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1.
Hum Immunol ; 77(2): 153-7, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26585776

RESUMEN

Human leukocyte antigen-G (HLA-G) plays an important role in tumor cell escape from immune surveillance and HLA-G polymorphisms might service as a potential risk factor for clinical outcomes in GAC (gastric adenocarcinoma). We investigated the association between HLA-G polymorphisms as well as soluble HLA-G level and accordance of GAC. This case-control study included 100 GAC patients and 102 unrelated Iranian individual's samples as control. The clinical stages ranged from I to IV. PCR-RFLP method was carried out in order to specify the genotypes of the HLA-G gene. Concentrations of sHLA-G in serum were determined with the sHLA-G-specific enzyme linked immunosorbent assay (ELISA) kit. The G*01:04:01 and G*01:01:02:01 alleles were the predominant alleles in GAC patients and healthy controls. The G*01:01:03:01 and G*01:01:08 allele distributions are significantly higher among controls comparing to cases and seem to have protective effect (P value=0.026 and 0.007 respectively). There is a substantial differences in G*01:01:02:01/G*01:04:01 genotype frequencies between cases and controls (OR=2.8, P value<0.001). The G*01:01:03:01/G*01:04:01 and G*01:01:02:01/G*01:01:08 genotypes frequency are higher among controls in comparison to patients (P value=0.028 and 0.007 respectively). The polymorphisms in HLA-G could affect GAC induction and its outcome. Also, increased sHLA-G levels in serum might be a useful biomarker for diagnosis.


Asunto(s)
Adenocarcinoma/genética , Antígenos HLA-G/genética , Neoplasias Gástricas/genética , Adenocarcinoma/inmunología , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Irán , Masculino , Persona de Mediana Edad , Polimorfismo Genético , Neoplasias Gástricas/inmunología , Escape del Tumor/genética
2.
Acta Biochim Pol ; 62(1): 83-8, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25750940

RESUMEN

Leukemia is a particular type of cancer characterized by the failure of cell death or disability in differentiation of hematopoietic cells. Chronic myelogenus leukemia (CML) is the most studied kind of this cancer. In this study, anti-cancer effect of dihydro-pyranochromenes derivatives were investigated in the human leukemia K562 cells. These compounds were found to be active cell proliferation inhibitors using MTT assay. Among these compounds, 3-NpC was determined as stronger compound with IC50 value of 100 ± 3.1 µM and was chosen for further studies. Induction of apoptosis was analyzed by AO/EtBr staining, DNA fragmentation assay, Annexin V/PI double staining and cell cycle analysis. Furthermore, Western Blot analysis showed that treatment of the cells with 3-NpC led to up-regulation and activation of caspase-3. The results of this investigation clearly indicated that dihydro-pyranochromenes derivatives induce apoptosis in the K562 cell line. This information signalizes also that these compounds may prepare a new therapeutic approach for the treatment of leukemia.


Asunto(s)
Apoptosis/efectos de los fármacos , Benzopiranos/farmacología , Proliferación Celular/efectos de los fármacos , Leucemia Mielógena Crónica BCR-ABL Positiva/patología , Humanos , Células K562
3.
Iran J Basic Med Sci ; 18(2): 145-52, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25810888

RESUMEN

OBJECTIVES: Multi-epitopic protein vaccines and direction of vaccine delivery to dendritic cells (DCs) are promising approaches for enhancing immune responses against mutable pathogens. Escherichia coli is current host for expression of recombinant proteins, and it is important to optimize expression condition. The aim of this study was the optimization of multi-epitopic HIV-1 tat/pol/gag/env recombinant protein (HIVtop4) expression by E. coli and conjugation of purified protein to anti DEC-205 monoclonal antibody as candidate vaccine. MATERIALS AND METHODS: In this study, expression was induced in BL21 (DE3) E. coli cells by optimization of induction condition, post induction incubation time, temperature and culture medium formula. Some culture mediums were used for cell culture, and isopropyl-beta-D-thiogalactopyranoside was used for induction of expression. Protein was purified by Ni-NTA column chromatography and confirmed against anti-His antibody in western-blotting. To exploit DCs properties for immunization purposes, recombinant protein chemically coupled to αDEC-205 monoclonal antibody and confirmed against anti-His antibody in western-blotting. RESULTS: The optimum condition for expression was 1 mM IPTG during 4 hr cultures in 2XYT medium, and final protein produced in soluble form. Conjugation of purified protein to αDEC-205 antibody resulted in smears of protein: antibodies conjugate in different molecular weights. CONCLUSION: The best cultivation condition for production of HIVtop4 protein is induction by 1 mM IPTG during 4 hr in 2XYT medium. The final concentration of purified protein was 500 µg/ml.

4.
Iran J Immunol ; 5(4): 207-11, 2008 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19098364

RESUMEN

BACKGROUND: HLA-G gene contains 15 alleles including a null allele, HLA-G*0105N. Previous studies have shown that HLA-G*0105N does not encode the complete HLA-G1 or HLA-G5 isoforms but encodes a functional HLA-G protein with the ability to inhibit NK cell cytolysis. Thus, although the biological functions of HLA-G1 and HLA-G5 proteins are abrogated, other isoforms such as HLA-G2 can replace their roles. Studies on the null allele of HLA-G gene could be useful in understanding the genetic variants of HLA-G alleles in ethnic groups. OBJECTIVE: The goal of this research was to determine the frequency of HLA-G*0105N null allele in Iranian healthy subjects. METHODS: The frequency of HLA-G*0105N null allele was evaluated in Iranian healthy subjects by PCR-RFLP method. Genomic DNA was isolated from the whole blood of 100 randomly selected, healthy, unrelated Iranian individuals using salting-out technique followed by PCR amplification of the exon 3 of HLA-G gene. PCR products were digested with PpUM-1 and the resulted fragments were analyzed using gel electrophoresis. RESULTS: In this study the restriction enzyme digestion confirmed homozygous HLA-G*0105N null allele for 9 % of the population. Furthermore obtained results indicated that the total frequency of HLA-G*0105N null allele was 20 % in the studied population of Iran. CONCLUSION: The final data analysis showed that the total frequency of this allele in Iranian people was higher than other ethnic groups that have been studied so far.


Asunto(s)
Frecuencia de los Genes/genética , Antígenos HLA/genética , Antígenos de Histocompatibilidad Clase I/genética , Alelos , Línea Celular Tumoral , Antígenos HLA-G , Humanos , Irán , Isoformas de Proteínas/genética
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