circulating in the county in the last few years of the study period, consistent with the anti-Yamagata influenza B virus antibodies detected in the serum samples of affected individuals residing in Israel in the year 2014. Interestingly, we found that elderly people were particularly vulnerable to Yamagata lineage influenza B virus infections.

BackgroundThe recent H1N1 influenza A pandemic was marked by multiple reports of illness and hospitalization in children, suggesting that children may have played a major role in the propagation of the virus. A comprehensive detailed analysis of the attack rates among children as compared with their contacts in various settings is of great importance for understanding their unique role in influenza pandemics.Methodology/principal findingsWe searched MEDLINE (PubMed) and Embase for published studies reporting outbreak investigations with direct measurements of attack rates of the 2009 pandemic H1N1 influenza A among children, and quantified how these compare with those of their contacts. We identified 50 articles suitable for review, which reported school, household, travel and social events. The selected reports and our meta-analysis indicated that children had significantly higher attack rates as compared to adults, and that this phenomenon was observed for both virologically confirmed and clinical cases, in various settings and locations around the world. The review also provided insight into some characteristics of transmission between children and their contacts in the various settings.Conclusion/significanceThe consistently higher attack rates of the 2009 pandemic H1N1 influenza A among children, as compared to adults, as well as the magnitude of the difference is important for understanding the contribution of children to disease burden, for implementation of mitigation strategies directed towards children, as well as more precise mathematical modeling and simulation of future influenza pandemics.

Vaccine preventable diseases have been responsible for a significant portion of childhood mortality in low-income countries, and have been re-emerging in medium- and high-income countries. The effectiveness of routine childhood immunization programs relies on multiple factors. Social determinants have the potential to affect immunization programs around the world, with globalization and ease of communication facilitating their effect. Exploring the types of social determinants affecting immunization efforts in various countries is of great importance to the ability of nations to address them, prevent the spread of disease and lower mortality rates. The social determinants affecting vaccination programs can vary among countries of different income levels, with some social determinants overlapping among these country groups. In this article we explore the various social determinants affecting routine immunization programs in low-, middle- and high-income countries and possible interventions to address them.

Mycobacterium tuberculosis CDC 1551, a highly immunogenic outbreak strain, previously reported to have unique surface distribution of capsular polysaccharide, was used to generate novel monoclonal antibodies (mAbs) to surface mycobacterial targets. Two Immunoglobulin G1 (IgG1) mAbs, 16a1 and 16a6 were generated. The mAbs originated from the same B cell, bound strongly to whole cell M. tuberculosis CDC1551 and to its cell wall, membrane and cytosol fractions recognizing a 90 kDa protein. Immunoprecipitation using mAb 16a1 isolated a protein with amino acid peptide sequences matching MPT51 from the cytosol. This immunogenic protein of unknown function, was previously reported only in culture filtrates of M. tuberculosis. Our findings suggest for the first time that this protein is found within the M. tuberculosis cell.

More than 100 years after the discovery of the tubercle bacillus by Robert Koch ([43][1]) and approximately 50 years after the introduction of effective antimycobacterial therapeutic agents ([3][2], [4][3], [46][4]), tuberculosis (TB) continues to be a leading cause of morbidity and mortality

CD1d is a non-polymorphic MHC class I-related protein that binds and presents glycolipid antigens to T cell antigen receptors expressed by NK-like T (NKT) cells. CD1d-dependent immune responses play critical roles in infectious disease, autoimmunity, allergy and cancer. We tested the hypothesis that B cell antigen receptor (BCR) targeting of a biotin-modified version of the CD1d-binding antigen alpha-galactosylceramide (biotin-alpha-GalCer) results in enhanced murine CD1d-mediated presentation as compared with presentation of non-targeted biotin-alpha-GalCer. Presentation of BCR-targeted antigen to NKT cells was enhanced 100- to 1000-fold compared with non-targeted antigen. CD1d presentation of BCR-targeted antigen was observed after 4 h treatment, consistent with a requirement for endosomal trafficking. Furthermore, unlike non-targeted antigen, BCR-targeted antigen was not loaded directly onto cell-surface CD1d. Blocking BCR signaling with the Syk tyrosine kinase inhibitor piceatannol inhibited presentation of BCR-targeted antigen but not non-targeted antigen. Piceatannol blocked transport of the BCR to CD1d-containing endosomes, showing that intersection of BCR-targeted antigen with endosomes is required for enhanced mCD1d antigen presentation. Our data suggest that the BCR facilitates capture of low quantities of mCD1d antigens to enhance CD1d-dependent immune responses.

A healthy 3.5-month-old infant developed septic arthritis with Haemophilus influenzae type f isolated from the knee aspirate and blood. The patient had no obvious risk factors and immunological evaluation revealed no abnormalities. To our knowledge, there are only two reported childhood cases of septic arthritis caused by non-type b H. influenzae , but this is the first one to be reported in a child without an underlying disorder. The importance of serotyping H. influenzae isolates in the H. influenzae type b vaccine era, and the need to look for predisposing factors in paediatric patients with invasive disease caused by non-type b H. influenzae are discussed.

Cell-mediated immunity is considered to be the major component of the host response against Mycobacterium tuberculosis, whereas antibody-mediated immunity historically has been considered inconsequential. In recent years, studies from several groups have challenged the traditional dogma and demonstrated that monoclonal antibodies can modify various aspects of mycobacterial infections. This review describes the experimental evidence supporting a role for antibodies in defense against mycobacterial infections and outlines future challenges to the field of antibody-mediated immunity against M. tuberculosis, with particular emphasis on the implications of these findings for a novel vaccine strategy.

Arabinomannan (AM) is a polysaccharide antigen of the mycobacterial capsule. However, it is uncertain whether AM constitutes an immunologically distinct fraction of Mycobacterium tuberculosis. In this study, we analyzed the repertoire and specificity of antibodies to AM by using AM-binding murine monoclonal antibodies (MAbs) and human serum samples. Murine MAbs were found to be diverse in their specificity to AM and cross-reactivity with other arabinose-containing mycobacterial polysaccharides, with MAb 9d8 binding exclusively to AM. Human antibodies to AM were detected in serum samples from patients with pulmonary tuberculosis (TB), as well as in those from healthy, purified protein derivative-negative controls, with significantly higher titers among patients. The binding of human antibodies to AM was inhibited by MAb 9d8 in three patients with TB but not in controls. MAb 5c11, which recognizes other mycobacterial arabinosecontaining carbohydrates in addition to AM, inhibited the binding of serum samples from 75% of patients and 76% of controls. Analysis of human antibodies with murine MAbs to human VH determinants demonstrated diversity among antibodies to AM with qualitative and quantitative differences compared with antibodies to lipoarabinomannan. In summary, our study suggests that antibodies to AM are diverse and heterogeneous with respect to antigen recognition and VH determinant expression, with human serum samples containing different subsets of antibodies to AM with the specificities of AM-binding murine MAbs. One MAb and a subset of human antibodies bind AM specifically, suggesting that this polysaccharide is antigenically distinct and is expressed in human infection. Tuberculosis (TB) is a leading cause of mortality worldwide, and it is estimated that 1.86 billion people are infected with Mycobacterium tuberculosis (13). Contributing factors to this severe health problem are the human immunodeficiency virus epidemic, the long and complicated course of treatment, drug resistance, and a lack of efficient diagnostic and preventive modalities. In addition, we lack a full understanding of the immunopathogenesis of TB. Our group is interested in the M. tuberculosis carbohydrate arabinomannan (AM), which is thought to be a component of the mycobacterial capsule. This interest arose from the generation of a monoclonal antibody (MAb), 9d8, that prolonged the survival of mice infected with M. tuberculosis (36) and binds AM. These findings suggested

Aerosolized delivery of virulent or hypervirulent Mycobacterium tuberculosis requires careful consideration of methodology and safety. To maximize safety, we installed a nose-only aerosol apparatus that can reproducibly deliver a low dose (M. tuberculosis in a carefully designed biohazard facility.

The outermost layer ofMycobacterium tuberculosiscontains two major polysaccharides, arabinomannan (AM) and glucan (GC). We studied the in vitro and in vivo expression of anM. tuberculosisAM antigen using monoclonal antibody (MAb) 9d8 (2a), an isotype-switched variant of the immunoglobulin G3 (IgG3) MAb 9d8. MAb 9d8 had been previously shown to bindM. tuberculosisAM and theM. tuberculosissurface. Our in vitro experiments showed that MAb 9d8(2a) bound strongly to whole-cellM. tuberculosisErdman but not to the CDC 1551 strain grown in medium for an extended period. However, AM antigen was detected in the culture supernatant of both strains, and its concentration increased in a time-dependent manner. The detection of AM antigen from both strains was decreased in the presence of Tween 80. In mice infected withM. tuberculosisErdman, AM antigen accumulated in organ homogenates concomitant to an increase in bacterial organ burden and an increase in IgG and IgM titer to AM. These results (i) indicate that the surface expression of AM during in vitro growth changes with culture age, is strain dependent, and is affected by the presence of Tween 80 in the culture media; (ii) show that AM is produced by bacteria growth in vivo; and (iii) demonstrate that the amount of in vivo-detected AM can be dependent on the number of bacteria in the infected organ.

Lipoarabinomannan (LAM) is a component of the mycobacterial surface which has been associated with a variety of deleterious effects on immune system function. Despite the importance of LAM to the pathogenesis of mycobacterial infection, there is no information available on its fate in vivo. In this study, we determined the pharmacokinetics and tissue distribution of exogenously administered LAM in mice. For measurements of serum and tissue LAM concentrations, we developed an enzyme-linked immunosorbent assay which used monoclonal antibodies of different isotypes to capture and detect LAM at concentrations of ≥0.4 μg/ml. Intravenous administration of LAM to mice resulted in transient serum levels with organ deposition in the spleen and in the liver. Immunohistochemical studies localized LAM to the spleen marginal zone macrophages and, to a lesser degree, to liver macrophages. When LAM was administered to mice previously given a LAM-binding immunoglobulin M (IgM), LAM was very rapidly cleared from circulation. In those mice, deposition of LAM in the spleen was significantly reduced while LAM deposition in the liver increased. Administration of LAM-binding IgM resulted in significant levels of IgM to LAM in bile consistent with an increased hepatobiliary excretion of LAM in the presence of specific antibody. Bile, liver extracts, and bile salts were found to rapidly inactivate the immunoreactivity of LAM. The results indicate that serum clearance and organ deposition of LAM in mice are affected by the presence of LAM-binding antibody and suggest a mechanism by which antibody could modify the course of mycobacterial infection.

The murine monoclonal antibody (MAb) 18B7 [immunoglobulin G1(κ)] is in preclinical development for treatment of Cryptococcus neoformans infections. In anticipation of its use in humans, we defined the serological and biological properties of MAb 18B7 in detail. Structural comparison to the related protective MAb 2H1 revealed conservation of the antigen binding site despite several amino acid differences. MAb 18B7 was shown by immunofluorescence and agglutination studies to bind to all four serotypes of C. neoformans , opsonize C. neoformans serotypes A and D, enhance human and mouse effector cell antifungal activity, and activate the complement pathway leading to deposition of complement component 3 (C3) on the cryptococcal capsule. Administration of MAb 18B7 to mice led to rapid clearance of serum cryptococcal antigen and deposition in the liver and spleen. Immunohistochemical studies revealed that MAb 18B7 bound to capsular glucuronoxylomannan in infected mouse tissues. No reactivity of MAb 18B7 with normal human, rat, or mouse tissues was detected. The results show that both the variable and constant regions of MAb 18B7 are biologically functional and support the use of this MAb in human therapeutic trials.

Three children with acquired immunodeficiency syndrome who had pneumonia develop were infected simultaneously with Pneumocystis carinii and Streptococcus pneumoniae. Such coexistence has not been previously reported in children. One patient received prophylactic treatment against Pneumocystis carinii before his illness.

Three monoclonal antibodies (MAbs) were generated from splenocytes of a BALB/c mouse immunized with heat-killed Mycobacterium tuberculosis. All three MAbs bound to surface epitopes of M. tuberculosis as shown by whole-cell enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence, and immunoelectron microscopy. One immunoglobulin M (IgM) MAb bound to lipoarabinomannan, the second IgM MAb bound to mycolyl-arabinogalactan-peptidoglycan complex, and the third MAb, an IgG3, bound to a surface epitope of an uncertain nature. The MAbs demonstrated different cross-reactivity patterns with other mycobacteria. Two of the MAbs were used to develop a modified ELISA spot assay for the detection of mycobacteria.

Background A major effort to introduce new vaccines into poor nations of the world was initiated in recent years with the help of the GAVI alliance. The first vaccines introduced have been the Haemophilus influenzae type B (Hib) and the hepatitis B (Hep B) vaccines. The introduction of these vaccines during the first phase of GAVI's operations demonstrated considerable variability. We set out to study the factors affecting the introduction of these vaccines. The African Region (AFRO), where new vaccines were introduced to a substantial number of countries during the first phase of GAVI's funding, was selected for this study. Methodology/Principal Findings GAVI-eligible AFRO countries with a population of 0.5 million or more were included in the study. Countries were analyzed and compared for new vaccine introduction, healthcare indicators, financial indicators related to healthcare and country-level Governance Indicators, using One Way ANOVA, correlation analysis and Qualitative Comparative Analysis (QCA). Introduction of new vaccines into AFRO nations was associated primarily with high country-level Governance Indicator scores. The use of individual Governance Indicator scores, as well as a combined Governance Indicator score we developed, demonstrated similar results. Conclusions/Significance Our study results indicate that good country-level governance is an imperative pre-requisite for the successful early introduction of new vaccines into poor African nations. Enhanced support measures may be required to effectively introduce new vaccines to countries with low governance scores. The combined governance score we developed may thus constitute a useful tool for helping philanthropic organizations make decisions regarding the type of support needed by different countries to achieve success.

Background The crude rate of early-onset Group B streptococcus disease (EOGBS) in Israel has been consistently under 0.5 for 1000 live births for the past 8 years. The Israeli Ministry of Health has adapted the risk factor based approach for preventing EOGBS and universal bacteriological screening for GBS is not recommended. In spite of this policy, there are indications that many pregnant women in Israel undergo bacteriological screening for GBS. The objective of this study is to assess the rate and characteristics of pregnant women who undergo screening for group B streptococcus (GBS) colonization in Israel. Methods Survey of expectant mothers who came to give birth in 29 delivery rooms throughout Israel during the month of July 2012 regarding GBS screening practice and demographics. Results A total of 2968 pregnant women participated in the assessment. Among them, 935 women (31.5 %) had been tested for GBS colonization. About 90 % of those women had no risk factors, only 542 women (60 %) underwent testing during the recommended gestational timing (35–37 weeks) and 23 % of the tested women reported being GBS carriers. GBS screening as part of the routine pregnancy follow- up was associated with: residence district, intermediate or high socioeconomic rank, being a member of certain health maintenance organization and being Jewish. Characteristics found to be significantly associated with being a GBS carrier were: low socioeconomic rank, and having a risk factor for GBS infection. Conclusions A substantial number of pregnant women in Israel undergo screening for GBS colonization despite the national policy against universal screening. While GBS colonization was more prevalent in women of lower socioeconomic status, screening is done more often in those of higher socioeconomic status, suggesting unnecessary monetary expenses.