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dc.contributor.authorMori, Masuko
dc.contributor.authorKurane, Ichiro
dc.contributor.authorJanus, Jurand
dc.contributor.authorEnnis, Francis A.
dc.date2022-08-11T08:09:31.000
dc.date.accessioned2022-08-23T16:34:10Z
dc.date.available2022-08-23T16:34:10Z
dc.date.issued1997-03-01
dc.date.submitted2009-03-10
dc.identifier.citation<p>J Leukoc Biol. 1997 Mar;61(3):338-45.</p>
dc.identifier.issn0741-5400 (Print)
dc.identifier.doi10.1002/jlb.61.3.338
dc.identifier.pmid9060457
dc.identifier.urihttp://hdl.handle.net/20.500.14038/38272
dc.description.abstractA number of studies suggest that cytokines may contribute to the pathogenesis of viral infections, including dengue. In this study, we developed a double immunocytochemical method and characterized cytokine-producing cells in the peripheral blood mononuclear cells (PBMC) of dengue virus-immune donors after in vitro stimulation with specific dengue antigens. We found that double immunostaining using immunoalkaline phosphatase (Vector blue) for cytokines [interferon-gamma (IFN-gamma), interleukin (IL) -2, -4, -1alpha, -1beta, and -6, tumor necrosis factor beta (TNF-beta), and TNF-alpha] and immunoperoxidase [diaminobenzidine (DAB)] for cell surface markers (CD3, CD4, CD8, CD20, and CD68) provided the best distinction of double-positive cells from single-positive or -negative cells. The number of IFN-gamma, IL-2, IL-4, and TNF-beta-positive cells increased 2 or 3 days after stimulation with specific dengue antigens. No or very few cytokine-producing cells were detected in the PBMC of non-immune donors stimulated with dengue antigens and the PBMC of immune donors stimulated with a control antigen. The analysis of cell surface markers showed that mainly CD4+ and CD8+ T cells produced these cytokines. The results obtained by immunocytochemistry were consistent with cytokine levels detected in the culture medium assayed by enzyme-linked immunosorbent assay. In conclusion, this double immunocytochemistry technique is suitable for the detection and characterization of cytokine-producing cells in PBMC. Furthermore, the results support the hypothesis that antigen-stimulated CD4+ and CD8+ T cells produce cytokines that may play a role in the pathogenesis of dengue virus infection.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=9060457&dopt=Abstract">Link to Article in PubMed</a></p>
dc.relation.urlhttps://doi.org/10.1002/jlb.61.3.338
dc.subjectAnimals
dc.subjectAntigens, Viral
dc.subjectCD4-Positive T-Lymphocytes
dc.subjectCD8-Positive T-Lymphocytes
dc.subjectCercopithecus aethiops
dc.subjectDengue Virus
dc.subjectHumans
dc.subjectInterferon Type II
dc.subjectInterleukin-2
dc.subjectInterleukin-4
dc.subjectInterleukins
dc.subjectLymphotoxin-alpha
dc.subjectMonocytes
dc.subjectVero Cells
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleCytokine production by dengue virus antigen-responsive human T lymphocytes in vitro examined using a double immunocytochemical technique
dc.typeArticle
dc.source.journaltitleJournal of leukocyte biology
dc.source.volume61
dc.source.issue3
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/oapubs/1145
dc.identifier.contextkey770123
html.description.abstract<p>A number of studies suggest that cytokines may contribute to the pathogenesis of viral infections, including dengue. In this study, we developed a double immunocytochemical method and characterized cytokine-producing cells in the peripheral blood mononuclear cells (PBMC) of dengue virus-immune donors after in vitro stimulation with specific dengue antigens. We found that double immunostaining using immunoalkaline phosphatase (Vector blue) for cytokines [interferon-gamma (IFN-gamma), interleukin (IL) -2, -4, -1alpha, -1beta, and -6, tumor necrosis factor beta (TNF-beta), and TNF-alpha] and immunoperoxidase [diaminobenzidine (DAB)] for cell surface markers (CD3, CD4, CD8, CD20, and CD68) provided the best distinction of double-positive cells from single-positive or -negative cells. The number of IFN-gamma, IL-2, IL-4, and TNF-beta-positive cells increased 2 or 3 days after stimulation with specific dengue antigens. No or very few cytokine-producing cells were detected in the PBMC of non-immune donors stimulated with dengue antigens and the PBMC of immune donors stimulated with a control antigen. The analysis of cell surface markers showed that mainly CD4+ and CD8+ T cells produced these cytokines. The results obtained by immunocytochemistry were consistent with cytokine levels detected in the culture medium assayed by enzyme-linked immunosorbent assay. In conclusion, this double immunocytochemistry technique is suitable for the detection and characterization of cytokine-producing cells in PBMC. Furthermore, the results support the hypothesis that antigen-stimulated CD4+ and CD8+ T cells produce cytokines that may play a role in the pathogenesis of dengue virus infection.</p>
dc.identifier.submissionpathoapubs/1145
dc.contributor.departmentCenter for Infectious Disease and Vaccine Research
dc.contributor.departmentDepartment of Medicine
dc.source.pages338-45


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