A study of MHC class I binding viral and host derived peptides and natural killer cell function

Natural killer (NK) cells are a key component of innate immunity and have been implicated in determining the outcome of HCV infection in both genetic and functional studies. The last two decades have seen significant advances in the understanding of NK cell regulation with the discovery of a multitude of activating and inhibitory receptors. CD94:NKG2A operates in tandem with the polymorphic killer cell immunoglobulin-like receptors (KIR) and Ly49 systems to inhibit NK cells, however it is not clear as to the benefits of having two distinct inhibitory receptor:ligand systems. Down regulation or modification of MHC class I expression is a key feature of NK cell recognition of virus infected cells. However, viruses can subvert this mechanism of NK cell surveillance by encoding peptides that can bind to MHC class I. The aim of this thesis is to further our understanding of the interaction between viral and host derived MHC class I binding peptides and their effect on NK cell inhibition. By using an MHC deficient cell line, we have shown that HCV core35-44 peptide is capable of enhancing cell surface expression of MHC class I (HLA-C and HLA-E). Although this peptide stabilises HLA-E, the HLA-E:HCV core35-44 complex alone is insufficient to inhibit at NKG2A positive NK cells. However, in the presence of HLA-E binding MHC class I signal peptides, HCV core35-44 has a synergistic effect in suppressing the NKG2A+ NK cell population. Peptides derived from other viruses such as EBV and HIV, and the stress related peptide derived from heat shock protein 60 also augment inhibition of NKG2A+ NK cells, but only when in the presence of MHC class I signal peptides. This augmentation is caused by recruitment of the non-signalling CD94 molecule to the immune synapse in the absence of its inhibitory signalling partner NKG2A. Thus CD94 can function independently as an enhancer of inhibition. The augmentation of inhibition of CD94:NKG2A by non-inhibitory peptides, contrasts with antagonism of inhibition of KIR by low affinity peptide:MHC complexes. We also show that KIR+ and NKG2A+ NK cells respond with differing stoichiometries to MHC class I down-regulation. Thus peptide selectivity and MHC class I sensitivity of natural killer cell receptors provides a rationale for the evolution of two distinct inhibitory systems for MHC class I.Open Acces

Report: Fatal case of disseminated BCG infection in an infant born to a mother taking infliximab for Crohn’s disease

Abstract We present the case of a 28 year old lady with refractory Crohn's Disease treated with infliximab throughout her pregnancy. Her baby was born healthy and received a Bacillus Calmette-Guérin (BCG) vaccine aged 3 months. Soon after this the infant became unwell and died aged 4.5 months. At post-mortem the cause of death was attributed to an unusual complication of the BCG vaccine, known as disseminated BCG. BCG vaccination is contraindicated in individuals who are receiving immunosuppressive drugs. We recommend physicians should exercise caution before such vaccines are used in infants born to mothers taking anti-TNF therapies or other potentially immunosuppressive IgG1 antibodies. © 2010 European Crohn's and Colitis Organisation. Published by Elsevier B.V. All rights reserved. Case report This case is of a 28 year old Caucasian lady diagnosed with pan-colitis and erythema nodosum secondary to Crohn's Disease (CD) in 2001. She was treated early on with mesalazine and azathioprine at a dose of 2 mg per kg bodyweight but experienced recurrent flare-ups requiring repeated courses of prednisolone with eventual loss of response. In 2004, infliximab 5 mg/kg was commenced with initial good symptomatic benefit but subsequent loss of response. Therefore, in 2006 the dose of infliximab was increased to 10 mg/kg every eight weeks. In 2008 she became pregnant. The pros and cons of continuing infliximab were discussed at length with the patient. Due the severity and corticosteroid refractory nature of her CD, the benefits of maintaining remission with infliximab was felt to outweigh the risks of foetus exposure to the drug. She consented to continue eightweekly infusions of infliximab 10 mg/kg as monotherapy for CD. Her disease remained in remission during pregnancy and in July 2008 she gave birth to a healthy baby boy. The baby was born at 36 + 3 gestation via spontaneous vaginal deliver

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.This work was supported by grants from the Wellcome Trust to KMJ and SIK (092675/Z/10/Z)

Safety of anti-rheumatic drugs for rheumatoid arthritis in pregnancy and lactation

Women with rheumatoid arthritis (RA) are often of childbearing age and therefore questions regarding reproductive health and the use of medications, including disease-modifying anti-rheumatic drugs (DMARDs) may arise during the clinical consultation. Each patient requires individual assessment in order to effectively manage the disease while minimizing any treatment-associated risks to the fetus. Although good-quality controlled trials are lacking, there is an increasing volume of evidence surrounding the use of immunosuppressive therapies in pregnancy and lactation. This review summarizes the currently available information which can be of benefit to clinicians guiding patients and their families through the risks and benefits of continuing RA therapy during pregnancy and lactation. Further studies and ongoing surveillance of drug safety in pregnancy are required to resolve the uncertainties that remain regarding synthetic and biologic DMARDs

Variations in killer-cell immunoglobulin-like receptor and human leukocyte antigen genes and immunity to malaria

Malaria is one of the deadliest infectious diseases in the world. Immune responses to Plasmodium falciparum malaria vary among individuals and between populations. Human genetic variation in immune system genes is likely to play a role in this heterogeneity. Natural killer (NK) cells produce inflammatory cytokines in response to malaria infection, kill intraerythrocytic Plasmodium falciparum parasites by cytolysis, and participate in the initiation and development of adaptive immune responses to plasmodial infection. These functions are modulated by interactions between killer-cell immunoglobulin-like receptors (KIR) and human leukocyte antigens (HLA). Therefore, variations in KIR and HLA genes can have a direct impact on NK cell functions. Understanding the role of KIR and HLA in immunity to malaria can help to better characterize antimalarial immune responses. In this review, we summarize the different KIR and HLA so far associated with immunity to malaria.This work was supported through the DELTAS Africa Initiative (Grant no. 107743), that funded Stephen Tukwasibwe through PhD fellowship award, and Annettee Nakimuli through group leader award. The DELTAS Africa Initiative is an independent funding scheme of the African Academy of Science (AAS), Alliance for Accelerating Excellence in Science in Africa (AESA) and supported by the New Partnership for Africa’s Development Planning and Coordinating Agency (NEPAD Agency) with funding from the Wellcome Trust (Grant no. 107743) and the UK government. Francesco Colucci is funded by Wellcome Trust grant 200841/Z/16/Z. The project received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (grant agreement No. 695551) for James Traherne and John Trowsdale. Jyothi Jayaraman is a recipient of fellowship from the Centre for Trophoblast Research

Effects of peptide on NK cell-mediated MHC I recognition

The inhibitory receptors for MHC class I have a central role in controlling natural killer (NK) cell activity. Soon after their discovery, it was found that these receptors have a degree of peptide selectivity. Such peptide selectivity has been demonstrated for all inhibitory killer cell immunoglobulin-like receptor (KIR) tested to date, certain activating KIR, and also members of the C-type lectin-like family of receptors. This selectivity is much broader than the peptide specificity of T cell receptors, with NK cell receptors recognizing peptide motifs, rather than individual peptides. Inhibitory receptors on NK cells can survey the peptide:MHC complexes expressed on the surface of target cells, therefore subsequent transduction of an inhibitory signal depends on the overall peptide content of these MHC class I complexes. Functionally, KIR-expressing NK cells have been shown to be unexpectedly sensitive to changes in the peptide content of MHC class I, as peptide:MHC class I complexes that weakly engage KIR can antagonize the inhibitory signals generated by engagement of stronger KIR-binding peptide:MHC class I complexes. This property provides KIR-expressing NK cells with the potential to recognize changes in the peptide:MHC class I repertoire, which may occur during viral infections and tumorigenesis. By contrast, in the presence of HLA class I leader peptides, virus-derived peptides can induce a synergistic inhibition of CD94:NKG2A-expressing NK cells through recruitment of CD94 in the absence of NKG2A. On the other hand, CD94:NKG2A-positive NK cells can be exquisitely sensitive to changes in the levels of MHC class I. Peptide antagonism and sensitivity to changes in MHC class I levels are properties that distinguish KIR and CD94:NKG2A. The subtle difference in the properties of NK cells expressing these receptors provides a rationale for having complementary inhibitory receptor systems for MHC class I

Natural killer cells and hepatitis C: action and reaction

In 1989, hepatitis C virus (HCV) was first identified as the infectious agent responsible for human non-A, non-B hepatitis. Two decades later, HCV remains a global public health problem with a suboptimal response rate to treatment and the absence of a protective vaccine. Recent work has highlighted the influence of the innate immune system, and in particular natural killer cells, on the outcome and pathology of HCV infection. These cells are considerably more complex than was originally thought and their role in viral infections is currently being unravelled. This review summarises our emerging understanding of natural killer cells in HCV infection