Estimating Point and Interval Frequency of Antigen-Specific CD4(+) T Cells Based on Short In Vitro Expansion and Improved Poisson Distribution Analysis (Articolo in rivista)

Type
Label
  • Estimating Point and Interval Frequency of Antigen-Specific CD4(+) T Cells Based on Short In Vitro Expansion and Improved Poisson Distribution Analysis (Articolo in rivista) (literal)
Anno
  • 2012-01-01T00:00:00+01:00 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi
  • 10.1371/journal.pone.0042340 (literal)
Alternative label
  • Di Lullo, Giulia; Ieva, Francesca; Longhi, Renato; Paganoni, Anna Maria; Protti, Maria Pia (2012)
    Estimating Point and Interval Frequency of Antigen-Specific CD4(+) T Cells Based on Short In Vitro Expansion and Improved Poisson Distribution Analysis
    in PloS one
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • Di Lullo, Giulia; Ieva, Francesca; Longhi, Renato; Paganoni, Anna Maria; Protti, Maria Pia (literal)
Pagina inizio
  • e42340 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume
  • 7 (literal)
Rivista
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#pagineTotali
  • 9 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroFascicolo
  • 8 (literal)
Note
  • ISI Web of Science (WoS) (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
  • Vita-Salute San Raffaele University; Vita-Salute San Raffaele University; Polytechnic University of Milan; Consiglio Nazionale delle Ricerche (CNR) (literal)
Titolo
  • Estimating Point and Interval Frequency of Antigen-Specific CD4(+) T Cells Based on Short In Vitro Expansion and Improved Poisson Distribution Analysis (literal)
Abstract
  • Methodology/Principal Findings: Purified blood CD4(+) T cells from healthy donors were cultured with autologous antigen presenting cells in several replicate wells in equal numbers in the absence (un-stimulated wells) or in the presence of synthetic peptides corresponding to viral antigens promiscuous HLA-DR epitopes (antigen-stimulated wells). At day 7 of culture low dose IL-2 was added and at day 14 IFN-gamma and IL-5 release in the supernatant was measured. A statistical analysis approach, based on Poisson distribution, was then implemented to calculate the frequency of viral-specific CD4(+) T cells. We first determined a patient-specific exceptionality threshold of cytokine release in the un-stimulated wells and then, based on this threshold, we counted the inactive/active wells within the antigen-stimulated wells. This number, along with the number of cells per well, allowed the point and interval estimates of frequencies. A ready-to-use Excel worksheet template with automatic calculations for frequencies estimate was developed and is provided as a supplemental file (Table S9). (literal)
  • Background: Knowledge of antigen-specific CD4(+) T cells frequencies is pivotal to the choice of the antigen to be used in anti-viral and anti-tumor vaccination procedures and for monitoring of immune responses. Methods that employ small cell numbers from patient samples, are easy to perform and do not require complex techniques/instrumentations and therefore standardization are desirable. (literal)
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