Serum cytokines associated with severity and complications of kala-azar

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2013-11-25T18:51:32Z No. of bitstreams: 1 Costa D Serum cytokines....pdf: 467811 bytes, checksum: 85431be177c9e68c0cfe228a632e4fbf (MD5)Made available in DSpace on 2013-11-25T18:51:32Z (GMT). No. of bitstreams: 1 Costa D Serum cytokines....pdf: 467811 bytes, checksum: 85431be177c9e68c0cfe228a632e4fbf (MD5) Previous issue date: 2013Federal University of Piauí. Maternal and Childhood Department. Piauí, Brasil / Institute of Tropical Medicine ‘Natan Portella’. Laboratory of Leishmaniasis. Piauí, BrasilFederal University of Minas Gerais. Department of Pediatrics. Minas Gerais, BH, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratorio de Imunoparasitologia. Salvador, BA, BrasilInstitute of Tropical Medicine ‘Natan Portella’. Laboratory of Leishmaniasis. Piauí, Brasil. / Federal University of Minas Gerais. Department of Community Medicine. Minas Gerais, BH, BrasilFederal University of Minas Gerais. Department of Community Medicine. Minas Gerais, BH, BrasilInstitute of Tropical Medicine ‘Natan Portella’. Laboratory of Leishmaniasis. Piauí, Brasil. / Federal University of Minas Gerais. Department of Community Medicine. Minas Gerais, BH, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Imunoregulação. Salvador, Bahia, BrasilFundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório de Imunoparasitologia. Salvador, BA, BrasilObjectives: Recent clinical data suggest that severe kala-azar (or visceral leishmaniasis) is an exaggerated innate immune response mediated by inflammatory cytokines, leading to a systemic inflammatory syndrome similar to what is observed in malaria, sepsis and other diseases. We tested this hypothesis by measuring serum cytokines in individuals with kala-azar. Methods: We compared patients with severe kala-azar (i.e. hemorrhagic manifestations, n538) with patients without evidence of hemorrhage (n596). We conducted a detailed clinical and laboratory evaluation, measuring serum IL-1beta, IL-6, IL-8, IL-10, IL-12, interferon-gamma, and TNF-alpha, and markers of disseminated intravascular coagulation (DIC). Results: Infants had higher levels of inflammatory cytokines, while HIV-infected patients had lower concentrations of IL-10 and interferon-gamma. Higher levels of IL-6, interferon-gamma, and IL-8 were found among deceased patients. IL-8 and interferon-gamma were independently associated with bleeding. Several cytokines were associated with different signs of severe clinical and laboratory manifestations, including DIC. IL-6 was highly positively and independently associated with IL-1beta, IL-8, IL-10, and negatively associated with TNF-alpha. IL-1beta and TNF-alpha were also highly independently associated with disease severity. Conclusion: In its severe form, kala-azar, a neglected tropical disease, initiates a systemic inflammatory response that leads to DIC and other manifestations. Children may have higher risk of death due to the more intense cytokine release. The data supports the notion that IL-6 is the central cytokine that is associated with lethal disease, but interferon-gamma, IL1beta, IL-8, and TNF-alpha are also involved with disease severity. Inhibition of IL-6 is a potential target of adjuvant therapy for severe or pediatric forms of this disease

Evaluation of the novel <i>Leishmania infantum</i> recombinant antigens for the diagnosis of visceral leishmaniasis in humans.

<p>A panel of human sera derived from individuals with confirmed visceral leishmaniasis was tested through an Enzyme-Linked ImmunoSorbent Assay (ELISA) with the recombinant antigens produced in this study, the rK39 and the total <i>L</i>. <i>infantum</i> lysate (LAg). The panel was composed of 50 serum samples from individuals with visceral leishmaniasis (VL), 50 serum samples from healthy individuals (Healthy), 23 serum samples from individuals with cutaneous leishmaniasis (CL) and 40 serum samples from individuals with Chagas disease (Chagas). Each symbol corresponds to the result obtained with an individual serum. The horizontal lines indicate cutoff values, calculated by the means of results obtained with serum samples from healthy donors plus three standard deviations.</p

Schematic representation of the various gene fragments and corresponding deduced recombinant proteins evaluated in this study.

<p>The maps were derived from the sequences produced after direct sequencing or from the coding genomic sequences available at TriTrypDb.</p

Performance of the recombinant antigens with dog sera.

<p>LAg represents the total <i>L</i>. <i>infantum</i> lysate used as positive control. C.I. stands for confidence interval. NE—Not Evaluated.</p

TriTrypDb accession numbers for the newly identified antigenic proteins.

<p>TriTrypDb accession numbers for the newly identified antigenic proteins.</p

Polyacrilamide gel electrophoresis showing the affinity purified polypeptides evaluated in this study.

<p>(A) His-tagged, recombinant fragments of Lci6, Lci7, Lci9, Lci10, Lci11 and Lci12 in denaturing 15% SDS-PAGE stained with Coomassie Blue. All lanes shown are from a single gel but selected regions were removed for clarity. For some of these polypeptides, especially the larger ones, bands of lower molecular weight can be generally seen, but these are likely degradation products due to proteolysis within the bacteria that vary in intensity between different batches of purification. (B) The same for Lci1 and Lci13. The numbers on the left indicate the sizes of molecular weight markers.</p

Reactivity of an antigenic mix with human and canine sera positive for visceral leishmaniasis.

<p>(A) Reactivity of an antigenic mix with human sera from individuals with confirmed visceral leishmaniasis (VL), cutaneous leishmaniasis (CL), Chagas’ disease (Chagas) and healthy controls (Healthy). (B) ROC curves displaying the performance with the human sera of the protein “Mix”, the individual recombinant proteins that were included in the “Mix”, rK39 and the total <i>L</i>. <i>infantum</i> lysate (LAg). (C) Serum reactivity of dogs with canine leishmaniasis (CanL) and healthy control animals (Healthy) with the same protein “Mix”. (D) SROC curve showing the performance with the canine sera of the protein “Mix”, the individual recombinant proteins that were included in the “Mix”, rK39 and the total <i>L</i>. <i>infantum</i> lysate.</p

Increased interregional virus exchange and nucleotide diversity outline the expansion of chikungunya virus in Brazil

Abstract The emergence and reemergence of mosquito-borne diseases in Brazil such as yellow fever, zika, chikungunya, and dengue have had serious impacts on public health. Concerns have been raised due to the rapid dissemination of the chikungunya virus across the country since its first detection in 2014 in Northeast Brazil. In this work, we carried out on-site training activities in genomic surveillance in partnership with the National Network of Public Health Laboratories that have led to the generation of 422 chikungunya virus genomes from 12 Brazilian states over the past two years (2021–2022), a period that has seen more than 312 thousand chikungunya fever cases reported in the country. These genomes increased the amount of available data and allowed a more comprehensive characterization of the dispersal dynamics of the chikungunya virus East-Central-South-African lineage in Brazil. Tree branching patterns revealed the emergence and expansion of two distinct subclades. Phylogeographic analysis indicated that the northeast region has been the leading hub of virus spread towards other regions. Increased frequency of C > T transitions among the new genomes suggested that host restriction factors from the immune system such as ADAR and AID/APOBEC deaminases might be driving the genetic diversity of the chikungunya virus in Brazil