16 research outputs found
Foreword (letter)
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Previous issue date: 2000Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.In 1997, Dr Sérgio Coutinho invited me to consider organizing, in 1998, a symposium to commemorate the ten years of flow cytomery in Brazil. The purpose of the meeting was to bring together several professionals who have contributed in someway to the development of this technology in our scientific milieu. The symposium took place between September 29-30, 1998 at Fundação Oswaldo Cruz, Fiocruz, Rio de Janeiro and consisted of round-tables emphasizing the applications of flow citomery to some important fields including cytokines, membrane receptors, cellular functions, cancer and Aids. The scientific content of the meeting made it both unique and very informative. The discussion generated by the presentations highlighted the importance of flow cytomery as a tool for studying several phenomena in clinical as well as basic biological research.
Sincere acknowledgments are indispensable to the co-organizers Wilson Savino, Geraldo Pereira, Rodrigo CorreaOliveria and especially to Marta Santiago for her invaluable assistance and extraordinary organizational acumen. We are equally grateful to all the speakers, administratrive assistans and participans Furthermore, none of this would have been possible without the generous financial and moral support provided by the Oswaldo Cruz Institute and Beckman-Colter, Brazil
Foreword
In 1997, Dr Srgio Coutinho invited me to consider organizing, in 1998,
a symposium to commemorate the ten years of flow cytomery in Brazil.
The purpose of the meeting was to bring together several professionals
who have contributed in someway to the development of this technology
in our scientific milieu. The symposium took place between September
29-30, 1998 at Fundao Oswaldo Cruz, Fiocruz, Rio de Janeiro and
consisted of round-tables emphasizing the applications of flow citomery
to some important fields including cytokines, membrane receptors,
cellular functions, cancer and Aids. The scientific content of the
meeting made it both unique and very informative. The discussion
generated by the presentations highlighted the importance of flow
cytomery as a tool for studying several phenomena in clinical as well
as basic biological research
Foreword
In 1997, Dr Srgio Coutinho invited me to consider organizing, in 1998,
a symposium to commemorate the ten years of flow cytomery in Brazil.
The purpose of the meeting was to bring together several professionals
who have contributed in someway to the development of this technology
in our scientific milieu. The symposium took place between September
29-30, 1998 at Fundao Oswaldo Cruz, Fiocruz, Rio de Janeiro and
consisted of round-tables emphasizing the applications of flow citomery
to some important fields including cytokines, membrane receptors,
cellular functions, cancer and Aids. The scientific content of the
meeting made it both unique and very informative. The discussion
generated by the presentations highlighted the importance of flow
cytomery as a tool for studying several phenomena in clinical as well
as basic biological research
The sample processing time interval as an influential factor in flow cytometry analysis of lymphocyte subsets
The objective of this paper is to propose a protocol to analyze blood
samples in yellow fever 17DD vaccinated which developed serious adverse
events. We investigated whether or not the time between sample
collection and sample processing could interfere in lymphocyte subset
percentage, for it is often impossible to analyze blood samples
immediately after collection due to transport delay from collection
places to the flow cytometry facility. CD4+CD38+ T, CD8+CD38+ T, CD3+
T, CD19+ B lymphocyte subsets were analyzed by flow cytometry in nine
healthy volunteers immediately after blood collection and after
intervals of 24 and 48 h. The whole blood lysis method and gradient
sedimentation by Histopaque were applied to isolate peripheral blood
mononuclear cells for flow cytometry analyses. With the lysis method,
there was no significant change in lymphocyte subset percentage between
the two time intervals (24 and 48 h). In contrast, when blood samples
were processed by Histopaque gradient sedimentation, time intervals for
sample processing influenced the percentage in T lymphocyte subsets but
not in B cells. From the results obtained, we could conclude that the
whole blood lysis method is more appropriate than gradient
sedimentation by Histopaque for immunophenotyping of blood samples
collected after serious adverse events, due to less variation in the
lymphocyte subset levels with respect to the time factor
Lymphocyte subset analyses in healthy adults vaccinated with yellow fever 17DD virus
In this study the kinetics of humoral and cellular immune responses in first-time vaccinees and re-vaccinees with the yellow fever 17DD vaccine virus was analyzed. Flow cytometric analyses were used to determine percentual values of T and B cells in parallel to the yellow fever neutralizing antibody production. All lymphocyte subsets analyzed were augmented around the 30th post vaccination day, both for first-time vaccinees and re-vaccinees. CD3+ T cells increased from 30.8% (SE ± 4%) to 61.15% (SE ± 4.2%), CD4+ T cells from 22.4% (SE ± 3.6%) to 39.17% (SE ± 2%) with 43% of these cells corresponding to CD4+CD45RO+ T cells, CD8+ T cells from 15.2% (SE ± 2.9%) to 27% (SE ± 3%) with 70% corresponding to CD8+CD45RO+ T cells in first-time vaccinees. In re-vaccinees, the CD3+ T cells increased from 50.7% (SE ± 3%) to 80% (SE ± 2.3%), CD4+ T cells from 24.9% (SE ± 1.4%) to 40% (SE ± 3%) presenting a percentage of 95% CD4+CD45RO+ T cells, CD8+ T cells from 19.7% (SE ± 1.8%) to 25% (SE ± 2%). Among CD8+CD38+ T cells there could be observed an increase from 15 to 41.6% in first-time vaccinees and 20.7 to 62.6% in re-vaccinees. Regarding neutralizing antibodies, the re-vaccinees presented high titers even before re-vaccination. The levels of neutralizing antibodies of first-time vaccinees were similar to those presented by re-vaccinees at day 30 after vaccination, indicating the success of primary vaccination. Our data provide a basis for further studies on immunological behavior of the YF 17DD vaccine
Detection of Intracytoplasmic Cytokines by Flow Cytometry
Flow cytometry has been used as a powerful technique for studying cell
surface antigen expression as well as intracellular molecules. Its
capability of analyzing multiple parameters simultaneously on a single
cell has allowed identification and studies of functional cell subsets
within heterogeneous populations. In this respect, several techniques
have been developed during the past few years to study
cytokine-producing cells by flow cytometry in humans and several animal
models
Immunophenotyping in Saliva as an Alternative Approach for Evaluation of Immunopathogenesis in Chronic Periodontitis
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Previous issue date: 2014Universidade do Estado do Amazonas (UEA). Escola de Ciências da Saúde. Faculdade de Odontologia. Manaus, AM, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Plataforma de Citometria de Fluxo – Purificação Celular. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas Leônidas e Maria Deane. Departamento de Biodiversidade em Saúde. Manaus, Am, Brasil.Universidade Federal do Amazonas (UFA). Instituto de Ciências Biológicas. Departamento de Parasitologia. Laboratório de Imunologia. Manaus, AM, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Plataforma de Citometria de Fluxo – Purificação Celular. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, Brasil.Universidade Federal do Amazonas (UFA). Instituto de Ciências Biológicas. Departamento de Parasitologia. Laboratório de Imunologia. Manaus, AM, Brasil.Background: To date, flow cytometric immunophenotyping has not been used to investigate immune patterns in saliva samples from individuals with inflammatory processes in the oral cavity, such as chronic periodontitis (CP). Saliva analysis could be a non-invasive method for evaluating oral health. The objective of this study is to determine the phenotype of leukocytes and total immunoglobulin A (IgA), IgG, and IgM titers in the saliva of individuals with CP.
Methods: Saliva samples were obtained from patients with CP (n = 12) and from a control group (n = 27) without oral diseases. Flow cytometry was performed to determine the frequency of T cells (CD4+ and CD8+), B cells, and natural killer (NK) cells as well as the total leukocyte population. Immunoglobulin titers were determined by dot enzyme-linked immunosorbent assay.
Results: Cell immunophenotyping revealed that patients with CP had a higher frequency of total leukocytes (47.94% ± 5.1%; P < 0.001), B cells (43.93% ± 6.2%; P = 0.006), NK cells (0.16% ± 0.04%; P = 0.03), and CD4+ T cells (38.99% ± 4.4%; P = 0.002) than individuals without oral pathologies (24.75% ± 2.2%, 20.60% ± 2.7%, 0.09% ± 0.03%, and 16.82% ± 3.5%, respectively). No significant differences in salivary total IgA, IgG, and IgM titers were found between the two cohorts studied. Nevertheless, higher total IgG levels were observed in patients with CP, which could indicate a possible correlation between clinical attachment level and salivary IgG (P = 0.07; r2 = 0.08).
Conclusion: These results show that cell phenotyping by flow cytometry could be an effective tool for determining leukocyte profiles in saliva samples from patients with CP and healthy individuals