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

Abstract

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 β1 (TGFβ1)] 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.

CCL11 increases the proportion of CD4+CD25+Foxp3+ Treg cells and the production of IL‑2 and TGF‑β by CD4+ T�cells via the STAT5 signaling pathway

“…Cell treatment. The peripheral blood mononuclear cells (PBMcs) were isolated from peripheral blood samples collected from each participant in tubes with K2 ethylenediaminetetraacetic acid (edTa) as anticoagulant as previously described (24). cd4 + T cells were isolated from the PBMcs of patients with Bc using the Magcellect Human cd4 + T cell isolation kit (cat.…”

CCL26 regulates the proportion of CD4+CD25+FOXP3+�Tregs and the production of inflammatory factors in peripheral blood mononuclear cells following acute ischemic stroke via the STAT5 pathway

“…Cell supernatant assay. PBMCs were isolated from peripheral blood samples collected from each subject in tubes containing K2 ethylenediamine tetraacetic acid as the anticoagulant, as previously described (16). They were treated with various concentrations of recombinant human CCL26 (ab243255, Abcam) (Rep CCL26; 0, 5, 25 and 100 ng/ml) or with 200 mg/l anti-CCL26 neutralizing antibody (ab109612, Abcam) in the absence or presence of 1 µM the STAT5 inhibitor for 24 h, STAT5-IN-1 (CAS: 285986-31-4, EMD Millipore).…”

Identification of a novel fully human anti-toxic shock syndrome toxin (TSST)-1 single-chain variable fragment antibody averting TSST-1-induced mitogenesis and cytokine secretion

“…The neutralizing activity of MS473 against the superantigenic activity of TSST-1 was examined on human PBMCs isolated from the whole blood of two healthy donors (men, 50 and 45 years) as described previously with some modifications [ 8 , 12 , 13 , 31 , 52 , 53 ]. To determine the effective proliferative dose of TSST-1, fresh human PBMCs (~ 10 6 cells/ml) in RPMI 1640 medium (Gibco; Grand Island, NY, USA) with L-glutamine (2 mM) (Gibco) supplemented with heat-inactivated fetal bovine serum (10%) (Gibco) were seeded in 96-well round-bottom tissue culture plates (JET BIOFIL, Guangzhou, China) and were incubated with the TSST-1 protein at concentrations of 1, 10, 25, 50, and 100 ng/ml (0.04, 0.4, 1.04, 2.1, and 4.2 nM, respectively) for 48 h at 37 °C, 5% CO2.…”