I realized there are some papers showing that adenovirus activated inflammasome in primary mice bone marrow macrophages. However, still there are some papers using adenovirus over-expression or shRNA, as transient transfection is very difficult in these cells. Is adenovirus accepted as a useful in vitro tool to study inflammation or Inflammasome?
I would think not but some people dedicated to the platform might chose to ignore this. Most viral delivery systems activate innate immune responses. Likely dependent on numerous factors such as replication, subsequent infectivity, delivery of extra viral genes not directly relevant to the purpose of the platform.
I'm over at CIM and we could chat about this sometime if you like. We have good protocols for lentiviral development of modified BMMs if you want them.
OK, I will try adenovirus first to see if there is any change comparing with non-virus control and then discuss with you whether lentivirus will be a better choice.
Hello Rongrong,
A late follow up question: So how did your macrophages react to the adenoviral shRNA delicery? Was there inflammasome activation and IL-1beta response to the negative control shRNA?
I'm curious cause I've had my share of problems with cationic lipid-based transfection reagents in primary macrophages.
I used adenovirus in RAW264.7 and PM cells which worked well, it seems IL1 b expression is increased slightly by adenovirus comparing with negative control. The inflammasome activation I don't know actually. For the BMDM adenovirus does not work, I used lentivirus instead which gives a pretty good transduction efficacy and less inflammatory response.
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