STAT4-associated natural killer cell tolerance following liver transplantation.

OBJECTIVE: Natural killer (NK) cells are important mediators of liver inflammation in chronic liver disease. The aim of this study was to investigate why liver transplants (LTs) are not rejected by NK cells in the absence of human leukocyte antigen (HLA) matching, and to identify a tolerogenic NK cell phenotype. DESIGN: Phenotypic and functional analyses on NK cells from 54 LT recipients were performed, and comparisons made with healthy controls. Further investigation was performed using gene expression analysis and donor:recipient HLA typing. RESULTS: NK cells from non-HCV LT recipients were hypofunctional, with reduced expression of NKp46 (p<0.05) and NKp30 (p<0.001), reduced cytotoxicity (p<0.001) and interferon (IFN)-γ secretion (p<0.025). There was no segregation of this effect with HLA-C, and these functional changes were not observed in individuals with HCV. Microarray and RT-qPCR analysis demonstrated downregulation of STAT4 in NK cells from LT recipients (p<0.0001). Changes in the expression levels of the transcription factors Helios (p=0.06) and Hobit (p=0.07), which control NKp46 and IFNγ expression, respectively, were also detected. Hypofunctionality of NK cells was associated with impaired STAT4 phosphorylation and downregulation of the STAT4 target microRNA-155. Conversely in HCV-LT NK cell tolerance was reversed, consistent with the more aggressive outcome of LT for HCV. CONCLUSIONS: LT is associated with transcriptional and functional changes in NK cells, resulting in reduced activation. NK cell tolerance occurs upstream of major histocompatibility complex (MHC) class I mediated education, and is associated with deficient STAT4 phosphorylation. STAT4 therefore represents a potential therapeutic target to induce NK cell tolerance in liver disease.

Investigating the frail elderly patient with lower bowel symptoms: what do we do now and can we improve?

AIMS: To assess the utility of flexible sigmoidoscopy (FS) and minimal preparation CT (MPCT) in investigating lower gastrointestinal (LGI) symptoms in elderly patients who are too frail to undergo colonoscopy or spiral CT. METHODS: All FS examinations performed in patients aged over 70 between 1 January and 31 December 2008 were analysed. Predictors of usefulness were determined using multivariable analysis. In patients who also underwent MPCT, we analyzed the correlation between FS and MPCT. RESULTS: 426 FS were performed. Bowel preparation was inadequate in 24% of procedures. Indications in which FS was useful were: radiological abnormality (odds ratio [OR] 9.32), history of polyps (OR 4.54) and rectal bleeding (OR 1.73). Indications for which FS was least useful were: change in bowel habit (OR 0.22), diarrhoea (OR 0.46) and constipation (OR 0.38). CONCLUSIONS: LGI investigation in frail elderly patients can be rationalised according to indication. Performing FS and MPCT together is not always necessary.

Salmonella enterica serovar Typhi-specific immunoglobulin A antibody responses in plasma and antibody in lymphocyte supernatant specimens in Bangladeshi patients with suspected typhoid fever.

Many currently available diagnostic tests for typhoid fever lack sensitivity and/or specificity, especially in areas of the world where the disease is endemic. In order to identify a diagnostic test that better correlates with typhoid fever, we evaluated immune responses to Salmonella enterica serovar Typhi (serovar Typhi) in individuals with suspected typhoid fever in Dhaka, Bangladesh. We enrolled 112 individuals with suspected typhoid fever, cultured day 0 blood for serovar Typhi organisms, and performed Widal assays on days 0, 5, and 20. We harvested peripheral blood lymphocytes and analyzed antibody levels in supernatants collected on days 0, 5, and 20 (using an antibody-in-lymphocyte-supernatant [ALS] assay), as well as in plasma on these days. We measured ALS reactivity to a serovar Typhi membrane preparation (MP), a formalin-inactivated whole-cell preparation, and serovar Typhi lipopolysaccharide. We measured responses in healthy Bangladeshi, as well as in Bangladeshi febrile patients with confirmed dengue fever or leptospirosis. We categorized suspected typhoid fever individuals into different groups (groups I to V) based on blood culture results, Widal titer, and clinical features. Responses to MP antigen in the immunoglobulin A isotype were detectable at the time of presentation in the plasma of 81% of patients. The ALS assay, however, tested positive in all patients with documented or highly suspicious typhoid, suggesting that such a response could be the basis of improved diagnostic point-of-care-assay for serovar Typhi infection. It can be important for use in epidemiological studies, as well as in difficult cases involving fevers of unknown origin.

Distinct mechanisms of action of V1 antagonists OPC-21268 and [d(CH2)5Tyr(Me)AVP] in mesangial cells.

The effects of two arginine vasopressin (AVP) V1 receptor antagonists, a peptide analogue [d(CH2)5Tyr(Me)AVP] and an orally active non-peptide OPC-21268, on AVP-stimulated Ca transients and 3H-AVP binding were examined in cultured rat mesangial cells. Although both [d(CH2)5Tyr(Me)AVP] and OPC-21268 dose-dependently inhibited AVP-stimulated Ca transients and 3H-AVP binding, their effects were quite different. Magnitude of inhibition of Ca transients by OPC-21268 was comparable when OPC-21268 was added together with or 5 min prior to AVP. However, the inhibition by [d(CH2)5Tyr(Me)AVP] was greater when it was added prior to AVP than when it was added together with AVP. Moreover, washing mesangial cells after OPC-21268 treatment, but not after [d(CH2)5Tyr(Me)AVP] treatment, completely restored AVP-stimulated Ca transients to the control level without antagonists. These results indicate that the cellular mechanisms for two V1 receptor antagonists are different: while the effect of OPC-21268 as a vasopressin antagonist appears to be competitive and reversible, the effect of the peptide antagonist [d(CH2)5Tyr(Me)AVP] seems to be rather irreversible in nature.

Expression of platelet activating factor receptor in renal tubular cell line (LLC-PK1).

Northern blot analyses of poly(A)+ RNA extracted from cloned renal tubule cells, LLC-PK1, MDCK and JTC-12 cells, were performed using an isolated cDNA of guinea pig platelet activating factor (PAF) receptor as a probe. The results showed the expression of single entity of PAF receptor mRNA in LLC-PK1, but not in MDCK or JTC-12 cells. PAF also induced a dose-dependent elevation of intracellular calcium in LLC-PK1 cells as assessed by Fura-2 fluorescence signals, which was completely blocked by a PAF receptor antagonist, WEB 2170. These results indicate a PAF receptor-effector system in a renal tubular cell line LLC-PK1, providing a possible model system for studying physiological roles of PAF receptors in renal tubules.