Entamoeba histolytica Calreticulin Induces the Expression of Cytokines in Peripheral Blood Mononuclear Cells Isolated From Patients With Amebic Liver Abscess.

Calreticulin (CRT) is a highly conserved protein in the endoplasmic reticulum that plays important roles in the regulation of key cellular functions. Little is known about the participation of E. histolytica CRT (EhCRT) in the processes of pathogenicity or in the modulation of the host immune response. The aim of this study was to evaluate the role of CRT in the proliferation and the cytokine profile in peripheral blood mononuclear cells (PBMCs) from patients with amebic liver abscess (ALA) during the acute phase (AP-ALA) of the disease compared to patients during the resolution phase (R-ALA). The PBMCs from each participant were cocultured with EhCRT and tested by the colorimetric method to evaluate their proliferation index (PI). The supernatants were subjected to an enzyme-linked immunosorbent assay (ELISA) to evaluate the concentration of cytokines. The mean values of all groups were compared using the independent t-test. When the PIs of individuals without diagnosis of liver abscess (NEG) were compared, there were no statistically significant differences in the proliferation of PBMCs between patients with AP-ALA and R-ALA when stimulated with EhCRT or concanavalin A (ConA). However, the levels of interleukins [IL-6, IL-10, granulocyte colony stimulating factor (GCSF), and transforming growth factor ß1 (TGFß1)] were higher in patients with AP-ALA, whereas in patients with R-ALA, higher levels of interferon gamma (IFNγ) were detected. These results suggest that EhCRT acts as a mitogen very similar to the activity of ConA. In addition, EhCRT is an excellent immunogen for the specific activation of PBMCs, inducing the differential expression of ILs depending on the outcome of disease, determining the type of immune response: a Th2 cytokine profile during the acute phase and a Th1 profile during the resolution phase.

Characterization of Single-Nucleotide Polymorphisms in the Tumor Necrosis Factor α Promoter Region and in Lymphotoxin α in Squamous Intraepithelial Lesions, Precursors of Cervical Cancer.

Development of cervical cancer is a long process of abnormal cancerous cell growth in the cervix and is primarily the result of infection with specific high-risk types of human papillomavirus (HPV). The cytokines tumor necrosis factor α (TNFα) and lymphotoxin α (LTA) have an important role in all stages of cervical cancer and have the ability to induce the regression or promote the development of human tumors. Biologically important single-nucleotide polymorphisms (SNPs) occur within the TNFα and LTA genes. Therefore, the purpose of this study was to investigate the SNPs in the TNFα promoter region (-163, -238, -244, -308, -376, -857, -863, and -1031) and in the first intron of LTA (+252) in women with precursor lesions of cervical cancer. Overall, we studied 396 women from Mexico City. A total of 191 patients with HPV infection and precursor cervical lesions were subdivided in two groups: those with low-grade squamous intraepithelial lesions (n = 132) and those with high-grade squamous intraepithelial lesions (n = 59). Women (n = 205) negative for HPV and without cervical lesions were also included in the study. DNA was extracted from peripheral white blood cells and from cervical samples, and detection of biallelic polymorphisms of TNFα and LTA was performed using the polymerase chain reaction-sequence-specific oligonucleotide probe and restriction fragment length polymorphism techniques, respectively. We demonstrated that risk is associated with the genotype G/A (odds ratio = 2.48) and that protection is associated with the genotype G/G of SNP TNFα -376 (odds ratio = 0.37).

La transfección de la línea celular TF-1 con el proto-oncogen bcl-2 humano proporciona supervivencia a corto plazo en ausencia de GM-CSF sin cambiar el fenotipo / Transfection of the tf-1 cell line with the human protooncogen bcl-2 favors short term cell survival and does not change phenotype

La muerte celular programada (apoptosis) es un proceso controlado genéticamente. El producto del gen bcl-2 (Bcl-2) es una proteína anti apoptótica integrada a la membrana externa mitocondrial. Objetivo. El objetivo del presente trabajo fue valorar los efectos de la transfección de bcl-2 sobre la supervivencia, proliferación, fenotipo y morfología en las líneas celulares TF-1, TF-1 neo (control negativo) y TB-1 (TF-1 transfectada con bcl-2). Metodología. Para ello utilizamos el método siguiente: examen del nivel de expresión de los antígenos de superficie CD13, CD34 y c-Kit, respuesta celular al GM-CSF y capacidad de supervivencia luego del retiro del factor de viabilidad. La apoptosis fue evaluada mediante la observación del blebbing membranal a diferentes tiempos luego del retiro del suero, el factor de viabilidad GM-CSF y la tolerancia a toxinas presentes en Justicia spicigera. Resultados. La expresión ectópica de bcl-2 en las células TB-1 previno la muerte celular por apoptosis sin inducir un cambio importante en el fenotipo y morfología en ausencia de GM-CSF, suero o en presencia del extracto de Justicia spicigera. Consistente con la función de Bcl-2, encontramos que la proliferación de la línea celular TB-1 fue muy similar a la línea parental TF-1 en respuesta al GM-CSF y por lo tanto Bcl-2 no altera dicha función. El retiro de suero y GM-CSF al medio de cultivo de las líneas TF-1 y TF-1 neo, induce apoptosis en 36 h. Por el contrario, la línea TB-1 entra en apoptosis a 96 h bajo las mismas condiciones. Conclusiones. Reuniendo los resultados, éstos sugieren que Bcl-2 es un inhibidor de la apoptosis a corto plazo en la línea celular TB-1, no cambia la respuesta al GM-CSF y no afecta el nivel de expresión de otros antígenos de superficie como CD13, CD34 y c-Kit.