Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4)
Background/Objectives: Equine rhinopneumonia, caused by equine herpesvirus types 1 and 4 (EHV-1 and EHV-4), continues to be a significant health and economic concern in the global equine industry, particularly in Kazakhstan. While vaccines targeting EHV-1 are available, there is currently no license...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2025-05-01
|
| Series: | Vaccines |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-393X/13/6/604 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849433866712907776 |
|---|---|
| author | Lespek Kutumbetov Balzhan Myrzakhmetova Aiganym Tussipova Gulzhan Zhapparova Talshyn Tlenchiyeva Karina Bissenbayeva Sergazy Nurabayev Aslan Kerimbayev |
| author_facet | Lespek Kutumbetov Balzhan Myrzakhmetova Aiganym Tussipova Gulzhan Zhapparova Talshyn Tlenchiyeva Karina Bissenbayeva Sergazy Nurabayev Aslan Kerimbayev |
| author_sort | Lespek Kutumbetov |
| collection | DOAJ |
| description | Background/Objectives: Equine rhinopneumonia, caused by equine herpesvirus types 1 and 4 (EHV-1 and EHV-4), continues to be a significant health and economic concern in the global equine industry, particularly in Kazakhstan. While vaccines targeting EHV-1 are available, there is currently no licensed monovalent vaccine for EHV-4, and existing formulations offer limited protection against this serotype. This study aimed to develop and evaluate a freeze-dried, live-attenuated EHV-4 vaccine with improved safety, stability, and immunogenicity. Methods: A field isolate of EHV-4 was attenuated through serial passaging in primary lamb testicle (LT-KK49) cell cultures. Viral biomass was concentrated and formulated with various stabilizers before freeze-drying. The most effective stabilizer composition—sucrose, gelatin, and lactalbumin hydrolysate—was selected based on viral titer retention. Safety and immunogenicity were assessed in mice, guinea pigs, rabbits, donkeys, and horses. A guinea pig reproductive challenge model was used to evaluate protective efficacy. Results: The optimized lyophilized vaccine retained infectivity (>6.0 log<sub>10</sub> TCID<sub>50/cm</sub><sup>3</sup>) for at least six months at 4 °C. No adverse clinical signs were observed in any test species. Immunization induced robust neutralizing antibody responses in both small animals and equines. In the guinea pig model, vaccinated females demonstrated 100% pregnancy retention and fetal viability following challenge with a virulent EHV-4 strain. Conclusions: This freeze-dried, live-attenuated EHV-4 vaccine candidate is safe, immunogenic, and thermostable. It offers a promising platform for the targeted prevention of EHV-4 infection, particularly in young horses and in regions with limited cold-chain infrastructure. |
| format | Article |
| id | doaj-art-e215bcb305b341478081cf4a61af82c6 |
| institution | Kabale University |
| issn | 2076-393X |
| language | English |
| publishDate | 2025-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj-art-e215bcb305b341478081cf4a61af82c62025-08-20T03:26:52ZengMDPI AGVaccines2076-393X2025-05-0113660410.3390/vaccines13060604Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4)Lespek Kutumbetov0Balzhan Myrzakhmetova1Aiganym Tussipova2Gulzhan Zhapparova3Talshyn Tlenchiyeva4Karina Bissenbayeva5Sergazy Nurabayev6Aslan Kerimbayev7Research Institute for Biological Safety Problems, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, Gvardeiskiy 080409, KazakhstanResearch Institute for Biological Safety Problems, Gvardeiskiy 080409, KazakhstanBackground/Objectives: Equine rhinopneumonia, caused by equine herpesvirus types 1 and 4 (EHV-1 and EHV-4), continues to be a significant health and economic concern in the global equine industry, particularly in Kazakhstan. While vaccines targeting EHV-1 are available, there is currently no licensed monovalent vaccine for EHV-4, and existing formulations offer limited protection against this serotype. This study aimed to develop and evaluate a freeze-dried, live-attenuated EHV-4 vaccine with improved safety, stability, and immunogenicity. Methods: A field isolate of EHV-4 was attenuated through serial passaging in primary lamb testicle (LT-KK49) cell cultures. Viral biomass was concentrated and formulated with various stabilizers before freeze-drying. The most effective stabilizer composition—sucrose, gelatin, and lactalbumin hydrolysate—was selected based on viral titer retention. Safety and immunogenicity were assessed in mice, guinea pigs, rabbits, donkeys, and horses. A guinea pig reproductive challenge model was used to evaluate protective efficacy. Results: The optimized lyophilized vaccine retained infectivity (>6.0 log<sub>10</sub> TCID<sub>50/cm</sub><sup>3</sup>) for at least six months at 4 °C. No adverse clinical signs were observed in any test species. Immunization induced robust neutralizing antibody responses in both small animals and equines. In the guinea pig model, vaccinated females demonstrated 100% pregnancy retention and fetal viability following challenge with a virulent EHV-4 strain. Conclusions: This freeze-dried, live-attenuated EHV-4 vaccine candidate is safe, immunogenic, and thermostable. It offers a promising platform for the targeted prevention of EHV-4 infection, particularly in young horses and in regions with limited cold-chain infrastructure.https://www.mdpi.com/2076-393X/13/6/604equine herpesvirus type 4lyophilized vaccineEHV-4 vaccinethermostabilityimmunogenicityvaccine development |
| spellingShingle | Lespek Kutumbetov Balzhan Myrzakhmetova Aiganym Tussipova Gulzhan Zhapparova Talshyn Tlenchiyeva Karina Bissenbayeva Sergazy Nurabayev Aslan Kerimbayev Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4) Vaccines equine herpesvirus type 4 lyophilized vaccine EHV-4 vaccine thermostability immunogenicity vaccine development |
| title | Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4) |
| title_full | Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4) |
| title_fullStr | Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4) |
| title_full_unstemmed | Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4) |
| title_short | Development and Preclinical Evaluation of a Lyophilized Vaccine Against Equine Herpesvirus Type 4 (EHV-4) |
| title_sort | development and preclinical evaluation of a lyophilized vaccine against equine herpesvirus type 4 ehv 4 |
| topic | equine herpesvirus type 4 lyophilized vaccine EHV-4 vaccine thermostability immunogenicity vaccine development |
| url | https://www.mdpi.com/2076-393X/13/6/604 |
| work_keys_str_mv | AT lespekkutumbetov developmentandpreclinicalevaluationofalyophilizedvaccineagainstequineherpesvirustype4ehv4 AT balzhanmyrzakhmetova developmentandpreclinicalevaluationofalyophilizedvaccineagainstequineherpesvirustype4ehv4 AT aiganymtussipova developmentandpreclinicalevaluationofalyophilizedvaccineagainstequineherpesvirustype4ehv4 AT gulzhanzhapparova developmentandpreclinicalevaluationofalyophilizedvaccineagainstequineherpesvirustype4ehv4 AT talshyntlenchiyeva developmentandpreclinicalevaluationofalyophilizedvaccineagainstequineherpesvirustype4ehv4 AT karinabissenbayeva developmentandpreclinicalevaluationofalyophilizedvaccineagainstequineherpesvirustype4ehv4 AT sergazynurabayev developmentandpreclinicalevaluationofalyophilizedvaccineagainstequineherpesvirustype4ehv4 AT aslankerimbayev developmentandpreclinicalevaluationofalyophilizedvaccineagainstequineherpesvirustype4ehv4 |