CHARACTERIZATION OF JAWSII CELL LINE AFTER LPS-STIMULATION IN DIFFERENT CULTURE CONDITIONS

MAYORDOMO ANDREA CONSTANZA; ARIAS JOSÉ LUIS; LEPORATI MARIANELA; SILVA JUAN EDUARDO; DI GENARO MARIA SILVIA

Villa de Merlo, San Luis

Congreso; XXXV Reunion Cientifica Anual de la Sociedad de Biología de Cuyo; 2017

Sociedad de Biologia de Cuyo

JAWS II is an immortalized dendritic cell (DC) line established from the bone marrow cultures of p53-/- C57BL/6 mice. The characterization of this cell line from scientific literature is controversial as depend on experimental settings. Therefore, we evaluated DC surface makers and IL-12/23p40 production after LPS-stimulation in different culture conditions. JAWS II cells were cultured in presence of GM-CSF or Flt3L, as DC differentiation factors, and they were stimulated with LPS. Surface markers and cytokine production were assessed by flow cytometry and ELISA, respectively. We corroborated that the cells cultured with GM-CSF or Flt3L express surface markers of DC. After LPS stimulation, they show maturation since increased MHC-II and CD86 expressions. Interestingly, these cells displayed the two TNF receptors (TNFRp55 and TNFRp75), and reduced TNFRp75 significantly after LPS-stimulation. In addition, the JAWS II cell line secreted TNF and IL-12/23p40 in response to LPS. Human TNF (hTNF) has previously demonstrated to bind to mouse TNFRp55 but not to TNFRp75. Because of this, JAWS II cells were stimulated with LPS in presence of hTNF, which significantly reduced IL-12/23p40 secretion only in presence of GM-CSF. In culture media with GM-CSF or Flt3L, IL-12/23p40 production decreased significantly by p38, ERK and mainly JNK MAPK inhibitions. We conclude that GM-CSF or Flt3L in the culture medium impact on LPS-induced responses of JAWS II cells. This cell line could be suitable as model to gain an insight into the mechanisms underlying the regulatory effect of TNF through TNFRp55 on IL-12/23p40 secretion.