Drug treated cells resembled those expressing Nef, however combined Nef expression and drug treatment caused even more pronounced disruption of F-actin structures.
Virus release could be restored by expressing Nef in trans.
This issue is critical for designing a vaccine expressing Nef.
DNA and MVA vectors expressing Nef of the CRF12_BF recombinant form of HIV-1 were generated and characterized.
We then compared this to cells expressing Nef:MHC-I BiFC and Rab11-dsRed, in order to focus specifically on the MHC-I molecules that are targeted by Nef (Fig. 2a: panel 1).
Phorbol 12-myristate 13-acetate and phytohemagglutinin stimulation of Jurkat cells expressing Nef fails to produce the usual translocation of thetaPKC from the cytosol to the particulate fraction; translocation of betaPKC and epsilonPKC was unaffected.
Fifty and twenty five micrograms of TSP extracted from leaves expressing Nef, Nef/AMCV-P19 or Nef/TBSV-P19 and Nef/PVX-P25 were loaded on the ELISA plate as four indipendent replicas.
To our knowledge, no attempt of transiently expressing Nef as a single gene has been reported in literature until now.
Plant extracts were prepared from 10 g of both transgenic N. tabacum plant leaves (line 64) expressing Nef [ 15] and N. benthamiana fresh leaves agroinfiltrated with Nef/AMCV-P19.
These analyses showed that infected T cells expressing Nef alleles obtained after the loss of CD4+ T cells also expressed increased levels of CD69.
