Expression of proinflammatory citokines of alpaca (Vicugna pacos) leukocytes induced by extract of macrocyst of Sarcocystis aucheniae

The aim of this research was to determine the expression of proinflammatory cytokines from alpaca leukocytes through the antigenic challenge with macrocyst extract of Sarcocystis aucheniae at various doses and exposure times. Leukocytes in a concentration of 500 000 cells/ml were exposed to concentrations of 0.5, 1, 50, 500 and 1000 ng of macrocyst extract of S. aucheniae and incubated for 1, 12 and 24 h. The total messenger RNA (mRNA) was extracted for each treatment using Trizol and used to perform realtime RTPCR with specific primers for cytokine TNFα and interleukins IL1α, IL1β and IL6. The generated mRNA levels of IL1α and TNF-α at 1 h were detectable and higher in comparison to the calibrator (leukocytes not exposed to the extract). IL1β increased at the 1 ng/ml concentration at 24 h showing a negative kinetic expression when compared with the untreated control group. IL6 was not evidenced by RTPCR real time. In addition, the leukocytes viability at 1, 12 and 24 h corroborated the toxic effect of the macrocyst extract of S. aucheniae at high concentrations.