Differences in the Distribution of Peripheral Blood Leukocyte and Lymphocyte Subsets in Pulmonary Tuberculosis Patients

Introduction: It has been suggested that individuals exposed to Mycobacterium tuberculosis can eliminate the infection through the innate immune response without the participation of the acquired immunity. However, T helper type 1 immune response is critical to control persistent infection. Objective: The purpose of this study was to evaluate the distribution of leukocyte and lymphocyte subsets in peripheral blood of individuals with pulmonary tuberculosis (TB) and positive Tuberculin Skin Test (TST). Methodology: In order to achieve that goal, we have collected blood sample from thirty-four treatment-naïve TB patients, fifty TST-positive and forty-one TST-negative individuals. Results: The evaluation has shown a significant reduction in the number of total lymphocytes, B cells, CD4+ and CD8+ T cells and percentage increase of NK cells, neutrophils and monocytes in TB patients, when compared to TSTpositive and TST-negative individuals. There was no statistical difference for leucocyte and lymphocyte subsets between TST-positive and TST-negative groups. Peripheral blood white cell counts change significantly at diagnosis. Conclusion: The quantification of these cells may support the diagnosis and monitoring of patients with pulmonary tuberculosis

Detection of IgG Anti-Leishmania Antigen by Flow Cytometry as a Diagnostic Test for Cutaneous Leishmaniasis

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2017-03-28T18:19:47Z No. of bitstreams: 1 Sampaio GP Detection of IgG....pdf: 2044916 bytes, checksum: c74411e57915fb616dd654af1a4d6359 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2017-03-29T16:19:14Z (GMT) No. of bitstreams: 1 Sampaio GP Detection of IgG....pdf: 2044916 bytes, checksum: c74411e57915fb616dd654af1a4d6359 (MD5)Made available in DSpace on 2017-03-29T16:19:14Z (GMT). No. of bitstreams: 1 Sampaio GP Detection of IgG....pdf: 2044916 bytes, checksum: c74411e57915fb616dd654af1a4d6359 (MD5) Previous issue date: 2016Grant-in-Aid for Scientific Research (C) 24593126 and 16K11836 from the Japan Society for the Promotion of Science, Tokyo, JapanUniversidade Federal da Bahia. Instituto de Ciências da Saúde. Laboratório de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Complexo Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Complexo Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, Brasil / Instituto Nacional de Ciências e Tecnologia - Doenças Tropicais. INCT-DT. Salvador, BA, BrasilUniversidade Federal da Bahia. Instituto de Ciências da Saúde. Laboratório de Imunologia. Salvador, BA, BrasilWeill Cornell Medical College. Division of Infectious Diseases. New York, USAUniversidade Federal da Bahia. Instituto de Ciências da Saúde. Laboratório de Imunologia. Salvador, BA, BrasilUniversidade Federal da Bahia. Complexo Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, Brasil / Instituto Nacional de Ciências e Tecnologia - Doenças Tropicais. INCT-DT. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Laboratório Avançado de Saúde Pública. Salvador, BA, BrasilUniversidade Federal da Bahia. Complexo Hospitalar Universitário Professor Edgard Santos. Serviço de Imunologia. Salvador, BA, Brasil / Instituto Nacional de Ciências e Tecnologia - Doenças Tropicais. INCT-DT. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Laboratório Avançado de Saúde Pública. Salvador, BA, BrasilDiagnosis of cutaneous leishmaniasis (CL) relies on clinical presentation, parasite isolation, histopathologic evaluation and positive Montenegro skin test. However, the low amounts of parasites in the lesion of these individuals make parasite isolation and histopatologic diagnosis unreliable, often leading to false-negative results. Also, 15% of people living in endemic areas have sub-clinical infection characterized by positive Montenegro skin test, which may contribute to misdiagnosis. Although the main Leishmania killing mechanism is through cell-mediated immune response, antibodies against Leishmania antigens are found in infected individuals. Here our goal was to develop a new serological technique using polystyrene microspheres sensitized with soluble Leishmania antigens as a tool for the detection of IgG in serum from CL patients by flow cytometry. To validate the assay we carried out a comparative test (ELISA) commonly used as a diagnostic test for parasitic diseases. To determine cross-reactivity we used serum from patients with Chagas disease, caused by a trypanosome that has several proteins with high homology to those of the Leishmania genus. We observed that the flow cytometry technique was more sensitive than the ELISA, but, less specific. Our results show that the flow cytometry serologic test can be used to confirm CL cases in L. braziliensis transmission areas, however, presence of Chagas disease has to be ruled out in these individuals

Evaluation of systemic inflammatory response and lung injury induced by Crotalus durissus cascavella venom.

This study investigated the systemic inflammatory response and mechanism of pulmonary lesions induced by Crotalus durissus cascavella venom in murine in the state of Bahia. In order to investigate T helper Th1, Th2 and Th17 lymphocyte profiles, we measured interleukin (IL) -2, IL-4, IL-6, IL-10, IL-17, tumor necrosis factor (TNF) and interferon gamma (IFN-γ) levels in the peritoneal fluid and macerated lungs of mice and histopathological alterations at the specific time windows of 1h, 3h, 6h, 12h, 24h and 48h after inoculation with Crotalus durissus cascavella venom. The data demonstrated an increase of acute-phase cytokines (IL-6 and TNF) in the first hours after inoculation, with a subsequent increase in IL-10 and IL-4, suggesting immune response modulation for the Th2 profile. The histopathological analysis showed significant morphological alterations, compatible with acute pulmonary lesions, with polymorphonuclear leukocyte (PMN) infiltration, intra-alveolar edema, congestion, hemorrhage and atelectasis. These findings advance our understanding of the dynamics of envenomation and contribute to improve clinical management and antiophidic therapy for individuals exposed to venom

Representative table and formulas used to calculate diagnostic tests performance.

<p>Representative table and formulas used to calculate diagnostic tests performance.</p

Number of individuals according to each parameter used for calculation of Flow cytometry test performance.

<p>Number of individuals according to each parameter used for calculation of Flow cytometry test performance.</p

Antibody titers assessed by ELISA.

<p>Anti-SLA IgG titers from healthy subjects (HS), cutaneous leishmaniasis patients (CL), Montenegro positive individuals (DTH+) and Chagas disease patients (CD), assessed by ELISA. Cut off was determined by a ROC curve containing absorbances from CL patients and healthy subjects. OD, optical density.</p

Gate strategy to determine beads population to be analyzed.

<p>A gate was performed based on FSC and SSC and than mean florescence intensity was assessed based on FITC staining.</p