Journal: BMC Veterinary Research
Article Title: Immune regulation and transcriptomic profiling of chicken CD8⁺ T cells in response to infection with the IBV strain K047-12 during in vitro culture
doi: 10.1186/s12917-026-05284-3
Figure Lengend Snippet: Immune cell composition in the chicken splenocytes during ConA + IL-2 stimulation and IBV infection. Chicken splenocytes were stimulated with Concanavalin A (ConA) and chicken IL-2 for 48 h, followed by IBV infection for an additional 48 h. A Flow cytometric analysis of immune cell phenotypes in splenocytes before and after ConA + IL-2 stimulation. Populations of CD3⁺ T cells, CD8⁺ T cells, CD4⁺ T cells, B cells (Bu-1⁺), and macrophages (Mac/Mono⁺ and MHC II⁺) are shown (n = 5). Statistical significance was determined with a paired t-test (*p < 0.05). B Cell morphology was monitored from 48 h (0 h post-IBV infection) to 96 h (48 h post- infection), and representative graphs show changes in cell confluency recorded every 12 h after IBV infection. The experiments are performed in duplicate. C Morphological changes in splenocytes were assessed at three time points—0 h (unstimulated), 48 h (stimulated with ConA + IL-2), and 96 h (stimulated with ConA + IL-2 for 48 h followed by IBV infection for 48 h) using Diff-Quik staining. Representative images are shown (n = 5) ( D ) Cell viability following IBV infection for 48 h, evaluated by flow cytometry using LIVE/DEAD™ fixable dye. ( E , F ) The cultured cells in the absence (Control) or presence of infectious bronchitis virus (IBV) were harvested and analyzed by flow cytometry for immune cell phenotypic changes. The frequencies of CD4⁺, CD8⁺, CD44⁺ CD4⁺, and CD44⁺ CD8⁺ cells within live cells were assessed in IBV-infected and uninfected control groups. (G) The frequencies of CD25high populations were assessed within CD4⁺ and CD44⁺ CD4⁺ T cells, as well as within CD8⁺ and CD44⁺ CD8⁺ T cells. The mean fluorescence intensity (MFI) of CD25 within CD44⁺ CD4⁺ and CD44⁺ CD8⁺ T cells was also evaluated. Representative flow cytometry plots are shown. Data are pooled from 5–6 independent experiments. Statistical significance was determined with a paired t-test (* p < 0.05, **p < 0.001)
Article Snippet: Chicken CD25 , FITC , AbD13504 , HCA173F , Bio-Rad (USA).
Techniques: Infection, Diff-Quik, Staining, Flow Cytometry, Cell Culture, Control, Virus, Fluorescence