Fig. 5

Single cells analysis of B cell distribution in bursal and FACS validation the subtype of bursary B cells change after IBDV infection. a t-SNE visualization of the major five cell types characterized with a different colour. The encircled green cells show the total B cell population. b t-SNE graph of the total B cell population divided into five clusters based on mRNA transcriptional profiling, shown with the unique colour difference. c Five B cell clusters distributed into normal and IBDV infected host groups shown in different tSNE graphs. S1 represents control 2-weeks-old-chick, S2 represents IBDV infected 2-weeks-old-chick, S3 represents control 3-weeks-old-chick and S4 represents IBDV infected-3-weeks-old chick. d Fraction of immunoglobulins of B cells (IgA, IgM and IgY) shown in different violin plots distributed into the five clusters of B cells. e Heatmap of the gene expression highly expressed in B cell population shown in five major cluster distribution. f t-SNE representation of the viral load (shown in stars) of IBDV strain BC6/85 segment A and B in IBDV infected hosts. g FACS analyses of bursal cells of two weeks old and three weeks old control and infected SPF chickens followed by intranasal inoculation of IBDV strain BC6/85 after 72 hpi. Bursal cells were isolated from each group and gated for Bu1 (marker of B cells), and immunoglobulins cells (IgM + and IgY +). h Immunoglobulins stained samples along with Bu1 were purified with MACS magnetic microbeads based on specific antibodies for viral load detection in each sample. The blank group contains the antibody-free and virus-free samples. Statistical significance was determined using one-way ANOVA. Significance difference was expressed as, *p < 0.05, ****p < 0.0001