Marine-Sulfated Polysaccharides Extracts Exhibit Contrasted Time-Dependent Immunomodulatory and Antiviral Properties on Porcine Monocytes and Alveolar Macrophages.

Porcine respiratory complex syndrome has a strong economic impact on the swine breeding sector, as well as a clear repercussion on the wellbeing of the animals, leading to overuse of antimicrobial molecules. Algal extracts used in short-term treatments are empirically recognized by farmers as having a positive effect on pigs' health, however, their mechanisms of action are not well known and more research is needed. Herein we studied the short and median term impact of three algal extracts, in vitro, on the pro-inflammatory and antiviral responses of porcine primary blood monocytes and alveolar macrophages, as well as the susceptibility of the treated cells to infection by Porcine Respiratory and Reproductive Syndrome Virus (PRRSV) and the Aujeszky's Disease Virus (ADV). All extracts presented a pro-inflammatory short-term effect, associated for two of them, with an inhibition of the PRRSV replication. Conversely, the three extracts presented an anti-inflammatory median term effect, with no impact on PRRSV replication. The observed immune modulation prompts us to test, in vivo, the anti-PRRSV action of algal extracts and strengthen the interest for this natural resource.

The Internal Conduit System of the Swine Inverted Lymph Node.

Lymph nodes (LN) are the crossroad where naïve lymphocytes, peripheral antigens and antigen presenting cells contact together in order to mount an adaptive immune response. For this purpose, LN are highly organized convergent hubs of blood and lymphatic vessels that, in the case of B lymphocytes, lead to the B cell follicles. Herein take place the selection and maturation of B cell clones producing high affinity antibodies directed against various antigens. Whereas the knowledge on the murine and human LN distribution systems have reached an exquisite precision those last years, the organization of the antigens and cells circulation into the inverted porcine LN remains poorly described. Using up to date microscopy tools, we described the complex interconnections between afferent lymphatics and blood vessels, perifollicular macrophages, follicular B cells and efferent blood vessels. We observed that afferent lymphatic sinuses presented an asymmetric Lyve-1 expression similar to the one observed in murine LN, whereas specialized perifollicular sinuses connect the main afferent lymphatic sinus to the B cell follicles. Finally, whereas it was long though that mature B cells egress from the inverted LN in the T cell zone through HEV, our observations are in agreement with mature B cells accessing the efferent blood circulation in the efferent, subcapsular area. This understanding of the inverted porcine LN circuitry will allow a more accurate exploration of swine pathogens interactions with the immune cells inside the LN structures. Moreover, the mix between similarities and differences of porcine inverted LN circuitry with mouse and human normal LN shall enable to better apprehend the functions and malfunctions of normal LN from a new perspective.

Porcine Reproductive and Respiratory Syndrome Virus Interferes with Swine Influenza A Virus Infection of Epithelial Cells.

Respiratory infections are still a major concern in pigs. Amongst the involved viruses, the porcine reproductive and respiratory syndrome virus (PRRSV) and the swine influenza type A virus (swIAV) have a major impact. These viruses frequently encounter and dual infections are reported. We analyzed here the molecular interactions between viruses and porcine tracheal epithelial cells as well as lung tissue. PRRSV-1 species do not infect porcine respiratory epithelial cells. However, PRRSV-1, when inoculated simultaneously or shortly before swIAV, was able to inhibit swIAV H1N2 infection, modulate the interferon response and alter signaling protein phosphorylations (ERK, AKT, AMPK, and JAK2), in our conditions. SwIAV inhibition was also observed, although at a lower level, by inactivated PRRSV-1, whereas acid wash treatment inactivating non-penetrated viruses suppressed the interference effect. PRRSV-1 and swIAV may interact at several stages, before their attachment to the cells, when they attach to their receptors, and later on. In conclusion, we showed for the first time that PRRSV can alter the relation between swIAV and its main target cells, opening the doors to further studies on the interplay between viruses. Consequences of these peculiar interactions on viral infections and vaccinations using modified live vaccines require further investigations.

Assessment of pulmonary tissue responses in pigs challenged with PRRSV Lena strain shows better protection after immunization with field than vaccine strains.

The porcine reproductive and respiratory syndrome virus (PRRSV) is plaguing porcine production. Previously piglets were immunized with a PRRSV-1 commercial modified live virus vaccine (MLV1), a PRRSV-2 MLV (MLV2) or a Western European strain (Finistere: Fini) to assess and compare the protection brought by these strains upon challenge with virulent Lena strain. Lena viremia was reduced in all the immunized groups with a slightly higher level of protection following immunization with Fini. Using lung samples collected from the same experiment, tissue response to Lena challenge was assessed at the acute and chronic stages of infection. A pre-immunization with any one of the three PRRSV strains globally exacerbated microscopic lung lesions. Ten days post-challenge (DPC), MLV1 group score was higher than unimmunized group score and 42 DPC, MLV2 group score was higher than in unimmunized group. Lowest lung Lena viral loads were measured in Fini group. Using principal component analysis, we showed 10 DPC that the lesion score was positively correlated with chemokine receptors and negatively correlated with viral load. Forty-two DPC, variables for lesion score, IL6, IL8, and CCL20 transcripts were positively correlated together and negatively correlated with CCL28, CXCL6, and CXCR4 transcripts suggesting a role for the latter ones in the tissue recovery process. In conclusions, our study shows a significant impact of the three immunizations on pulmonary tissue with the best protection against Lena challenge conferred by Fini strain. Furthermore, it gives insight into the interactions between vaccine and Fini strains and the lung upon Lena challenge.

First demonstration of the circulation of a pneumovirus in French pigs by detection of anti-swine orthopneumovirus nucleoprotein antibodies.

The presence of pneumoviruses in pigs is poorly documented. In this study, we used the published sequence of the nucleoprotein (N) of the recently identified Swine Orthopneumovirus (SOV) to express and purify SOV N as a recombinant protein in Escherichia coli. This protein was purified as nanorings and used to set up an enzyme-linked immunosorbent assay, which was used to analyse the presence of anti-pneumovirus N antibodies in swine sera. Sera collected from different pig farms in the West of France and from specific pathogen free piglets before colostrum uptake showed indirectly that a pneumovirus is circulating in pig populations with some variations between animals. Piglets before colostrum uptake were sero-negative for anti-pneumovirus antibodies while most of the other pigs showed positivity. Interestingly, in two farms presenting respiratory clinical signs and negative or under control for some common respiratory pathogens, pigs were detected positive for anti-pneumovirus antibodies. Globally, anti-pneumovirus N antibody concentrations were variable between and within farms. Further studies will aim to isolate the circulating virus and determine its potential pathogenicity. SOV could potentially become a new member of the porcine respiratory complex, important on its own or in association with other viral and bacterial micro-organisms.