Is IFN expression by NK cells a hallmark of severe COVID-19?

Natural Killer cells (NK) are crucial in host defense against viruses. There are many unanswered questions about the immune system in COVID-19, especially the mechanisms that contribute to the development of mild or severe forms of the disease. Although NK cells may have an essential role in the pathogenesis of COVID-19, the mechanisms involved in this process are not yet fully elucidated. Here, we demonstrate that CD3-CD56+ NK cells frequency in the volunteers who recovered from mild COVID-19 (Mild CoV) presented a significant increase compared to the healthy control (HC) and individuals recovering from severe COVID-19 (Severe CoV) groups. Furthermore, distinct IFN profiles in recovered COVID-19 patients with mild or severe clinical forms of the disease were observed in the total NK cells (CD3-CD56+). In the first group, NK cells express increased levels of IFN-α compared to the severe CoV, while higher production of IFN-γ in severe CoV was found. Moreover, NK cells in mild CoV express more cytolytic granules depicted by granzyme B and perforin. Compared to HC, PBMCs from mild CoV presented higher Ki-67 and TIM-3 production after Pool CoV-2 and Pool Spike CoV-2 peptides stimulus. In addition, non-stimulated PBMCs in the mild CoV group had higher NK TIM-3+ frequency than severe CoV. In the mild CoV group, Pool Spike CoV-2 and Pool CoV-2 peptides stimuli elicited higher granzyme B and perforin coexpression and IFN-α production by PBMCs. However, in severe CoV, Pool Spike CoV-2 reduced the coexpression of granzyme B, perforin, and CD107a suggesting a decrease in the cytotoxic activity of NK cells. Therefore, our study shows that NK cells may have a crucial role in COVID-19 with the involvement of IFN-α and cytotoxic properties that aid in developing qualified immune responses. Furthermore, the data suggest that higher amounts of IFN-γ may be linked to the severity of this disease.

Serological and molecular detection of Neospora caninum and Toxoplasma gondii in human umbilical cord blood and placental tissue samples.

Neosporosis primarily affects cattle and dogs and is not currently considered a zoonotic disease. Toxoplasmosis is a zoonosis with a worldwide distribution that is asymptomatic in most cases, but when acquired during pregnancy, it can have serious consequences. The seropositivity rates determined by the indirect fluorescent antibody test for Neospora caninum (N. caninum) and Toxoplasma gondii (T. gondii) were 24.3% (49 samples) and 26.8% (54 samples), respectively. PCR positivity for N. caninum was observed in two samples of cord blood (1%) using the Nc5 and ITS1 gene, positivity for T. gondii was observed in 16 samples using the primer for the B1 gene (5.5% positivity in cord blood and 2.5% positivity in placental tissue). None of the samples showed structures characteristic of tissue cysts or inflammatory infiltrate on histopathology. Significant associations were observed only between N. caninum seropositivity and the presence of domestic animals (p = 0.039) and presence of dogs (p = 0.038) and between T. gondii seropositivity and basic sanitation (p = 0.04). This study obtained important findings regarding the seroprevalence and molecular detection of N. caninum and T. gondii in pregnant women; however, more studies are necessary to establish a correlation between risk factors and infection.

Amblyomma scalpturatum Neumann, 1906 (Acari: Ixodidae): confirmation in Acre State, Brazil, and description of parasitism in a human / Amblyomma scalpturatum Neumann, 1906 (Acari: Ixodidae): confirmação no Estado do Acre, Brasil, e descrição de parasitismo em um humano

Abstract Amblyomma scalpturatum is a tick species that is unique to South America. It is commonly associated with the Amazon biome and has been reported in some Brazilian states. This tick species exhibits host specificity: it parasitizes tapirs and suidae. Its role in transmitting pathogens to humans is still unknown. Amblyomma scalpturatum is known to be a human-biting tick; however, there is only one report showing that humans make suitable hosts for this species. The knowledge of tick fauna is lacking in the Acre State. This study collected free-living ticks with the aim of finding new records in Acre State. Collections were carried out in Amazon forest fragments in Rio Branco municipality. An A. scalpturatum specimen was identified and submitted sequencing of the ITS-2 gene. This study presents the first molecular confirmation of A. scalpturatum collected in situ in Acre State, North Region, Brazil. This study also presents the first record of a successfully completed feeding by an A. scalpturatum nymph on a human host in the North region of Brazil. Further investigation is needed to ascertain the complete life cycle of this tick species, its seasonality in the environment, and its relationship to pathogens and competence to transmit them.

Resumo Amblyomma scalpturatum é uma espécie de carrapato que ocorre apenas na América do Sul. Está comumente associada ao bioma da Amazônia e tem sido relatada em alguns estados brasileiros. Esta espécie de carrapato apresenta especificidade a hospedeiros: parasita antas e suídeos. Sua competência em transmitir agentes patogênicos a seres humanos é ainda desconhecida. Amblyomma scalpturatum é também conhecido por picar seres humanos; entretanto, há apenas um relato que evidencie que seres humanos sejam capazes de serem hospedeiros adequados. Ainda existem lacunas no conhecimento sobre a ixodofauna no estado do Acre. Neste estudo foram coletados carrapatos de vida livre com o objetivo de encontrar novos registros para o estado do Acre. Coletas de carrapatos foram realizadas em fragmentos de floresta Amazônica no município de Rio Branco. Um exemplar de A. scalpturatum foi identificado e submetido a sequenciamento do gene ITS-2. Este estudo apresenta a primeira confirmação molecular de A. scalpturatum coletado in situ no estado do Acre, região Norte, Brasil. Este estudo também apresenta o primeiro relato de parasitismo completo de uma ninfa de A. scalpturatum em um hospedeiro humano na região Norte do Brasil. Mais investigações são necessárias para elucidar o ciclo de vida completo dessa espécie de carrapato, a sazonalidade de seus estádios no meio ambiente, sua relação a agentes patogênicos e competência em transmiti-los.

Analysis of Bm86 conserved epitopes: is a global vaccine against Cattle Tick Rhipicephalus microplus possible? / Análise de epítopos conservados da Bm86: é possível uma vacina global contra Carrapato-do-Boi Rhipicephalus microplus?

Abstract The cattle tick Rhipicephalus microplus causes significant economic losses in agribusiness. Control of this tick is achieved mainly through the application of chemical acaricides, often resulting in contamination of animal food products and of the environment. Another major concern associated with acaricide use is the increasing reports of resistance of this tick vector against the active ingredients of many commercial products. An alternative control method is vaccination. However, the commercially available vaccine based on a protein homologous to Bm86 exhibits variations in efficacy relative to the different geographical locations. This study aimed to identify antigenic determinants of the sequences of proteins homologous to Bm86. Phylogenetic analyses were performed to determine the extent of divergence between different populations of R. microplus to identify the sequence that could be used as a universal vaccine against the multiple geographically distinct populations of R. microplus and related tick species. Considering the extensive sequence and functional polymorphism observed among strains of R. microplus from different geographical regions, we can conclude that it may be possible to achieve effective vaccination against these cattle ticks using a single universal Bm86-based antigen.

Resumo O carrapato Rhipicephalus microplus é responsável por perdas significativas no agronegócio. O controle deste carrapato é feito principalmente por meio da aplicação de acaricidas químicos, geralmente resultando na contaminação de produtos de origem animal e do meio ambiente. Outra preocupação importante associada ao uso de acaricidas é o crescente aumento de relatos sobre a resistência deste carrapato a princípios ativos de vários produtos comerciais. Uma alternativa de controle é por meio de vacinação. Porém, a vacina comercializada contendo proteína homóloga à Bm86, apresenta variações de eficácia em relação às diferentes localizações geográficas. Este estudo buscou identificar determinantes antigênicos das sequencias de proteínas homólogas a Bm86. As análises filogenéticas foram feitas para determinar a extensão da divergência entre diferentes populações de R. microplus com o objetivo de identificar a sequência que poderia ser usada como vacina universal contra as múltiplas populações geograficamente distintas de R. microplus e espécies de carrapatos relacionados. Considerando-se a extensa sequência e o polimorfismo observados entre linhagens de R. microplus de diferentes regiões geográficas, podemos concluir que pode ser possível obter uma vacinação efetiva contra esses carrapatos bovinos utilizando um único antígeno universal baseado em Bm86.

Molecular characterization of the recombinant protein RmLTI-BmCG-LTB: Protective immunity against Rhipicephalus (Boophilus) microplus.

The bovine tick Rhipicephalus (Boophilus) microplus is found in several tropical and subtropical regions of the world. This parasite transmits pathogens that cause disease, such as babesiosis (Babesia bovis and B. bigemina) and anaplasmosis (Anaplasma marginale). Tick infestations cause enormous livestock losses, and controlling tick infestations and the transmission of tick-borne diseases remains a challenge for the livestock industry. Because the currently available commercial vaccines offer only partial protection against R. (B.) microplus, there is a need for more efficient vaccines. Several recombinant antigens have been evaluated using different immunization strategies, and they show great promise. This work describes the construction and immunological characterization of a multi-antigen chimera composed of two R. (B.) microplus antigens (RmLTI and BmCG) and one Escherichia coli antigen (B subunit, LTB). The immunogenic regions of each antigen were selected and combined to encode a single polypeptide. The gene was cloned and expressed in E. coli. For all of the experiments, two groups (treated and control) of four Angus heifers (3-6 months old) were used. The inoculation was performed via intramuscular injection with 200 µg of purified recombinant chimeric protein and adjuvated. The chimeric protein was recognized by specific antibodies against each subunit and by sera from cattle inoculated with the chimera. Immunization of RmLTI-BmCG-LTB cattle reduced the number of adult female ticks by 6.29% and vaccination of cattle with the chimeric antigen provided 55.6% efficacy against R. (B.) microplus infestation. The results of this study indicate that the novel chimeric protein is a potential candidate for the future development of a more effective vaccine against R. (B.) microplus.

New insights from molecular characterization of the tick Rhipicephalus (Boophilus) microplus in Brazil / Novos pontos de vista sobre a caracterização molecular em carrapatos Rhipicephalus (Boophilus) microplus, no Brasil

Abstract The Rhipicephalus (Boophilus) microplus complex currently consists of five taxa, namely R. australis, R. annulatus, R. (B.) microplus clade A sensu, R. microplus clade B sensu, and R. (B.) microplus clade C sensu. Mitochondrial DNA-based methods help taxonomists when they are facing the morpho-taxonomic problem of distinguishing members of the R. (B.) microplus complex. The purpose of this study was to perform molecular characterization of ticks in all five regions of Brazil and infer their phylogenetic relationships. Molecular analysis characterized 10 haplotypes of the COX-1 gene. Molecular network analysis revealed that haplotype H-2 was the most dispersed of the studied populations (n = 11). Haplotype H-3 (n = 2) had the greatest genetic differentiation when compared to other Brazilian populations. A Bayesian phylogenetic tree of the COX-1 gene obtained strong support. In addition, it was observed that the population of R. (B.) microplus haplotype H-3 exhibited diverging branches among the other Brazilian populations in the study. The study concludes that the different regions of Brazil have R. (B.) microplus tick populations with distinct haplotypes.

Resumo Carrapatos do complexo R. (B.) microplus se distribuem em cinco taxa: R. australis, R. annulatus, R. (B.) microplus clado A sensu R. microplus clado B sensue e R. (B.) microplus clado C sensu. Métodos baseados no DNA mitocondrial podem auxiliar taxonomistas quando há dificuldades em estabelecer diferenças morfológicas para distinguir membros do complexo R. (B.) microplus. O objetivo deste estudo foi a caracterização molecular e a inferência de relações filogenéticas em carrapatos de todas as cinco regiões geográficas do Brasil. Para o gene COX-1, a análise molecular caracterizou 10 haplótipos. Na análise molecular em rede foi observado que o haplótipo H-2 é o mais disperso entre as populações (n=11). O haplótipo H-3 (n=2) foi o que obteve maior diferenciação genética ao ser comparado com outras populações brasileiras. A árvore filogenética Bayesiana de gene COX-1 gerou suporte robusto e foi observado que a população de R. (B.) microplus haplótipo H-3 apresentou ramificação com divergência entre as outras populações brasileiras apresentadas neste estudo. Conclui-se que as populações brasileiras possuem diversidade haplotípica com divergência entre as diversas populações de R. (B.) microplus no Brasil.

Geographical distribution of Trypanosoma cruzi in triatomine vectors in the State of Mato Grosso do Sul, Brazil

Introduction This work presents the initial findings of a molecular epidemiological investigation of Trypanosoma cruzi in triatomine insects in State of Mato Grosso do Sul. Methods A total of 511 triatomines from different regions of the state were examined. Deoxyribonucleic acid (DNA) was extracted from the intestinal contents of the insects using phenol-chloroform-isoamyl alcohol (25:24:1). Polymerase chain reaction (PCR) using primers 121/122 targeting DNA kinetoplast (kDNA) was then performed to identify T. cruzi, and positive samples were subjected to PCR using the primer pair TcSC5D-F/R followed by restriction fragment length polymorphism (RFLP) with the restriction enzymes SphI and HpaI (1 U/reaction), cloning and sequencing. Results One hundred samples were positive for T. cruzi, and three discrete typing units (DTUs) were identified (TcI, TcII, and TcBat). Triatoma sordida had the highest T. cruzi occurrence (83.3%), and DTUs were found in three samples: 58.3% of the samples were TcI, 33.3% were TcII and 8.3% were TcBat. There was a clear geographical distribution of the DTUs throughout the state, with TcI, TcII and TcBat located in the center, TcI located in the east, and TcII located in the west. Conclusions This study showed the occurrence ...