Pseudomonas aeruginosa is member of the Gamma Proteobacteria class of Bacteria. It is a Gram-negative, aerobic rod belonging to the bacterial family Pseudomonadaceae. Since the revisionist taxonomy based on conserved macromolecules (e.g. 16S ribosomal RNA) the family includes only members of the genus Pseudomonas which are cleaved into eight groups. Pseudomonas aeruginosa is the type species of its group. which contains 12 other members. Pseudomonas aeruginosa is an opportunistic pathogen, meaning that it exploits some break in the host defenses to initiate an infection. In fact, Pseudomonas aeruginosa is the epitome of an opportunistic pathogen of humans. The bacterium almost never infects uncompromised tissues, yet there is hardly any tissue that it cannot infect if the tissue defenses are compromised in some manner. It causes urinary tract infections, respiratory system infections, dermatitis, soft tissue infections, bacteremia, bone and joint infections, gastrointestinal infections and a variety of systemic infections, particularly in patients with severe burns and in cancer and AIDS patients who are immunosuppressed. Pseudomonas aeruginosa infection is a serious problem in patients hospitalized with cancer, cystic fibrosis, and burns. The case fatality rate in these patients is near 50 percent (Textbook of Bacteriology).
4. Microbial Pathogenesis
P. aeruginosa infections are widely varied. The bacteria can colonize wounds and enter the blood stream, they can colonize the lungs, and they can also colonize the a scratched cornea. P. aeruginosa has a variety of virulence factors that contribute to its widely varied pathogenesis (Salyers and Whitt., 2002).
5. Host Ranges and Animal Models
Pseudomonas aeruginosa is an opportunistic human pathogen. The burned mouse model is used to model pseudomonas infection in burn patients. For lung infections guinea pig and rat models are used. C. elegans is also a model used to study host-pathogen interactions (Salyers and Whitt., 2002).
6. Host Protective Immunity
Most strains of P. aeruginosaare resistant to killing in serum alone, but the addition of polymorphonuclear leukocytes results in bacterial killing. Killing is most efficient in the presence of type-specific opsonizing antibodies, directed primarily at the antigenic determinants of LPS. This suggests that phagocytosis is an important defense and that opsonizing antibody is the principal functional antibody in protecting from P. aeruginosa infections. Once P. aeruginosa infection is established, other antibodies, such as antitoxin, may be important in controlling disease. Cell-mediated immunity does not seem to play a major role in resistance or defense against Pseudomonas infections (Textbook of Bacteriology).
>NP_252234.1 alginate production protein AlgE [Pseudomonas aeruginosa PAO1]
MNSSRSVNPRPSFAPRALSLAIALLLGAPAFAANSGEAPKNFGLDVKITGESENDRDLGTAPGGTLNDIG
IDLRPWAFGQWGDWSAYFMGQAVAATDTIETDTLQSDTDDGNNSRNDGREPDKSYLAAREFWVDYAGLTA
YPGEHLRFGRQRLREDSGQWQDTNIEALNWSFETTLLNAHAGVAQRFSEYRTDLDELAPEDKDRTHVFGD
ISTQWAPHHRIGVRIHHADDSGHLRRPGEEVDNLDKTYTGQLTWLGIEATGDAYNYRSSMPLNYWASATW
LTGDRDNLTTTTVDDRRIATGKQSGDVNAFGVDLGLRWNIDEQWKAGVGYARGSGGGKDGEEQFQQTGLE
SNRSNFTGTRSRVHRFGEAFRGELSNLQAATLFGSWQLREDYDASLVYHKFWRVDDDSDIGTSGINAALQ
PGEKDIGQELDLVVTKYFKQGLLPASMSQYVDEPSALIRFRGGLFKPGDAYGPGTDSTMHRAFVDFIWRF
Molecule Role :
Protective antigen
Molecule Role Annotation :
Furthermore, we suggest to include AlgE in a multicomponent experimental vaccine for potential protection of non-colonized CF patients from colonization with mucoid P. aeruginosa.(Rehm et al., 1994)
>NP_250468.1 outer membrane porin F [Pseudomonas aeruginosa PAO1]
MKLKNTLGVVIGSLVAASAMNAFAQGQNSVEIEAFGKRYFTDSVRNMKNADLYGGSIGYFLTDDVELALS
YGEYHDVRGTYETGNKKVHGNLTSLDAIYHFGTPGVGLRPYVSAGLAHQNITNINSDSQGRQQMTMANIG
AGLKYYFTENFFAKASLDGQYGLEKRDNGHQGEWMAGLGVGFNFGGSKAAPAPEPVADVCSDSDNDGVCD
NVDKCPDTPANVTVDANGCPAVAEVVRVQLDVKFDFDKSKVKENSYADIKNLADFMKQYPSTSTTVEGHT
DSVGTDAYNQKLSERRANAVRDVLVNEYGVEGGRVNAVGYGESRPVADNATAEGRAINRRVEAEVEAEAK
Molecule Role :
Protective antigen
Molecule Role Annotation :
The oprF gene was cloned into plasmid vector pVR1020, and the plasmid vaccines were delivered to mice by biolistic (gene gun) intradermal inoculation. Following the initial immunization and two consecutive boosts, each at 2-week intervals, protection was demonstrated in a mouse model of chronic pulmonary infection by P. aeruginosa. Eight days postchallenge, both lungs were removed and examined. A significant reduction in the presence of severe macroscopic lesions, as well as in the number of bacteria present in the lungs, was seen (Price et al., 2001). Our data showed that mice immunized with OprF/OprI or OprF/OprI and flagellin B are significantly protected from infection caused by mucoid and nonmucoid strains of P. aeruginosa.(Hassan et al., 2018).
Molecule Role Annotation :
Recombinant OprI protein was used to immunize mice, and was found to be protective against a challenge with a four- to fivefold 50% lethal dose of P. aeruginosa (Finke et al., 1990). Our data showed that mice immunized with OprF/OprI are significantly protected from infection caused by mucoid and nonmucoid strains of P. aeruginosa.(Hassan et al., 2018)
>NP_250397.1 type III secretion protein PcrV [Pseudomonas aeruginosa PAO1]
MEVRNLNAARELFLDELLAASAAPASAEQEELLALLRSERIVLAHAGQPLSEAQVLKALAWLLAANPSAP
PGQGLEVLREVLQARRQPGAQWDLREFLVSAYFSLHGRLDEDVIGVYKDVLQTQDGKRKALLDELKALTA
ELKVYSVIQSQINAALSAKQGIRIDAGGIDLVDPTLYGYAVGDPRWKDSPEYALLSNLDTFSGKLSIKDF
LSGSPKQSGELKGLSDEYPFEKDNNPVGNFATTVSDRSRPLNDKVNEKTTLLNDTSSRYNSAVEALNRFI
QKYDSVLRDILSAI
Molecule Role :
Protective antigen
Molecule Role Annotation :
Burned Pseudomonas aeruginosa-infected mice immunized against PcrV, a type III virulence system translocating protein, showed significantly enhanced survival compared to controls (Holder et al., 2001). The results clearly show that, with PcrV as the vaccine antigen, all three adjuvants tested, FA, alum and CpG ODN, were effective in inducing protective immunity against infection with P. aeruginosa. (Hamaoka et al., 2017)
Vaccination Protocol:
At 14-day intervals, groups of 30 mice were inoculated a total of three times in the abdomen via biolistic particle injection (Helios Gene Gun kit; Bio-Rad, Richmond, Calif.) on days 0, 14, and 28 with 2 μg of either pVR1020 (control) or pVR1020/oprF, which was used to coat 1-μm-diameter gold beads (Price et al., 2001).
Challenge Protocol:
Two weeks after the final immunization, the mice were challenged with agar beads containing the P. aeruginosa FD immunotype 4 strain. The mice were first anesthetized with an intraperitoneal injection of sodium pentobarbital and then inoculated via a tracheal incision with 50 μl of an agar bead slurry encasing approximately 7 × 10^2 CFU of P. aeruginosa (Price et al., 2001).
Efficacy:
A significant reduction in the presence of severe macroscopic lesions, as well as in the number of bacteria present in the lungs, was seen in immunized mice (Price et al., 2001).
Description:
The immunoprotective potential of the pVR1020/oprF vaccine was tested in a mouse model of chronic pulmonary infection (Price et al., 2001).
Efficacy:
Monovalent DNA vaccination targeting OprF/OprI could protect 80% of mice 4 days p.i. On the other hand, 70% of mice immunized with the control plasmid, PilA vaccine, or multivalent vaccine via GG were dead within 5 days p.i. (p < 0.01) (Saha et al., 2006).
7. P. aeruginosa DNA vaccine pGACAG-OprF/OprI + pGACAG-PcrV + pGACAG-PilA
Efficacy:
All mice vaccinated with multivalent DNA via imEPT survived for more than 10 days p.i. with PAK. On the other hand, 70% of mice immunized with the control plasmid, PilA vaccine, or multivalent vaccine via GG were dead within 5 days p.i. (p < 0.01) (Saha et al., 2006).
Efficacy:
Monovalent DNA vaccination targeting PcrV could protect 80% of mice 4 days p.i. On the other hand, 70% of mice immunized with the control plasmid, PilA vaccine, or multivalent vaccine via GG were dead within 5 days p.i. (p < 0.01) (Saha et al., 2006).
Description:
100 μl of 1.5% Al(OH)3 (Finke et al., 1990).
g. Immunization Route
Intraperitoneal injection (i.p.)
h.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
Female BALB/c mice, 12 to 16 weeks old, received 100 μl (27 ,ug) of OprI suspended in 100 μl of 1.5% Al(OH)3 intraperitoneally on days 0, 14, 35, and 63. Controls received Al(OH)3 only (Finke et al., 1990).
Challenge Protocol:
mice were challenged intraperitoneally 10 days after the last immunization with 200 μl of a P. aeruginosa serogroup 6 suspension. Mice vaccinated with recombinant OprI received 6 x 10^6 to 1 x 10^8 living organisms each. (Finke et al., 1990).
Efficacy:
Recombinant OprI protein was used to immunize mice, and was found to be protective against a challenge with a four- to fivefold 50% lethal dose of P. aeruginosa (Finke et al., 1990).
Description:
PcrV was produced as a lipopolysaccharide-free histidine-tagged infusion protein in pET16b and was purified by nickel chromatography (Holder et al., 2001).
Description:
0.1 ml of incomplete Freund's adjuvant (Holder et al., 2001).
g. Immunization Route
Intramuscular injection (i.m.)
h.
Mouse Response
Host Strain:
CF-1
Vaccination Protocol:
On day 0, groups of 10 female CF-1 mice weighing 22 to 25 g were immunized intramuscularly in the hind leg (10 μg of immunogen in 0.1 ml of incomplete Freund's adjuvant), followed by a booster dose (10 μg in saline) without adjuvant on day 14 (Holder et al., 2001).
Challenge Protocol:
Using the burned mouse model, mice were challenged by one of three strains of P. aeruginosa (Holder et al., 2001).
Efficacy:
Burned Pseudomonas aeruginosa-infected mice immunized against PcrV, a type III virulence system translocating protein, showed significantly enhanced survival compared to controls (Holder et al., 2001).
Attenuated Salmonella enterica serovar Typhimurium SL3261 expressing P. aeruginosa serogroup O11 O antigen (DiGiandomenico et al., 2004).
f. Immunization Route
Intramuscular injection (i.m.)
g.
Mouse Response
Vaccination Protocol:
For oral vaccination, mice were fed 100 μl of either PBS or the Salmonella vector and vaccine strains (1 × 10^9 to 5 × 10^9 CFU) by intragastric gavage. Oral inoculation was repeated once per week for a total of 4 weeks. For i.p. vaccination, mice were inoculated with a single dose of either PBS or each Salmonella strain (10^6 CFU) (DiGiandomenico et al., 2004).
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
Mice were challenged with P. aeruginosa strains 9882-80 (serogroup O11) and 6294 (serogroup O6) (DiGiandomenico et al., 2004).
Efficacy:
Orally vaccinated mice with an O11 strain (9882-80) at 6 and 12 times the 50% lethal dose showed increased survival in mice that received the vaccine compared to phosphate-buffered saline (PBS)- and vector-treated controls; no difference in survival was seen with a heterologous strain, 6294 (serogroup O6). In addition, significant protection against 9882-80 was not observed in i.p. vaccinated animals (DiGiandomenico et al., 2004).
IV. References
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