Molecular cloning and immunogenicity evaluation of rotavirus structural proteins as candidate vaccine

El‐Senousy, Waled M.; Shahein, Yasser E.; Barakat, Ahmed B.; Ghanem, Hossam E.; El-Hakim, Amr E.; Ameen, Saeed M.

doi:10.1016/j.ijbiomac.2013.04.003

Cited by 9 publications

(22 citation statements)

References 26 publications

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“…Recombinant vector protein rVP6F, chimeric protein that carrying VP4 epitope, P [4]223, P [4]56, or P [8]296, and reference rotavirus Wa, SA11 were used as immunogens. Four guinea pigs were used for each immunogen inoculation.…”

Section: Animals and Immunizationmentioning

confidence: 99%

“…So far, 27 G genotypes and 35 P genotypes in RVAs have been defined (27), and at least 73 G/P genotype combinations of RVAs infecting humans have been described (25). In humans, 12 G and 15 P types have been reported in the RVA (28), and G1P [8], G2P [4], G3P [8], G4P [8], G9P [8], and to a lesser extent G12P [8] are currently the most important genotypes worldwide (24)(25)(26)33,36). It is estimated that RVAs cause 453,000 deaths annually (39).…”

Section: Introductionmentioning

confidence: 99%

“…There are two oral live RV vaccines are currently available, Rotarix, a monovalent vaccine derived from a clinical human isolate of serotype G1P1A [8], and Rotateq, a pentavalent vaccine containing human RV (HRV) VP7 proteins of serotypes G1, G2, G3, and G4 and genotype P1A [8] on a G6P [5] bovine RV background (WC3 strain) (35,41). Clinical trials of the current RV vaccines reported at least 85% protection rates against severe diarrhea (35,41,42).…”

Section: Introductionmentioning

confidence: 99%

“…VP6 was reported to be involved in RV cell entry via its binding to the cellular heat shock protein (hsp70) (15); it might be related to the presence of neutralizing epitopes in VP6. Even a short fragment of VP6 could provide significant reduction in virus infectivity in vitro (8).…”

Section: Introductionmentioning

confidence: 99%

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A New Rotavirus VP6-Based Foreign Epitope Presenting Vector and Immunoreactivity of VP4 Epitope Chimeric Proteins

Teng

Zhao²,

Pan³

et al. 2014

Viral Immunology

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The VP6, the group antigenic rotavirus (RV), is highly conserved and the most abundant, constituting about 39% of the viral structure proteins by weight. The high degree of identity ( > 87%-99%) in the primary amino acid sequences suggests VP6-based vaccines could potentially provide heterotypic protection. Although some efforts have been made toward producing recombinant rotavirus VP6 vaccines, the native VP6 is still unsatisfactory as an optimal vaccine. The major neutralizing antigenic epitopes that exist on VP4 or VP7 are not on the native VP6, and as a vector the native VP6 lacks insertion sites that can be used for insertion of foreign epitopes. In this study, a new foreign epitope presenting system using VP6 as a vector (VP6F) was constructed on the outer surface of the vector six sites that could be used for insertion of the foreign epitopes created. Using this system, three VP6-based VP4 epitope chimeric proteins were constructed. Results showed that these chimeric proteins reacted with anti-VP6 and -VP4 antibodies, and elicited antibodies against VP6 and VP4 in guinea pigs. Antibodies against VP6F or antibodies against the chimeric proteins neutralized RV Wa and SA11 infection in vitro. It is optimistic that the limitation for using the native VP6 as a vaccine candidate or vector will be solved with our proposed approach. It is expected that this VP6-based epitope presenting system and the VP6-based VP4 epitope chimeric proteins will be valuable for and contribute to the development of novel RV vaccines and vaccine vectors.

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Section: Animals and Immunizationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A New Rotavirus VP6-Based Foreign Epitope Presenting Vector and Immunoreactivity of VP4 Epitope Chimeric Proteins

Teng

Zhao²,

Pan³

et al. 2014

Viral Immunology

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“…VP6, as the group antigen of RV, is the major structural protein forming the middle layer in the triple-layered viral capsid. For the last few years, studies have shown that VP6 could stimulate a protective immune response (Bugli et al, 2014;Li et al, 2014;Marashi et al, 2014;Pastor et al, 2014;Shoja et al, 2015) and a short fragment of VP6 could provide significant reduction in virus infectivity in vitro (El-Senousy et al, 2013). Previous experiments demonstrated that anti-VP6 llama-derived single-chain antibody fragments (VHH) had neutralizing activity against VP6 in vitro (Garaicoechea et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Immunogenicity and antigenicity of a recombinant chimeric protein containing epitopes of poliovirus type 1

Xx¹,

Wang²,

Wy³

et al. 2016

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Summary. -To design a vaccine that simultaneously prevents both rotavirus (RV) and poliovirus (PV), a PV type 1 (PV1) chimeric protein using RV VP6 as a vector (VP6F) was constructed, expressed in Escherichia coli expression system and characterized by SDS-PAGE, Western blot, immunofluorescence assay and neutralization test. The results showed that the chimeric protein reacted with anti-VP6F and anti-PV1 antibodies and elicited production of serum antibodies against the chimeric protein in guinea pigs. Antibodies against the chimeric protein neutralized RV Wa and PV1 infection in vitro. The results provided a relevant possibility of developing novel approaches in the rational design of vaccines effective against both RV and PV.Keywords: chimeric protein; neutralizing activity; rotavirus; poliovirus epitope * Corresponding author. E-mail: chenyd@imbcams.com.cn; phone: +86-871-68335402. Abbreviations: CPE = cytopathic effect; PV = poliovirus; RV = rotavirus; TCID 50 = 50% tissue culture infectivity doses

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Prevalence of Rotaviruses Groups A and C in Egyptian Children and Aquatic Environment

2015

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The objective of this study is to compare the prevalence of rotaviruses groups A and C in Egyptian children and aquatic environment. From 110 stool specimens of children with acute diarrhea and using RT-PCR, 35 samples (31.8 %) were positive for human rotavirus group A and 15 samples (13.6 %) were positive for human rotavirus group C. From 96 samples collected from Zenin wastewater treatment plant over a 2-year period (November 2009-October 2011) and using RT-PCR, rotavirus group A was detected in (4/24) 16.7 %, (5/24) 20.8 %, (4/24) 16.7 %, and (4/24) 16.7 %, while rotavirus group C was detected in (2/24) 8.3 %, (3/24) 12.5 %, (3/24) 12.5 %, and (0/24) 0 % in raw sewage, after primary sedimentation, after secondary sedimentation, and after final chlorination, respectively. Moreover, from 96 samples collected from El-Giza water treatment plant over a 2-year period (November 2009-October 2011), rotavirus group A was detected in (7/24) 29.2 %, (6/24) 25 %, (5/24) 20.8 %, and (3/24) 12.5 %, while rotavirus group C was detected in (3/24) 12.5 %, (1/24) 4.2 %, (1/24) 4.2 %, and (0/24) 0 % in raw Nile water, after sedimentation, after sand filtration, and after final chlorination, respectively. Using SYBR Green real-time RT-PCR, the number of human rotavirus group A genome or infectious units was higher than rotavirus group C. VP6 sequence analysis of the RT-PCR positive rotavirus group C samples revealed that four clinical specimens and three environmental samples showed similar sequences clustered with Moduganari/Human Nigerian strain AF 325806 with 98 % homology, and two clinical specimens and one environmental sample showed similar sequences clustered with Dhaka CB/Human Bangladesh strain AY 754826 with 97 % homology.

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Molecular cloning and immunogenicity evaluation of rotavirus structural proteins as candidate vaccine

Cited by 9 publications

References 26 publications

A New Rotavirus VP6-Based Foreign Epitope Presenting Vector and Immunoreactivity of VP4 Epitope Chimeric Proteins

A New Rotavirus VP6-Based Foreign Epitope Presenting Vector and Immunoreactivity of VP4 Epitope Chimeric Proteins

Immunogenicity and antigenicity of a recombinant chimeric protein containing epitopes of poliovirus type 1

Prevalence of Rotaviruses Groups A and C in Egyptian Children and Aquatic Environment

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