Interaction of Type 1 Porcine Reproductive and Respiratory Syndrome Virus With In Vitro Derived Conventional Dendritic Cells

Abstract

The present study delineates the interaction of a typical PRRSV1.1 isolate 3267 (moderate virulence) with in vitro derived pig conventional dendritic cells, cDC1, cDC2, and a CD14+ population (designated as CD14+ DCs). cDC1 and cDC2 were not susceptible to 3267 infection, but a fraction of CD14+ DCs were infected. After exposure to the virus, all three DC types remained immature as determined by no increase of maturation molecules (MHC-I, MHC-II, CD80/86, CCR7), no release of cytokines, no modification of antigen presentation abilities, and no alteration of endocytic/phagocytic capabilities. However, when infected MARC-145 cells were used as a source of viral antigens, cDC2 and CD14+ DCs showed a significant increase in the expression of maturation molecules and substantial release of cytokines, notably IL-12/IL-23p40 (by both DC types) and IL-10 (by CD14+ DCs). To address the impact of PRRSV1 3267 on TLR3- and TLR7-mediated activation, cDC1, cDC2, and CD14+ DCs were inoculated by the virus (live or UV-inactivated) for 6 h prior to or simultaneously with the addition of poly I:C (TLR3 ligand) or gardiquimod (TLR7 ligand; not used for cDC1). Compared with using TLR ligand alone, combination with the virus did not result in any alteration to the maturation markers on all DC types but changed the cytokine response to either TLR3 or TLR7 ligand. Pre-exposure of cDC2 or CD14+ DCs to the live virus resulted in an increased production of IFN-α upon poly I:C stimulation, while pre-exposure to UV-inactivated virus tended to enhance the release of IL-10 upon gardiquimod stimulation. Simultaneous addition of the live virus and the TLR ligand either had no effect (mainly in cDC2) or impaired most of the cytokine release after gardiquimod stimulation (in CD14+ DCs). When used as antigen presenting cells, cDC2 pre-inoculated by the live virus before addition of gardiquimod impaired the proliferation of CD4–CD8– T cells. In the case of CD14+ DCs, pre-exposure to the live virus or simultaneously added with TLR3 or TLR7 ligand largely decreased the proliferation of CD4–CD8+ and CD4–CD8+ T-cell subsets. For cDC1, no significant changes were observed in cytokine responses or T-cell proliferation after poly I:C stimulation. Of note, cDC1 had a short life during in vitro culturing, for which the results obtained might be biased. Overall, exposure to PRRSV1 did not induce maturation of cDC1, cDC2, or CD14+ DCs, but modified TLR3 and TLR7-associated responses (except for cDC1), which may affect the development of adaptive immunity during PRRSV1 infection. Moreover, the sensing of infected cells was different from that of the free virus.

Swine Dendritic Cell Response to Porcine Reproductive and Respiratory Syndrome Virus: An Update

“…Exposure of pDCs to PRRSV did not significantly inhibit the natural capability of these cells to produce IFN-α except for some PRRSV2 strains, including the highly pathogenic PRRSV2 that emerged in China in 2006 and the highly virulent PRRSV1.3 Lena strain. When pDCs were exposed to infected macrophages, the IFN-α production was higher than that obtained when using cell-free virions ( 73 ), similar to what has been reported recently for cDCs ( 56 ). Efficient sensing of infected macrophages by pDCs required integrin-mediated intercellular contact and intact actin filaments in the macrophages ( 73 ).…”

“…Recently, Li and Mateu ( 56 ), using in vitro derived cDCs, also proved the non-susceptibility of cDC1 and cDC2. This work further revealed that cDC1 and cDC2 remained immature upon PRRSV inoculation concerning the expression of maturation molecules, cytokine production, and the endocytosis/phagocytosis capabilities.…”

“…Moreover, Bordet et al (29) did not observe IL-10 or TGF-b induced by highly virulent PRRSV1.3 Lena although Tregs were not assessed (29). Also, Li and Mateu (56) did not observe a significant proliferation of Tregs induced by in vitro derived allogeneic PRRSV1-stimulated cDCs (56). Given the diversity of PRRSV, further studies are required to determine whether PRRSV1 differed from PRRSV2 in inducing Tregs.…”

“…Taken together, cDCs are assumed to poorly participate in launching the adaptive immune response against PRRSV, which, however, remains to be confirmed by further experiments. Interestingly, Li and Mateu ( 56 ) reported that sensing of PRRSV by cDC was enhanced when infected cells were used as a source of antigen and hypothesized that the effective sensing of PRRSV by cDCs might not happen at the early stage of infection ( 56 ).…”

“…However, the lack of inclusion of groups where moDCs were stimulated with TLR ligands alone makes difficult to produce a definitive conclusion. In a study by Li and Mateu ( 56 ) the authors showed that simultaneous addition of a TLR7 ligand and PRRSV resulted in impaired response, particularly in cytokine production. Therefore, the use of TLR7 ligand as the adjuvant of PRRSV vaccines should be further examined.…”