Nonimmune Host Research Articles

Case Report of Tuberculous Meningitis in an Intravenous Drug Dependent Patient

Tubercle bacilli can cause tuberculous meningitis by hematogenous spread from a primary focus, usually located in the lung, peribronchial and peritracheal regions. In the non-immune host, small emboli carrying mycobacteria usually spread from primary pulmonary foci to the meninges and other organs. Tuberculous meningitis can also develop by hematogenous spread from a primary focus. A 49-year-old foreign national male patient was admitted to the emergency department with complaints of joint pains, fever and malaise. He complained of dry cough and physical examination revealed hyperemic oropharynx, normal respiratory sounds and other system examinations. After the tests performed, the patient was diagnosed with viral upper respiratory tract infection. The patient re-presents to the emergency room the next day with the complaint of inability to urinate. Physical examination reveals no acute pathologic findings except oropharyngeal hyperemia and suprapubic tenderness. The patient's blood parameters were different from the previous day with WBC: 16000 and CRP: 115. The patient was again discharged from the emergency department after oral antibiotics were prescribed and outpatient follow-up was recommended. On his 3rd admission to the emergency department, he presented with complaints of weakness, numbness in the legs, spasm and pain in the neck. As a result of the tests performed, the patient was transferred to the intensive care unit with a preliminary diagnosis of atypical meningitis and was diagnosed with tuberculous meningitis and followed up and treated. In patients presenting with atypical neurologic symptoms with this case report, anamnesis and examination should be very detailed and rare infections such as atypical meningitis should be considered in the differential diagnosis.

Evidence for Aerosol Transfer of SARS-CoV-2-Specific Humoral Immunity.

Infectious particles can be shared through aerosols and droplets formed as the result of normal respiration. Whether Abs within the nasal/oral fluids can similarly be shared between hosts has not been investigated. The circumstances of the SARS-CoV-2 pandemic facilitated a unique opportunity to fully examine this provocative idea. The data we show from human nasal swabs provides evidence for the aerosol transfer of Abs between immune and nonimmune hosts.

A B C Fractional Derivative for Varicella-Zoster Virus Using Two-Scale Fractal Dimension Approach with Vaccination

Chickenpox or varicella is an infectious disease caused by the varicella-zoster virus V Z V . This virus is the cause of chickenpox (usually a primary infection in the nonimmune host) and herpes zoster. In this paper, a compartmental model for the dynamics of V Z transmission with the effect of vaccination is solved using the A B C fractional derivative. The possibility of using the fractal dimension as a biomarker to identify different diseases is being investigated. The problem is investigated in two different levels of research using two scale dimensions. To ascertain the existence and uniqueness of the solution, we qualitatively evaluate the model. We have used the Euler method to compute the numerical solution for the system. At the end, we provide the graphical results showing the effectiveness of two-scale dimension and fractional calculus in the current model.

Angiotensin Converting Enzyme 2 (ACE2) Expression in the Aged Brain and Visual System

Multiple lines of evidence currently indicate that the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) gains entry into human host cells via a high-affinity interaction with the angiotensin-converting enzyme 2 (ACE2) transmembrane receptor. Research has further shown the widespread expression of the ACE2 receptor on the surface of many different immune, non-immune and neural host cell types, and that SARS-CoV-2 has the remarkable capability to attack many different types of human-host cells simultaneously. One principal neuroanatomical region for high ACE2 expression patterns occurs in the brainstem, an area of the brain containing regulatory centers for respiration, and this may in part explain the predisposition of many COVID-19 patients to respiratory distress. Early studies also indicated extensive ACE2 expression in the whole eye and the brain’s visual circuitry in aged humans. In this study we analyzed ACE2 receptor expression at the mRNA and protein level in multiple cell types involved in human vision, including cell types of the external eye and several deep brain regions known to be involved in the processing of visual signals. Here we provide evidence: (i) that many different optical and neural cell types of the human visual system provide receptors essential for SARS-CoV-2 invasion; (ii) of the remarkable ubiquity of ACE2 presence in cells of the eye and anatomical regions of the brain involved in visual signal processing; (iii) that ACE2 receptor expression in different ocular cell types and visual processing centers of the brain provide multiple compartments for SARS-CoV-2 infiltration; and (iv) of a gradient of increasing ACE2 expression from the anterior surface of the eye to the visual signal processing areas of the occipital lobe and the primary visual neocortex. A gradient of ACE2 expression from the eye surface to the occipital lobe may provide the SARS-CoV-2 virus a novel pathway from the outer eye into deeper anatomical regions of the brain involved in vision. These findings may explain, in part, the many recently reported neuro-ophthalmic manifestations of SARS-CoV-2 infection in COVID-19 affected patients.

Evaluation of intermolecular forces between lipopolysaccharides and monoclonal antibodies using atomic force microscopy

Understanding of interactions between a bacterium and an immune or non-immune host organism at the cellular and subcellular level is important in order to improve new and existing immunobiological tools for the treatment of bacterial infections (including pseudotuberculosis). The aim of this work was to quantify the interaction force between Yersinia pseudotuberculosis and monoclonal antibodies (mAbs) in the model system “lipopolysaccharide (LPS) - mAbs” by atomic force microscopy (AFM). Our research findings provided the methodical approaches to force measurements between an AFM probe, which was functionalized with Y. pseudotuberculosis LPS, and mica coated by different mAbs. Based on the criteria for force estimation there was shown a greater binding force in the system “LPS – complementary mAbs” than in the system “LPS – heterologous mAbs”. In both cases binding force increase followed by increase a contact time between the functionalized AFM probe and mica from 1 to 5 s. It has been shown that single bonds between LPS and complementary mAbs molecules also included a clearly defined non-specific component along with immunochemically specific one. The evidence suggests a significant proportion of applied force exerted to unfolding of high-molecular aggregates whose length may attain many hundreds of nanometers.

Evasion of host defenses by intracellular Staphylococcus aureus.

Staphylococcus aureus is one of the leading causes of hospital and community-acquired infections worldwide. The increasing occurrence of antibiotic resistant strains and the high rates of recurrent staphylococcal infections have placed several treatment challenges on healthcare systems. In recent years, it has become evident that S. aureus is a facultative intracellular pathogen, able to invade and survive in a range of cell types. The ability to survive intracellularly provides this pathogen with yet another way to evade antibiotics and immune responses during infection. Intracellular S. aureus have been strongly linked to several recurrent infections, including severe bone infections and septicemias. S. aureus is armed with an array of virulence factors as well as an intricate network of regulators that enable it to survive, replicate and escape from a number of immune and nonimmune host cells. It is able to successfully manipulate host cell pathways and use it as a niche to multiply, disseminate, as well as persist during an infection. This bacterium is also known to adapt to the intracellular environment by forming small colony variants, which are metabolically inactive. In this review we will discuss the clinical evidence, the molecular pathways involved in S. aureus intracellular persistence, and new treatment strategies for targeting intracellular S. aureus.

Detection of canine parvovirus types 2b and 2c in canine faecal samples contaminating urban thoroughfares in Brazil.

Canine parvovirus type 2 (CPV-2) is a highly contagious virus that causes acute gastroenteritis in dogs all over the world. Because of its stability in the environment, CPV-2 can remain infective for a long time, especially if protected in organic matter. To demonstrate CPV-2's potential as an environmental hazard for nonimmunized susceptible hosts, we investigated 50 faecal samples collected from public areas in a municipality of Paraná state, Brazil. Seven samples tested positive for CPV by a PCR assay targeting the partial VP2 gene, with three strains being confirmed as CPV-2b variant and one as CPV-2c variant by sequence analysis. These findings were supported by phylogenetic analysis, and the species identity of faecal samples source was confirmed by canine mitochondrial DNA amplification and sequencing. Our results demonstrate the presence of CPV in canine faeces contaminating urban thoroughfares and reinforce the importance of environmental control to reduce the potential exposure risks to susceptible hosts.

Necroptosis mediators RIPK3 and MLKL suppress intracellular Listeria replication independently of host cell killing.

RIPK3, a key mediator of necroptosis, has been implicated in the host defense against viral infection primary in immune cells. However, gene expression analysis revealed that RIPK3 is abundantly expressed not only in immune organs but also in the gastrointestinal tract, particularly in the small intestine. We found that orally inoculated Listeria monocytogenes, a bacterial foodborne pathogen, efficiently spread and caused systemic infection in Ripk3-deficient mice while almost no dissemination was observed in wild-type mice. Listeria infection activated the RIPK3-MLKL pathway in cultured cells, which resulted in suppression of intracellular replication of Listeria Surprisingly, Listeria infection-induced phosphorylation of MLKL did not result in host cell killing. We found that MLKL directly binds to Listeria and inhibits their replication in the cytosol. Our findings have revealed a novel functional role of the RIPK3-MLKL pathway in nonimmune cell-derived host defense against Listeria invasion, which is mediated through cell death-independent mechanisms.

Tuberculosis verrucosa cutis: case report of a diagnostic challenge

<p class="abstract">Cutaneous tuberculosis occurs by either exogenous inoculation in a previously sensitized or non-immune host or endogenous spread from an internal focus by contiguous, hematogenous or lymphatic route. Inoculation occurs at sites of minor wounds or abrasions, sometimes from the patient’s own sputum. Cutaneous tuberculosis includes lupus vulgaris and tuberculosis verrucosa cutis (TVC) at one end and scrofuloderma and tuberculosis cutis orificialis at the other end with decrease in cell-mediated immunity across the spectrum. Of various cutaneous forms, we report a case of tuberculosis verrucosa cutis in a 14 year old girl who presented with a hyperpigmented verrucous plaque over foot since three years. Histopathological characteristics, GeneXpert and response to antitubercular therapy confirmed the diagnosis.</p><p class="abstract"> </p>

Bordetella parapertussis Circumvents Neutrophil Extracellular Bactericidal Mechanisms.

B. parapertussis is a whooping cough etiological agent with the ability to evade the immune response induced by pertussis vaccines. We previously demonstrated that in the absence of opsonic antibodies B. parapertussis hampers phagocytosis by neutrophils and macrophages and, when phagocytosed, blocks intracellular killing by interfering with phagolysosomal fusion. But neutrophils can kill and/or immobilize extracellular bacteria through non-phagocytic mechanisms such as degranulation and neutrophil extracellular traps (NETs). In this study we demonstrated that B. parapertussis also has the ability to circumvent these two neutrophil extracellular bactericidal activities. The lack of neutrophil degranulation was found dependent on the O antigen that targets the bacteria to cell lipid rafts, eventually avoiding the fusion of nascent phagosomes with specific and azurophilic granules. IgG opsonization overcame this inhibition of neutrophil degranulation. We further observed that B. parapertussis did not induce NETs release in resting neutrophils and inhibited NETs formation in response to phorbol myristate acetate (PMA) stimulation by a mechanism dependent on adenylate cyclase toxin (CyaA)-mediated inhibition of reactive oxygen species (ROS) generation. Thus, B. parapertussis modulates neutrophil bactericidal activity through two different mechanisms, one related to the lack of proper NETs-inducer stimuli and the other one related to an active inhibitory mechanism. Together with previous results these data suggest that B. parapertussis has the ability to subvert the main neutrophil bactericidal functions, inhibiting efficient clearance in non-immune hosts.

IDENTIFICATION OF FUNCTIONALLY AND STRUCTURALLY IMPORTANT RESIDUES OF OUTER MEMBRANE PROTEIN P1 IN HAEMOPHILUSINFLUENZAE

Haemophilusinfluenzae (formerly called Pfeiffer's bacillus or Bacillus influenzae) is a Gram-negative, coccobacillary, facultatively anaerobic pathogenic bacterium belonging to the Pasteurellaceae family. The presence of the capsule in encapsulated type b (Hib), a serotype causing conditions such as epiglottitis, is known to be a major factor in virulence. Their capsule allows them to resist phagocytosis and complement-mediated lysis in the nonimmune host. The unencapsulated strains are almost always less invasive. Haemophilusinfluenzae infections cause some severe and fatal, especially in children such as meningitis and epiglottitis. The property of an antigen to bind specifically complementary antibodies is known as the antigen’s antigenicity; likewise, the ability of an antigen to induce an immune response is called its immunogenicity. Attempts should be made to discover peptides that could mimic protein epitopes and possess the same immunogenicity as the whole protein. Subsequently, theoretical methods for epitope prediction have been developed leading to synthesis of such peptides that are important for development of immunodiagnostic tests and vaccines. The present study was designed to in silico resolving the major obstacles in the control or in prevention of the diseases caused by Haemophilusinfluenzae . We exploitedbioinformatic tools to better understanding and characterizing the Outer Membrane Protein P1 structure in Haemophilusinfluenzae and select appropriate regions as effectiveB cell epitops.

Cathelicidins positively regulate pancreatic β-cell functions.

Cathelicidins are pleiotropic antimicrobial peptides largely described for innate antimicrobial defenses and, more recently, immunomodulation. They are shown to modulate a variety of immune or nonimmune host cell responses. However, how cathelicidins are expressed by β cells and modulate β-cell functions under steady-state or proinflammatory conditions are unknown. We find that cathelicidin-related antimicrobial peptide (CRAMP) is constitutively expressed by rat insulinoma β-cell clone INS-1 832/13. CRAMP expression is inducible by butyrate or phenylbutyric acid and its secretion triggered upon inflammatory challenges by IL-1β or LPS. CRAMP promotes β-cell survival in vitro via the epidermal growth factor receptor (EGFR) and by modulating expression of antiapoptotic Bcl-2 family proteins: p-Bad, Bcl-2, and Bcl-xL. Also via EGFR, CRAMP stimulates glucose-stimulated insulin secretion ex vivo by rat islets. A similar effect is observed in diabetes-prone nonobese diabetic (NOD) mice. Additional investigation under inflammatory conditions reveals that CRAMP modulates inflammatory responses and β-cell apoptosis, as measured by prostaglandin E2 production, cyclooxygenases (COXs), and caspase activation. Finally, CRAMP-deficient cnlp(-/-) mice exhibit defective insulin secretion, and administration of CRAMP to prediabetic NOD mice improves blood glucose clearance upon glucose challenge. Our finding suggests that cathelicidins positively regulate β-cell functions and may be potentially used for intervening β-cell dysfunction-associated diseases.

Novel disease susceptibility factors for fungal necrotrophic pathogens in Arabidopsis.

Host cells use an intricate signaling system to respond to invasions by pathogenic microorganisms. Although several signaling components of disease resistance against necrotrophic fungal pathogens have been identified, our understanding for how molecular components and host processes contribute to plant disease susceptibility is rather sparse. Here, we identified four transcription factors (TFs) from Arabidopsis that limit pathogen spread. Arabidopsis mutants defective in any of these TFs displayed increased disease susceptibility to Botrytis cinerea and Plectosphaerella cucumerina, and a general activation of non-immune host processes that contribute to plant disease susceptibility. Transcriptome analyses revealed that the mutants share a common transcriptional signature of 77 up-regulated genes. We characterized several of the up-regulated genes that encode peptides with a secretion signal, which we named PROVIR (for provirulence) factors. Forward and reverse genetic analyses revealed that many of the PROVIRs are important for disease susceptibility of the host to fungal necrotrophs. The TFs and PROVIRs identified in our work thus represent novel genetic determinants for plant disease susceptibility to necrotrophic fungal pathogens.

Optimizing recellularization of whole decellularized heart extracellular matrix.

RationalePerfusion decellularization of cadaveric hearts removes cells and generates a cell-free extracellular matrix scaffold containing acellular vascular conduits, which are theoretically sufficient to perfuse and support tissue-engineered heart constructs. However, after transplantation, these acellular vascular conduits clot, even with anti-coagulation. Here, our objective was to create a less thrombogenic scaffold and improve recellularized-left ventricular contractility by re-lining vascular conduits of a decellularized rat heart with rat aortic endothelial cells (RAECs).Methods and ResultsWe used three strategies to recellularize perfusion-decellularized rat heart vasculature with RAECs: retrograde aortic infusion, brachiocephalic artery (BA) infusion, or a combination of inferior vena cava (IVC) plus BA infusion. The re-endothelialized scaffolds were maintained under vascular flow in vitro for 7 days, and then cell morphology, location, and viability were examined. Thrombogenicity of the scaffold was assessed in vitro and in vivo. Both BA and IVC+BA cell delivery resulted in a whole heart distribution of RAECs that proliferated, retained an endothelial phenotype, and expressed endothelial nitric oxide synthase and von Willebrand factor. Infusing RAECs via the combination IVC+BA method increased scaffold cellularity and the number of vessels that were lined with endothelial cells; re-endothelialization by using BA or IVC+BA cell delivery significantly reduced in vitro thrombogenicity. In vivo, both acellular and re-endothelialized scaffolds recruited non-immune host cells into the organ parenchyma and vasculature. Finally, re-endothelialization before recellularization of the left ventricular wall with neonatal cardiac cells enhanced construct contractility.ConclusionsThis is the first study to re-endothelialize whole decellularized hearts throughout both arterial and venous beds and cavities by using arterial and venous delivery. The combination (IVC+BA) delivery strategy results in enhanced scaffold vessel re-endothelialization compared to single-route strategies. Re-endothelialization reduced scaffold thrombogencity and improved contractility of left ventricular-recellularized constructs. Thus, vessel and cavity re-endothelialization creates superior vascularized scaffolds for use in whole-organ recellularization applications.

Vaccination with recombinant Brugia malayi cystatin proteins alters worm migration, homing and final niche selection following a subcutaneous challenge of Mongolian gerbils (Meriones unguiculatus) with B. malayi infective larvae

BackgroundCysteine protease inhibitors of Brugia malayi have been ascribed to be involved in parasite development as well as to immunomodulate the host’s immune response. In Onchocerca volvulus, Onchocystatin has been shown to induce partial protection in the mouse diffusion chamber vaccination model. In the present study we investigated the impact of vaccination with recombinant Bm-CPI-1 and Bm-CPI-2 proteins on protection against a subcutaneous challenge of B. malayi third stage larvae in gerbils.FindingsVaccination with E. coli derived recombinant B. malayi cysteine protease inhibitors (Bm-CPI-1 or -2) did not confer protection against B. malayi L3 challenge infection in gerbils but altered the homing of a significant number of adult worms from the lymphatics to the heart and lungs.ConclusionBm-CPI vaccination-induced alteration in worm migration is consistent with our previous observations in gerbils vaccinated with B. pahangi excretory-secretory (ES) proteins, which resulted in delayed migration of the L3s and altered the final location of adult worms. Similar observations have also been made in dogs vaccinated with Ancylostoma caninum proteins; an increased number of worms were recovered in the colon and not the expected small intestine. A change in the final niche was also reported in immune versus non-immune hosts of two other gut dwelling nematodes. Vaccination induced alteration of the parasite’s final homing might be a rare or a common phenomenon, which unfortunately is rarely recorded. The reason for the alteration in the final niche selection by adult nematode worms following vaccination is unknown and necessitates further investigation.

Fish immune responses against endoparasitic nematodes – experimental models

Vertebrates mount a series of immune reactions when invaded by helminths but antihelmintic immune strategies allow, in many cases, the first invaders of the non-immune host to survive for prolonged periods, whereas subsequent larval invaders of the same parasite species face increased host resistance and thereby decreased colonization success. This concomitant immunity may represent a trade-off between adverse side effects (associated with killing of large helminths in the host tissue) and the need for future protection against invasion. Encapsulation and isolation of large live endoparasitic larvae may be associated with less pathology compared to coping with excess dead parasite tissue in host organs. Likewise, live adult nematodes may be accepted in tissues at a certain activity level for the same reasons. Various host cell receptors bind helminth molecules after which signal-transducing events lead to mobilization of specific reaction patterns depending on the combination of receptors and ligands involved. Both innate and adaptive responses (humoral and cellular) are prominent actors, but skewing of the Th1 lymphocyte response towards a Th2 type is a characteristic element of antihelminthic responses in mammalian hosts. Similar patterns may be expected also to occur in at least some fish species, such as salmonids, producing relevant cytokines, MHCII and CD4+ cells required for these lymphocyte subpopulations. Atlantic cod, Gadus morhua L., is without these immunological elements that indicate that alternative reaction pathways exist in at least some fish groups. Recent achievements within teleost immunology have made it possible to track these host responses in fish and the present work outlines the main immune reactions in fish against helminths and suggests three experimental fish models for exploration of these immune pathways in fish infected with nematodes.

Antecedent Avian Immunity Limits Tangential Transmission of West Nile Virus to Humans

BackgroundWest Nile virus (WNV) is a mosquito-borne flavivirus maintained and amplified among birds and tangentially transmitted to humans and horses which may develop terminal neuroinvasive disease. Outbreaks typically have a three-year pattern of silent introduction, rapid amplification and subsidence, followed by intermittent recrudescence. Our hypothesis that amplification to outbreak levels is contingent upon antecedent seroprevalence within maintenance host populations was tested by tracking WNV transmission in Los Angeles, California from 2003 through 2011.MethodsPrevalence of antibodies against WNV was monitored weekly in House Finches and House Sparrows. Tangential or spillover transmission was measured by seroconversions in sentinel chickens and by the number of West Nile neuroinvasive disease (WNND) cases reported to the Los Angeles County Department of Public Health.ResultsElevated seroprevalence in these avian populations was associated with the subsidence of outbreaks and in the antecedent dampening of amplification during succeeding years. Dilution of seroprevalence by recruitment resulted in the progressive loss of herd immunity following the 2004 outbreak, leading to recrudescence during 2008 and 2011. WNV appeared to be a significant cause of death in these avian species, because the survivorship of antibody positive birds significantly exceeded that of antibody negative birds. Cross-correlation analysis showed that seroprevalence was negatively correlated prior to the onset of human cases and then positively correlated, peaking at 4–6 weeks after the onset of tangential transmission. Antecedent seroprevalence during winter (Jan – Mar) was negatively correlated with the number of WNND cases during the succeeding summer (Jul–Sep).ConclusionsHerd immunity levels within after hatching year avian maintenance host populations <10% during the antecedent late winter and spring period were followed on three occasions by outbreaks of WNND cases during the succeeding summer. Because mosquitoes feed almost exclusively on these avian species, amplification was directly related to the availability of receptive non-immune hosts.

Pathogenesis of Helicobacter Pylori Infection: Colonization, Virulence Factors of the Bacterium and Immune and Non-immune Host Response

Helicobacter pylori (Hp), a gram-negative, spiral-shaped bacterium is one of the most widely spread pathogens in humans, as it concerns half of the world population. Mechanisms that allow Hp to cause a life-long infection involve modulation of the immune response and host cellular processes which include activation of the innate immune response, resistance to phagocytosis, modulation of dendritic cell activity and regulatory T cells, and production of proinflammatory cytokines. This is accomplished via virulence factors such as colonization factors (a variety of adhesins), factors that allow it to evade host defence (flagella and motility, urease system, induction of hypochlorhydria) and factors that are responsible for tissue injury (heat shock proteins A and B, vacuolating cytotoxin A, neutrophil activating protein of Hp, and cytotoxin-associated gene A). The interaction between bacterial effectors, environmental factors (genetic susceptibility to infection) and factors that modulate the host's response, such as polymorphisms in genes encoding cytokines or cytokine receptors, have been shown to influence the clinical outcome of Hp infection either towards peptic ulcer and/or cancer. Future studies, directed toward understanding interactions between Hp and immune cells in vivo, may lead to the development of novel therapeutic approaches for eradication of Hp.

Strategies for Control of Pandemic Influenza: Active and Passive Immunization

Pandemic influenza poses a serious threat to global health and the world economy. Highly pathogenic avian influenza A virus (HPAIV) of the H5N1 subtype that has emerged since 2004, resulted in more than 430 cases in 15 countries with a 50% case-fatality rate. Novel vaccine strategies for early response include the use of attenuated vaccines/vectors administered via novel mucosal immunization routes, therapeutic anti-virals and passive immunization with virus-specific antibodies (Abs). The current influenza vaccines designed for inducing antibody (Ab) responses against viral surface antigens (i.e., hemagglutinin [HA] and neuraminidase [NA]) are limited to seasonal use because of the ability of the virus to mutate these major antigenic glycoproteins. Vaccines that target determinants conserved among influenza A viruses (IAV) to generate broad protection against infection with different influenza A subtypes (i.e., heterosubtypic immunity [HSI]) remain elusive. We have currently developed a recombinant adenovirus (Ad) vector co-encoding HA (H5 subtype) and a conserved ectodomain of matrix protein 2 (M2e) (AdH5/M2e) for induction of protective immunity to H5N1 and other subtypes. Another approach based on the use of influenza virus carrying a deletion in the nonstructural NS1 gene is being explored. Since NS1 enables the virus to disarm the host type 1 IFN response, such deletion leads to attenuation of the viruses and enhanced host antiviral response. Therefore, vaccines based on NS1 deleted viruses (DelNS1) may provide better protection than inactivated vaccines and could induce HSI to infection with different influenza virus A subtypes. Sub-lingual immunization has been found to be a safe and effective route for induction of protective immune responses in systemic and mucosal compartments including respiratory tract. We found that sublingual immunization with either AdH5/M2e or DelNS1 induces broad protective immunity to H5 viruses and other influenza virus A subtypes including H1N1. Passive immunization (the transfer of specific immunoglobulins/Abs to a previously nonimmune recipient host) could offer an alternative strategy to prevent and treat influenza virus infection and an additional therapeutic option to antiviral drugs that are limited by widespread drug resistance among influenza virus strains. Even after targeted vaccines become available, passive immunization could still have prophylactic effects and provides an additional countermeasure against influenza, especially for individuals who do not respond well to the vaccines. Attempts to develop monoclonal Abs (mAbs) have been made. However, passive immunization based on mAbs may require a cocktail of mAbs with broader specificity in order to provide full protection since mAbs are generally specific for single epitopes. Because the recent epidemic of highly pathogenic avian influenza virus (HPAIV) strain H5N1 has resulted in serious economic losses to the poultry industry, many countries including Vietnam have introduced mass vaccination of poultry with H5N1 virus vaccines. We found that eggs obtained from chicken farms and supermarkets in Vietnam contain H5N1-specific immunoglobulins (IgY) that provide protection against infections with HPAIV H5N1 and related H5N2 strains in mice. When administered intranasally before or after lethal infection with HPAIV H5N1, H5N1-specific IgY prevent disease or significantly reduce viral replication resulting in complete recovery from the disease, respectively. In addition, we generated H1N1 virusspecific IgY by immunization of hens with inactivated H1N1 A/PR/8/34 as a model virus for current pandemic H1N1/09 and found that such H1N1-specific IgY protect mice from lethal influenza virus infection. These results underscore the usefulness of recombinant Ad vectors encoding surface glycoprotein (HA) and conserved protein (M2e) and NS1 deleted viruses (DelNS1) as vaccine candidates for control of pre-pandemic H5N1 and newly emerging subtypes. Data on antiviral efficacy of IgY provide a proof-of-concept for the approach using virus-specific IgY as affordable, safe, and effective alternative for the control of influenza outbreaks, including the potential H5N1 and current H1N1 pandemic.

How selection forces dictate the variant surface antigens used by malaria parasites

Red blood cells infected by the malaria parasite Plasmodium falciparum express variant surface antigens (VSAs) that evade host immunity and allow the parasites to persist in the human population. There exist many different VSAs and the differential expression of these VSAs is associated with the virulence (damage to the host) of the parasites. The aim of this study is to unravel the differences in the effect key selection forces have on parasites expressing different VSAs such that we can better understand how VSAs enable the parasites to adapt to changes in their environment (like control measures) and how this may impact the virulence of the circulating parasites. To this end, we have built an individual-based model that captures the main selective forces on malaria parasites, namely parasite competition, host immunity, host death and mosquito abundance at both the within- and between-host levels. VSAs are defined by the net growth rates they infer to the parasites and the model keeps track of the expression of, and antibody build-up against, each VSA in all hosts. Our results show an ordered acquisition of VSA-specific antibodies with host age, which causes a dichotomy between the more virulent VSAs that reach high parasitaemias but are restricted to young relatively non-immune hosts, and less virulent VSAs that do not reach such high parasitaemias but can infect a wider range of hosts. The outcome of a change in the parasite's environment in terms of parasite virulence depends on the exact balance between the selection forces, which sets the limiting factor for parasite survival. Parasites will evolve towards expressing more virulent VSAs when the limiting factor for parasite survival is the within-host parasite growth and the parasites are able to minimize this limitation by expressing more virulent VSAs.