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This paper highlights the career of an exceptional woman virologist, Dr. Ilaria Capua. It recollects her major achievements, awards and noteworthy events that have shaped her scientific and political career. It retraces Dr. Capua’s major contributions to the study of viral zoonoses, in particular influenza virus, and her strong commitment to an open, more ethical science at the service of society in its broadest sense. It describes how she became the long-term champion of “Open Access” and “Data Sharing” for virus genetic sequences and introduces her new concept of “Circular Health”, where health becomes a circular system that represents a central and vital connection hub between humans and nature. This paper features Dr. Capua’s value as a role model for young women scientists and their empowerment.

For decades, scientists have used column chromatography to purify an array of analytes. The same chromatography system has also been deployed for the isolation and purification of antibodies to increase sensitivity and specificity of detection assays such as western blot, enzyme-linked immunosorbent assay (ELISA), and immunohistochemistry. However, even with the combination of these modalities the detection of specific antibodies developed in response to treatment, like vaccination, remains difficult due to physiological differences among species, sample types evaluated, and differing physiological states. Therefore, we developed a high-throughput antibody isolation protocol to measure influenza-specific antibodies in pregnant and lactating pigs, because the samples, in particular serum, milk and colostrum, contain components that cause high background. We developed a high-throughput 96-well plate-based method using a modified column chromatography technique to specifically isolate swine immunoglobulin (Ig) isotypes. This method utilizes isotype-specific reagents to isolate the IgG and IgA antibody isotypes from various fluids collected at multiple time points from individual animals following immunization. After sample processing and antibody isolation, the results showed a rapid and consistent yield of specific IgG and IgA, with comparable outcomes between single-column chromatography and our 96-well plate system, the latter offering a time-saving advantage. Specifically, the standard single-column chromatography required 2 to 3 hours to isolate 12 samples, whereas our method enabled the isolation of 192 samples in just 8 to 9 hours making this the ideal method for an immunogenicity study utilizing a variety of animal samples from multiple timepoints.

Selenium is an essential trace mineral important for the maintenance of homeostasis in animals and humans. It evinces a strong antioxidant, anti-inflammatory and potential antimicrobial capacity. Selenium biological function is primarily achieved by its presence in selenoproteins as a form of selenocysteine. Selenium deficiency may result in an array of health disorders, affecting many organs and systems; to prevent this, dietary supplementation, mainly in the forms of organic (i.e., selenomethionine and selenocysteine) inorganic (i.e., selenate and selenite) sources is used. In pigs as well as other food animals, dietary selenium supplementation has been used for improving growth performance, immune function, and meat quality. A substantial body of knowledge demonstrates that dietary selenium supplementation is positively associated with overall animal health especially due to its immunomodulatory activity and protection from oxidative damage. Selenium also possesses potential antiviral activity and this is achieved by protecting immune cells against oxidative damage and decreasing viral replication. In this review we endeavor to combine established and novel knowledge on the beneficial effects of dietary selenium supplementation, its antioxidant and immunomodulatory actions, and the putative antimicrobial effect thereof. Furthermore, our review demonstrates the gaps in knowledge pertaining to the use of selenium as an antiviral, underscoring the need for further in vivo and in vitro studies, particularly in pigs. Graphical abstract

Selenium (Se) is a trace mineral with antioxidant and anti-inflammatory properties. Se deficiency increases oxidative stress and immunosuppression. In swine, dietary Se supplementation enhances immunity and growth, and previous studies suggest it protects immune cells during viral infection. Porcine reproductive and respiratory syndrome virus (PRRSV) causes severe respiratory and reproductive failure in swine, resulting in annual losses of 1.2 billion USD. Vaccine efficacy is hampered by the virus’s high mutation rate, requiring alternative approaches. This study examines the effects of organic (DL-Selenomethionine, L-Selenomethionine, yeast-selenium) and inorganic (sodium selenite) Se on PRRSV infection in vitro. Porcine alveolar macrophages, the primary target of PRRSV in the lung, were isolated from healthy animals and infected with PRRSV-2 with or without Se. Mitochondrial function, gene expression, oxidative stress, and viral load were assessed post-infection. DL-selenomethionine showed increased glycolytic and mitochondrial ATP production relative to other compounds, suggesting improved mitochondrial function. No antiviral activity against PRRSV was observed. Transcriptome analysis revealed infection-driven modulation, with upregulation of IL6, IL8, IL1B1, MX1, and TXNRD1, but Se had no significant effect. While Se did not exhibit antiviral activity in vitro, its enhancement of mitochondrial function offers additional insight supporting its potential immunomodulatory benefits observed in previous in vivo studies.

The placenta is a dynamic, embryo-derived organ essential for fetal growth and development. While all eutherian mammals have placentas composed of fetal-derived trophoblasts that mediate maternal–fetal exchange, their anatomical and histological structures vary across species due to evolutionary divergence. Despite the cellular heterogeneity of porcine trophoblasts in vivo, understanding the mechanisms driving porcine placental development has been limited by the lack of in vitro models replicating this heterogeneity. In this study, we derived swine trophoblast organoids (sTOs) from full-term porcine placentas, retaining key transcriptional signatures of in vivo trophoblasts. To identify conserved cell populations, we integrated Visium spatial transcriptomics from mid-gestation porcine placentas with single-cell transcriptomics from sTOs. Spatial transcriptomics revealed novel markers of the porcine uterus and placenta, enabling precise separation of histological structures at the maternal–fetal interface. The integration of tissue and sTO transcriptomics showed that sTOs spontaneously differentiate into distinct trophoblast populations, with conserved gene expression and cell communication programs. These findings demonstrate that sTOs recapitulate porcine placental trophoblast populations, offering a powerful model for advancing placentation research. Our work also provides a spatially resolved whole-transcriptome dataset of the porcine maternal–fetal interface, opening new avenues for discoveries in placental development, evolution, and health across mammals.

Asplenia is an important cause of morbidity and mortality in humans. However, there are only very rare examples of this condition reported in domesticated species. Here we present a case of asplenia, diagnosed at necropsy, in a crossbred adult female pig from an influenza vaccine study. The humoral antibody response, including immune response to an influenza A virus vaccine, was characterized and compared to a parity-matched pig from the same study. The antibody profiles, lower total IgM with similar levels of IgG, were remarkably similar to those described in human patients with asplenia. However, in response to vaccination, the asplenic pig showed a robust hemagglutinin-specific IgM response with lower levels of IgG and IgA. These results were mirrored in the passively transferred antibody profiles of the asplenic dam’s piglets. This constitutes the first case of congenital asplenia described in the pig.

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically devastating viruses for the global swine industry. PRRSV has a known tropism for lung macrophages, where it causes impaired immune responses. This study evaluated the metabolic and immune profiles of primary porcine alveolar macrophages (PAMs) and pulmonary intravascular macrophages (PIMs) infected with different strains of PRRSV-2 isolated from North Carolina (NC) pig herds (NC134, NC18-9-7 referred to as NC174, and NC20-1 referred to as NC144), and VR2232, a PRRSV-2 prototype strain. Primary enriched mononuclear phagocytes were infected with NC134 and NC174, sorted, and processed. The total RNA was used for a transcriptomic approach; additionally, gene expression was further validated using RT-qPCR and NanoString technology. Complementary functional assays with additional NC strains were used to further investigate the mitochondrial and metabolic dysfunction, as well as the oxidative stress induced by PRRSV-2 infection. PAMs infected with both NC PRRSV-2 strains NC174 and NC134 showed similar transcriptomic profiles during the early stage of infection, with downregulation of genes involved in the oxidative phosphorylation and electron transport chain pathways. PIMs infected with both NC174 and NC134 strains showed limited alteration in the transcriptomic profiles compared to uninfected cells. Genetic reprogramming matched the PRRSV-2-induced mitochondrial impairment observed in functional assays performed using Seahorse technology. Mitochondrial respiration displayed slightly different profiles between PIMs and PAMs infected with the different PRRSV-2 strains, with PAMs showing a more substantial decrease in mitochondrial fitness compared to control cells. When reactive oxygen species (ROS) and nitric oxide (NO) production were evaluated, no differences were observed between PRRSV-2-infected PAMs and PIMs and control cells. These results provide valuable insights into the pathogenetic mechanism of different NC PRRSV-2 strains by focusing on the alteration in mitochondrial function in lung macrophages during early infection and highlighting differences in lung macrophage responses to distinct PRRSV-2 strains.

Background/Objectives: The porcine respiratory disease complex (PRDC) is a multifaceted, polymicrobial syndrome resulting from a combination of environmental stressors, primary infections (e.g., PRRSV) and secondary infectious agents (viruses and bacteria). PRDC causes severe lung pathology, leading to reduced performance, increased mortality rates, and higher production costs in the global pig industry. Our goal was to conduct a comprehensive study correlating both the anti-PRRSV immune response and 21 secondary infectious agents with PRDC severity. Methods: To this end, PRRSV-negative weaners were vaccinated with a PRRSV-2 MLV and put into a farm with a history of PRDC. Subsequently, anti-PRRSV cellular and antibody responses were monitored pre-vaccination, at 28 days post vaccination (dpv) and during PRDC outbreak (49 dpv). NanoString was used to quantify 21 pathogens within the bronchoalveolar lavage (BAL) at the time of necropsy (51 dpv). PRRSV-2 was present in 53 out of 55 pigs, and the other five pathogens (PCMV, PPIV, B. bronchiseptica, G. parasuis, and M. hyorhinis) were detected in BAL samples. Results: Although the uncontrolled settings of field trials complicated data interpretation, multivariate correlation analyses highlighted valuable lessons: (i) high weaning weight predicted animal resilience to disease and high weight gains correlated with the control of the PRRSV-2 field strain; (ii) most pigs cleared MLV strain within 7 weeks, and the field PRRSV-2 strain was the most prevalent lung pathogen during PRDC; (iii) all pigs developed a systemic PRRSV IgG antibody response which correlated with IgG and IgA levels in BAL; (iv) the induction of anti-field strain-neutralizing antibodies by MLV PRRSV-2 vaccination was both late and limited; (v) cellular immune responses were variable but included strong systemic IFN-γ production against the PRRSV-2 field strain; (vi) the most detected lung pathogens correlated with PRRSV-2 viremia or lung loads; (vii) within the six detected pathogens, two viruses, PRRSV-2 and PCMV, significantly correlated with the severity of the clinical outcome. Conclusions: While a simple and conclusive answer to the multifaceted nature of PRDC remains elusive, the key lessons derived from this unique study provide a valuable framework for future research on porcine respiratory diseases.

Porcine respiratory disease complex (PRDC) is a polymicrobial syndrome that results from a combination of infectious agents, such as environmental stressors, population size, management strategies, age, and genetics. PRDC results in reduced performance as well as increased mortality rates and production costs in the pig industry worldwide. This review focuses on the interactions of two enveloped RNA viruses—porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza virus (SwIV)—as major etiological agents that contribute to PRDC within the porcine cellular innate immunity during infection. The innate immune system of the porcine lung includes alveolar and parenchymal/interstitial macrophages, neutrophils (PMN), conventional dendritic cells (DC) and plasmacytoid DC, natural killer cells, and γδ T cells, thus the in vitro and in vivo interactions between those cells and PRRSV and SwIV are reviewed. Likewise, the few studies regarding PRRSV-SwIV co-infection are illustrated together with the different modulation mechanisms that are induced by the two viruses. Alterations in responses by natural killer (NK), PMN, or γδ T cells have not received much attention within the scientific community as their counterpart antigen-presenting cells and there are numerous gaps in the knowledge regarding the role of those cells in both infections. This review will help in paving the way for future directions in PRRSV and SwIV research and enhancing the understanding of the innate mechanisms that are involved during infection with these viruses.

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause severe reproductive and respiratory pathologies resulting in immense monetary and welfare costs for the swine industry. The vaccines against PRRSV are available; but they struggle with providing protection against the plethora of heterologous PRRSV strains. To improve PRRSV vaccine development, the aim of this study was to provide an in-depth analysis of the crucial heterologous T-cell response to type-2 PRRSV. Following PRRSV modified live virus (MLV) vaccination or infection using one high- or one low-pathogenic PRRSV-strain, this nine-week study evaluated the T-cell response to different PRRSV strains. Our results demonstrate an important role for T cells in this homo- and heterologous response. Specifically, the T-helper cells were the main responders during viremia. Their peak response at 28 dpi correlated with a reduction in viremia, and their homing receptor expression indicated the additional importance for the anti-PRRSV response in the lymphatic and lung tissue. The cytotoxic T lymphocyte (CTL) response was the strongest at the site of infection—the lung and bronchoalveolar lavage. The TCR-γδ T cells were the main responders post viremia and PRRSV induced their expression of the lymph node homing the chemokine receptor, CCR7: This indicates a crucial role for TCR-γδ T cells in the anti-PRRSV response in the lymphatic system.