Rotavirus is a genus of double-stranded RNA virus in the family Reoviridae. Rotavirus is a leading cause of severe acute gastroenteritis (severe diarrhoea) in infants and young children, with over 95% of these children infected by the time they are 5 years old. The most severe cases occur among infants and young children between 6 months and 24 months of age. By the age of five, nearly every child in the world has been infected with rotavirus at least once. However, with each infection, immunity develops, subsequent infections are less severe,[3] and adults are rarely affected. There are seven species of this virus, referred to as A, B, C, D, E, F and G. Rotavirus A, the most common, causes more than 90% of infections in humans (Wiki: Rotavirus).
4. Microbial Pathogenesis
Rotavirus is transmitted by the faecal-oral route and possibly by the respiratory route. The diarrhoea is caused by multiple activities of the virus. Malabsorption occurs because of the destruction of gut enterocytes. The toxic rotavirus protein NSP4 induces age- and calcium ion-dependent chloride secretion, and disrupts SGLT1 transporter-mediated reabsorption of water. A recurrence of mild diarrhoea often follows the reintroduction of milk into the child's diet, due to bacterial fermentation of the disaccharide lactose in the gut (Wiki: Rotavirus).
5. Host Ranges and Animal Models
In addition to humans, rotaviruses infect and cause diarrhoea in young animals. They have been shown to infect mammals (for example, apes, cattle, pigs, sheep, rats,, cats, and dogs, mice, horses, rabbits) and birds (chickens and turkeys). These rotaviruses are a potential reservoir for genetic exchange with human rotaviruses. There is evidence that animal rotaviruses can infect humans, either by direct transmission of the virus or by contributing one or several RNA segments to reassortants with human strains (Wiki: Rotavirus).
6. Host Protective Immunity
Protective immune responses against rotavirus infections have been correlated with production of rotavirusspecific fecal IgA in vivo in human and porcine studies as well as in mice (Herrmann et al., 1999).
Molecule Role Annotation :
Both inbred BALB/c and outbred CD-1 mice were immunized with rotavirus VP2/6-rVLPs (2/6-VLPs) combined with CT-E29H, orally or intranasally (i.n.), and the comparative efficacy of different formulations was then determined. Two immunizations with 2-6VLPs and CT-E29H were sufficient to protect BALB/c mice, regardless of the route of administration (Siadat-Pajouh and Cai, 2001).
Molecule Role Annotation :
Rotavirus structural proteins VP4, VP6 and VP7 from Bovine Rotavirus Strain C486 were cloned and expressed in a baculovirus expression system. Challenge of neonates born to animals immunized with VP4 protein on assembled particles or in cell lysates showed protection against challenge with both homologous (bovine C486) and heterologous (SA-11) strains of rotavirus (Redmond et al., 1993).
Molecule Role Annotation :
Plasmid DNA vaccines encoding for murine rotaviral proteins VP4, VP6, and VP7 were tested in adult BALB/c mice for their ability to induce immune responses and provide protection against rotavirus challenge. Vaccines encoding VP4 were effective in protecting mice against infection after rotavirus (100 ID50) challenge (Herrmann et al., 1996).
Molecule Role Annotation :
Intranasal or oral delivery of the chimeric rotavirus VP6 protein MBP::VP6 to mice elicited >90% reductions in fecal rotavirus shedding after murine rotavirus challenge (McNeal et al., 2007).
Molecule Role Annotation :
VP6 derived from Group A rotavirus conferred in vivo partial protection against rotavirus challenge in a neonatal mouse model (Garaicoechea et al., 2008).
Molecule Role Annotation :
Rotavirus structural proteins VP4, VP6 and VP7 from Bovine Rotavirus Strain C486 were cloned and expressed in a baculovirus expression system. Challenge of neonates born to animals immunized with VP6 protein showed partial protection (Redmond et al., 1993).
Molecule Role Annotation :
Rotavirus structural proteins VP4, VP6 and VP7 from Bovine Rotavirus Strain C486 were cloned and expressed in a baculovirus expression system. Neonates of animals immunized with virus-like particles composed of VP7 assembled on VP6 spherical particles were protected against challenge with the homotypic virus and significantly protected from a heterotypic challenge. Unassembled VP7 protein provided partial protection against challenge (Redmond et al., 1993).
Molecule Role Annotation :
Plasmid DNA vaccines encoding for murine rotaviral proteins VP4, VP6, and VP7 were tested in adult BALB/c mice for their ability to induce immune responses and provide protection against rotavirus challenge. Vaccines encoding VP7 were effective in protecting mice against infection after rotavirus (100 ID50) challenge (Herrmann et al., 1996).
Molecule Role Annotation :
IFN-gamma plays a critical role in Th1 type immune response. It is important for protection against infections by various viruses and intracellular bacteria.
Additional Molecule Role :
Vaximmutor
Additional Molecule Role Annotation :
The experimental data demonstrated that three time vaccinations with BCG in BALB/c mice induced strong TB Ag-specific IFN-gamma immune responses in splenocytes (Wang et al., 2009).
Description:
HSV-1 amplicon vectors encoding individual or multiple structural RV proteins from a polycistronic transgene cassette in mammalian cells (Laimbacher et al., 2012).
Description:
HSV-1 amplicon vectors encoding individual or multiple structural RV proteins from a polycistronic transgene cassette in mammalian cells (Laimbacher et al., 2012).
Description:
HSV-1 amplicon vectors encoding individual or multiple structural RV proteins from a polycistronic transgene cassette in mammalian cells (Laimbacher et al., 2012).
Herpes simplex virus type 1 (HSV-1) amplicon vectors were constructed to coexpress the rotavirus (RV) structural genes VP2, VP6, and VP7 (Laimbacher et al., 2012).
i. Immunization Route
Intramuscular injection (i.m.)
j.
Mouse Response
Vaccination Protocol:
intramuscular injection of mice with the amplicon vectors as a two-dose regimen without adjuvants (Laimbacher et al., 2012).
Vaccine Immune Response Type:
VO_0001030
Challenge Protocol:
Immunized mice were challenged with live wild-type (wt) rotavirus infection (Laimbacher et al., 2012).
Efficacy:
Immunized mice were partially protected at the mucosal level (Laimbacher et al., 2012).
A live, attenuated rotavirus vaccine derived from the human 89-12 strain which belongs to G1P[8] type.
k. Preparation
ROTARIX is a live, attenuated rotavirus vaccine derived from the human 89-12 strain which belongs to G1P[8] type. The rotavirus strain is propagated on Vero cells. The lyophilized vaccine contains amino acids, dextran, Dulbecco’s Modified Eagle Medium (DMEM), sorbitol, and sucrose. The liquid diluent contains calcium carbonate, sterile water, and xanthan. The diluent includes an antacid component (calcium carbonate) to protect the vaccine during passage through the stomach and prevent its inactivation due to the acidic environment of the stomach. ROTARIX contains no preservatives (FDA: ROTARIX).
l. Storage
Store the vials of lyophilized ROTARIX refrigerated at 2° to 8°C (36° to 46°F). The diluent may be stored at a controlled room temperature 20° to 25°C (68° to 77°F). Do not freeze.
m . Approved Age for Licensed Use
6 to 24 months
n. Contraindication
History of uncorrected congenital malformation of the gastrointestinal tract that would predispose the infant to intussusception.
o. Description
Indication: Prevention of rotavirus gastroenteritis caused by G1 and non-G1 types (G3, G4, and G9) in infants and children when administered as a two-dose series between the ages of 6-24 weeks (FDA: ROTARIX).
p.
Human Response
Immune Response:
Seroconversion was defined as the appearance of anti-rotavirus IgA antibodies (concentration ≥20 U/mL) post-vaccination in the serum of infants previously negative for rotavirus. In one clinical trial, it was recorded that 86.5% of 787 recipients of ROTARIX seroconverted compared with 6.7% of 420 placebo recipients and 76.8% of 393 recipients of ROTARIX seroconverted compared with 9.7% of 341 placebo recipients (FDA: ROTARIX).
Side Effects:
Side effects of vaccination include: fussiness/irritability, cough/runny nose, fever, loss of appetite, and vomiting.
q.
Human Response
Vaccination Protocol:
A randomized, double-blind, placebo-controlled study was conducted in 6 European countries. A total of 3,994 infants were enrolled to receive ROTARIX or placebo. Analyses were also done to evaluate the efficacy of ROTARIX against rotavirus gastroenteritis among infants who received at least one vaccination (FDA: ROTARIX).
Efficacy:
In this study, the efficacy of ROTARIX against any grade of severity of rotavirus gastroenteritis through one rotavirus seasons was 87.1% and for ROTARIX against any grade of severity of rotavirus gastroenteritis through two rotavirus seasons was 78.9% (FDA: ROTARIX).
5 live reassortant rotaviruses: Four reassortant rotaviruses express one of the outer capsid proteins (G1, G2, G3, or G4) from the human rotavirus parent strain and the attachment protein (serotype P7) from the bovine rotavirus parent strain. The fifth reassortant virus expresses the attachment protein, P1A (genotype P[8]), herein referred to as serotype P1A[8], from the human rotavirus parent strain and the outer capsid protein of serotype G6 from the ovine rotavirus parent strain (FDA: ROTARIX).
k. Preparation
RotaTeq is a live, oral pentavalent vaccine that contains 5 live reassortant rotaviruses. The reassortants are propagated in Vero cells using standard cell culture techniques in the absence of antifungal agents. The reassortants are suspended in a buffered stabilizer solution. RotaTeq contains no preservatives (FDA: ROTARIX).
l. Storage
Store and transport refrigerated at 2-8°C (36-46°F).
m. Contraindication
A demonstrated history of hypersensitivity to any component of the vaccine. Infants who develop symptoms suggestive of hypersensitivity after receiving a dose of ROTATEQ should not receive further doses of ROTATEQ.
n. Description
Indication: Prevention of rotavirus gastroenteritis in infants and children caused by the serotypes G1, G2, G3, and G4 when administered as a 3-dose series to infants between the ages of 6 to 32 weeks (FDA: ROTARIX).
o.
Human Response
Vaccination Protocol:
71,725 infants were evaluated in 3 placebo-controlled clinical trials including 36,165 infants who received RotaTeq and 35,560 infants who received placebo (FDA: RotaTeq).
Side Effects:
Possible side effects of vaccinations include: intussusception, vomiting, diarrhea, irrittability
p.
Human Response
Vaccination Protocol:
72,324 infants were randomized in 3 placebo-controlled, phase 3 studies conducted in 11 countries on 3 continents. The vaccine was given as a three-dose series to healthy infants with the first dose administered between 6 and 12 weeks of age and followed by two additional doses administered at 4- to 10-week intervals (FDA: RotaTeq).
Immune Response:
92.9% to 100% of 439 recipients of RotaTeq achieved a 3-fold or more rise in serum anti-rotavirus IgA after a three-dose regimen.
Efficacy:
Primary efficacy against any grade of severity of rotavirus gastroenteritis caused by naturally occurring serotypes G1, G2, G3, or G4 through the first rotavirus season after vaccination was 74.0% (FDA: RotaTeq).
Vaccination Protocol:
Mice were lightly anaesthetized in a closed vessel with Metafane (methoxyflurane, Pitman–Moore Inc., Mundelein, IL). Vaccine formulations containing chimeric VP6 admixed with different adjuvants were then delivered intranasally or orally into groups of mice. Two i.n. doses (60 μl per dose, 30 μl per nostril) or two oral doses (100 μl per dose), separated by 2 weeks, were delivered. Each dose of immunogen consisted of 9 μg of purified chimeric VP6, which was mixed with LT(R192G) (10 μg), CTA1-DD (10 μg), Adjumer® (50 μg), CpG ODN (10 μg) or QS-21 (20 μg) just before immunization. The amounts of adjuvants used were based on the amounts determined for other immunogens. A group of unimmunized mice served as the control in each experiment (Choi et al., 2002).
Immune Response:
Mice immunized with Adjumer®, CTA1-DD or LT(R192G), also developed low titers of fecal rotavirus IgA. Use of Adjumer as an adjuvant had a large IgG1 titer but no increase in IgG2a titer (Choi et al., 2002).
Challenge Protocol:
Four weeks after the last immunization, mice were orally (gavage) challenged with 4×10^4 FFUs (100 μl, 100 SD50) of passage-9 EDIM rotavirus (Choi et al., 2002).
Efficacy:
Mice immunized with chimeric VP6 and Adjumer had a 35% reduction in shedding of virus compared to unimmunized mice (Choi et al., 2002).
Description:
A detoxified version of cholera toxin, CT-E29H (Siadat-Pajouh and Cai, 2001).
f. Immunization Route
Orally
g.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
Four-week-old BALB/c mice were vaccinated twice at Weeks 0 and 2 either orally or i.n. with 100 and 10 ug of 2/6-VLPs, respectively; each vaccine dose was formulated with 10 mg of CT-E29H. A third group of BALB/c mice (n = 4) received 2/6-VLPs with CT-E29H i.n. followed by an oral booster immunization (i.e., mixed group). Control mice in this experiment were inoculated with CT-E29H, 13 TNC, and 2/6-VLPs alone. Each mouse was inoculated i.n. with 20 uL of inoculums , 2 uL at a time into alternating nares at 1-min intervals (Siadat-Pajouh and Cai, 2001).
Challenge Protocol:
All animals were challenged by gavage with 10 SD50 of wild-type EDIM rotavirus at week 13 (BALB/c mice). The trypsin-activated challenge virus (100 uL) was administered following oral inoculation of 100 uL of 4% sodium bicarbonate solution to neutralize gastric acidity (Siadat-Pajouh and Cai, 2001).
Efficacy:
In BALB/c mice, intranasal vaccination with 2/6-VLPs and CT-E29H proved protective (PRAS 5 98.7%), in contrast to intranasal immunization with 2/6-VLPs alone (PRAS 5 39%), demonstrating the significant augmentation in protective immune responses due to CT-E29H. BALB/c mice in all three immunization groups showed nearly complete protection from the challenge. PRAS was 99.6, 98.8, and 98.8% for the oral, intranasal, and the mixed groups, respectively. The unimmunized control group shed significantly more viral antigen than the three immunized groups (Siadat-Pajouh and Cai, 2001).
Vaccination Protocol:
For primary immunization, each immunogen (based on 10μg of VP6) was emulsified with an equal volume of Freund's complete adjuvant. For the secondary 'booster' immunizations, each immunogen was emulsified with an equal volume of Freund's incomplete adjuvant. Eight groups of mice (Strain CD-1 ; ten animals/group) were immunized intramuscularly with the immunogen preparations. The primary immunization was given to the mice at 7 weeks of age and was followed by two booster immunizations at 2 week intervals. The mice were bred and litters were born to the dams at 12 to 14 weeks of age (Redmond et al., 1993).
Challenge Protocol:
Pups were allowed to suckle their dams and ten pups/group were challenged at 7 days of age with one of four rotavirus isolates. The challenge dose for each isolate was ~ 10^6 p.f.u./mouse, suspended in MEM in 100 microliter volume. This dose had been demonstrated to produce diarrhoea in 100% of neonatal mice born to rotavirus antibody-free dams 26. The virus challenge preparations were administered by intubation of the stomach with a soft flexible plastic feeding tube. The appearance of diarrhoea was scored clinically for up to 72 h postchallenge (Redmond et al., 1993).
Efficacy:
The highest antibody and virus neutralization titres were achieved following vaccination with VP4 cell-lysate. The significance of VP4 in the neutralization of virus was clearly defined by the high virus neutralization titres achieved following vaccination with any preparation containing VP4 (Redmond et al., 1993).
Description:
Two adjuvants were used in this study and were either mixed with MBP::VP6 prior to immunization or were administered separately. One was the attenuated E. coli heat-labile toxin LT(R192G). LT(R192G) carries a mutation in the trypsin cleavage site of the A subunit at arginine 192 (replaced by glycine) which abrogates cleavage and attenuates the toxicity of the protein. The other adjuvant was CTA1-DD which is composed of the enzymatically active A1 subunit of cholera toxin combined with a dimer of an immunoglobulin binding element from S. aureus protein A. Pre-clinical evaluations of CTA1-DD have reported it to be nontoxic (McNeal et al., 2007).
f. Adjuvant: cholera toxin A1-subunit-ProteinA D-fragment fusion protein
Description:
Two adjuvants were used in this study and were either mixed with MBP::VP6 prior to immunization or were administered separately. One was the attenuated E. coli heat-labile toxin LT(R192G). LT(R192G) carries a mutation in the trypsin cleavage site of the A subunit at arginine 192 (replaced by glycine) which abrogates cleavage and attenuates the toxicity of the protein. The other adjuvant was CTA1-DD which is composed of the enzymatically active A1 subunit of cholera toxin combined with a dimer of an immunoglobulin binding element from S. aureus protein A. Pre-clinical evaluations of CTA1-DD have reported it to be nontoxic (McNeal et al., 2007).
g. Immunization Route
Intraperitoneal injection (i.p.)
h.
Mouse Response
Host Strain:
BALB/c
Vaccination Protocol:
Under sedation, mice were immunized intrarectally by gently inserting a micropipette tip ca. 0.5 cm and instilling a 10 μl volume. The total volume for all immunogens used was 20 μl split in two doses of 10 μl. Groups of mice received two intrarectal immunizations of 3 μg MBP::VP6 and either LT(R192G) or CTA1-DD (10 μg of either adjuvant). This quantity of MBP::VP6 was used, based on the concentration of the MBP::VP6, to keep the volume under 20 μl to avoid loss from the anus. Groups of control mice were either mock immunized with phosphate-buffered saline (PBS) only or with MBP::VP6, LT(R192G), or CTA1-DD only. Immunizations were separated by 2 weeks (McNeal et al., 2007).
Challenge Protocol:
Four weeks after the second immunization, animals were either challenged with 10^5 shedding dose-50 (SD50) of wt EDIM by oral gavage to measure protection against fecal rotavirus shedding or they were sacrificed and their spleen cells were isolated and used for in vitro stimulation studies (McNeal et al., 2007).
Efficacy:
Intranasal or oral delivery of the chimeric rotavirus VP6 protein MBP::VP6 to mice elicited >90% reductions in fecal rotavirus shedding after murine rotavirus challenge (McNeal et al., 2007).
Host Gene Response of
Ifng (Interferon gamma)
Gene Response:
Splenic lymphocytes obtained 42 days after first immunization were stimulated for 18 hours with MBP::VP6. LT(R192G)+MBP:VP6 and CTA1-DD+MBP:VP6 produced significantly more IFN-gamma than did mock immunized (PBS-vaccinated) mice (McNeal et al., 2007).
Gene Response:
Splenic lymphocytes obtained 42 days after first immunization were stimulated for 18 hours with MBP::VP6. LT(R192G)+MBP:VP6 produced significantly more IL-17 than did mock immunized (PBS-vaccinated) mice (McNeal et al., 2007).
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2. Choi et al., 2002: Choi AH, McNeal MM, Flint JA, Basu M, Lycke NY, Clements JD, Bean JA, Davis HL, McCluskie MJ, VanCott JL, Ward RL. The level of protection against rotavirus shedding in mice following immunization with a chimeric VP6 protein is dependent on the route and the coadministered adjuvant. Vaccine. 2002; 20(13-14); 1733-1740. [PubMed: 11906760].
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13. Siadat-Pajouh and Cai, 2001: Siadat-Pajouh M, Cai L. Protective efficacy of rotavirus 2/6-virus-like particles combined with CT-E29H, a detoxified cholera toxin adjuvant. Viral immunology. 2001; 14(1); 31-47. [PubMed: 11270595].
