Efficient delivery of the immunodominant genes of African swine fever virus by adeno-associated virus serotype 2.

Background and Aim: Adeno-associated virus serotype 2 (AAV2) represents a promising basis for developing a virus-vector vaccine against African swine fever (ASF). This study aimed to create genetic constructs based on AAV2 to deliver the immunodominant genes of ASF virus (ASFV) and to evaluate their functionality in vitro. The efficiency and specificity of transgene expression, as well as their non-toxicity in cells of target animals, were evaluated. Materials and Methods: Bioinformatics analysis methods were used to identify the immunodominant genes of ASFV. The target genes B646L, E183L, CP204L, and CP530R were identified and subsequently cloned into the pAAV-MCS vector. Assembly of recombinant AAV2 (rAAV2) was performed by cotransfection of AAV293 cells with the following plasmids: pAAV-MCS with the gene of interest, envelope, and packaging. Quantitative polymerase chain reaction was used to determine the AAV2 titer. The functionality of the constructs was evaluated in HEK293 and SPEV cells by determining the presence of mature proteins in the cell lysate and the expression levels of messenger RNA. The specificity of the target proteins in cell lysates was confirmed by Western blotting. Results: The proposed AAV2 assembly protocol makes it possible to achieve a concentration of mature viral particles of at least 280 billion/mL of virus-containing material. The rAAV2 could effectively transduce host SPEV cells. The expression of both cistrons was detectable during the transduction of cells; therefore, the combined expression of immunogens in the cells of target animals should be possible using this method. Conclusion: This study demonstrated the potential of using genetic constructs based on AAV2 for the delivery of ASFV genes in vitro.

Viral Vector Vaccines Against ASF: Problems and Prospectives.

African swine fever (ASF) is a highly contagious viral disease affecting pigs, with mortality rates a primary focus as they can reach up to 100%. The widespread and colossal economic losses from ASF have impacts on the development of animal husbandry practices in most countries within Africa, Asia, and Europe. Currently, a variety of approaches toward the development of vaccines against ASF are being employed. A promising new concept centered around more economical and time-consuming vaccine production is based on the use of viral vectors to deliver selected immunogens. This review discusses the results obtained from testing various viral vectors as carriers of targeted ASF virus genes. The safety and prospects of viral vectors, the possibilities around modulating cellular and humoral immune responses by choosing genes expressing immunodominant antigens, and the degree of protection in experimental animals from infection with a lethal dose of virulent ASF virus strains have been shown and discussed.