Enterovirus and Parechovirus meningitis in children: a review of the epidemiology, diagnostic challenges, and significance of on-site CSF virology tests in tropical paediatric patients’ care

Enteroviruses and Parechoviruses are increasingly recognized as the cause of aseptic meningitis, especially in the paediatric age group. However, because of indistinguishable clinical features with bacterial meningitis, many clinicians cannot make a clear distinction in disease presentation, and a large number of cases go undiagnosed. Although polymerase chain reaction is the current standard diagnostic approach, it takes many hours or days to get a result and these tests are not available at primary and secondary levels of care in many resource-poor countries. Furthermore, diagnosis is often difficult in children due to nonspecific cellular and biochemical cerebrospinal fluid findings. Some affected children may develop neurologic or/and systemic complications, resulting in prolonged hospital admission, increasing the risk of avoidable deaths, and healthcare expenditures. This review focuses on epidemiology, presentation, and diagnosis of Enterovirus and Parechovirus meningitis, highlighting the challenges in diagnosis and the potential roles of on-site CSF virology tests in improving the quality of paediatric patient’s care. The information provided should help early case detection, thereby ensuring avoidance of unnecessary antibiotics, minimal complications, a short period of hospital stays, and a reduction in healthcare-associated costs. Keywords: Aseptic meningitis; Enterovirus; Parechovirus; Diagnostic challenge; On-site virology test; Children   FrenchTitle: Méningite à Entérovirus et Parechovirus chez les enfants: un examen de l’épidémiologie, des défis diagnostiques et de l’importance du tests virologique sur site du LCR dans les soins aux patients pédiatriques tropicaux   Les entérovirus et les parechovirus sont de plus en plus reconnus comme la cause de la méningite aseptique, en particulier dans le groupe d'âge pédiatrique. Cependant, en raison des caractéristiques cliniques indiscernables de la méningite bactérienne, de nombreux cliniciens ne peuvent pas faire une distinction claire dans la présentation de la maladie, et un grand nombre de cas ne sont pas diagnostiqués. Bien que la réaction en chaîne par polymérase soit l'approche diagnostique standard actuelle, il faut plusieurs heures ou jours pour obtenir un résultat et ces tests ne sont pas disponibles aux niveaux de soins primaires et secondaires dans de nombreux pays pauvres en ressources. En outre, le diagnostic est souvent difficile chez les enfants en raison de découvertes non spécifiques du liquide céphalo-rachidien cellulaire et biochimique. Certains enfants atteints peuvent développer des complications neurologiques ou systémiques, entraînant une hospitalisation  prolongée, augmentant le risque de décès évitables et les dépenses de santé. Cette revue se concentre sur l'épidémiologie, la présentation et le diagnostic de la méningite à entérovirus et parechovirus, mettant en évidence les défis du diagnostic et les rôles potentiels des tests virologiques sur place dans le LCR dans  l'amélioration de la qualité des soins aux patients pédiatriques. Les informations fournies devraient contribuer à la détection précoce des cas, garantissant ainsi d'éviter les antibiotiques inutiles, des complications minimales, une courte période d'hospitalisation et une réduction des coûts associés aux soins de santé. Mots clés: méningite aseptique; Entérovirus; Parechovirus; Défi diagnostique; Test de virologie sur place;  Enfant

Preparation of Cerium Oxide Nanoparticles and Their Cytotoxicity Evaluation In vitro and In vivo

Background: Nanotechnology plays a significant role in medicine, especially in diagnosis and treatment as a carrier to drugs and vaccinology. Several studies were conducted using nanoparticles as an adjuvant. The main aim of this study was in vivo and in vitro toxicity evaluation of synthesized Cerium Nanoparticles (CeNPs).Methods: In the present study, cerium nanoparticles were prepared using the wet chemical method. The formation of cerium nanoparticles was confirmed by scanning electron microscopy, transmission electron microscopes, x-ray diffraction analysis, dynamic light scattering. In vivo and in vitro toxicity of synthesized nanoparticles was evaluated in three different amounts of cerium nanoparticles (30 µg, 50 µg, & 100 µg) in mice and human fibroblast cell lines, respectively.Results: Cerium nanoparticles were successfully synthesized, and the identity was confirmed by x-ray diffraction analysis. The shape and size of nanoparticles were spherical and <100 nm, respectively. The prepared nanoparticles had a charge of -26.6 mV and a hydrodynamic radius of 446 nm. MTT assay indicated that none of the concentration of cerium was toxic, and in vivo toxicity also clarified the safety of cerium nanoparticles in mice; no significant un-normal behavioral and physical symptoms were observed in mice after CeNP administrationConclusion: Cerium nanoparticles have special properties, especially low toxicity, unique capabilities in stimulating the immune system. Cerium nanoparticles can be considered an effective and safe candidate in vaccines

Intra-peritoneal and intra-rectal immunogenicity induced by rotavirus virus like particles 2/6/7 in mice

We previously developed virus like particles of rotavirus (RV) with VP2, VP6, and VP7 proteins (VLP2/6/7) using stable High-five cell line. To evaluate the immunogenicity of our construct, we assessed the humoral and cytokine responses induced by VLP2/6/7 in BALB/c mice immunized intra-peritoneally and intra-rectally. Enzyme-linked immunosorbent assay (ELISA) and Relative quantitative (RQ) Real-time PCR were used to evaluate the antibody (IgG and IgA) levels in serum and mRNA levels of IL-6, IL-10 and IFN-gamma in spleen cells, respectively. Our results showed that VLP2/6/7 is capable of intra-peritoneal (I.P.) and intra-rectal (I.R.) induction of serum IgG and IgA responses. IgA was detected in fecal samples of immunization groups by I.P. and I.R. routes. Interestingly, I.R. route induced higher IgA titer compared with I.P. route which was statistically significant. Moreover, mRNA levels of IL-6 and IFN-gamma were significantly elevated in mice immunized intra- peritoneally with VLP2/6/7 compared to control group. As such, the mean change was 7.4 (P < 0.05) and 14.8 (P < 0.001) for IFN-gamma and IL-6, respectively. Likewise, the same pattern was found when mice were immunized intra-rectally. Although elevated, the difference in the mean change for IL-10 was not statistically significant when compared to control group. Our findings indicated that VLPs constructed via a stable insect cell line are able to induce both humoral and cellular responses, a similar pattern as observed after immunization with live RVs. (C) 2014 Elsevier Ltd. All rights reserved

Vaccine-associated Paralytic Poliomyelitis in Immunodeficient Children, Iran, 1995–2008

To determine the prevalence of vaccine-associated paralytic poliomyelitis (VAPP) in immunodeficient infants, we reviewed all documented cases caused by immunodeficiency-associated vaccine-derived polioviruses in Iran from 1995 through 2008. Changing to an inactivated polio vaccine vaccination schedule and introduction of screening of neonates for immunodeficiencies could reduce the risk for VAPP infection

Evaluation of therapeutic potency of human papillomavirus-16 E7 DNA vaccine alone and with interleukin-18 as a genetic adjuvant = Avaliação da potência terapêutica da vacina de DNA do papilomavírus humano-16 E7 isolada e com interleucina-18 como adjuvante genético

OBJETIVOS: Apesar da existência de vacinas preventivas eficazes contra o papilomavírus humano (HPV), são necessárias vacinas terapêuticas que desencadeiem respostas imunes mediadas por células para tratar infecções e malignidades estabelecidas. O objetivo deste estudo foi avaliar a potência terapêutica da vacina de ácido desoxirribonucleico (DNA) HPV-16 E7 isolada e com interleucina (IL)-18. MÉTODOS: Expressões in vitro de IL-18 foram realizadas em células renais embrionárias humanas 293 e confirmadas por Western blotting. A vacina de DNA foi disponibilizada em um estudo anterior. Um total de 45 camundongos fêmeas C57BL/6 divididos em cinco grupos (vacina de DNA, vacina de DNA adjuvada com IL-18, pcDNA3. 1 e solução salina tamponada com fosfato) e foram inoculados com linhagem murina-1 de carcinoma relacionado ao HPV, expressando antígenos E6 / E7 do HPV-16. Os animais foram então imunizados por via subcutânea duas vezes no intervalo de sete dias. A imunidade antitumoral e antígeno-celular específica foi avaliada pela proliferação de linfócitos (ensaio de brometo de 3- [4,5-dimetiltiazol-2-il] -2,5-difeniltetrazólio: MTT), ensaio de liberação de lactato desidrogenase, ensaio de IL-4 e ensaio de interferon-gama [IFN-γ]. O tamanho do tumor foi seguido por 62 dias. RESULTADOS: O ensaio MTT, que mede a proliferação de linfócitos em resposta ao antígeno específico, aumentou nos grupos de coadministração e de vacina de DNA em comparação aos grupos controle e adjuvante genético (p <0,001). Os camundongos imunizados com a coadministração geraram significativamente mais IFN-γ e IL-4 do que os outros camundongos imunizados (p<0,001) A redução do tamanho do tumor nos grupos de coadministração e de vacina de DNA foi significativamente mais acentuada do que nos grupos controle e adjuvante genético (p<0,001), mas não houve nenhuma diferença estatisticamente significativa entre os grupos vacina de DNA e coadministração (p=0,15). CONCLUSÕES: A IL-18 como adjuvante genético e a vacina de DNA E7 aumentaram as respostas imunes em sistemas modelo de camundongos contra o câncer cervical. No entanto, o uso de IL-18 como adjuvante genético com a vacina de DNA E7 não teve efeito sinérgico significativo nas respostas imunes in viv

Establishment of national primary immunodeficiency network, requisite of health organization and final stage of polio eradication: Review article

Primary immunodeficiency diseases (PIDs) is a diverse group of diseases, characterized by a defect in the immune system. These patients are susceptible to recurrent respiratory infections, gastrointestinal problems, autoimmune diseases, and malignancies. In most cases, patients with primary immunodeficiency disorders have genetic defects and are monogenic disorders that follow a simple Mendelian inheritance, however, some PIDs recognize a more complex polygenic origin. Overall, almost 70 to 90 percent of patients with primary immunodeficiency are undiagnosed. Given that these patients are exposing to respiratory infectious agents and some live-attenuated vaccines, thus they have a high risk to some clinical complications. The administration of oral polio vaccine in patients with PIDs especially can increase the possibility of acute flaccid paralysis. These patients will excrete the poliovirus for a long time through their feces, even though they are not paralyzed. Long-term virus proliferation in the vaccinated individuals causes a mutation in the poliovirus and creates a vaccine-derived polioviruses (VDPVs), which is a major challenge to the final stages of the worldwide eradication of polio. To increase the diagnosis and identification of patients with immunodeficiency and carrying out a national plan for screening patients with immunodeficiency from the fecal excretion of the poliovirus, a possible polio epidemic can be prevented during post-eradication. Development of laboratory facilities in provincial and city centers, improvement of communications among physicians regarding medical consultation and establishment of referring systems for patients by national network lead to improve status of diagnosis and treatment of patients with primary immunodefiicencies. In this context, launching and activating the national network of immunodeficiency diseases is essential for improving the health of children and reducing the cost of the health system of the country. A national network of immunodeficiency can lead to increase awareness of physiciansregarding primary immunodeficiency disorders, improve collaboration among physicians about genetic consultation and establish a practical referral system in Iran that resultsin increased diagnosis and improve treatment of patients with primary immunodeficiency disorders. © 2020 Tehran University of Medical Sciences. All rights reserved

Efficient inhibition of human immunodeficiency virus replication using novel modified microRNA-30a targeting 3'-untranslated region transcripts

RNA interference (RNAi)-based gene therapy is currently considered to be a combinatorial anti-human immunodeficiency virus-1 (HIV-1) therapy. Although arti­ficial polycistronic microRNAs (miRs) can reduce HIV-1 escape mutant variants, this approach may increase the risk of side effects. The present study aimed to optimize the efficiency of anti-HIV RNAi gene therapy in order to reduce the cell toxicity induced by multi-short hairpin RNA expression. An artificial miR-30a-3'-untranslated region (miR-3'-UTR) obtained from a single RNA polymerase II was used to simultaneously target all viral transcripts. The results of the present study demonstrated that HIV-1 replication was signifi­cantly inhibited in the cells with the miR-3'-UTR construct, suggesting that miR-3'-UTR may serve as a promising tool for RNAi-based gene therapy in the treatment of HIV-1. © 2016, Spandidos Publications. All Rights Reserved

Enhanced synergistic antitumor effect of a DNA vaccine with anticancer cytokine, MDA-7/IL-24, and immune checkpoint blockade

MDA-7/IL-24 cytokine has shown potent antitumor properties in various types of cancer without exerting any significant toxicity on healthy cells. It has also been proved to encompass pro-immune Th1 cytokine-like behavior. Several E7 DNA vaccines have developed against human papillomavirus (HPV)-related cervical cancer. However, the restricted immunogenicity has limited their clinical applications individually. To address this deficiency, we investigated whether combining the E7 DNA vaccine with MDA-7/IL-24 as an adjuvant would elicit efficient antitumor responses in tumor-bearing mouse models. Next, we evaluated how suppression of immunosuppressive IL-10 cytokine would enhance the outcome of our candidate adjuvant vaccine. Methods For this purpose, tumor-bearing mice received either E7 DNA vaccine, MDA-7/IL-24 cytokine or combination of E7 vaccine with MDA-7/IL-24 adjuvant one week after tumor challenge and boosted two times with one-week interval. IL-10 blockade was performed by injection of anti-IL-10 mAb before each immunization. One week after the last immunization, mice were sacrificed and the treatment efficacy was evaluated through immunological and immunohistochemical analysis. Moreover, the condition of tumors was monitored every two days for six weeks intervals from week 2 on, and the tumor volume was measured and compared within different groups. Results A highly significant synergistic relationship was observed between the E7 DNA vaccine and the MDA-7/IL-24 cytokine against HPV-16+ cervical cancer models. An increase in proliferation of lymphocytes, cytotoxicity of CD8+ T cells, the level of Th1 cytokines (IFN-γ, TNF-α) and IL-4, the level of apoptotic markers (TRAIL and caspase-9), and a decrease in the level of immunosuppressive IL-10 cytokine, together with the control of tumor growth and the induction of tumor regression, all prove the efficacy of adjuvant E7&IL-24 vaccine when compared to their individual administration. Surprisingly, vaccination with the DNA E7&IL-24 significantly reduced the population of Regulatory T cells (Treg) in the spleen of immunized mice compared to sole administration and control groups. Moreover, IL-10 blockade enhanced the effect of the co-administration by eliciting higher levels of IFN-γ and caspase-9, reducing Il-10 secretion and provoking the regression of tumor size. Conclusion The synergy between the E7 DNA vaccine and MDA-7/IL-24 suggests that DNA vaccines’ low immunogenicity can be effectively addressed by coupling them with an immunoregulatory agent. Moreover, IL-10 blockade can be considered a complementary treatment to improve the outcome of conventional or novel cancer therapies

Update on Immunodeficiency-Associated Vaccine-Derived Polioviruses - Worldwide, July 2018-December 2019.

Since establishment of the Global Polio Eradication Initiative* in 1988, polio cases have declined >99.9% worldwide; extensive use of live, attenuated oral poliovirus vaccine (OPV) in routine childhood immunization programs and mass campaigns has led to eradication of two of the three wild poliovirus (WPV) serotypes (types 2 and 3) (1). Despite its safety record, OPV can lead to rare emergence of vaccine-derived polioviruses (VDPVs) when there is prolonged circulation or replication of the vaccine virus. In areas with inadequate OPV coverage, circulating VDPVs (cVDPVs) that have reverted to neurovirulence can cause outbreaks of paralytic polio (2). Immunodeficiency-associated VDPVs (iVDPVs) are isolated from persons with primary immunodeficiency (PID). Infection with iVDPV can progress to paralysis or death of patients with PID, and excretion risks seeding cVDPV outbreaks; both risks might be reduced through antiviral treatment, which is currently under development. This report updates previous reports and includes details of iVDPV cases detected during July 2018-December 2019 (3). During this time, 16 new iVDPV cases were reported from five countries (Argentina, Egypt, Iran, Philippines, and Tunisia). Alongside acute flaccid paralysis (AFP) surveillance (4), surveillance for poliovirus infections among patients with PID has identified an increased number of persons excreting iVDPVs (5). Expansion of PID surveillance will facilitate early detection and follow-up of iVDPV excretion among patients with PID to mitigate the risk for iVDPV spread. This will be critical to help identify all poliovirus excretors and thus achieve and maintain eradication of all polioviruses