Increased expression and phosphorylation of the two S100A9 isoforms in mononuclear cells from patients with systemic lupus erythematosus: A proteomic signature for circulating low-density granulocytes

Esther J. Pavón^a^,¹,
Sonia García-Rodríguez^a,
Esther Zumaquero^a^,²,
Rubén Perandrés-López^a,
Antonio Rosal-Vela^a,
Antonio Lario^b,
Victoria Longobardo^b,
Montserrat Carrascal^c,
Joaquín Abián^c,
José-Luis Callejas-Rubio^d,
Norberto Ortego-Centeno^d,
Mercedes Zubiaur^a,
Jaime Sancho^a^,^,

^a Departamento de Biología Celular e Inmunología, Instituto de Parasitología y Biomedicina “López-Neyra”, IPBLN-CSIC, Armilla, Spain
^b Unidad de Proteómica, Instituto de Parasitología y Biomedicina “López-Neyra”, CSIC, Armilla, Spain
^c CSIC/UAB Proteomics Laboratory, Instituto de Investigaciones Biomédicas de Barcelona-Consejo Superior de Investigaciones Científicas (IIBB-CSIC/IDIBAPS), Building M, Barcelona Autonomous University, E-08193 Bellaterra, Spain
^d Unidad de Enfermedades Autoinmunes Sistémicas, Hospital Universitario San Cecilio, Granada, Spain

Received 19 July 2011. Accepted 14 December 2011. Available online 30 December 2011.

http://dx.doi.org/10.1016/j.jprot.2011.12.020, How to Cite or Link Using DOI

Abstract

Proteins differentially expressed in peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients versus Normal controls were identified by 2-DE and MALDI-MS. Thus, S100A9 expression was significantly increased in SLE PBMCs relative to Normal PBMCs at both mRNA and protein levels. Increased S100A9 levels in SLE PBMCs correlated positively with the abnormal presence of low-density granulocytes (LDGs) detected by flow-cytometry in the mononuclear cell fractions. Another set of proteins that were differentially expressed in SLE PBMCs formed S100A9-independent clusters, suggesting that these differences in protein expression are in fact reflecting changes in the abundance of specific cell types. In SLE PBMCs spots of the two S100A9 isoforms, S100A9-l and S100A9-s, and their phosphorylated counterparts were identified and confirmed to be phosphorylated at Thr¹¹³ by MS/MS analyses. In addition, the phorbol ester PMA alone or in combination with ionomycin induced a stronger increase in threonine phosphorylation of S100A9 in SLE than in Normal PBMCs, while the same stimuli caused the opposite effect on phosphorylation and activation of Erk1/2, suggesting the existence of an abnormal S100A9 signaling in SLE PBMCs. Therefore, the expansion and activation of LDGs in SLE seems to underlie this prominent S100A9 signature.