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While B cell development in the birds' primary B cell organ, the bursa Fabricius, is relatively well understood, very little is known about post bursal B cell differentiation into plasma and memory cells though these cells are essential for a protecting antibody response and so far, no specific markers for these cells were available. Since immunoglobulin class switch is one part of the B cell differentiation process, our objective was to conduct a first detailed investigation of class-switched chicken B cells. As only very few IgY and IgA expressing cells were detected in lymphoid organs of young chickens, we used CD40L and IL-10 to establish a prolonged in vitro culture system, which induces B cell proliferation, class switch to IgY and IgA and enhanced antibody secretion. This enabled a phenotypic analysis of differentiating B cells. Importantly, these cells lost surface expression of the B cell markers chB6 and BAFF-R. B cell receptor surface expression remained unchanged, showing that while differentiating toward plasma cells, B cells can be addressed by L chain staining. Newly generated potential plasma cell markers CD138 and TACI showed only a transient expression on cultured cells and rather act as markers for B cell activation than plasma/memory cells in general. CD57 upregulation was connected to activation and blast formation but not to class switch. We also examined potential changes in class-switched cells in different age groups and post vaccination. Surprisingly, bursa involution, laying and age had no distinct effects on the presence of class-switched cells, but we detected significantly more class-switched B cells post vaccination. Hence, we are now able to generate class-switched plasmablasts in vitro for a more detailed characterization and can address them under different conditions in chickens for further analysis of their B cell response. [ABSTRACT FROM AUTHOR]

Cell cultures are models in biological and medical research to understand physiological and pathological processes. Cell lines are not always available depending on cell type and required species. In addition, the immortalization process often affects cell biology. Primary cells generally maintain a greater degree of similarity in short-term culture to the cells in tissue. Goal of this study was to verify the suitability of chicken primary epithelial caecal cells (PECCs) for in vitro investigations of host‒pathogen interactions. Epithelial nature of PECCs was confirmed by detection of tight and adherens junctions and cobblestone-like cell morphology. Sialic acids distribution was similar to that in caecal cyrosections. To understand the capacity of PECCs to respond to microbial challenges, the Toll-like receptors (TLRs) repertoire was determined. Exposure of PECCs to polyinosinic-polycytidylic acid (poly(I:C)) or lipopolysaccharide (LPS) led to upregulation of type I and III interferon (IFN) as well as interleukin (IL-) 1β, IL-6 and IL-8 mRNA expression. Overall, the PECCs showed properties of polarized epithelial cells. The presence of TLRs, their differential expression, as well as pattern recognition receptor dependent immune responses enable PECCs to act as suitable in vitro model for host‒pathogen interaction studies, which are difficult to conduct under in vivo conditions. [ABSTRACT FROM AUTHOR]

Campylobacter hepaticus (C. hepaticus) was recently discovered as the causative agent of Spotty Liver Disease (SLD). SLD affects laying hens and causes significant economic losses in egg production in several countries throughout the world. Field observations reveal that cases of SLD appear with a high risk of reoccurrence, specifically in free-range and organic brown-feathered layer lines. Possible factors contributing to the development of SLD still have to be elucidated. In this field study, one free range (Flock 1) and one organic flock (Flock 2) of brown laying hens kept on farms with a history of clinical SLD were monitored for C. hepaticus colonization, clinical signs, and egg production from 16 to 79 wk of age on the first farm and from 17 to 83 wk of age on the other. The flocks showed a significant drop in egg production at 32 to 39 or 56 wk of age, respectively, which was associated with macroscopically visible liver lesions typical for SLD. Interestingly, in both cases observed clinical disease was linked to a stressful event: heat stress for Flock 1 and respiratory symptoms for Flock 2. C. hepaticus was detected by PCR during the acute phase of the disease in Flock 1. At 50 wk after the initial clinical outbreak had waned, C. hepaticus was still able to be isolated by culture in this flock. This clearly demonstrates that C. hepaticus persists either in the birds or their environment. We speculate that this long persistence may favor chronic SLD in affected flocks and the reoccurrence of SLD in subsequent flocks. Clinically less severe SLD outbreaks may be observed after re-exposure of clinically recovered flocks. Investigación sobre la necrosis hepática focal y Campylobacter hepaticus en parvadas de ponedoras: Un estudio de campo. Campylobacter hepaticus (C. hepaticus) se descubrió recientemente como el agente causante de la necrosis hepática focal (SLD por sus siglas en inglés). La necrosis hepática focal afecta a las gallinas de postura y provoca importantes pérdidas económicas en la producción de huevo en varios países del mundo. Las observaciones de campo revelan que los casos de necrosis hepática focal aparecen con un alto riesgo de recurrencia, específicamente en las líneas de ponedoras de plumaje marrón bajo condiciones de pastoreo y de producción orgánica. Aún deben dilucidarse los posibles factores que contribuyen al desarrollo de la necrosis hepática focal. En este estudio de campo, en una parvada mantenida en pastoreo (parvada 1) y en una parvada bajo producción orgánica (parvada 2) de gallinas de postura de color marrón criadas en granjas con antecedentes de necrosis hepática focal se monitorearon la colonización por C. hepaticus, los signos clínicos y la producción de huevos desde las 16 a las 79 semanas de edad en la primera granja y de las 17 a las 83 semanas de edad en la segunda granja. Las parvadas mostraron una caída significativa en la producción de huevo entre las 32 a 39 semanas, o a las 56 semanas de edad, respectivamente, que se asociaron con lesiones hepáticas macroscópicamente visibles y típicas de la necrosis hepática focal. Curiosamente, en ambos casos, la enfermedad clínica observada se vinculó con un evento estresante: estrés por calor en la parvada 1 y síntomas respiratorios en la parvada 2. Se detectó C. hepaticus mediante cultivo o por PCR durante la fase aguda de la enfermedad en la parvada 1. A las 50 semanas después de que el brote clínico inicial se había disminuido, todavía se pudo aislar C. hepaticus mediante cultivo en esta parvada. Esto demuestra claramente que C. hepaticus persiste tanto en las aves como en su entorno. Se especula que esta larga persistencia puede favorecer a la necrosis hepática focal crónica en las parvadas afectadas y la reaparición de este problema en parvadas posteriores. Se pueden observar brotes de necrosis hepática focal clínicamente menos severos después de la reexposición de parvadas clínicamente recuperadas. [ABSTRACT FROM AUTHOR]

Vaccination is crucial for health protection of poultry and therefore important to maintaining high production standards. Proper vaccination requires knowledge of the key players of the well-orchestrated immune system of birds, their interdependence and delicate regulation, and, subsequently, possible modes of stimulation through vaccine antigens and adjuvants. The knowledge about the innate and acquired immune systems of birds has increased significantly during the recent years but open questions remain and have to be elucidated further. Despite similarities between avian and mammalian species in their composition of immune cells and modes of activation, important differences exist, including differences in the innate, but also humoral and cell-mediated immunity with respect to, for example, signaling transduction pathways, antigen presentation, and cell repertoires. For a successful vaccination strategy in birds it always has to be considered that genotype and age of the birds at the time point of immunization as well as their microbiota composition may have an impact and may drive the immune reactions into different directions. Recent achievements in the understanding of the concept of trained immunity will contribute to the advancement of current vaccine types helping to improve protection beyond the specificity of an antigen-driven immune response. The fast developments in new omics technologies will provide insights into protective B- and T-cell epitopes involved in cross-protection, which subsequently will lead to the improvement of vaccine efficacy in poultry.