PEGylation of bacteriophages increases blood circulation time and reduces T-helper type 1 immune response.
The increasing occurrence of antibiotic-resistant pathogens is of growing concern, and must be counteracted by alternative antimicrobial treatments. Bacteriophages represent the natural enemies of bacteria. However, the strong immune response following application of phages and rapid clearance from the blood stream are hurdles which need to be overcome. Towards our goal to render phages less immunogenic and prolong blood circulation time, we have chemically modified intact bacteriophages by conjugation of the non-immunogenic polymer monomethoxy-polyethylene glycol (mPEG) to virus proteins. As a proof of concept, we have used two different polyvalent and strictly virulent phages of the Myoviridae, representing typical candidates for therapeutical approaches: Felix-O1 (infects Salmonella) and A511 (infects Listeria). Loss of phage infectivity after PEGylation was found to be proportional to the degree of modification, and could be conveniently controlled by adjusting the PEG concentration. When injected into naïve mice, PEGylated phages showed a strong increase in circulation half-life, whereas challenge of immunized mice did not reveal a significant difference. Our results suggest that the prolonged half-life is due to decreased susceptibility to innate immunity as well as avoidance of cellular defence mechanisms. PEGylated viruses elicited significantly reduced levels of T-helper type 1-associated cytokine release (IFN-γ and IL-6), in both naïve and immunized mice. This is the first study demonstrating that PEGylation can increases survival of infective phage by delaying immune responses, and indicates that this approach can increase efficacy of bacteriophage therapy.
Institute of Food Science and Nutrition, ETH Zurich, Schmelzbergstrasse 7, 8092 Zurich, Switzerland.
Kwang-Pyo
KP
Kim
Jeong-Dan
JD
Cha
Eun-Hye
EH
Jang
Jochen
J
Klumpp
Steven
S
Hagens
Wolf-Dietrich
WD
Hardt
Kyung-Yeol
KY
Lee
Martin J
MJ
Loessner
D000818:Animals; D001691:Biological Therapy / Q000379:methods*; D001775:Blood Circulation; D004195:Disease Models, Animal; D006801:Humans; D008087:Listeria / Q000502:physiology / Q000821:virology; D008088:Listeriosis / Q000276:immunology / Q000382:microbiology / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D017900:Myoviridae / Q000737:chemistry* / Q000276:immunology* / Q000502:physiology; D011092:Polyethylene Glycols / Q000737:chemistry; D012480:Salmonella Infections / Q000276:immunology / Q000382:microbiology / Q000628:therapy*; D012486:Salmonella typhimurium / Q000502:physiology / Q000821:virology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology*; D013997:Time Factors
D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol
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