Microbial Translocation Induces an Intense Proinflammatory Response in PatientsWith Visceral Leishmaniasis and HIV Type 1 Coinfection

Made available in DSpace on 2015-08-19T13:49:27Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) carmen_grippetal_IOC_2013.pdf: 1114513 bytes, checksum: 1f42f1059dc7ace46b709ed310bae90c (MD5) Previous issue date: 2013Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório Interdisciplinar de Pesquisas Médicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa sobre o Timo. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio Grande do Norte. Departamento de Bioquímica. Natal, RN, Brasil.Universidade Federal do Mato Grosso do Sul. Hospital Universitário. 7Hospital-Dia Profa Esterina Corsini. Campo Grande, MS, Brasil.Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo. São Paulo, SP, Brasil.Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo. São Paulo, SP, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas (IPEC). Rio de Janeiro, RJ, Brasil.Fundação de Medicina Tropical Dr. Heitor Vieira Dourado. Manaus, AM, BrasilFundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas (IPEC). Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio Grande do Norte. Departamento de Bioquímica. Natal, RN, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de AIDS e Imunologia Molecular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório Interdisciplinar de Pesquisas Médicas. Rio de Janeiro, RJ, Brasil / Universidade do Estado do Rio de Janeiro, Faculdade de Ciências Médicas. Disciplina de Parasitologia. Rio de Janeiro, RJ, Brasil.Background. Leishmania infection is a cofactor in the heightened cellular activation observed in patients with American visceral leishmaniasis and human immunodeficiency virus type 1 (HIV) infection, with or without progression to AIDS (AVL/HIV). Thus, the persistence of a high parasite load despite antileishmanial therapy could be responsible for the continued immune stimulation. Methods. CD8+ T cells expressing CD38, parasite load, lipopolysaccharide (LPS), soluble CD14, macrophage migration inhibitory factor (MIF), intestinal fatty acid–binding protein (IFABP), and proinflammatory cytokines (interleukin 1β, interleukin 6, interleukin 8, interleukin 17, interferon γ, and tumor necrosis factor) were measured in 17 patients with AVL/HIV, 16 with HIV, and 14 healthy subjects (HS). Results. Lower Leishmania parasitemia was observed after antileishmanial and antiretroviral therapies. However, higher levels of CD38+ on CD8+ T cells were observed in both clinical phases of leishmaniasis, compared with HIV cases. AVL/HIV and HIV patients showed higher levels of LPS and IFABP than HS. Proinflammatory cytokine levels were significantly augmented in patients with active coinfection, as well as those with remission of Leishmania infection. LPS levels and Leishmania infection were positively correlated with CD38 expression on CD8+ T cells and with IL-6 and IL-8 levels. Conclusions. LPS levels along with the immune consequences of Leishmania infection were associated with elevated cellular activation in coinfected patients. As a consequence, secondary chemoprophylaxis for leishmaniasis or even the use of antiinflammatory drugs or antibiotics may be considered for improving the prognosis of AVL/HIV

Improved Performance of ELISA and Immunochromatographic Tests Using a New Chimeric A2-Based Protein for Human Visceral Leishmaniasis Diagnosis

Summary. Human visceral leishmaniasis (VL) is a major public health problem worldwide, leading to significant mortality rates if not properly treated and controlled. Precise identification of infected patients is essential to establish treatment and control measures. Although several VL serological diagnosis advances have been accomplished lately, mainly using recombinant antigens and immunochromatographic tests (ICTs), improvements may still be achieved using multiepitope chimeric proteins in different test platforms. Here, we reported on the evaluation of ELISA and an ICT developed with a new chimeric protein, named DTL-4, based on repetitive antigenic sequences, including those present in the A2 protein. Methods. A total of 1028 sera samples were used for the development and validation of ELISA (321 samples from L. infantum-infected patients, 62 samples from VL/AIDS coinfected patients, 236 samples from patients infected with other diseases, and 409 samples from healthy donors). A total of 520 sera samples were used to develop and validate ICT (249 samples from L. infantum-infected patients, 46 samples from VL/AIDS coinfected patients, 40 samples from patients infected with other diseases, and 185 samples from healthy donors). Findings. Using the validation sera panels, DTL-4-based ELISA displayed an overall sensitivity of 94.61% (95% CI: 89.94-97.28), a specificity of 99.41% (95% CI: 96.39-99.99), and an accuracy of 97.02% (95% CI: 94.61-98.38), while for ICT, sensitivity, specificity, and accuracy values corresponded to 91.98% (95% CI: 86.65-95.39), 100.00% (95% CI: 96.30-100.00), and 95.14% (95% CI: 91.62-97.15), respectively. When testing sera samples from VL/AIDS coinfected patients, DTL-4-ELISA displayed a sensitivity of 77.42% (95% CI: 65.48-86.16), a specificity of 99.41% (95% CI: 96.39-99.99), and an accuracy of 93.51% (95% CI: 89.49%-96.10%), while for DTL-4-ICT, sensitivity was 73.91% (95% CI: 59.74-84.40), specificity was 90.63% (95% CI: 81.02-95.63), and accuracy was 82.00% (95% CI: 73.63-90.91). Conclusion. DTL-4 is a promising candidate antigen for serodiagnosis of VL patients, including those with VL/AIDS coinfection, when incorporated into ELISA or ICT test formats

Comparative genomics of two Leptospira interrogans serovars reveals novel insights into physiology and pathogenesis.

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2016-02-04T17:03:16Z No. of bitstreams: 1 Nascimento ATLO Comparative genomics of two....pdf: 540725 bytes, checksum: 25afb9fa02dcbdfe748ffdb0fedba3c5 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2016-02-12T13:21:56Z (GMT) No. of bitstreams: 1 Nascimento ATLO Comparative genomics of two....pdf: 540725 bytes, checksum: 25afb9fa02dcbdfe748ffdb0fedba3c5 (MD5)Made available in DSpace on 2016-02-12T13:21:56Z (GMT). No. of bitstreams: 1 Nascimento ATLO Comparative genomics of two....pdf: 540725 bytes, checksum: 25afb9fa02dcbdfe748ffdb0fedba3c5 (MD5) Previous issue date: 2004Instituto Butantan, Centro de Biotecnologia. São Paulo, SP, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Hospital Universitário Professor Edgard Santos, Serviço de Imunologia. Salvador, BA, BrasilWeill Medical College of Cornell University. Division of International Medicine and Infectious Disease. New York, NYInstituto Butantan, Centro de Biotecnologia. São Paulo, SP, BrasilUniversidade de São Paulo. Escola Superior de Agricultura Luiz de QueirozInstituto Butantan, Centro de Biotecnologia. São Paulo, SP, BrasilUniversity of California. Los Angeles School of Medicine. New York, New York / Veterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, CaliforniaVeterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, CaliforniaKIT (Koninklijk Instituut voor de Tropen / Royal Tropical Institute), KIT Biomedical Research. Amsterdam, The NetherlandsInstituto de Ciências Biomédicas. Universidade de São Paulo, SP, BrasilInstituto de Biociências. Universidade de São Paulo, SP, BrasilInstituto de Ciências Biomédicas. Universidade de São Paulo, SP, BrasilInstituto Butantan, Centro de Biotecnologia. São Paulo, SP, BrasilInstituto Butantan, Centro de Biotecnologia. São Paulo, SP, BrasilInstituto Butantan, Centro de Biotecnologia. São Paulo, SP, BrasilInstituto Oswaldo Cruz. Departamento de Bioquímica e Biologia Molecular. Rio de Janeiro, RJ, Brasil 11Universidade Federal de Pelotas. Centro de Biotecnologia. Pelotas, RGS, BrasilVeterans Affairs Greater Los Angeles Healthcare System. Division of Infectious Diseases. Los Angeles, California BrasilInstituto de Ciências Biomédicas. Universidade de São Paulo, SP, BrasilUniversidade de São Paulo. Faculdade de Ciências Agrárias eUniversidade de São Paulo. Faculdade de Ciências Agrárias eInstituto Butantan, Centro de Biotecnologia. São Paulo, SP, BrasilUniversidade Federal de São Carlos, Centro de Ciências Agrárias.Universidade Estadual de Feira de Santana (UEFS). Laboratório de Pesquisa em Microbiologia (LAPEM). Departamento de Ciências Biológicas. Feira de Santana, Bahia, BrasilFaculdade de Ciencias Farmaceuticas de Ribeirão Preto. Ribeirão Preto, SP, BrasilUniversidade de São Paulo. Faculdade de Filosofia, Ciências e Letras. Ribeirão Preto São Paulo, SP, BrasilUniversidade de São Paulo. Faculdade de Filosofia, Ciências e Letras. Ribeirão Preto São Paulo, SP, Brasil 17Instituto Biológico. Universidade de São Paulo, SP, BrasilUniversidade Federal do Rio Grande do Norte. Centro de Biociências. Natal, RGN, BrasilInstituto Butantan, Centro de Biotecnologia. São Paulo, SP, BrasilInstituto de Ciências Biomédicas. Universidade de São Paulo, SP, BrasilFaculdade de Ciências Agronômicas. São Paulo, SP, BrasilUniversidade de São Paulo. Faculdade de Ciências Agrárias eUniversidade de São Paulo. Faculdade de Ciências Agrárias eInstituto de Biociências. São Paulo, SP, BrasilUniversidade Estadual Paulista, Botucatu, Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, BrasilUniversidade Estadual Paulista, Botucatu, Núcleo Integrado de Biotecnologia, Universidade de Mogi das Cruzes, Mogi das Cruzes, SP, BrasilDepartamento de Genética e Evolução. Mogi das Cruzes, SP, BrasilInstituto de Computação. São Paulo, SP, BrasilUniversidade Federal da Bahia. Hospital Universitário Professor Edgard Santos, Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal do Rio Grande do Norte. Centro de Biociências. Natal, RGN, BrasilCentro de Energia Nuclear na Agricultura. São Paulo, SP, BrasilLudwig Institute for Cancer Research. New York, NYUniversidade de São Paulo. Faculdade de Ciências Agrárias e Veterinárias. São Paulo, SP, Brasil. Universidade Estadual Paulista, Jaboticabal. Piracicaba, SP, BrasilUniversidade de São Paulo. Escola Superior de Agricultura Luiz de Queiroz. São Paulo, SP, BrasilInstituto de Computação. São Paulo, SP, BrasilInstituto de Biociências. Universidade de São Paulo, SP, BrasilLeptospira species colonize a significant proportion of rodent populations worldwide and produce life-threatening infections in accidental hosts, including humans. Complete genome sequencing of Leptospira interrogans serovar Copenhageni and comparative analysis with the available Leptospira interrogans serovar Lai genome reveal that despite overall genetic similarity there are significant structural differences, including a large chromosomal inversion and extensive variation in the number and distribution of insertion sequence elements. Genome sequence analysis elucidates many of the novel aspects of leptospiral physiology relating to energy metabolism, oxygen tolerance, two-component signal transduction systems, and mechanisms of pathogenesis. A broad array of transcriptional regulation proteins and two new families of afimbrial adhesins which contribute to host tissue colonization in the early steps of infection were identified. Differences in genes involved in the biosynthesis of lipopolysaccharide O side chains between the Copenhageni and Lai serovars were identified, offering an important starting point for the elucidation of the organism's complex polysaccharide surface antigens. Differences in adhesins and in lipopolysaccharide might be associated with the adaptation of serovars Copenhageni and Lai to different animal hosts. Hundreds of genes encoding surface-exposed lipoproteins and transmembrane outer membrane proteins were identified as candidates for development of vaccines for the prevention of leptospirosis