NK Cells and Their Role in Disease and Cancer Treatment

Natural killer (NK) cells are a crucial part of the human immune system. They play an important role in fighting and controlling viral infections as well as killing cells that have transformed into tumors. NK cells are involved in the pathogenesis of a number of diseases and in this thesis we will explore NK cells and their role in disease and cancer treatment. First, we will discuss antibody dependent cell mediated cytotoxicity (ADCC) and how deficiencies in ADCC play a role in human disease. We will also cover improvements we have made to quantifying and measuring ADCC through an improved chromium 51 assay. Then we will look at chronic fatigue syndrome (CFS) and the role innate immunity and impairment of NK cells function contribute to the disease. We report the results of our research on patients with CFS and compare their innate immunity to their healthy family members. Finally, we will discuss NK cells and their role in cancer treatment as well as ongoing work involving development of an “off the shelf” NK cell therapy for cancer treatment through CRISPR/Cas9 gene editing and deletion of MHC I. The last chapter provides my insights into potential research directions for the study of human NK cells

The oxytocin receptor antagonist, Atosiban, activates pro-inflammatory pathways in human amnion via Gαi signalling

Oxytocin (OT) plays an important role in the onset of human labour by stimulating uterine contractions and promoting prostaglandin/inflammatory cytokine synthesis in amnion via oxytocin receptor (OTR) coupling. The OTR-antagonist, Atosiban, is widely used as a tocolytic for the management of acute preterm labour. We found that in primary human amniocytes, Atosiban (10 μM) signals via PTX-sensitive Gαi to activate transcription factor NF-κB p65, ERK1/2, and p38 which subsequently drives upregulation of the prostaglandin synthesis enzymes, COX-2 and phospho-cPLA2 and excretion of prostaglandins (PGE2) (n = 6; p < 0.05, ANOVA). Moreover, Atosiban treatment increased expression and excretion of the inflammatory cytokines, IL-6 and CCL5. We also showed that OT-simulated activation of NF-κB, ERK1/2, and p38 and subsequent prostaglandin and inflammatory cytokine synthesis is via Gαi−2 and Gαi−3 but not Gαq, and is not inhibited by Atosiban. Activation or exacerbation of inflammation is not a desirable effect of tocolytics. Therefore therapeutic modulation of the OT/OTR system for clinical management of term/preterm labour should consider the effects of differential G-protein coupling of the OTR and the role of OT or selective OTR agonists/antagonists in activating proinflammatory pathways

Three-Terminal Si-Based Negative Differential Resistance Circuit Element with Adjustable Peak-to-Valley Current Ratios Using a Monolithic Vertical Integration

Si-based resonant bipolar transistors are demonstrated by the monolithic vertical integration of Si-based resonant interband tunnel diodes atop the emitter of Si/SiGe heterojunction bipolar transistors ~HBTs! on a silicon substrate. In the common emitter configuration, IC versus VCE shows negative differential resistance characteristics. The resulting characteristics are adjustable peak-to-valley current ratios, including infinite and negative values, and tailorable peak current densities by the control of the HBT base current under room temperature operation. With the integrated RITD-HBT combination, latching properties which are the key operating principle for high-speed mixed-signal, memory, and logic circuitry, are experimentally demonstrated

Prostaglandin F2α requires activation of calcium-dependent signalling to trigger inflammation in human myometrium

IntroductionPreterm birth is one of the major causes of neonatal morbidity and mortality across the world. Both term and preterm labour are preceded by inflammatory activation in uterine tissues. This includes increased leukocyte infiltration, and subsequent increase in chemokine and cytokine levels, activation of pro-inflammatory transcription factors as NF-κB and increased prostaglandin synthesis. Prostaglandin F2α (PGF2α) is one of the myometrial activators and stimulators.MethodsHere we investigated the role of PGF2α in pro-inflammatory signalling pathways in human myometrial cells isolated from term non-labouring uterine tissue. Primary myometrial cells were treated with G protein inhibitors, calcium chelators and/or PGF2α. Nuclear extracts were analysed by TranSignal cAMP/Calcium Protein/DNA Array. Whole cell protein lysates were analysed by Western blotting. mRNA levels of target genes were analysed by RT-PCR.ResultsThe results show that PGF2α increases inflammation in myometrial cells through increased activation of NF-κB and MAP kinases and increased expression of COX-2. PGF2α was found to activate several calcium/cAMP-dependent transcription factors, such as CREB and C/EBP-β. mRNA levels of NF-κB-regulated cytokines and chemokines were also elevated with PGF2α stimulation. We have shown that the increase in PGF2α-mediated COX-2 expression in myometrial cells requires coupling of the FP receptor to both Gαq and Gαi proteins. Additionally, PGF2α-induced calcium response was also mediated through Gαq and Gαi coupling.DiscussionIn summary, our findings suggest that PGF2α-induced inflammation in myometrial cells involves activation of several transcription factors – NF-κB, MAP kinases, CREB and C/EBP-β. Our results indicate that the FP receptor signals via Gαq and Gαi coupling in myometrium. This work provides insight into PGF2α pro-inflammatory signalling in term myometrium prior to the onset of labour and suggests that PGF2α signalling pathways could be a potential target for management of preterm labour

The latency-associated transcript locus of herpes simplex virus 1 is a virulence determinant in human skin

Herpes simplex virus 1 (HSV-1) infects skin and mucosal epithelial cells and then travels along axons to establish latency in the neurones of sensory ganglia. Although viral gene expression is restricted during latency, the latency-associated transcript (LAT) locus encodes many RNAs, including a 2 kb intron known as the hallmark of HSV-1 latency. Here, we studied HSV-1 infection and the role of the LAT locus in human skin xenografts in vivo and in cultured explants. We sequenced the genomes of our stock of HSV-1 strain 17syn+ and seven derived viruses and found nonsynonymous mutations in many viral proteins that had no impact on skin infection. In contrast, deletions in the LAT locus severely impaired HSV-1 replication and lesion formation in skin. However, skin replication was not affected by impaired intron splicing. Moreover, although the LAT locus has been implicated in regulating gene expression in neurones, we observed only small changes in transcript levels that were unrelated to the growth defect in skin, suggesting that its functions in skin may be different from those in neurones. Thus, although the LAT locus was previously thought to be dispensable for lytic infection, we show that it is a determinant of HSV-1 virulence during lytic infection of human skin

Video-Assisted Thoracoscopic Surgery for Correction of Adolescent Idiopatic Scoliosis: Comparison of 4.5 mm versus 5.5 mm Rod Constructs

PURPOSE: The purpose of this study is to report the comparative results of thoracoscopic correction achieved via cantilever technique using a 4.5 mm thin rod and the poly-axial reduction screw technique using a 5.5 mm thick rod in Lenke type 1 adolescent idiopathic scoliosis (AIS). MATERIALS AND METHODS: Radiographic data, Scoliosis Research Society (SRS) patient-based outcome questionnaires, and operative records were reviewed for forty-nine patients undergoing surgical treatment of scoliosis. The study group was divided into a 4.5 mm thin rod group (n = 24) and a 5.5 mm thick rod group (n = 25). The radiographic parameters that were analyzed included coronal curve correction, the most caudal instrumented vertebra tilt angle correction, coronal balance, and thoracic kyphosis. RESULTS: The major curve was corrected from 49.8 degrees and 47.2 degrees pre-operatively to 24.5 degrees and 18.8 degrees at the final follow-up for the thin and thick rod groups, respectively (50.8% vs. 60.2% correction). There were no significant differences between the two groups in terms of kyphosis, coronal balance, or tilt angle at the time of the final follow-up. The mean number of levels fused was 6.2 in the thin rod group, compared with 5.9 levels in the thick rod group. There were no major intraoperative complications in either group. CONCLUSION: Significant correction loss was observed in the thin rod system at the final follow-up though both groups had comparable correction immediately post-operative. Therefore, the thick rod with poly axial screw system helps to maintain post-operative correctionope

Four Martian years of nightside upper thermospheric mass densities derived from electron reflectometry: Method extension and comparison with GCM simulations

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/95498/1/jgre2766.pd