Contraceptive efficacy of recombinant fusion protein comprising zona pellucida glycoprotein-3 fragment and gonadotropin releasing hormone

Arukha, Ananta Prasad; Minhas, Vidisha; Shrestha, Anmol Purna; Gupta, Seema

doi:10.1016/j.jri.2016.01.004

Cited by 13 publications

(19 citation statements)

References 66 publications

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“…3a, b ). These results were similar to other studies in which immunization with GnRH caused a significant reduction in FSH and E2 concentration [ 21 – 27 ]. Lower concentration of FSH and E2 possibly resulted in ovary dysfunction and infertility.…”

Section: Discussionsupporting

confidence: 92%

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The preclinical evaluation of immunocontraceptive vaccines based on canine zona pellucida 3 (cZP3) in a mouse model

Wang¹,

Li²,

Zhang³

et al. 2018

Reprod Biol Endocrinol

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BackgroundStray dogs are the reservoirs and carriers of rabies and are definitive hosts of echinococcosis. To control the overpopulation of stray dogs, zona pellucida 3 (ZP3), a primary receptor for sperm, is a potential antigen for developing contraceptive vaccines.To enhance the immune responses and contraceptive effects of canine ZP3 (cZP3), dog gonadotropin-releasing hormone (GnRH) and a T cell epitope of chicken ovalbumin (OVA) were selected to construct two fusion proteins with cZP3, ovalbumin-GnRH-ZP3 (OGZ) and ovalbumin-ZP3 (OZ), and their contraceptive effects were evaluated in mice.MethodsThe synthesized DNA sequences of OGZ and OZ were cloned into plasmid pET-28a respectively. The fusion proteins OGZ and OZ were identified by SDS-PAGE and Western blot. Mice were immunized with OGZ, OZ and cZP3, and the infertility rates were monitored. Mice immunized with mouse ZP3 (mZP3) or adjuvant alone were used as positive control and negative control, respectively. cZP3- and GnRH-specific antibodies (Abs) were detected by ELISA. The bindings of the Abs to oocytes were detected by indirect immunofluorescence assay. The paraffin sections of mice ovaries were observed under microscope for analyzing pathological characteristics.ResultsSDS-PAGE and Western blot analyses showed that the two fusion proteins OGZ and OZ were correctly expressed. ELISA results showed that OGZ vaccine induced both cZP3- and GnRH-specific Abs, and OZ vaccine induced cZP3-specific Ab, which lasted for up to 168 days. The levels of follicle stimulating hormone (FSH) and estradiol (E2) in sera were significantly decreased in OGZ immunized mice. Indirect immunofluorescence results showed that Abs induced by cZP3 and mZP3 could bind to the mouse ZP and dog ZP each other. Compared with the adjuvant group, all vaccine immunized groups significantly decreased the fertility rate and mean litter size. Interestingly, the fertility rate in OGZ-immunized group is the lowest, and only 1 mouse out of 10 mice is fertile. Histological analysis of murine ovarian sections indicated that most of the infertile mice in the immunized groups lacked mature follicles as well as accompanied by inflammatory infiltration. Meanwhile, immunization with OGZ decreased the number of corpora lutea in the infertile mice.ConclusionsThe fusion protein OGZ resulted in the lowest fertility rate and the least mean litter size in the immunized mice. OGZ might be a promising antigen for developing a new contraceptive vaccine for stray dog controlling.

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Section: Discussionsupporting

confidence: 92%

“…2b ), and this was probably due to the differences in the coating antigen. The Ab levels detected by ELISA with fusion proteins as coating antigens are usually higher than synthetic peptides as coating antigens, and this difference is similar to the results of previous studies [ 21 , 22 ].…”

Section: Discussionsupporting

confidence: 90%

The preclinical evaluation of immunocontraceptive vaccines based on canine zona pellucida 3 (cZP3) in a mouse model

Wang¹,

Li²,

Zhang³

et al. 2018

Reprod Biol Endocrinol

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“…Based on these results, we combined the S1‐WT and S1‐Mut proteins to form a bivalent vaccine, and our bivalent vaccine showed cross‐protection against both wild type and variants of SARS‐CoV‐2, which could be a candidate as a universal vaccine against SARS‐CoV‐2 and its mutants. Besides, fusion protein with different antigens has been reported as a cross‐protective bivalent vaccine to prevent several other viruses 24–26 . Therefore, we are delving into cross‐protective bivalent fusion vaccines composed of wild‐type and mutant spike protein fragments, which might be a promising strategy to prevent SARS‐CoV‐2 and mutant strains.…”

Section: Discussionmentioning

confidence: 99%

A bivalent recombinant vaccine targeting the S1 protein induces neutralizing antibodies against both SARS‐CoV‐2 variants and wild‐type of the virus

Yang

et al. 2021

MedComm

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The emerging variants of severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2) in pandemic call for the urgent development of universal corona virus disease 2019 (COVID‐19) vaccines which could be effective for both wild‐type SARS‐CoV‐2 and mutant strains. In the current study, we formulated protein subunit vaccines with AS03 adjuvant and recombinant proteins of S1 subunit of SARS‐CoV‐2 (S1‐WT) and S1 variant (K417N, E484K, N501Y, and D614G) subunit (S1‐Mut), and immunized transgenic mice that express human angiotensin‐converting enzyme 2 (hACE2). The S1 protein‐specific antibody production and the neutralization capability for SARS‐CoV‐2 and B.1.351 variant were measured after immunization in mice. The results revealed that the S1‐Mut antigens were more effective in inhibiting the receptor‐binding domain and ACE2 binding in B.1.351 variant than in wild‐type SARS‐CoV‐2. Furthermore, the development of a bivalent vaccine exhibited the ideal neutralization properties against wild‐type and B.1.351 variant, as well as other variants. Our findings may provide a rationale for the development of a bivalent recombinant vaccine targeting the S1 protein that can induce the neutralizing antibodies against both SARS‐CoV‐2 variants and wild‐type of the virus and may be of importance to explore the potential clinical use of bivalent recombinant vaccine in the future.

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“…Some of the methods used are by using multiple copies of GnRH arranged linearly, and use of toxin receptors such as tetanus toxoid [27]. The use of diphtheria toxin conjugated to zona pellucida protein has shown to improve the in vitro contraceptive efficacy, this suggests that the use of B cell epitopes of particular zona proteins may enhance contraceptive efficacy [31,32]. In this study, the contraceptive potential of a recombinant fusion protein GVAC08 containing the three genes GnRH, GnRH receptor, and ZP3 was explored.…”

Section: Discussionmentioning

confidence: 99%

“…The reason could be that antibody titer peak during a time period and then subside and eventually become undetected in the blood. With respect to contraceptive vaccine, the biggest challenge is to improve the efficacy to inhibit fertility [ 32 ].…”

Section: Discussionmentioning

confidence: 99%

Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein

Sandam

Prakash

Thimmareddy³

2021

Journal of Genetic Engineering and Biotechnology

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Background The management of stray dog population has been of utmost importance due to their overpopulation, increase in dog bites incidence, and rabies. Contraceptive vaccines, a non-surgical alternative to spaying and neutering are viewed as a valuable option for the management of dog population. In this study, the contraceptive potential of a recombinant fusion protein containing the three genes GnRH, GnRH receptor, and ZP3 was explored. Results The gene fragment encoding GnRH, GnRHR, and ZP3 along with the antigenic epitopes of canine distemper virus and tetanus toxoid was assembled, synthesized, and cloned into pET28a expression vector. The resulting construct GVAC08 was successfully transformed into BL21DE3 strain of E. coli and confirmed by colony PCR. The recombinant GVAC08 protein was expressed and purified using Ni-NTA and was confirmed to be a 50-KDa protein by SDS PAGE and Western blot. Mice were immunized with the GVAC08 protein using Freund’s complete adjuvant followed by a booster using Freund’s incomplete adjuvant. This induced a high antibody titer against GnRH, GnRH receptor, and ZP3 which was determined by ELISA. Conclusion Mating studies showed that the GVAC08 recombinant protein was able to reduce the litter size in immunized mice showing improved efficacy. However, the vaccine candidate with further improvements will be a viable contraceptive vaccine.

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Contraceptive efficacy of recombinant fusion protein comprising zona pellucida glycoprotein-3 fragment and gonadotropin releasing hormone

Cited by 13 publications

References 66 publications

The preclinical evaluation of immunocontraceptive vaccines based on canine zona pellucida 3 (cZP3) in a mouse model

The preclinical evaluation of immunocontraceptive vaccines based on canine zona pellucida 3 (cZP3) in a mouse model

A bivalent recombinant vaccine targeting the S1 protein induces neutralizing antibodies against both SARS‐CoV‐2 variants and wild‐type of the virus

Immunocontraceptive potential of a GnRH receptor-based fusion recombinant protein

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