RESUMO
Most human tumors, including cervical cancer, are characterized by telomerase activation (cell proliferation activation enzyme). Such activation is implemented in the elongation of the terminal segments (telomeres) of the telomerase chromosome. The gene of the enzyme is RNA-encoded, the RNA in tumors being observed in a few isoforms. The hTERT RNA role in cell activation and control was simulated using cervical cancer, as well as its pretumoral states (CIN), as a model object. The goal of this work was to clone of the human hTERT isoforms (normal, α-, ß-, and α+ß-splice-variants). The genetic constructions containing normal hTERT sequence, α- and ß-deletion variants based on the lentivirus vector pR780 were obtained. The α- and ß-deletion variants were not obtained in this variant because of methodological problems. In further research, we plan to implement splice-variants of hTERT in eukaryotic human cells.
Assuntos
Telomerase/genética , Processamento Alternativo , Domínio Catalítico , Linhagem Celular , Clonagem Molecular , Feminino , Vetores Genéticos , Humanos , Lentivirus/genética , Subunidades Proteicas , RNA Mensageiro , Deleção de Sequência , Telomerase/metabolismo , Neoplasias do Colo do Útero/enzimologia , Neoplasias do Colo do Útero/genéticaRESUMO
Fixed erythema--a kind of clinical and histopathologic reaction, fixed drug eruption. The purpose of the study--the study of characteristics of the cytokine profile in patients with erythema and the dynamics of the basal levels of proinflammatory and antiinflammatory cytokines during immunotherapy. All 41 patients with fixed erythema at baseline and after treatment was carried out determination of levels of pro-, anti-inflammatory and regulatory cytokines in the serum by ELISA using test systems "Biosource" (Austria). In patients with fixed erythema Immunovac treatment increased serum IFN-gamma (p < 0.05), IL-1beta (p > 0.05), IL-6. While Kagocel led to an increase in IFN-gamma (p < 0.05), IL-1beta, IL-6 and reduction of TGF-beta (p < 0.05). At the same time in patients with fixed erythema basic therapy contributed to the significant increase in TGF-â and decrease in IL-10. Immunovac-VP-4 had the highest activity for the induction of IFN-gamma. Inclusion in the range of therapeutic and prophylactic measures in patients with fixed erythema immunomodulators promotes activation links innate and adaptive immunity triggers mechanisms, thus increasing the antiviral response in patients with erythema.
Assuntos
Antígenos de Bactérias/uso terapêutico , Antivirais/uso terapêutico , Eritema Nodoso/sangue , Eritema Nodoso/terapia , Fatores Imunológicos/uso terapêutico , Imunoterapia , Imunidade Adaptativa/efeitos dos fármacos , Adulto , Idoso , Antígenos de Bactérias/farmacologia , Antivirais/farmacologia , Ensaio de Imunoadsorção Enzimática , Eritema Nodoso/imunologia , Feminino , Humanos , Imunidade Inata/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Interferon gama/sangue , Interferon gama/imunologia , Interleucina-10/sangue , Interleucina-10/imunologia , Interleucina-1beta/sangue , Interleucina-1beta/imunologia , Interleucina-6/sangue , Interleucina-6/imunologia , Masculino , Pessoa de Meia-Idade , Fator de Crescimento Transformador beta/sangue , Fator de Crescimento Transformador beta/imunologiaRESUMO
An Escherichia coli strain producing human tumor necrosis factor (TNF-alpha) was obtained using a semisynthetic gene partially optimized in respect of codon composition and a phage T7 promoter. The expression product was accumulated in cells as inclusion bodies in a yield of 50-70 mg/l of culture medium. The recombinant TNF-alpha in the form of inclusion bodies was used for immunization of rats to give a polyclonal antiserum. The resulting antibodies were specific under the immunoblotting conditions to the antigen used for the immunization. A dilution-based refolding procedure was developed; it provided a yield of soluble protein exceeding 85%.