Streptococcus pneumoniae, or pneumococcus, is a Gram-positive, alpha-hemolytic diplococcus aerotolerant anaerobe and a member of the genus Streptococcus. S. pneumoniae is the most common cause of both pneumonia overall and fatal pneumonia. Antibiotic resistance has developed worldwide and is most frequent in pneumococcal serotypes that are most prevalent in children (types/groups 6, 14, 19, and 23). The incidence of pneumococcal disease is the highest in children < 2 years of age and in adults > 65 years of age. Other important risk factors are chronic heart and lung disease, cigarette smoking, and asplenia (Ortqvist et al., 2005). Other than pneumonia, it also causes acute sinusitis, otitis media, meningitis, bacteremia, osteomyelitis, septic arthritis, endocarditis, peritonitis, pericarditis, cellulitis, and brain abscess.
4. Microbial Pathogenesis
S. pneumoniae is normally found in the nasopharynx. It attaches to nasopharyngeal cells through interaction of bacterial surface adhesins. It can cause otitis media or sinusitis if it enters areas such as the Eustachian tube or nasal sinuses. Pneumonia occurs if the organisms are inhaled into the lungs and not eliminated. S. pneumoniae can activate complement, stimulate cytokine production, and attracts neutrophils and other white blood cells. The organism's polysaccharide capsule makes it resistant to phagocytosis. S. pneumoniae can spread to other areas and cause various diseases. S. pneumoniae has several virulence factors, including the polysaccharide capsule, pneumococcal surface proteins, and IgA1 protease (Wiki: S. pneumoniae).
5. Host Ranges and Animal Models
Pneumococci spontaneously cause disease in humans, monkeys, rabbits, horses, mice and guinea pigs. Nasopharyngeal colonization occurs in approximately 40% of the population (Textbook of Bacteriology).
6. Host Protective Immunity
The bacteria invade and grow primarily due to their resistance to the host phagocytic response. The cell wall components directly activate multiple inflammatory cascades, including the alternative pathway of complement activation, the coagulation cascade and the cytokine cascade, inducing interleukin-1, interleukin-6 and tumor necrosis factor (TNF) from macrophages and other cells (Textbook of Bacteriology).
Protein Note :
equivalent gene in S.pneumoniae R6 = spr1995 This gene is also called SpsA, pspC (pneumococcal surface protein C)and Hic; identified by match to protein family HMM PF01473; match to protein family HMM PF04650; match to protein family HMM PF05062; match to protein family HMM TIGR01168
>YP_817402.1 choline binding protein A [Streptococcus pneumoniae D39]
MFASKSERKVHYSIRKFSIGVASVAVASLVMGSVVHATENEGSTQAATSSNMAKTEHRKAAKQVVDEYIE
KMLREIQLDRRKHTQNVALNIKLSAIKTKYLRELNVLEEKSKDELPSEIKAKLDAAFEKFKKDTLKPGEK
VAEAKKKVEEAKKKAEDQKEEDRRNYPTNTYKTLELEIAEFDVKVKEAELELVKEEAKESRNEGTIKQAK
EKVESKKAEATRLENIKTDRKKAEEEAKRKADAKLKEANVATSDQGKPKGRAKRGVPGELATPDKKENDA
KSSDSSVGEETLPSSSLKSGKKVAEAEKKVEEAEKKAKDQKEEDRRNYPTNTYKTLDLEIAESDVKVKEA
ELELVKEEAKEPRDEEKIKQAKAKVESKKAEATRLENIKTDRKKAEEEAKRKAAEEDKVKEKPAEQPQPA
PATQPEKPAPKPEKPAEQPKAEKTDDQQAEEDYARRSEEEYNRLTQQQPPKTEKPAQPSTPKTGWKQENG
MWYFYNTDGSMATGWLQNNGSWYYLNANGAMATGWLQNNGSWYYLNANGSMATGWLQNNGSWYYLNANGA
MATGWLQYNGSWYYLNSNGAMATGWLQYNGSWYYLNANGDMATGWLQNNGSWYYLNANGDMATGWLQYNG
SWYYLNANGDMATGWVKDGDTWYYLEASGAMKASQWFKVSDKWYYVNGSGALAVNTTVDGYGVNANGEWV
N
Molecule Role :
Protective antigen
Molecule Role Annotation :
Mice were actively immunized with a fragment of choline binding protein A (CbpA; also known as PspC, Hic, and SpsA). CbpA was able to protect mice from intraperitoneal challenge with medium to very high doses of a highly virulent capsular type 2 pneumococcal strain, D39 (Ogunniyi et al., 2001).
Molecule Role Annotation :
The protective effects of ATP-dependent caseinolytic protease (ClpP) was evaluated. Vaccinated mice were intraperitoneally and/or intranasally challenged with different pneumococcal strains. In intraperitoneal challenge models with pneumococcal strain D39 (serotype 2), clpP elicited some protection in mice (Wu et al., 2010).
Molecule Role Annotation :
All the control mice died within 2 days of post-infection, while 70% of animals immunized with DnaJ survived the lethal challenge by S. pneumoniae. The study reveals that immunization of mice with DnaJ elicits protective immunity against S. pneumoniae infection.(Khan et al., 2006)
Molecule Role Annotation :
The results of colonization experiment showed that compared with the control group, the PepO, PsaA, and combined immunization groups showed a significant reduction in the colonization of Streptococcus pneumoniae (CMCC31693 and CMCC31207) in the nasopharynx and lung (P<0.05).(Zhang et al., 2017)
Molecule Role Annotation :
Glutamyl tRNA synthetase (GtS) has been found to be among the Streptococcus pneumoniae cell wall-derived proteins that have age-dependent immunogenicity in children. Here, GtS was cloned, expressed, and purified and then was used to immunize 7-week-old BALB/c OlaHsd mice. Thirty-nine percent of rGtS-immunized mice survived a lethal bacterial challenge, whereas no control mice survived (Mizrachi et al., 2007).
Molecule Role Annotation :
BALB/c mice were immunized with recombinant 6-Phosphogluconate dehydrogenase (r6PGD) and boosted after 3 weeks. Immunized mice were challenged intranasally with a lethal dose of S. pneumoniae. Immunization of mice with r6PGD protected 60% (P < 0.001) of mice for 5 days and 40% (P < 0.05) of the mice for 21 days following intranasal lethal challenge (Daniely et al., 2006).
Molecule Role Annotation :
The production of a recombinant vaccine was achieved following PCR amplification of putative lipoate protein ligase (Lpl), with subsequent cloning and expression of the genes in E. coli. The putative lipoate protein ligase (Lpl) , resulted in a reduced CFU titre and a trend towards reduced mortality in an animal sepsis model (Morsczeck et al., 2008).
Molecule Role Annotation :
Seven proteins induced protective responses, in terms of reduced lung burdens of the serotype 3 pneumococci. Three of the protective proteins were histidine triad protein family members (PhtB, PhtD and PhtE). Four other proteins, all bearing LPXTG linkage domains, also had activity in this model (PrtA, NanA, PavB and Eng).(Anderson et al., 2016)
Molecule Role Annotation :
Seven proteins induced protective responses, in terms of reduced lung burdens of the serotype 3 pneumococci. Three of the protective proteins were histidine triad protein family members (PhtB, PhtD and PhtE). Four other proteins, all bearing LPXTG linkage domains, also had activity in this model (PrtA, NanA, PavB and Eng).(Anderson et al., 2016)
Molecule Role Annotation :
Vaccination with PcpA significantly reduced the bacterial burdens in MEF at both 24 (p=0.02) and 48hpi (p=0.004)(Xu et al., 2017)
Molecule Role Annotation :
Immunization with rPhpA-79 protein adjuvanted with monophosphoryl lipid A (for subcutaneous immunization) or a mutant cholera toxin, CT-E29H (for intranasal immunization), protected CBA/N mice against death and bacteremia, as well as reduced nasopharyngeal colonization, following intranasal challenge with a heterologous pneumococcal strain (Zhang et al., 2001).
Molecule Role Annotation :
Seven proteins induced protective responses, in terms of reduced lung burdens of the serotype 3 pneumococci. Three of the protective proteins were histidine triad protein family members.(Anderson et al., 2016)
Molecule Role Annotation :
Seven proteins induced protective responses, in terms of reduced lung burdens of the serotype 3 pneumococci. Three of the protective proteins were histidine triad protein family members.(Anderson et al., 2016) Vaccination with PhtD reduced BG7322 bacterial burdens in MEF at both 24 (p=0.05) and 48hpi (p=0.16).(Xu et al., 2017)
Molecule Role Annotation :
Seven proteins induced protective responses, in terms of reduced lung burdens of the serotype 3 pneumococci. Three of the protective proteins were histidine triad protein family members (PhtB, PhtD and PhtE). Four other proteins, all bearing LPXTG linkage domains, also had activity in this model (PrtA, NanA, PavB and Eng).(Anderson et al., 2016)
Molecule Role Annotation :
Mice immunized with recombinant PiaA were protected against systemic challenge to a degree similar to those immunized with an existing protein vaccine candidate, PdB (a genetically modified pneumolysin toxoid). Immunization with a combination of both PiuA and PiaA resulted in additive protection and was highly protective against systemic infection with S. pneumoniae (Brown et al., 2001).
Molecule Role Annotation :
Immunization with a combination of both PiuA and PiaA resulted in additive protection and was highly protective against systemic infection with S. pneumoniae.(Brown et al., 2001)
Protein Note :
equivalent gene in S.pneumoniae TIGR4 = SP1923; equivalent gene in S.pneumoniae R6 = spr1739; identified by match to protein family HMM PF01289
Molecule Role Annotation :
Immunization with the pneumococcal proteins pneumolysin (Ply), choline binding protein A (CbpA), or pneumococcal surface protein A (PspA) elicits protective responses against invasive pneumococcal disease in animal models.(Chen et al., 2015)
Additional Molecule Role :
Virmugen
Additional Molecule Role Annotation :
The protective effects a pneumolysin mutant (DeltaA146 Ply) against pneumococcal infections in mice was evaluated. Vaccinated mice were challenged with pneumococcal strain D39 at 1,500 times the LD50 (1.5 x 105 CFU) and the vaccinated mice survived significantly longer than control mice after challenge (Wu et al., 2010).
Molecule Role Annotation :
Results have shown that PlyD1 immunization protected mice against lethal intranasal (i.n.) challenge with pneumococci and lung injury mediated by PLY challenge. (Salha et al., 2012) PlyD1 significantly reduced bacterial burden in MEF at 48 hpi (p = 0.02)(Xu et al., 2017)
Molecule Role Annotation :
Seven proteins induced protective responses, in terms of reduced lung burdens of the serotype 3 pneumococci. Three of the protective proteins were histidine triad protein family members (PhtB, PhtD and PhtE). Four other proteins, all bearing LPXTG linkage domains, also had activity in this model (PrtA, NanA, PavB and Eng).(Anderson et al., 2016)
Protein Name :
ABC transporter substrate-binding protein - manganese transport
Protein pI :
5.19
Protein Weight :
33579.58
Protein Length :
421
Protein Note :
Metal binding protein PsaA. These proteins have been shown to function as initial receptors in ABC transport of Mn2+ and as surface adhesins in some eubacterial species. They belong to the TroA superfamily of periplasmic metal binding proteins that...; cd01137
>AAL00298.1 ABC transporter substrate-binding protein - manganese transport [Streptococcus pneumoniae R6]
MKKLGTLLVLFLSAIILVACASGKKDTTSGQKLKVVATNSIIADITKNIAGDKIDLHSIVPIGQDPHEYE
PLPEDVKKTSEADLIFYNGINLETGGNAWFTKLVENAKKTENKDYFAVSDGVDVIYLEGQNEKGKEDPHA
WLNLENGIIFAKNIAKQLSAKDPNNKEFYEKNLKEYTDKLDKLDKESKDKFNKIPAEKKLIVTSEGAFKY
FSKAYGVPSAYIWEINTEEEGTPEQIKTLVEKLRQTKVPSLFVESSVDDRPMKTVSQDTNIPIYAQIFTD
SIAEQGKEGDSYYSMMKYNLDKIAEGLAK
Molecule Role :
Protective antigen
Molecule Role Annotation :
Vaccination with an RASV synthesizing full-length PsaA induced high titers of anti-PsaA antibodies in both systemic (IgG in serum) and mucosal (IgA in vaginal washes, nasal washes, and lung homogenates) sites. BALB/c (haplotype H2(d)) or C57BL/6 (haplotype H2(b)) mice vaccinated either orally or intranasally exhibited a significant reduction in colonization of nasopharyngeal tissues after intranasal challenge with S. pneumoniae strains compared to controls, although protection was not observed with all challenge strains. None of the vaccine constructs provided protection against intraperitoneal challenge with S. pneumoniae strain WU2 (serotype 3) (Wang et al., 2010). The results of colonization experiment showed that compared with the control group, the PepO, PsaA, and combined immunization groups showed a significant reduction in the colonization of Streptococcus pneumoniae (CMCC31693 and CMCC31207) in the nasopharynx and lung (P<0.05).(Zhang et al., 2017)
>YP_815641.1 surface protein A [Streptococcus pneumoniae D39]
MNKKKMILTSLASVAILGAGFVASQPTVVRAEESPVASQSKAEKDYDAAKKDAKNAKKAVEDAQKALDDA
KAAQKKYDEDQKKTEEKAALEKAASEEMDKAVAAVQQAYLAYQQATDKAAKDAADKMIDEAKKREEEAKT
KFNTVRAMVVPEPEQLAETKKKSEEAKQKAPELTKKLEEAKAKLEEAEKKATEAKQKVDAEEVAPQAKIA
ELENQVHRLEQELKEIDESESEDYAKEGFRAPLQSKLDAKKAKLSKLEELSDKIDELDAEIAKLEDQLKA
AEENNNVEDYFKEGLEKTIAAKKAELEKTEADLKKAVNEPEKPAPAPETPAPEAPAEQPKPAPAPQPAPA
PKPEKPAEQPKPEKTDDQQAEEDYARRSEEEYNRLTQQQPPKAEKPAPAPKTGWKQENGMWYFYNTDGSM
ATGWLQNNGSWYYLNSNGAMATGWLQYNGSWYYLNANGAMATGWAKVNGSWYYLNANGAMATGWLQYNGS
WYYLNANGAMATGWAKVNGSWYYLNANGAMATGWLQYNGSWYYLNANGAMATGWAKVNGSWYYLNANGAM
ATGWVKDGDTWYYLEASGAMKASQWFKVSDKWYYVNGLGALAVNTTVDGYKVNANGEWV
Molecule Role :
Protective antigen
Molecule Role Annotation :
Groups of BALB/c mice immunized with choline binding protein PspA were challenged with either D39 or WCH16. PspA elicited protection by using moderate challenge doses of S. pneumoniae (Ogunniyi et al., 2007).
Molecule Role Annotation :
neutralization with purified recombinant PspK-R3 or rabbit anti-UD:R3 IgG inhibited binding of NESp to lung epithelial cells in vitro.(Jang et al., 2017)
Molecule Role Annotation :
Mice immunized with recombinant (r)PsrP(BR) had significantly less bacteria in their blood and improved survival versus controls following intranasal challenge (Shivshankar et al., 2009).
Protein Note :
equivalent gene in S.pneumoniae TIGR4 = SP1218; equivalent gene in S.pneumoniae R6 = spr1098; identified by match to protein family HMM PF04203; match to protein family HMM TIGR01076
Molecule Role Annotation :
Intraperitoneal immunization with recombinant SrtA was found to have conferred to mice protection against S. pneumoniae intraperitoneal challenge and the passive transfer of immune serum before intraperitoneal challenge was also protective. Moreover, by using the intranasal challenge model, significant reduction of bacteremia when mice were intraperitoneally immunized with SrtA was observed, while a moderate decrease of lung infection was achieved by intranasal immunization, even though no influence on nasopharynx colonization was seen (Gianfaldoni et al., 2009).
Vaccination Protocol:
Groups of at least six mice were inoculated intramuscularly with 50 μl of 10 μM cardiotoxin into each tibialis anterior muscle, 5 days before immunization with 50 μg vaccine vector in PBS (100 μl). Mice received a booster after 3 weeks with the same dose of plasmid DNA. For immunization with recombinant protein, mice were injected subcutaneously with 5 μg rPspA3NS, using aluminium hydroxide as the adjuvant, and also boosted 3 weeks after priming (Ferreira et al., 2006).
Vaccine Immune Response Type:
VO_0000286
Immune Response:
Animals immunized with a vector expressing secreted PspA developed higher levels of antibody than mice immunized with the vector expressing the antigen in the cytosol (Ferreira et al., 2006).
Challenge Protocol:
Immunized mice were challenged by intraperitoneal injection of 200 c.f.u. S. pneumoniae strain St 679/99 (PspA clade 3, serotype 6B) in 0·5 ml saline 6 weeks after priming. Animals were then observed for 2 weeks (Ferreira et al., 2006).
Efficacy:
All the immunized groups showed significant protection when compared with control groups. Animals immunized with pSec-pspA3NS showed a tendency towards increased survival when compared with animals primed with rPspA3NS (Ferreira et al., 2006).
Vaccination Protocol:
Mice were immunized i.n. with 0.5 or 2.0 μg of PNC-1 alone or mixed with either LT-K63 or LT-R72 (Jakobsen et al., 1999).
Immune Response:
Administration of PNC-1 i.n. with either LT-K63 or LT-R72 elicited significantly higher antibody responses than the s.c. route for both 0.5- and 2.0-μg doses of PNC-1 in saline (Jakobsen et al., 1999).
Challenge Protocol:
Mice were challenged with 50 μl of pneumococcal suspension i.n. and were allowed to aspirate it into the lungs for 10 min (Jakobsen et al., 1999).
Efficacy:
Immunization i.n. with PNC-1 and LT-K63 or LT-R72 conferred 100% clearance of lung infection caused by serotype 1 pneumococci (Jakobsen et al., 1999).
It consists of a mixture of highly purified capsular polysaccharides from the 23 most prevalent or invasive pneumococcal types of Streptococcus pneumoniae, including the six serotypes that most frequently cause invasive drug-resistant pneumococcal infections among children and adults in the United States (FDA: PNEUMOVAX 23).
k. Storage
Store unopened and opened vials at 2-8°C (36-46°F).
l. Contraindication
Hypersensitivity to any component of the vaccine.
m. Description
Pneumococcal polysaccharide vaccine provides at least 85% protection in those under 55 years of age for five years or longer. Immunization is suggested for those at highest risk of infection, including those 65 years or older. The vaccine is generally given with a single lifetime dose (Wiki: S. pneumoniae). The most common adverse experiences reported with PNEUMOVAX 23 in clinical trials were: Local reaction at injection site including soreness, erythema, warmth, swelling and induration, and fever <102°F. In post-marketing experience, injection site cellulitis-like reactions were reported rarely; between 1989 and 2002, when approximately 43 million doses were distributed, the annual reporting rate was <2/100,000 doses (Merck: PNEUMOVAX 23).
n.
Human Response
Immune Response:
It has been established that the purified pneumococcal capsular polysaccharides induce antibody production and that such antibody is effective in preventing pneumococcal disease (Merck: PNEUMOVAX 23).
Side Effects:
Side effects of vaccination include: redness and soreness of the injection site, fever, and chills (Merck: PNEUMOVAX 23).
Efficacy:
In many clinical studies on the efficacy of the vaccine, it has been found to have a 70-90% efficacy rate (FDA: PNEUMOVAX 23).
A sterile solution of saccharides of the capsular antigens of Streptococcus pneumoniae serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F individually conjugated to diphtheria CRM197 protein (FDA: Prevnar).
k. Preparation
Each serotype is grown in soy peptone broth. The individual polysaccharides are purified through centrifugation, precipitation, ultrafiltration, and column chromatography.
l. Storage
DO NOT FREEZE. Store refrigerated at 2°C TO 8°C.
m . Approved Age for Licensed Use
2-15 months of age.
n. Contraindication
Hypersensitivity to any component of the vaccine, including diphtheria toxoid, is a contraindication to use of this vaccine.
o. Description
In the USA, a heptavalent pneumococcal conjugate vaccine vaccine (PCV 7) (e.g. Prevnar) is recommended since 2000 for all children aged 2–23 months and for at-risk children aged 24–59 months. The normally 4-doses series is given at 2, 4, 6 & 12–14 months of age. Protection is good against deep pneumococcal infections (especially septicemia and meningitis) (Wiki: S. pneumoniae).
p.
Human Response
Vaccination Protocol:
Efficacy was assessed in a randomized, double-blinded clinical trial in a multiethnic population at Northern California Kaiser Permanente (NCKP) from October 1995 through August 20, 1998, in which 37,816 infants were randomized to receive either Prevnar or a control vaccine at 2, 4, 6, and 12-15 months of age (FDA: Prevnar).
Immune Response:
There were no reports of streptococcus pneumoniae in the group vaccinated with PREVNAR whereas in the control group, there were between 17-22 cases reported after vaccination (FDA: Prevnar).
Side Effects:
Side effects of vaccination include: fever, irratibility, drowsiness, decreased appetite, rash and soreness/redness of the injection site.
Vaccination Protocol:
Five male BALB/c mice were inoculated into each muscle tibialis anterior as previously described with 100 μl of 10 μM cardiotoxin and after 5 days with 25 μg of plasmid DNA in PBS (50 μl) (Miyaji et al., 2001).
Vaccine Immune Response Type:
VO_0000286
Immune Response:
Analysis of sera from mice immunized with purified PsaA showed the induction of mostly IgG1, that is, a Th2 like response. Furthermore, mice immunized with pCI-psaA developed readily detectable serum antibody titers 1 month after priming (Miyaji et al., 2001).
Efficacy:
Immunization of mice with purified PsaA was shown to be protective against challenge with an heterologous S. pneumoniae strain (Miyaji et al., 2001).
Description:
Inject Alum adjuvant (Daniely et al., 2006).
g. Immunization Route
Intraperitoneal injection (i.p.)
h.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
Six-week-old BALB/c female mice were immunized intraperitoneally with 25 µg of r6PGD and 75 µl of Inject Alum adjuvant on days 0 (primary immunization) and 21 (booster). Control mice were sham-immunized with adjuvant only (Daniely et al., 2006).
Challenge Protocol:
For respiratory challenge r6PGD immunized and control mice were anaesthetized with pentobarbital sodium (0·6 mg/kg) and inoculated intranasally with 1 × 10^8 S. pneumoniae strain WU2 (in 25 µl PBS). This inoculum's size was used as it was found to be the lowest that causes 100% mortality in our mouse model system within 96 h. Survival was monitored daily (Daniely et al., 2006).
Efficacy:
Immunization of mice with r6PGD protected 60% (P < 0.001) of mice for 5 days and 40% (P < 0.05) of the mice for 21 days following intranasal lethal challenge (Daniely et al., 2006).
Description:
Monophospholipid A (MPL) and aluminium phosphate (AlPO4) (Ogunniyi et al., 2001).
h. Immunization Route
Intraperitoneal injection (i.p.)
i.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
5- to 6-week-old male BALB/c mice (13 to 15 per group) were immunized intraperitoneally with either adjuvant alone, adjuvant plus CbpA, or CbpA alone. Each mouse received three doses of 10 μg of each protein antigen in either formulation (AlPO4 or MPL plus AlPO4) at 12- to 14-day intervals (Ogunniyi et al., 2001).
Challenge Protocol:
Intraperitoneal-challenge experiments were carried out 2 weeks after the third immunization using a highly virulent capsular type 2 strain (D39). Groups of immunized BALB/c mice were infected with either 1.3 × 10^5 or 1.3 × 10^7 CFU of the challenge strain. The challenge dose was equivalent to approximately 10^3 or 10^5 times the 50% lethal dose (LD50), respectively, for BALB/c mice. The mice were closely monitored for 21 days, and the survival time of each mouse was recorded (Ogunniyi et al., 2001).
Efficacy:
CbpA was able to protect mice from intraperitoneal challenge with medium to very high doses of a highly virulent capsular type 2 pneumococcal strain, D39 (Ogunniyi et al., 2001).
Vaccination Protocol:
Female BALB/c mice, weighing 16 to 18 g, were immunized three times at 14-day intervals with 10 µg of protein in alum adjuvant (3:1 [vol/vol]) of either ClpP protein in alum adjuvant or control protein in alum. Mice were boosted intraperitoneally with the same doses on days 14 and 28 (Wu et al., 2010).
Challenge Protocol:
For the intraperitoneal challenge models, at 2 weeks after the last immunization BALB/c mice were administrated the virulent pneumococcal strain D39 at 1,500 times the 50% lethal dose (LD50; 1.5 x 10^5 CFU) or 150 times the LD50 (2.0 x 10^4 CFU). In the focal pneumonia models, BALB/c mice were intranasally inoculated with pneumococcal strain 31614 (serotype 14, 1.5 x 10^6 CFU) or 31693 (serotype 19F, 10^7 CFU). For intranasal challenge with pneumococcal strain D39, mice inspired the virulent pneumococcal strain D39 at 104 (7.5 x 10^7 CFU) or 103 (6.0 x 10^6 CFU) times the LD50 in a volume of 40 µl under injection anesthesia (Wu et al., 2010).
Efficacy:
Compared to the control protein, ClpP reduced the median bacterial load nearly 100-fold in the focal pneumonia model (Wu et al., 2010).
11. S. pneumoniae detoxified PlyD1 Protein Vaccine
Recombinant PLY was produced from Escherichia coli containing the entire gene sequence of wild-type pneumolysin from strain R36A; the protein was column purified. PlyD1, a highly detoxified PLY variant, was generated by site-directed mutagenesis and differs from the wild-type form by three amino acid substitutions of T65C, G293C, and C428A. Recombinant PlyD1 protein was expressed in E. coli as soluble protein and column purified.(Salha et al., 2012)
f. Immunization Route
Intramuscular injection (i.m.)
12. S. pneumoniae DnaJ Protein Vaccine
a. Type:
Recombinant vector vaccine
b. Status:
Research
c. Host Species for Licensed Use:
Baboon
d. Host Species as Laboratory Animal Model:
Mouse
e. Gene Engineering of
DnaJ
Type:
Recombinant protein preparation
Description:
PCR amplified DnaJ was ligated in pQE-30 expression vector and subsequently transformed in E. coli DH5alpha strain.(Khan et al., 2006)
Vaccination Protocol:
Mice were immunized intraperitoneally with 25 μg of rGtS and 75 μL of Imject Alum adjuvant on days 0 and 21. Control mice were sham immunized with adjuvant only (Mizrachi et al., 2007).
Challenge Protocol:
For lethal respiratory challenge with S. pneumoniae, rGtS‐immunized and control mice were anesthetized with isoflurane and inoculated intranasally with S. pneumoniae serotype 3 strain WU2 (in 25 μL of PBS). The size of this inoculum was found to be the lowest that causes 100% mortality in control mice within 96 h. Survival was monitored daily (Mizrachi et al., 2007).
Efficacy:
Thirty-nine percent of rGtS-immunized mice survived a lethal bacterial challenge, whereas no control mice survived (Mizrachi et al., 2007).
Description:
100 μg of monophosphoryl lipid A (MPL; Corixa, Hamilton, Mont.) per dose (Zhang et al., 2001).
g. Immunization Route
subcutaneous injection
h.
Mouse Response
Host Strain:
CBA/N
Vaccination Protocol:
CBA/N mice, at 10 animals per group, were immunized with 5 μg of either rPhpA-79 or rPhpA-20. For parenteral immunization, rPhpA-79 or rPhpA-20 protein was mixed with 100 μg of monophosphoryl lipid A (MPL; Corixa, Hamilton, Mont.) per dose to a final volume of 200 μl in saline and then injected s.c. into mice. All groups received a booster with the same dose and by the same route 3 and 5 weeks after the primary immunization. Control mice were injected with MPL alone (Zhang et al., 2001).
Challenge Protocol:
Mice were challenged with either serotype 3 or serotype 14 streptomycin-resistant S. pneumoniae (Zhang et al., 2001).
Efficacy:
Immunization with either rPhpA-79–MPL provided 100% protection against death caused by type 3 S. pneumoniae infection (Zhang et al., 2001).
Vaccination Protocol:
For the active protection experiments, each mouse was immunized by intraperitoneal (i.p.) injection of 100 μl of each protein preparation in alum adjuvant on days 0, 10, and 20 (Brown et al., 2001).
Challenge Protocol:
On day 34 mice were challenged i.p. with ca. 10^6 CFU of S. pneumoniae strain D39 organisms (Brown et al., 2001).
Efficacy:
After 14 days the mortality of the group of mice immunized PiuA was 52%, showing that immunization with these proteins protects against systemic infection with S. pneumoniae (Brown et al., 2001).
Vaccination Protocol:
The mice received three doses of 10 μg of each antigen alone (or in combination) in 100 μg of alum adjuvant via Intraperitoneal injection (i.p.) at 14-day intervals or a placebo (Ogunniyi et al., 2007).
Challenge Protocol:
Mice were challenged i.p. 2 weeks after the third immunization with either the highly virulent capsular type 2 strain D39 or a serotype 6A (WCH16) strain (Ogunniyi et al., 2007).
Efficacy:
PspA elicited protection by using moderate challenge doses of S. pneumoniae (Ogunniyi et al., 2007).
Streptococcus pneumoniae is one of the leading causes of bacterial pneumonia and acute otitis media, a type of meningitis. It is becoming increasingly antibiotic resistant and the current available vaccine does not elicit protection in children. (Langermann et al., 1994)
j.
Mouse Response
Host Strain:
BALB/c and C3H/HeJ mice
Vaccination Protocol:
A single intraperitoneal inoculation with a live BCG bacterial vaccine expressing PspA as a secreted mature protein in or a chimeric exported lipoprotein (rBCG-PspA), followed by a booster at 17 weeks(Langermann et al., 1994)
Vaccine Immune Response Type:
VO_0003057
Challenge Protocol:
BALB/c and C3H/HeJ mice immunized with rBCG-PspA vaccine strains were challenged intraperitoneally with 10^4 CFU of the highly lethal S. pneumoniae strain WU2 (Langermann et al., 1994)
Challenge Detail
No.
Pathogen Name
Dose
Route
Age
Interval
1
S. pneumoniae strain WU2
10000 CFU in volume ml
Intraperitoneal injection (i.p.)
day
day
Efficacy:
Mice experienced passive protection with the use of rBCG against lethal challenge with the highly virulent, encapsulated WU2 strain of Streptococcus pneumoniae and two additional virulent strains exhibiting heterologous PspA and capsular sero- types(Langermann et al., 1994)
Vaccination Protocol:
Mice were inoculated intranasally with 10 µl or orally with 20 µl of BSG containing 1 x 10^9 CFU of the RASV or control strain. In some experiments, the mice were boosted at week 6 with the same dose by using the same route as that used for primary immunization (Wang et al., 2010).
Challenge Protocol:
At week 10, mice were challenged either by intraperitoneal injection with 2 x 10^4 CFU of S. pneumoniae WU2 (equal to 100x the 50% lethal dose [LD50]) or intranasally with 20 µl containing 5 x 10^6 CFU S. pneumoniae strain L82016 or E134 or 1 x 10^7 CFU of strain A66.1 or D39 (Wang et al., 2010).
Efficacy:
BALB/c (haplotype H2(d)) or C57BL/6 (haplotype H2(b)) mice vaccinated either orally or intranasally exhibited a significant reduction in colonization of nasopharyngeal tissues after intranasal challenge with S. pneumoniae strains compared to controls, although protection was not observed with all challenge strains (Wang et al., 2010).
Vaccination Protocol:
Specific-pathogen-free BALB/c mice (Charles River) received three intraperitoneal administrations, 2 weeks apart, of 20 μg of SrtA along with Freund's adjuvant. Controls received the same course of saline plus adjuvant (Gianfaldoni et al., 2009).
Challenge Protocol:
Three weeks after the last immunization, the animals were challenged intraperitoneally with one of the following S. pneumoniae strains: TIGR4, 1.4 × 10^2 CFU/mouse; D39, 10^3 CFU/mouse; or 35B-SME15, 7 × 10^3 CFU/mouse (Gianfaldoni et al., 2009).
Efficacy:
Intraperitoneal immunization with recombinant SrtA was found to have conferred to mice protection against S. pneumoniae intraperitoneal challenge and the passive transfer of immune serum before intraperitoneal challenge was also protective. Moreover, by using the intranasal challenge model, significant reduction of bacteremia when mice were intraperitoneally immunized with SrtA was observed, while a moderate decrease of lung infection was achieved by intranasal immunization, even though no influence on nasopharynx colonization was seen (Gianfaldoni et al., 2009).
20. Synflorix
a. Product Name:
Pneumococcal conjugate vaccine (Non-Typeable Haemophilus influenzae (NTHi) protein D, diphtheria or tetanus toxoid conjugates) adsorbed
Infants and children from 6 weeks up to 2 years of age (GSK: Synflorix).
n. Description
SYNFLORIX™ [(pneumococcal conjugate vaccine (Non-Typeable Haemophilus influenzae (NTHi) protein D, diphtheria or tetanus toxoid conjugates) adsorbed] is a 10- valent pneumococcal polysaccharide conjugate vaccine using protein D derived from Non-Typeable Haemophilus influenzae as a carrier protein for 8 out of the 10 serotypes (1, 4, 5, 6B, 7F, 9V, 14 and 23F). Serotypes 18C and 19F are conjugated to tetanus toxoid and to diphtheria toxoid, respectively. All conjugates are adsorbed onto aluminum phosphate (GSK: Synflorix).
V. References
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