Filamentous Bacteriophage Produced

by Pseudomonas aeruginosa Alters the

Inflammatory Response and Promotes Noninvasive
Infection In Vivo

Pseudomonas aeruginosa is an opportunistic pathogen found as biofilm-

like aggregates in nonresolving inflammation and on medical devices. This
bacteria produce filamentous bacteriophage in vitro and in vivo, suggesting
that it may play a role in infection pathogenesis. When the gene
responsible of the creation of bacteriophage was deleted, it was showed
that the patogenicity of P. aeruginosa was not afected, contrary to other
studies. This study looks into for the interaction between bacteriophage and
the host immune response, showing that P. aeruginosa could escape host
defense mechanism through resistance to phagocytosis and tempered
inflammatory response.

For research P. aeruginosa PAO1 was infected by a strain Pf4,

superinfective causing lytin Pf4 replication. PAO1 produce very low
amounts of bacteriophages in broth cultures, but in the lungs of mice wild-
type PAO1 and PAO1+Pf4 produced equals quantities of phage. Half of the
mice infected with PAO1 died in the first 48 hours, with signs of
hemorrhage and edema in the survivors, while none of the mice with
PAO1+Pf4 showed signs of infection. The number of macrophages
increased in bouth groups, almost equally, but in the PAO1+Pf4 infected
mice were more CD45+ leukocytes, neutrophil chemokine, IL-1B, IL-17.
Also, an immunosuppressive factor IL-10 was higher in PAO1 infected
mice.

In both groups there was no difference in bacterial number at 24 hours post

infection, wich suggest that reduced lung injury in PAO1+Pf4 was not due
to lower bacterial loads, but after 48 hours there were fewer bacteria in
PAO1+Pf4. In BAL fluid there was always fewer bacteria in mice group
infected with PAO1+Pf4, wich suggest that PAO1+Pf4 was moreadherent
to the lung mucosa. Also speens from PAO1+Pf4 infected mice were
sterile, suggesting that the production of filamentous phage by P.
aeruginosa reduces inflammation and dissemination of the bacteria.
Motility of the two bacteria was the same, suggesting that there is no
reduced swimming motility of PAO1+Pf4. This type of bacteria showed
minimal twitching motility because type IV pili responsible of twitching are
receptors for Pf4. Pf phage increases the viscosity of polymers, such as
mucin and promotes beacterial adhesion to abiotic surfaces, an experiment
showing that there are more PAO1+Pf4 bacteria after washing a mucin
coated glass that initialy contained both bacteria.

Another explanation of decreased dissemination of PAO1+Pf4 is that both

PAO1 and PAO1+Pf4 escape equally the lungs, but PAO1+Pf4 grew more
slowly. To test this idea, another filamentous phage (fd) was used to infect
P. aeruginos, but this bacteriophage does not reproduce inside of PAO1.
Purified fd phage was mixed with PAO1 and added to the apical surface of
human tracheal epithelial cell cultures and was observed that bacteria was
less cytotoxic and less invasive.

There are two types of macrophages: M1 - proinflamatory and M2 –

antiinflamatory. Pf4 influeces macrophage polarization because when it is
plesent alongside with PAO1 it stimulates the differentiation of M2 and
inhibits the differentiation of M1.

In conclusion, this study show that filamentous Pf phage cause P.

aeruginosa to be less invasive, less inflammatory and more resistant to
phagocytosis, wich reduce its virulence.