PMID- 27332822
OWN - NLM
STAT- MEDLINE
DCOM- 20170719
LR  - 20201215
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 11
IP  - 6
DP  - 2016
TI  - Recombinant AAV Vectors for Enhanced Expression of Authentic IgG.
PG  - e0158009
LID - 10.1371/journal.pone.0158009 [doi]
LID - e0158009
AB  - Adeno-associated virus (AAV) has become a vector of choice for the treatment of a 
      variety of genetic diseases that require safe and long-term delivery of a missing 
      protein. Muscle-directed gene transfer for delivery of protective antibodies 
      against AIDS viruses and other pathogens has been used experimentally in mice and 
      monkeys. Here we examined a number of variations to AAV vector design for the 
      ability to produce authentic immunoglobulin G (IgG) molecules. Expression of 
      rhesus IgG from a single single-stranded AAV (ssAAV) vector (one vector approach) 
      was compared to expression from two self-complementary AAV (scAAV) vectors, one 
      for heavy chain and one for light chain (two vector approach). Both the one 
      vector and the two vector approaches yielded considerable levels of expressed 
      full-length IgG. A number of modifications to the ssAAV expression system were 
      then examined for their ability to increase the efficiency of IgG expression. 
      Inclusion of a furin cleavage sequence with a linker peptide just upstream of the 
      2A self-cleaving sequence from foot-and-mouth disease virus (F2A) increased IgG 
      expression approximately 2 fold. Inclusion of these sequences also helped to 
      ensure a proper sequence at the C-terminal end of the heavy chain. Inclusion of 
      the post-transcriptional regulatory element from woodchuck hepatitis virus (WPRE) 
      further increased IgG expression 1.5-2.0 fold. IgG1 versions of the two rhesus 
      IgGs that were examined consistently expressed better than the IgG2 forms. In 
      contrast to what has been reported for AAV2-mediated expression of other 
      proteins, introduction of capsid mutations Y445F and Y731F did not increase 
      ssAAV1-mediated expression of IgG as determined by transduction experiments in 
      cell culture. Our findings provide a rational basis for AAV vector design for 
      expression of authentic IgG.
FAU - Fuchs, Sebastian P
AU  - Fuchs SP
AD  - Department of Pathology, Miller School of Medicine, University of Miami, Miami, 
      Florida, United States of America.
AD  - Institut fur Klinische und Molekulare Virologie, Friedrich-Alexander-Universitat 
      Erlangen-Nurnberg, Erlangen, Germany.
FAU - Martinez-Navio, Jose M
AU  - Martinez-Navio JM
AD  - Department of Pathology, Miller School of Medicine, University of Miami, Miami, 
      Florida, United States of America.
FAU - Gao, Guangping
AU  - Gao G
AD  - Gene Therapy Center, University of Massachusetts Medical School, Worcester, 
      Massachusetts, United States of America.
FAU - Desrosiers, Ronald C
AU  - Desrosiers RC
AD  - Department of Pathology, Miller School of Medicine, University of Miami, Miami, 
      Florida, United States of America.
LA  - eng
GR  - P01 AI100263/AI/NIAID NIH HHS/United States
GR  - P30 AI073961/AI/NIAID NIH HHS/United States
GR  - R01 AI098446/AI/NIAID NIH HHS/United States
GR  - U19 AI095985/AI/NIAID NIH HHS/United States
PT  - Journal Article
DEP - 20160622
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (DNA, Recombinant)
RN  - 0 (Immunoglobulin G)
SB  - IM
MH  - DNA, Recombinant/*metabolism
MH  - Dependovirus/*metabolism
MH  - Genetic Vectors/*metabolism
MH  - HEK293 Cells
MH  - Humans
MH  - Immunoglobulin G/*genetics
MH  - Staining and Labeling
MH  - Transduction, Genetic
MH  - Transgenes
PMC - PMC4917256
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2016/06/23 06:00
MHDA- 2017/07/20 06:00
PMCR- 2016/06/22
CRDT- 2016/06/23 06:00
PHST- 2016/03/15 00:00 [received]
PHST- 2016/06/08 00:00 [accepted]
PHST- 2016/06/23 06:00 [entrez]
PHST- 2016/06/23 06:00 [pubmed]
PHST- 2017/07/20 06:00 [medline]
PHST- 2016/06/22 00:00 [pmc-release]
AID - PONE-D-16-10918 [pii]
AID - 10.1371/journal.pone.0158009 [doi]
PST - epublish
SO  - PLoS One. 2016 Jun 22;11(6):e0158009. doi: 10.1371/journal.pone.0158009. 
      eCollection 2016.