Expression at mRNA level of cytokines and A238L gene in porcine blood-derived macrophages infected in vitro with African swine fever virus (ASFV) isolates of different virulence.
Porcine macrophage cultures were infected with two ASFV isolates of variable virulence and mRNA levels of several relevant macrophage-derived cytokines were quantified by real time PCR. At six hours post infection, a clear enhancement of mRNA expression of TNFalpha, IL6, IL12 and IL15 was observed in macrophages infected with the low virulent ASFV/NH/P68 (NHV) when compared to those infected with the highly virulent ASFV/L60 (L60). The sequence of the A238L gene homologue to the cellular IkappaB was found identical in both viral isolates and its expression at mRNA level was higher in macrophages infected with NHV when compared to macrophages infected with L60. Furthermore our results suggest a negative correlation between the mRNA expression of A238L gene and the mRNA expression of the above mentioned cytokines (with the exception of IL10) in L60 infected macrophages in opposition to the positive correlation (with exception of the IL1) suggested in NHV infection. Overall, our data strongly emphasize that virulence of ASFV isolates may depend on their capacity to regulate the expression of macrophage-derived cytokines relevant for the development of host protective responses by yet unknown mechanisms triggered by the virus at early stages of the cellular infection.
Gil
S
S
Laboratório Doenças Infecciosas, CIISA, Faculdade de Medicina Veterinária, Lisboa, Portugal.
Spagnuolo-Weaver
M
M
Canals
A
A
Sepúlveda
N
N
Oliveira
J
J
Aleixo
A
A
Allan
G
G
Leitão
A
A
Martins
C L V
CL
D000357:African Swine Fever / Q000276:immunology* / Q000821:virology; D000358:African Swine Fever Virus / Q000145:classification / Q000235:genetics / Q000472:pathogenicity*; D000818:Animals; D016207:Cytokines / Q000235:genetics*; D007375:Interleukin-1 / Q000235:genetics; D016753:Interleukin-10 / Q000235:genetics; D018664:Interleukin-12 / Q000235:genetics; D019409:Interleukin-15 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D008264:Macrophages / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis*; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*; D014774:Virulence
C113585:A238L protein, African swine fever virus; D016207:Cytokines; D007375:Interleukin-1; D019409:Interleukin-15; D015850:Interleukin-6; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D014764:Viral Proteins; D016753:Interleukin-10; D018664:Interleukin-12