Replication of small ruminant lentiviruses in aluminum hydroxide-induced granulomas in sheep: a potential new factor for viral dissemination.

Aluminum (Al)-based salts are widely used adjuvants in ruminants and other species strengthening the immune response elicited against vaccine antigen/s. However, they can lead to the formation of long-lasting granulomas composed of abundant activated macrophages. Small ruminant lentiviruses (SRLV) are widely distributed macrophage-tropic retroviruses that cause persistent infections in sheep and goats. Infected monocytes/macrophages and dendritic cells establish an inflammatory microenvironment that eventually leads to clinical manifestations. The aim of this work was to study the effect of Al-induced granulomas in the replication and pathogenesis of SRLV. Eleven adult, naturally SRLV-infected sheep showing clinical arthritis were distributed in Vaccine (n=6), Adjuvant-only (n=3) and Control (n=2) groups and inoculated with commercial Al-based vaccines, Al hydroxide adjuvant or phosphate buffered saline, respectively. In vitro studies demonstrated viral replication in Al-induced granulomas in 5 out of 10 sheep. Immunohistochemistry (IHC) evinced granular, intracytoplasmic SRLV presence in macrophages within granulomas. Viral sequences obtained from granulomas, blood monocytes and other tissues were highly similar in most animals, suggesting virus circulation among body compartments. However, notable differences between isolated strains in granulomas and other tissues in specific animals were also noted. Interestingly, the B2 subtype was the most commonly found SRLV genotype reaching a wider body distribution than previously described. Recombination events between genotypes B2 and A3 along gag region were identified in two sheep. Our results indicate that Al-hydroxide derived granulomas may represent an ideal compartment for SRLV replication, perhaps altering natural SRLV infection by providing a new, suitable target tissue.IMPORTANCE Granulomas are inflammation-derived structures elicited by foreign bodies or certain infections. Aluminum adjuvants included in vaccines induce granulomas in many species. In sheep, these are persistent and consist of activated macrophages. Small Ruminant Lentiviruses (SRLV), which are macrophage-tropic lentivirus, cause a chronic wasting disease affecting animal welfare and production. Here, we studied the occurrence of SRLV in post-vaccination granulomas retrieved from naturally infected ewes after vaccination or inoculation with aluminum only. SRLV infection was confirmed in granulomas by identification of viral proteins, genomic fragments and enzymatic activity. The infecting SRLV strain, previously found exclusively in carpal joints, reached the central nervous system, suggesting that occurrence of SRLV in post-vaccination granulomas may broaden tissue tropism. SRLV recombination was found in inoculated animals, a rare event in sheep lentiviruses. Potentially, virus-host interactions within granulomas may modify viral pathogenesis and lead to more widespread infection.