The effects of subcutaneous and intraocular administration of class B ODN CpG in chicken on the expression of TLR21, IFN-γ and IL-1ß.

The synthetic unmethylated oligodeoxynucleotides with CpG motifs (CpG ODN) were shown to activate Toll-like receptor 21 (TLR21) and stimulate the innate and adaptive immune system. In this study we tested the expression of TLR21, interferon (IFN)-γ and interleukin (IL)-1ß mRNA in the blood after subcutaneous and intraocular application of the class B CpG ODN in chicken. The relative expression of mRNA of TLR21, IFN-γ and IL-1ß were quantified at 3, 6, 12, 24 and 72 h post-stimulation. The study revealed that IFN-γ mRNA expression was significantly upregulated 12 h after subcutaneous stimulation with a high and low dose of ODN CpG, whereas the IL-1ß mRNA expression levels were significantly upregulated 3 and 72 h after subcutaneous administration. After intraocular administration, the IL-1ß mRNA levels were the highest at 24 h post-application, albeit not specifically. This data indicates that class B CpG ODN has the ability to induce TLR21 response in blood when administered parenterally in chicken. In contrast, intraocular administration of CpG ODN was not able to produce a significant increase in cytokine mRNA expression in blood. The data suggest that additional stimulus, e.g. the antigen, may be needed on the site of mucosal administration to activate systemic immune response.

Impact of CpG oligodeoxynucleotide stimulation on percentage of T and B cells in chicken.

TLR stimulation in chickens has been shown to play a role in the initiation and regulation of innate and adaptive immune responses. The aim of this study was to use flow cytometry to establish the percentage of T and B subset in blood and lymphoid organs in chicks after CpG oligodeoxynucleotide (ODN) stimulation. It was demonstrated that the percentages of CD3+, CD4+, TCRgamma delta+ cells and Bu-1+MHC class II+ cells in blood 24 h post-injection were significantly higher than in the control groups. It was also shown that the percentages of CD3+ and CD4+ cells in the spleen at 48 h post-injection were significantly higher than in control groups. The percentage of Bu-1+ cells in the bursa of Fabricius after CpG ODN stimulation (98.38 +/- 0.84) was significantly higher than that found in the non-CpG ODN control group (94.54 +/- 2.51) (p < or = 0.05). The results indicate that class B CpG ODN increases the percentage of both T (especially CD4+ cells) and B cells.

New insight into the structure, development, functions and popular disorders of bursa Fabricii.

Humoral immune responses in birds, contrary to mammals, depend on the normal functioning of bursa Fabricii. Recent studies have delivered new information about the structure, development and origin of cells that compose the bursa environment. Several viral infections affect bursa, causing lymphocyte depletion or excessive proliferation. This review summarizes data on the development and histology of healthy bursa and introduces some common disorders that affect this organ.

The influence of antibiotics on B-cell number, percentage, and distribution in the bursa of Fabricius of newly hatched chicks.

Antibiotics are commonly used to prevent and treat poultry microbial infections, but certain antibiotic families depress humoral immunity, such as antibody production. Poultry humoral immunity depends on the normal functioning of the bursa of Fabricius and the B lymphocytes that mature in that gland. In this study, recommended therapeutic doses of enrofloxacin, florfenicol, or ceftiofur were administered to 2-d-old chicks. On d 7 post-hatch, bursae were sampled for histological, immunohistochemical, and flow cytometric determination of Bu-1-positive (Bu-1+) cell number, percentage, and distribution. The bursa of Fabricius from all treatment and control groups had normal morphology. The administration of antibiotics significantly decreased the number of Bu-1+ cells in the bursal medulla, with a simultaneous increase of these cells in the cortex. Flow cytometry revealed a significant decrease in the percentage of bursal Bu-1+ cells from all of the studied antibiotics: enrofloxacin (93.91 ± 3.27), florfenicol (87.84 ± 7.14), and ceftiofur (89.16 ± 5.68) compared with that of the control (96.48 ± 2.60). The combination of reduced percentages of Bu-1+ cells and a decrease in these cells in the medullary region suggests lower B cell maturation.