RESUMO
Phenytoin (PHE) and carbamazepine (CBZ) are first rank causative drugs that can induce drug rash with eosinophilia and systemic symptoms (DRESS) and Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN). Identification of anti-epileptic drugs as a culprit drug has been problematic; hence, in vitro tests could be promising methods to define causative drugs without clinical risk. The aim of this study is to evaluate the efficacy of lymphocyte transformation tests (LTT) and cytokine detection assays in identifying PHE and CBZ as culprit drugs. Peripheral blood mononuclear cells were collected from normal, PHE/CBZ tolerance and PHE/CBZ hypersensitivity cohorts and utilized for cell-culture assays. LTT was performed and culture supernatants were subjected to multiple cytokine detection assays. Our study showed that LTT correlated with outcomes of Naranjo's assessment with statistical significance (râ¯=â¯0.614). Various sensitivities of LTT and cytokine detection assays were demonstrated. Although both assays provided excellent specificity, LTT yielded higher sensitivity as compared to those of cytokine detection assays in both DRESS and SJS/TEN. Regardless whether specificity is, this is the first report to demonstrate that IL-4 detection assay could enhance sensitivity to identify culprit drug when it was combined with LTT for SJS/TEN patients or it was combined with IL-2/IFN-γ detection assays for DRESS ones.
Assuntos
Anticonvulsivantes/efeitos adversos , Carbamazepina/efeitos adversos , Síndrome de Hipersensibilidade a Medicamentos/imunologia , Ativação Linfocitária/efeitos dos fármacos , Fenitoína/efeitos adversos , Síndrome de Stevens-Johnson/imunologia , Adulto , Citocinas/imunologia , Epilepsia/tratamento farmacológico , Epilepsia/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The simple and less expensive technique based on magnetic nanoparticles (MNPs) was developed for separation of malaria parasites containing specific antigens. The carboxylated MNPs were chemically bound with anti-P. falciparum IgG antibodies (Ab-MNPs) purified from the plasma of malaria patients and then used for removal of P. falciparum malaria-infected erythrocytes from other non-infected blood cells in malaria culture at a given percent parasitemia. The results from optical microscope showed that all blood stages parasites, i.e., ring, trophozoite and schizont, could be separated from other blood components with high purity (> or = 95%) and yield of 33.5% (the early stages of ring and trophozoite:the schizont stage were 1:1.34). Highly specific interaction between Ab-MNPs and the P. falciparum malaria infected erythrocytes was confirmed by scanning electron microscope. When compared to the centrifugation with Percoll gradient and depletion by sorbitol lysis which are specific to the mature and the ring stages, respectively, our technique would be more useful for production of high quality of parasites to use in malaria pathogenesis or immunological studies, and in detection techniques.