The use of an avian macrophage-like cell line (HD11) transduced with recombinant lentiviruses to dissect the immunomodulatory role of Marek's disease virus gene products

Authors: Alvarez Narvaez, Sonsiray; Kim, Taejoong; Spatz, Stephen; Conrad, Steven; Dunn, John

Submitted to: Virus Genes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/22/2026
Publication Date: 3/27/2026
Citation: Alvarez Narvaez, S., Kim, T.N., Spatz, S.J., Conrad, S.J., Dunn, J.R. 2026. The use of an avian macrophage-like cell line (HD11) transduced with recombinant lentiviruses to dissect the immunomodulatory role of Marek's disease virus gene products. Virus Genes. https://doi.org/10.1007/s11262-026-02224-z.
DOI: https://doi.org/10.1007/s11262-026-02224-z

Interpretive Summary: Marek's disease is a sickness chickens can get from Marek's disease virus (MDV). This virus is highly contagious and can cause tumors. Right now, we use shots (vaccines) to protect chickens. But MDV is tricky. It keeps changing, and new, stronger versions can pop up that the vaccines don't work against. So, we need better vaccines. When a chicken gets a virus, its body has a first line of defense called the innate immune system. Special helpers called type I interferons (IFN-I) are a big part of this defense. Like other viruses in its family, MDV has ways to stop the chicken's body from making IFN-I. If we can turn off these parts of MDV, we could make weaker versions of the virus to use as vaccines. These new vaccines would help the chicken's body make more IFN-I. To do this, we need a way to find which parts of MDV stop the chicken's body from making IFN-I. We made a new test that measures this. We tested five MDV genes that we thought might stop IFN-I from being made. With our new test, we showed that all five genes inhibited the chicken cells from making IFN-I.

Technical Abstract: Marek’s disease (MD) is caused by the highly contagious Marek’s disease virus (MDV), a tumorigenic alphaherpesvirus. The disease is presently managed through vaccination. Nevertheless, the dynamic nature of MDV presents challenges, as new virulent strains may arise that can potentially overcome existing vaccine immunity. To address this, new and more effective vaccines are crucial. The immune system's first line of defense against infections is the innate immune response, where type I interferons (IFN-I) play a key role. As with other alphaherpesviruses, MDV encodes numerous genes capable of modulating IFN-I production during infection. Ablation of these genes in MDV could result in attenuation, thereby facilitating the creation of novel vaccine strains with augmented IFN-I induction during infection. Hence, a system to identify immunomodulatory MDV gene products that impede IFN-I production in avian innate immune cells is required. Herein, we present an innovative screening method that quantifies the expression of Interferon Omega 1 (IFN'1) in an avian macrophage-like cell line expressing MDV genes via recombinant lentivirus (rLV) integration. Five MDV gene products hypothesized to inhibit IFN-I production (Meq, US3, R-LORF4, UL46, and UL48) were employed to evaluate our selection system. Stable expression of all MDV genes was obtained when rLV was utilized to insert these genes into HD11 cells. Additionally, we established the optimal conditions for activation of the IFN-I response in HD11 cells using a 2kb double-stranded DNA segment as a stimulant. With this approach, we demonstrated that these MDV gene products significantly decreased the gene expression of IFN-I when activated by transfection with double-stranded DNA.