I try to do CTL analysis in outbred animals that were immunized by a vaccine, such as CD1 mice, rabbits and dogs. It's hard to find such a protocol.
One reference established rabbit fibroblast cell line of each rabbit for the CTL assay.
Do you have any alternative suggestion on this?
I plan to use the same setting as the ELISPOT assay and measure the LDH releasing. However, with this setting, I can't show the different E:T ratio.
Does anyone has the experience on this?
Thanks a lot!
Any assay NOT using a cell line derived from the outbred animal will have the complicating factor of a being an MLR in addition to an CTL. It would be possible to do a partial splenectomy, prepare a single cell suspension, and freeze that suspension (potentially in aliquots) to use as target cells after the subject has been immunized. This of course has the potentially problematic factor that now the subject is no longer in it's naive state missing part of it's spleen . . . Alternatively repeated bleeds (especially with larger animals) might yield enough cells to use as targets.
You can please refer to the links below to obtain certain valuable informations:-
1. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2852241/
2. https://books.google.co.in/books?id=LY3L4Dm4-1YC&pg=PA118&lpg=PA118&dq=CTL+assay+in+outbred+animals&source=bl&ots=NbOBcxkWV6&sig=MWkcMObUB12ibfybdm_hh0_nOs0&hl=en&sa=X&ved=0CC8Q6AEwBGoVChMIlPCds_uZyAIV2AeOCh3VrQdr#v=onepage&q=CTL%20assay%20in%20outbred%20animals&f=false
Thank you, Hans and Sandip! It seems there is no simple method for CTL in out bred animals. Thans so much for you suggestions!
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