Data di Pubblicazione:
2004
Citazione:
Analisi della espressione di citochine in monociti bovini isolati / V. Pocacqua, A. Giordano, M. Rinaldi, F. Ceciliani, P. Sartorelli - In: Atti I Congresso Nazionale AIPVet[s.l] : Associazione Italiana Patologi Veterinari, 2004. (( convegno I Congresso Nazionale AIPVet tenutosi a Gargnano (BS).
Abstract:
Cells of monocyte lineage partecipate
in immune response by processing and presenting
antigens and by producing cytokines, membrane
receptors and growth factors that modulate
immune response. In order to study cellular
interactions in the immune responses of cattles, it
is necessary to set up reliable methods to a) obtain
highly enriched mononuclear phagocytes
populations and b) evaluate the expression ratio of
the cyotkines involved in immune response.
The present communication describes a method
for purify the CD14+ cells from bovine blood, and
a procedure to evaluate and quantify by Real
Time PCR the espression of cytokines and
membrane receptors that are most involved in the
modulation of immune response, including IL-1,
TNF-alpha, IL-8 receptor, IL-1 beta antagonist,
CD18, using GAPDH as housekeeping gene.
CD14+ cells have been purified by means of
magnetic sorting. The expression of cytokines has
been determined after mRNA extraction and
reverse trascription using commercial standard
procedures and Oligo dT primer. Real Time PCR
has been carried out by means of SYBR GREEN
technique on iCycler iQ Real-Time PCR
Detection System.
Relative quantitation of cytokines was calculated
as an n-fold difference in transcription of the
respective cytokines from control samples (2 -
ΔΔCt), using GAPDH as housekeeping gene.
Tipologia IRIS:
03 - Contributo in volume
Elenco autori:
V. Pocacqua, A. Giordano, M. Rinaldi, F. Ceciliani, P. Sartorelli
Link alla scheda completa:
Titolo del libro:
Atti I Congresso Nazionale AIPVet