PMID- 25781939
OWN - NLM
STAT- MEDLINE
DCOM- 20160204
LR  - 20181113
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 10
IP  - 3
DP  - 2015
TI  - Mycoplasma gallisepticum inactivated by targeting the hydrophobic domain of the 
      membrane preserves surface lipoproteins and induces a strong immune response.
PG  - e0120462
LID - 10.1371/journal.pone.0120462 [doi]
LID - e0120462
AB  - An innovative approach for inactivation of Mycoplasma gallisepticum using the 
      hydrophobic photoinduced alkylating probe 1, 5-iodonaphthylazide (INA) is 
      described. Treatment of washed M. gallisepticum mid-exponential culture (0.2 mg 
      cell protein /mL) with INA followed by irradiation with far-ultraviolet light 
      (310-380 nm) completely abolished viability. Transmission electron microscopy 
      showed that the majority of the inactivated M. gallisepticum were comparable in 
      size to intact cells, but that part of the INA-treated M. gallisepticum 
      preparation also contained low density cells and membrane vesicles. Confocal 
      microscopy revealed that untreated M. gallisepticum cells were internalized by 
      chicken red blood cells (c-RBCs), whereas the INA-inactivated cells remained 
      attached to the outer surface of the c-RBCs. INA treatment of M. gallisepticum 
      resulted in a complete inactivation of F0F1 -ATPase and of the L-arginine uptake 
      system, but the cytoplasmatic soluble NADH2 dehydrogenase was only partially 
      affected. Western blot analysis of the lipoprotein fraction showed that the 
      INA-treated M. gallisepticum retained their lipoproteins. Following subcutaneous 
      injection of M. gallisepticum INA-bacterin, 100% and 68.8% of chickens were 
      positive by the rapid serum agglutination test and enzyme-linked immunosorbent 
      assay respectively, 2 weeks post-injection. These data suggest that the 
      photoinducible alkylating agent INA inactivates M. gallisepticum but preserves 
      its surface lipoproteins and thus has the potential to be used as a general 
      approach for the inactivation of mycoplasmas for vaccine development.
FAU - Atalla, Hazem
AU  - Atalla H
AD  - Department of Microbiology and Molecular Genetics, The Hebrew University-Hadassah 
      Medical School, Jerusalem, Israel.
FAU - Lysnyansky, Inna
AU  - Lysnyansky I
AD  - Division of Avian and Aquatic Diseases, Kimron Veterinary Institute, Beit Dagan, 
      Israel.
FAU - Raviv, Yossef
AU  - Raviv Y
AD  - SAIC-Frederick Inc, National Cancer Institute, Frederick, Maryland, United States 
      of America.
FAU - Rottem, Shlomo
AU  - Rottem S
AD  - Department of Microbiology and Molecular Genetics, The Hebrew University-Hadassah 
      Medical School, Jerusalem, Israel.
LA  - eng
PT  - Journal Article
DEP - 20150317
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Bacterial Proteins)
RN  - 0 (Bacterial Vaccines)
RN  - 0 (Lipoproteins)
SB  - IM
MH  - Animals
MH  - Bacterial Proteins/chemistry/*immunology
MH  - Bacterial Vaccines/chemistry/immunology
MH  - Chickens/*immunology
MH  - Hydrophobic and Hydrophilic Interactions
MH  - Lipoproteins/chemistry/*immunology
MH  - Male
MH  - Mycoplasma gallisepticum/chemistry/*immunology
MH  - Protein Structure, Tertiary
PMC - PMC4363144
COIS- Competing Interests: During the course of the study Dr. Y. Raviv was an employee 
      of SAIC-Frederick. This does not alter the authors' adherence to PLOS ONE 
      policies on sharing data and materials.
EDAT- 2015/03/18 06:00
MHDA- 2016/02/05 06:00
PMCR- 2015/03/17
CRDT- 2015/03/18 06:00
PHST- 2014/10/02 00:00 [received]
PHST- 2015/01/22 00:00 [accepted]
PHST- 2015/03/18 06:00 [entrez]
PHST- 2015/03/18 06:00 [pubmed]
PHST- 2016/02/05 06:00 [medline]
PHST- 2015/03/17 00:00 [pmc-release]
AID - PONE-D-14-43925 [pii]
AID - 10.1371/journal.pone.0120462 [doi]
PST - epublish
SO  - PLoS One. 2015 Mar 17;10(3):e0120462. doi: 10.1371/journal.pone.0120462. 
      eCollection 2015.