A simple and rapid method for generating antibodies against bovine alphaherpesvirus 1 viral proteins through immunization with virus-infected murine cells

Bovine alphaherpesvirus 1 (BoAHV-1) infects cattle and typically results in significant economic losses for the cattle industry worldwide. Currently, antibodies targeting only a limited number of viral proteins are commercially available. It has been reported that BoAHV-1 is capable of infecting numerous tumor cell lines. Based on the rationale of immune tolerance, we hypothesized that virus-infected murine cells could be directly used to immunize mice, thereby generating antibodies against viral proteins. In this study, we found that BoAHV- 1 can infect murine cell lines including LA795 and MC38, as determined using both Western blot and immu nofluorescence analyses. Immunizing mice with virus-infected cells, either through subcutaneous or intraperi toneal injection, stimulates the production of high levels of antibodies that specifically recognize the viral proteins synthesized in bovine kidney (MDBK) cells, as characterized by both Western blot and/or immunoflu orescence. Furthermore, our findings suggest that intraperitoneal immunization could more effectively elicit antibodies against a wider array of viral proteins. As a homemade antibody generation method, this approach bypasses the complex and time-consuming steps of producing and purifying recombinant proteins as antigen, which are typically performed in conventional methods for antibody generation. Thus, we present a simple, rapid, and cost-effective method for generating virus-specific antibodies.