Co-Expression of the B-Cell Key Transcription Factors Blimp-1 and IRF4 Identifies Plasma Cells in the Pig

Antibody-secreting plasma cells (PCs) have remained largely uncharacterized for years in the field of porcine immunology. For an in-depth study of porcine PCs, we identified cross-reactive antibodies against three key transcription factors: PR domain zinc finger protein-1 (Blimp-1), interferon regulatory factor 4 (IRF4), and paired box 5 (Pax5). A distinct Blimp-1(+)IRF4(+) cell population was found in cells isolated from blood, spleen, lymph nodes, bone marrow, and lung of healthy pigs. These cells showed a downregulation of Pax5 compared to other B cells. Within Blimp-1(+)IRF4(+) B cells, IgM-, IgG-, and IgA-expressing cells were identified and immunoglobulin-class distribution was clearly different between the anatomical locations, with IgA(+) PCs dominating in lung tissue and IgM(+) PCs dominating in the spleen. Expression patterns of Ki-67, MHC-II, CD9, and CD28 were investigated in the different organs. A high expression of Ki-67 was observed in blood, suggesting a plasmablast stage. Blimp-1(+)IRF4(+) cells showed an overall lower expression of MHC-II compared to regular B cells, confirming a progressive loss in B-cell differentiation toward the PC stage. CD28 showed slightly elevated expression levels in Blimp-1(+)IRF4(+) cells in most organs, a phenotype that is also described for PCs in mice and humans. This was not seen for CD9. We further developed a FACS-sorting strategy for live porcine PCs for functional assays. CD3(-)CD16(-)CD172a(-) sorted cells with a CD49d(high)FSC-A(high) phenotype contained Blimp-1(+)IRF4(+) cells and were capable of spontaneous IgG production, thus confirming PC identity. These results reveal fundamental phenotypes of porcine PCs and will facilitate the study of this specific B-cell subset in the future.

Automated summary

Antibody-secreting plasma cells in pigs have been understudied. This study identified cross-reactive antibodies against three transcription factors and characterized a distinct population of Blimp-1(+)IRF4(+) cells in multiple organs. The cells showed different immunoglobulin-class distribution based on anatomical location. A FACS-sorting strategy was developed for functional assays. These findings provide fundamental knowledge on porcine plasma cells and facilitate future studies.