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Vaccine Comparison

E. coli heat-labile enterotoxin B-subunit (LB-T) Vaccine E. coli O157:H7 subunit vaccine expressing Esps and Tir E. coli vaccine using intimin polypeptide
Vaccine Information Vaccine Information Vaccine Information
  • Vaccine Ontology ID: VO_0004144
  • Type: Subunit vaccine
  • Antigen: The antigen for this vaccine is a plant-optimized synthetic gene encoding for the E. Coli heat-labile enterotoxin B-subunit (LT-B) (Mason et al., 1998).
  • Preparation: DNA plasmids were created, and the expression cassettes were purified. Potato plants were transformed by the leaf disc cocultivation method. Transformed lines were analyzed, and the best lines were clonally propogated. These plantlets were transferred to soil and grown. The tubers produced were the vaccine (Mason et al., 1998).
  • Vaccine Ontology ID: VO_0000459
  • Type: Subunit vaccine
  • Antigen: The antigen for this vaccine is supernatant proteins (containing Type III proteins Esps and Tir) prepared from E. Coli O157-H7 (Potter et al., 2004).
  • Tir gene engineering:
    • Type: Protein purification
    • Description: The E. coli strain O157:H7 was used for the production of Type III secreted proteins Tir and Esps (Potter et al., 2004).
    • Detailed Gene Information: Click Here.
  • Adjuvant: VSA-3
  • Preparation: Supernatent proteins from E. coli O157-H7 were combined with the VSA3 to form protein concentrations of either 25 or 100 micrograms/ml. Each dose was 50 micrograms/ml (Potter et al., 2004).
  • Virulence: Not noted.
  • Tradename: None
  • Vaccine Ontology ID: VO_0000478
  • Type: Subunit vaccine
  • Antigen: E. coli Intimin polypeptide(van et al., 2007).
  • eae gene engineering:
    • Type: Preparation of recombinant protein
    • Description: The portion of the eae gene that encodes the carboxyl-terminal 280 amino acids of intimin was amplified by polymerase chain reaction from EHEC O26:H- strain 193 (Int280-β) and EHEC O157:H7 strain EDL933 (Int280-γ) using a conserved forward primer (Int-LIC-for: 5′-GAC GAC GAC AAG ATT ACT GAG ATT AAG GCT G-3′) and subtype-specific reverse primers (O26Int-LIC-rev: 5′-GAG GAG AAG CCC GGT TTA TTT TAC ACA AAC AG-3′ and O157Int-LIC-rev: 5′-GAG GAG AAG CCC GGT TTA TTC TAC ACA AAC CG-3′). The products were cloned in pET30-Ek/Lic (Novagen®) by a ligation-independent method as amino-terminal 6×His-S-tag fusions. Proteins were expressed in E. coli K-12 strain BL21 (DE3) Star cells which lack RNaseE to stabilise mRNA. The Overnight Express™ Autoinduction System I (Novagen®) was used to induce Int280-γ and Int280-β expression. Cell extracts were prepared using BugBuster® (Novagen®) and the supernatant fraction mixed with His-Mag™ Agarose Beads (Novagen®) for affinity purification of the Int280 proteins as described by the manufacturer
      (van et al., 2007).
    • Detailed Gene Information: Click Here.
  • Adjuvant: aluminum hydroxide vaccine adjuvant
    • VO ID: VO_0000127
    • Description: Aluminium hydroxide oil-based adjuvant (Alu-Oil; Intervet International BV, Boxmeer, The Netherlands) (van et al., 2007).
  • Preparation: Proteins were expressed in E. coli K-12 strain BL21 (DE3) Star cells which lack RNaseE to stabilise mRNA. The Overnight Express™ Autoinduction System I (Novagen®) was used to induce Int280-γ and Int280-β expression. Cell extracts were prepared using BugBuster® (Novagen®) and the supernatant fraction mixed with His-Mag™ Agarose Beads (Novagen®) for affinity purification of the Int280 proteins as described by the manufacturer (van et al., 2007).
  • Virulence: Not noted.
  • Description: Enterohaemorrhagic Escherichia coli (EHEC) are zoonotic enteric pathogens of worldwide importance. EHEC strains produce intimin, an outer membrane adhesin encoded by the eae gene located in a chromosomal pathogenicity island termed the locus of enterocyte effacement (LEE). Intimin mediates intimate bacterial attachment to enterocytes by binding to Tir, a bacterial protein which is translocated into host cells by a LEE-encoded type III secretion system. Intimin can also bind in vitro to β1-integrins and cell-surface localised nucleolin and these proteins can be detected proximal to adherent EHEC O157:H7 in vivo. Intimin is a key colonisation factor for EHEC O157:H7 in neonatal calves, young and weaned calves, and adult cattle and sheep. In addition, intimin influences the carriage and virulence of EHEC O157:H7 in streptomycin pre-treated mice, infant rabbits, and gnotobiotic and neonatal piglets (van et al., 2007).
Host Response Host Response Host Response

Mouse Response

  • Host Strain: Female BALB/c mice of 16 to 18 g (Charles River Laboratories, Inc.).
  • Vaccination Protocol: NT-1 cells or transgenic NT-1 cell clones that expressed Int261 were grown in 40-ml suspension cultures to confluence. Five grams of NT-1 cell material was divided into aliquots, and 0.5 g of sucrose was added to each sample. A 7.5-µg dose of purified cholera toxin (CT) (Sigma) was also added to appropriate samples to serve as an oral adjuvant. Mice were made to fast overnight before they were allowed to eat the plant material ad libitum. Mice immunized i.p. with purified His-tagged Int261 plus TiterMax served as the positive control (Judge et al., 2004).
  • Persistence: Not noted.
  • Side Effects: Not noted.
  • Challenge Protocol: Mice were made to fast overnight and fed a total inoculum of 108 to 109 CFU of E. coli O157:H7 strain 86-24 Strr or 86-24 Strr eae10 in each of two doses administered 4 h apart (Judge et al., 2004).
  • Efficacy: Parenteral priming of mice with intimin purified from transgenic plant cells can assist in the development of an intimin-specific fecal immune response when these mice are subsequently boosted with oral feeding of the same intimin-expressing transgenic plant material. Mice that were parenterally primed and then given an oral booster showed a statistically significant decrease in the duration of colonization by wild-type E. coli O157:H7 upon challenge. Mice immunized entirely by oral feeding did exhibit a reduction in the duration of colonization versus unimmunized mice, but the reduction was not statistically significant. These results suggest that a combination of vaccination strategies with a vaccine antigen produced in and delivered by transgenic plants can function in inducing beneficial, specific immune responses (Judge et al., 2004).
  • Description: An oral inoculation system was sought to facilitate induction of mucosal antibodies and for ease of administration. A transgenic plant cell system for intimin expression was used, with the ultimate goal of moving the antigen into whole-plant expression and delivery systems. Transgenic plants offer the flexibility to function as low-cost, efficient, and practical vaccine antigen oral delivery systems to stimulate mucosal immunity or to boost and shift initial immunity to a mucosal antibody response. Transgenic plants have already been used as successful vaccine antigen production and delivery systems. Carboxy-terminal third of intimin-expressing plant cells were created. Capacity of this transgenic material to induce adherence-blocking antibodies and to reduce levels and/or time of E. coli O157:H7 fecal shedding in a mouse model of intimin-dependent colonization were then evaluated (Judge et al., 2004).

Cattle Response

  • Vaccination Protocol: The authors used two independent transgenic lines of potato tubers, THllO-8 and THllO-51, and nontransformed tubers of line FL1607. Potato tubers were peeled and sliced before offering to mice as food. BALB/c mice were fasted overnight before feeding raw tuber slices. Tuber feedings were performed at weekly intervals for 3 weeks (days 0, 7 and 14). Mice were divided into four groups of five animals each. Each animal in Groups 1-3 received 5 g of FL1607, THllO-8 or THllO-51 tubers per feeding, respectively (Mason et al., 1998).
  • Immune Response: Mice that ingested three doses of transformed tubers developed anti-LT-B faecal IgA and anti-LT-B serum IgG antibody responses equivalent to or greater than responses developed by mice gavaged orally with 5 micrograms purified LT-B. THllO-51 tubers, in contrast to THllO-8 tubers or purified LT-B, produced significantly higher anti-LT-B faecal IgA responses when ingested by mice. Control mice that were fed nontransformed potatoes had no detectable anti-LT-B faecal IgA or anti-LT-B serum IgG responses (Mason et al., 1998).
  • Challenge Protocol: Mice were challenged by oral administration of 25 micrograms LT (heat-labile enterotoxin) (Mason et al., 1998).
  • Efficacy: Compared to mice that were fed non-transformed tubers, mice that were immunized by gavage with purified bacteria LT-B had the greatest reduction in gut/carcass ratio on subsequent challenge. Mice that were fed transformed potatoes had somewhat less but still a significant reduction in fluid. No mouse was completely protected. Control mice fed nontransformed potatoes developed no anti-LT-B faecal or serum antibodies (Mason et al., 1998).

Cattle Response

  • Vaccination Protocol: Groups of 8 seronegative 6-month-old calves were immunized with 2 ml of vaccine delivered subcutaneously in the neck. A control group that received only the adjuvant was included (Potter et al., 2004).
  • Persistence: Not noted.
  • Immune Response: The group that received the vaccine showed a 13-fold increase in specific antibody titre to type III secreted proteins after a single immunization, and after a booster vaccination, the group demonstrated a 45-fold increase in specific antibody titre (Potter et al., 2004; Potter et al., 2004)
  • Challenge Protocol: Animals were challenged two weeks following the final vaccination with 108 CFU of E. coli O157-H7 by oral-gastic intubation (Potter et al., 2004).
  • Efficacy: On each of the post-challenge days, fewer vaccinated animals shed bacteria compared to the placebo group (Potter et al., 2004).

Cattle Response

  • Host Strain: Calves
  • Vaccination Protocol: In Trial 1, on day 0 and day 28 calves were vaccinated i.m. with Int280-γ. In Trial 2, calves were vaccinated with Int280-β on days 0 and 28 (van et al., 2007).
  • Persistence: Not noted.
  • Side Effects: Not noted.
  • Challenge Protocol: In Trial 1, on day 42 oral challenge was administered with 2.9 ± 0.78 × 1010 colony forming units (CFU) of EHEC O157:H7 strain EDL933 nalR. In Trial 2, on day 42 oral challenge was performed using 2.8 ± 0.67 × 1010 CFU EHEC O26:H- strain STM2H2 (van et al., 2007).
  • Efficacy: Subunit vaccines based on intimin polypeptides induced serum IgG and variable salivary IgA responses following parenteral immunisation of cattle. However, such responses did not confer significant resistance to intestinal colonisation by EHEC strains expressing the homologous antigens, even after boosting of such animals by the mucosal route (van et al., 2007).
  • Description: While it has been shown that i.n. immunisation of cattle with a carboxyl-terminal 64 kDa intimin polypeptide adjuvated with a low-toxicity derivative of E. coli heat-labile toxin induces antigen-specific serum IgG1 and salivary IgA, the protective efficacy of intimin-based subunit vaccines in cattle has yet to be tested. The present study assessed the protective efficacy of subunit vaccines comprising of intimin polypeptides against intestinal colonisation of cattle by EHEC strains of serotypes O157:H7 and O26:H- following parenteral and mucosal immunisation (van et al., 2007).
References References References
Mason et al., 1998: Mason HS, Haq TA, Clements JD, Arntzen CJ. Edible vaccine protects mice against Escherichia coli heat-labile enterotoxin (LT): potatoes expressing a synthetic LT-B gene. Vaccine. 1998; 16(13); 1336-1343. [PubMed: 9682399 ].
Potter et al., 2004: Potter AA, Klashinsky S, Li Y, Frey E, Townsend H, Rogan D, Erickson G, Hinkley S, Klopfenstein T, Moxley RA, Smith DR, Finlay BB. Decreased shedding of Escherichia coli O157:H7 by cattle following vaccination with type III secreted proteins. Vaccine. 2004; 22(3-4); 362-369. [PubMed: 14670317].
Judge et al., 2004: Judge NA, Mason HS, O'Brien AD. Plant cell-based intimin vaccine given orally to mice primed with intimin reduces time of Escherichia coli O157:H7 shedding in feces. Infection and immunity. 2004 Jan; 72(1); 168-75. [PubMed: 14688094 ].
van et al., 2007: van Diemen PM, Dziva F, Abu-Median A, Wallis TS, van den Bosch H, Dougan G, Chanter N, Frankel G, Stevens MP. Subunit vaccines based on intimin and Efa-1 polypeptides induce humoral immunity in cattle but do not protect against intestinal colonisation by enterohaemorrhagic Escherichia coli O157:H7 or O26:H-. Veterinary immunology and immunopathology. 2007 Mar 15; 116(1-2); 47-58. [PubMed: 17258324].