Interleukin-21 Drives Proliferation and Differentiation of Porcine Memory B Cells into Antibody Secreting Cells.

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PloS one, ISSN: 1932-6203, Vol: 12, Issue: 1, Page: e0171171

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10.1371/journal.pone.0171171; 10.1371/journal.pone.0171171.g003; 10.1371/journal.pone.0171171.g001; 10.1371/journal.pone.0171171.g005; 10.1371/journal.pone.0171171.g006; 10.1371/journal.pone.0171171.g004; 10.1371/journal.pone.0171171.g002
Michael C. Rahe; Michael P. Murtaugh; Gourapura J. Renukaradhya
Public Library of Science (PLoS); Figshare
Biochemistry, Genetics and Molecular Biology; Agricultural and Biological Sciences; Cell Biology; Physiology; Biotechnology; Immunology; Biological Sciences not elsewhere classified; Developmental Biology; Cancer; Hematology; Infectious Diseases; Virology; surface immunoglobulin receptor; 40L; IL -21; memory cell differentiation; Antibody Secreting Cells Immunological prevention; Interleukin -21 Drives Proliferation; IgG antibody secreting cells; Porcine Memory B Cells; ïve age-matched pigs; B cells; Mature B cells; BAFF; ELISA; memory B cell culture system; IL -21-driven expansion; ASC; PRRSV non-structural protein 7 ELISPOT; memory B cells; 110309 Infectious Diseases; 60506 Virology; 69999 Biological Sciences not elsewhere classified
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Immunological prevention of infectious disease, especially viral, is based on antigen-specific long-lived memory B cells. To test for cellular proliferation and differentiation factors in swine, an outbred model for humans, CD21+ B cells were activated in vitro with CD40L and stimulated with purported stimulatory cytokines to characterize functional responses. IL-21 induced a 3-fold expansion in total cell numbers with roughly 15% of all B cells differentiating to IgM or IgG antibody secreting cells (ASCs.) However, even with robust proliferation, cellular viability rapidly deteriorated. Therefore, a proliferation inducing ligand (APRIL) and B cell activating factor (BAFF) were evaluated as survival and maintenance factors. BAFF was effective at enhancing the viability of mature B cells as well as ASCs, while APRIL was only effective for ASCs. Both cytokines increased approximately two-fold the amount of IgM and IgG which was secreted by IL-21 differentiated ASCs. Mature B cells from porcine reproductive and respiratory virus (PRRSV) immune and naïve age-matched pigs were activated and treated with IL-21 and then tested for memory cell differentiation using a PRRSV non-structural protein 7 ELISPOT and ELISA. PRRSV immune pigs were positive on both ELISPOT and ELISA while naïve animals were negative on both assays. These results highlight the IL-21-driven expansion and differentiation of memory B cells in vitro without stimulation of the surface immunoglobulin receptor complex, as well as the establishment of a defined memory B cell culture system for characterization of vaccine responses in outbred animals.