title: Molecular signatures distinguishing active from latent tuberculosis in peripheral blood mononuclear cells, after in vitro antigenic stimulation with purified protein derivative of tuberculin (PPD) or Candida: a preliminary report.
abstract: Purified protein derivative (PPD) or tuberculin skin testing is used to identify infected individuals with Mycobacterium tuberculosis (Mtb) and to assess cell-mediated immunity to Mtb. In the present study, we compared PBMC cultures in the presence of tuberculin or Candida antigens using cytokine bead arrays and RNA microarrays. Measurements of different cytokines and chemokines in supernatants of PMBC cultures in the presence of PPD showed increased levels of interferon (IFN)-gamma in active tuberculosis infection (ATBI) and latent TB infected (LTBI) compared to controls, and increased levels of TNF-alpha in ATBI compared with LTBI. Also, we found increase of IL-6 in cultures of PPD positive and controls but not in the cultures with Candida. We also report the molecular signature of tuberculosis infection, in ATBI patients, the following genes were found to be up-regulated and absent in LTBI individuals: two kinases (JAK3 and p38MAPK), four interleukins (IL-7, IL-2, IL-6, and IFNbeta1), a chemokine (HCC-4) a chemokine receptor (CxCR5), two interleukin receptors (IL-1R2 and IL-18R1), and three additional ones (TRAF5, Smad2, CIITA, and NOS2A). By contrast, IL-17 and IGFBP3 were significantly up-regulated in LTBI. And, STAT4, GATA3, Fra-1, and ICOS were down-regulated in ATBI but absent in LTBI. Conversely, TLR-10, IL-15, DORA, and IKK-beta were down-regulated in LTBI but not in ATBI. Interestingly, the majority of the up-regulated genes found in ATBI were found in cultures stimulated with tuberculin (PPD) or Candida antigens, suggesting that these pathogens stimulate similar immunological pathways. We believe that the molecular signature distinguishing active from latent tuberculosis infection may require using cytokine bead arrays along with RNA microarrays testing cell cultures at different times following in vitro proliferation assays using several bacterial antigens and PPD.
authors.affiliation: Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA 02115, USA. jstern@fas.harvard.edu
authors.forename: Joel N H
authors.initials: JN
authors.lastname: Stern
authors.forename: Derin B
authors.initials: DB
authors.lastname: Keskin
authors.forename: Viviana
authors.initials: V
authors.lastname: Romero
authors.forename: Joaquin
authors.initials: J
authors.lastname: Zuniga
authors.forename: Liliana
authors.initials: L
authors.lastname: Encinales
authors.forename: Changlin
authors.initials: C
authors.lastname: Li
authors.forename: Carlos
authors.initials: C
authors.lastname: Awad
authors.forename: Edmond J
authors.initials: EJ
authors.lastname: Yunis
mesh_terms: D000942:Antigens, Bacterial / Q000276:immunology; D002175:Candida / Q000276:immunology* / Q000472:pathogenicity; D002177:Candidiasis / Q000097:blood / Q000175:diagnosis / Q000276:immunology / Q000473:pathology; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D003937:Diagnosis, Differential; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D053616:Janus Kinase 3 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D055985:Latent Tuberculosis / Q000097:blood / Q000175:diagnosis / Q000276:immunology* / Q000473:pathology; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D046228:Microarray Analysis; D009169:Mycobacterium tuberculosis / Q000276:immunology* / Q000472:pathogenicity; D015398:Signal Transduction; D014157:Transcription Factors / Q000276:immunology; D014373:Tuberculin / Q000276:immunology; D014374:Tuberculin Test; D014397:Tuberculosis, Pulmonary / Q000097:blood / Q000175:diagnosis / Q000276:immunology* / Q000473:pathology; D014774:Virulence; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D016207:Cytokines; D014157:Transcription Factors; D014373:Tuberculin; C507933:JAK3 protein, human; D053616:Janus Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases