Anti-IL23A (N-terminal) polyclonal antibody

Objective. The aim of this study was to assess the possible association between the interleukin-12B (IL12B) and interleukin-12 receptor beta 1 (IL12RB1) gene polymorphisms with systemic lupus erythematosus (SLE). In addition, we have undertaken a systematic search for genetic variants of interleukin 23 (IL23A). Methods. The study was conducted on 559 SLE patients and 603 ethnically matched healthy controls. Genotyping of the IL12B [IL12Bpro and IL12B 3' untranslated region (UTR)] and IL12RB1 (641A -> G, 1094T -> C and 1132G -> C) polymorphisms was performed with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and PCR-fluorescent methods, whereas IL23A genetic variants were realized with direct sequencing. Results. No statistically significant differences in the distribution of the IL12B and the IL12RB1 genotypes and alleles were observed when comparing SLE patients and control subjects. Additionally, no differences in the genotype and allele distribution were found when SLE patients were stratified according to the presence or absence of lupus nephritis. Despite an extensive analysis in 30 individuals, variations located in the exons and in the 5' and 3' UTR regions of IL23A gene were not found in any case. Conclusions. These results suggest that polymorphisms located in IL12B, IL12RB1 and IL23A genes may not play a relevant role in the susceptibility or severity of SLE in the Spanish population.

Background: CCAAT/enhancer binding protein beta ( C/ EBP beta) is a transcription factor that regulates the expression of important pro-inflammatory genes in microglia. Mice deficient for C/ EBPa show protection against excitotoxic and ischemic CNS damage, but the involvement in this neuroprotective effect of the various C/ EBP beta-expressing cell types is not solved. Since C/ EBP beta-deficient microglia show attenuated neurotoxicity in culture, we hypothesized that specific C/ EBPa deficiency in microglia could be neuroprotective in vivo. In this study, we have tested this hypothesis by generating mice with myeloid C/ EBP beta deficiency. Methods: Mice with myeloid C/ EBP beta deficiency were generated by crossing LysMCre and C/ EBP beta(fl/) (fl) mice. Primary microglial cultures from C/ EBP beta fl/ fl and LysMCre- C/ EBP beta fl mice were treated with lipopolysaccharide interferon. ( IFN gamma.) for 6 h, and gene expression was analyzed by RNA sequencing. Gene expression and C/ EBP beta deletion were analyzed in vivo in microglia isolated from the brains of C/ EBP beta(f1/f1) fl and LysMCre- C/ EBP beta(f1/f1) fl mice treated systemically with lipolysaccharide or vehicle. Mice of LysMCre- C/ EBP beta fl or control genotypes were subjected to experimental autoimmune encephalitis and analyzed for clinical signs for 52days.One- or two-way ANOVA or Kruskal- Wallis with their appropriate post hoc tests were used. Results: LysMCre- C/ EBP beta(f1/f1) fl mice showed an efficiency of C/ EBP beta deletion in microglia of 100 and 90% in vitro and in vivo, respectively. These mice were devoid of female infertility, perinatal mortality and reduced lifespan that are associated to full C/ EBP beta deficiency. Transcriptomic analysis of C/ EBP beta deficient primary microglia revealed C/ EBP beta dependent expression of 1068 genes, significantly enriched in inflammatory and innate immune responses GO terms. In vivo, microglial expression of the pro-inflammatory genes Cybb, Ptges, Il23a, Tnf and Csf3 induced by systemic lipopolysaccharide injection was also blunted by C/ EBP beta deletion. CNS expression of C/ EBP beta was upregulated in experimental autoimmune encephalitis and in multiple sclerosis samples. Finally, LysMCre- C/ EBP beta(F1/F1) mice showed robust attenuation of clinical signs in experimental autoimmune encephalitis. Conclusion: This study provides new data that support a central role for C/ EBP beta in the biology of activated microglia, and it offers proof of concept for the therapeutic potential of microglial C/ EBP beta inhibition in multiple sclerosis.