RESUMEN
BAY 41-2272 increases guanosine 3', 5'-cyclic monophosphate (cGMP) levels by stimulating soluble guanylate cyclase (sGC). In this study, we evaluated the effect of BAY 41-2272 on human T lymphocyte functions. Pretreating T cells for 24â¯h with BAY 41-2272 at 3⯵M and 30⯵M, followed by activation with 90â¯nM phorbol myristate acetate (PMA), inhibited interferon-gamma (IFN-γ) production, with 3⯵M and 30⯵M BAY causing 16.5-fold and 12.1-fold inhibition, respectively, compared to PMA alone (pâ¯<â¯0.05, one-way ANOVA followed by Tukey's test). We also observed suppressive effects on the expression of CD69, with 30⯵M BAY causing 3.55-fold lower expression than PMA/ionomycin (pâ¯<â¯0.001 one-way ANOVA followed by Tukey's test), and T-bet, with 30⯵M BAY causing 1.47-fold lower expression than PMA/ionomycin (pâ¯<â¯0.05, one-way ANOVA test followed by Tukey's test). Additionally, T lymphocyte proliferation was reduced 2.13-fold and 4.3-fold, respectively, by 3⯵M BAY and 30⯵M BAY compared to PMA/ionomycin (pâ¯<â¯0.01, pâ¯<â¯0.001, one-way ANOVA followed by Tukey's test). BAY 41-2272 inhibits human T lymphocyte function and may be explored as an immunomodulatory drug in patients with autoimmune/inflammatory diseases and lymphoproliferative syndromes.
Asunto(s)
Pirazoles/farmacología , Piridinas/farmacología , Linfocitos T/efectos de los fármacos , Antígenos CD/metabolismo , Antígenos de Diferenciación de Linfocitos T/metabolismo , Proliferación Celular/efectos de los fármacos , Guanilato Ciclasa/metabolismo , Humanos , Factores Inmunológicos/metabolismo , Interferón gamma/metabolismo , Ionomicina/farmacología , Lectinas Tipo C/metabolismo , Linfocitos T/metabolismo , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
BAY 41-2272 is a guanylyl cyclase (GC) stimulator derived from YC-1 (3-[(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole]). Previous studies by our group showed that BAY 41-2272 activates human monocytes via soluble guanylyl cyclase (sGC) and cGMP. In this study, we investigated the effect of BAY 41-2272 on human neutrophil function and found that 30⯵M BAY 41-2272 inhibits neutrophil migration (1.82-fold lower than FMLP, Pâ¯<â¯0.05 by one-way ANOVA followed by Tukey's test), oxidative burst (1.70-fold lower than PMA, Pâ¯<â¯0.05 by one-way ANOVA followed by Tukey's test), and IL-8 cytokine production (1.80-fold lower than PMA, Pâ¯<â¯0.05 by one-way ANOVA followed by Tukey's test). Our results suggest that these effects are independent of the sGC pathway but dependent instead on cGMP production, as the response induced by 30⯵M BAY 41-2272 was 6.40-fold greater than that observed in our negative control (Pâ¯<â¯0.05 by parametric t-test). 1H-[1, 2, 4] oxadiazolo [4,3-a] quinoxalin-1-one (ODQ), which is an irreversible inhibitor of sGC, was unable to reverse the effects of BAY 41-2272 on human neutrophils, indicating that this drug acts independently of sGC. Our results confirm the immunomodulatory effect of BAY 41-2272 on human neutrophils.
Asunto(s)
Factores Inmunológicos/farmacología , Neutrófilos/efectos de los fármacos , Pirazoles/farmacología , Piridinas/farmacología , Movimiento Celular/efectos de los fármacos , Células Cultivadas , GMP Cíclico/metabolismo , Humanos , Interleucina-8/metabolismo , Neutrófilos/fisiología , Especies Reactivas de Oxígeno/metabolismo , Estallido Respiratorio/efectos de los fármacos , Superóxidos/metabolismoRESUMEN
OBJECTIVE: HLA-G has well recognized tolerogenic properties in physiological and nonphysiological conditions. The 3' untranslated region (3'UTR) of the HLA-G gene has at least 3 polymorphic sites (14-bpINS/DEL, +3142C/G, and +3196C/G) described as associated with posttranscriptional influence on messenger RNA production; however, only the 14-bpINS/DEL and +3142C/G sites have been studied in systemic lupus erythematosus (SLE). METHODS: We investigated the HLA-G 3'UTR polymorphic sites (14-bpINS/DEL, +3003C/T, +3010C/G, +3027A/C, +3035C/T, +3142C/G, +3187A/G, and +3196C/G) in 190 Brazilian patients with SLE and 282 healthy individuals in allele, genotype, and haplotype analyses. A multiple logistic regression model was used to assess the association of the disease features with the HLA-G 3'UTR haplotypes. RESULTS: Increased frequencies were observed of the 14-bpINS (p = 0.053), +3010C (p = 0.008), +3142G (p = 0.006), and +3187A (p = 0.013) alleles, and increased frequencies of the 14-bpINS-INS (p = 0.094), +3010 C-C (p = 0.033), +3142 G-G (p = 0.021), and +3187 A-A (p = 0.035) genotypes. After Bonferroni correction, only the +3142G (p = 0.05) and +3010C (p = 0.06) alleles were overrepresented in SLE patients. The UTR-1 haplotype (14-bpDEL/+3003T/+3010G/+3027C/+3035C/+3142C/+3187G/+3196C) was underrepresented in SLE (pcorr = 0.035). CONCLUSION: These results indicate that HLA-G 3'UTR polymorphic sites, particularly +3142G and +3010C alleles, were associated with SLE susceptibility, whereas UTR-1 was associated with protection against development of SLE.