T-Cell Cytokine Response in Salmonella Typhimurium-Vaccinated versus Infected Pigs.

Vaccination with the live attenuated vaccine Salmoporc is an effective measure to control Salmonella Typhimurium (STM) in affected swine populations. However, the cellular immune response evoked by the Salmoporc vaccine including differences in vaccinated pigs versus non-vaccinated pigs upon STM infection have not been characterized yet. To investigate this, tissue-derived porcine lymphocytes from different treatment groups (vaccination-only, vaccination and infection, infection-only, untreated controls) were stimulated in vitro with heat-inactivated STM and abundances of IFN-γ, TNF-α and/or IL-17A-producing T-cell subsets were compared across organs and treatment groups. Overall, our results show the induction of a strong CD4+ T-cell response after STM infection, both locally and systemically. Low-level induction of STM-specific cytokine-producing CD4+ T cells, notably for the IFN-γ/TNF-α co-producing phenotype, was detected after vaccination-only. Numerous significant contrasts in cytokine-producing T-cell phenotypes were observed after infection in vaccinated and infected versus infected-only animals. These results suggest that vaccine-induced STM-specific cytokine-producing CD4+ T cells contribute to local immunity in the gut and may limit the spread of STM to lymph nodes and systemic organs. Hence, our study provides insights into the underlying immune mechanisms that account for the efficacy of the Salmoporc vaccine.

Vaccination and Infection of Swine With Salmonella Typhimurium Induces a Systemic and Local Multifunctional CD4+ T-Cell Response.

The gram-negative facultative intracellular bacteria Salmonella Typhimurium (STM) often leads to subclinical infections in pigs, but can also cause severe enterocolitis in this species. Due to its high zoonotic potential, the pathogen is likewise dangerous for humans. Vaccination with a live attenuated STM strain (Salmoporc) is regarded as an effective method to control STM infections in affected pig herds. However, information on the cellular immune response of swine against STM is still scarce. In this study, we investigated the T-cell immune response in pigs that were vaccinated twice with Salmoporc followed by a challenge infection with a virulent STM strain. Blood- and organ-derived lymphocytes (spleen, tonsils, jejunal and ileocolic lymph nodes, jejunum, ileum) were stimulated in vitro with heat-inactivated STM. Subsequently, CD4+ T cells present in these cell preparations were analyzed for the production of IFN-γ, TNF-α, and IL-17A by flow cytometry and Boolean gating. Highest frequencies of STM-specific cytokine-producing CD4+ T cells were found in lamina propria lymphocytes of jejunum and ileum. Significant differences of the relative abundance of cytokine-producing phenotypes between control group and vaccinated + infected animals were detected in most organs, but dominated in gut and lymph node-residing CD4+ T cells. IL-17A producing CD4+ T cells dominated in gut and gut-draining lymph nodes, whereas IFN-γ/TNF-α co-producing CD4+ T cells were present in all locations. Additionally, the majority of cytokine-producing CD4+ T cells had a CD8α+CD27- phenotype, indicative of a late effector or effector memory stage of differentiation. In summary, we show that Salmonella-specific multifunctional CD4+ T cells exist in vaccinated and infected pigs, dominate in the gut and most likely contribute to protective immunity against STM in the pig.

T-helper cells from naive to committed.

T-helper cells play a central role in the onset and regulation of the antigen-specific immune response. In swine, two subpopulations of CD4 positive T-helper cells could be defined in extra-thymic compartments. Both differ in the surface antigen expression of distinct antigens. Besides the CD4(+)CD8(-) T-helper cell subpopulation representing the phenotype of T-helper cells known from other species, in swine CD4(+) T-helper cells exist showing expression of CD8alpha and MHCII molecules.In primary in vitro immune reactions after simulation with SEB the main response is distributed to the CD4(+)CD8(-) T-cell subpopulation, which contains the majority of naive T-helper cells. During the immune response CD8alpha as well as MHCII molecules are expressed on the surface of activated T-helper cells. This in vitro maturation is combined with a down-regulation of CD45RC. Thus, activated T-helper cells represent a cell phenotype with high similarity to the second porcine T-helper cell subpopulation. This CD4(+)CD8alpha(+) subpopulation contains in its majority MHCII(+) and CD45RC(-) cells. In a secondary in vitro immune response against classical swine fever virus only the CD4(+)CD8(+) T-helper subpopulation is able to respond. Therefore, T-helper memory cells can be distributed to this T-lymphocyte subpopulation. In summary, naive CD4(+)CD8(-)MHCII(-) porcine T-helper cells show an extra-thymic maturation to committed CD4(+)CD8alpha(+)MHCII(+) T-helper cells.

Influence of age on the effectiveness of PCV2 vaccination in piglets with high levels of maternally derived antibodies.

Two field studies were conducted to investigate the influence of age on the efficacy of vaccination against Porcine Circovirus Diseases (PCVD) in animals with high levels of maternally derived antibodies (MDA). A total of 416 piglets (Study 1) and 600 piglets (Study 2) were randomly allocated to one of three groups. Two groups in each study received a single dose of a PCV2 subunit vaccine, one group at 1 week old and the other at 3 weeks of age. The third group was left untreated. Animals vaccinated at 3 weeks of age showed a significantly higher average daily weight gain and significantly reduced viraemia following PCV2 infection than the respective control groups. This difference was not observed in pigs vaccinated at 1 week of age. Furthermore, only animals vaccinated at 3 weeks of age showed an increased serological response and a higher frequency of IgM-positive animals compared with controls. The data indicated that PCV2 vaccination in the presence of high MDA levels is efficacious when used in 3-week old but not in 1-week old pigs. As the range of MDA titres of pigs vaccinated at both 1 and 3 weeks of age were comparable, the data suggest that PCV2 vaccine efficacy was independent of the level of MDA. It appears that other age-related factors affecting the active and passive transfer of immunity may perhaps have interfered with the efficacy of the vaccine in 1-week old piglets. These findings have implications for future PCV2 vaccine testing and administration strategies.

Respiratory function and pulmonary lesions in pigs infected with porcine reproductive and respiratory syndrome virus.

Pulmonary dysfunction was evaluated in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV, isolate VR-2332) and compared to clinical and pathological findings. Infected pigs developed fever, reduced appetite, respiratory distress and dullness at 9 days post-inoculation (dpi). Non-invasive pulmonary function tests using impulse oscillometry and rebreathing of test gases (He, CO) revealed peripheral airway obstruction, reduced lung compliance and reduced lung CO-transfer factor. PRRSV-induced pulmonary dysfunction was most marked at 9-18 dpi and was accompanied by a significantly increased respiratory rate and decreased tidal volume. Expiration was affected more than inspiration. On histopathological examination, multifocal areas of interstitial pneumonia (more severe and extensive at 10 dpi than 21 dpi) were identified as a possible structural basis for reduced lung compliance and gas exchange disturbances.

The effect of vaccination against porcine circovirus type 2 in pigs suffering from porcine respiratory disease complex.

A field study was conducted to investigate the effect of vaccination against porcine circovirus type 2 (PCV2) in pigs suffering from porcine respiratory disease complex (PRDC). A total of 1542 pigs were allocated randomly into two treatment groups at approximately 20 days of age. Groups received either a Baculovirus-expressed recombinant PCV2 Open Reading Frame (ORF) 2 vaccine or placebo by single intramuscular injection. Median onset of PCV2 viraemia and respiratory signs occurred when animals were 18 weeks old. Vaccination reduced the mean PCV2 viral load by 55-83% (p < 0.0001) and the mean duration of viraemia by 50% (p < 0.0001). During the period of study (from 3 to 25 weeks of age) vaccinated animals exhibited a reduced mortality rate (6.63% vs. 8.67%, difference -2.04%; p = 0.1507), an improved average daily weight gain (649 g/day vs. 667 g/day; difference +18 g/day; p < 0.0001) and a reduced time to market (164.8 days vs. 170.4 days; difference -5.6 days; p < 0.0001). The effects on performance were greatest in the 8-week period between the onset of PCV2 viraemia and the end of finishing. These data demonstrate that vaccination against PCV2 alone can significantly improve the overall growth performance of pigs in a multi-factorial, late occurring disease complex such as PRDC.

Reduction of PMWS-associated clinical signs and co-infections by vaccination against PCV2.

The effects of a single-dose recombinant Porcine circovirus type 2 (PCV2) open reading frame 2 (ORF2) subunit vaccine were studied in a post-weaning multisystemic wasting syndrome (PMWS)-affected pig herd. A total of 1519 3-week-old piglets were allocated randomly into two treatment groups and either vaccinated against PCV2 or treated with a placebo. Study animals were followed from the time of vaccination until the end of finishing. Onset of PCV2 viraemia and clinical signs of PMWS (wasting, cough, dyspnoea, pallor and lethargy) were observed when animals were approximately 9-10 weeks old. Compared to placebo-treated animals, vaccinated animals had a significantly reduced PCV2 viral load and duration of viraemia (p < 0.0001). This reduction in viraemia was not affected by the level of maternal anti-PCV2 antibodies present at the time of vaccination. During the period of viraemia (10-26 weeks of age) vaccinated animals exhibited a 53% reduction in mortality rate (p = 0.0010), a 4.84 kg higher body weight gain (p < 0.0001) and a significant reduction in clinical signs (p < or = 0.0004). Furthermore, lung samples of vaccinated animals had a considerably reduced number of co-infections with PRRSV and Mycoplasma hyorhinis than lung samples of placebo-treated animals. These data indicate that vaccination against PCV2 alone protects pigs from clinical signs and co-infections associated with PMWS.