Engineered T cell therapies.

Alongside advancements in gene therapy for inherited immune disorders, the need for effective alternative therapeutic options for other conditions has resulted in an expansion in the field of research for T cell gene therapy. T cells are easily obtained and can be induced to divide robustly ex vivo, a characteristic that allows them to be highly permissible to viral vector-mediated introduction of transgenes. Pioneering clinical trials targeting cancers and infectious diseases have provided safety and feasibility data and important information about persistence of engineered cells in vivo. Here, we review clinical experiences with γ-retroviral and lentiviral vectors and consider the potential of integrating transposon-based vectors as well as specific genome editing with designer nucleases in engineered T cell therapies.

Myelodysplastic syndrome with trisomy 8 associated with Behçet syndrome: an immunologic link to a karyotypic abnormality.

Myelodysplastic syndrome (MDS) in children is often associated with chromosomal anomalies and trisomy 8 is a characteristic karyotypic feature in up to 20% of the cases. Behçet disease is a rare multisystem inflammatory disorder characterized by recurrent mouth and genital ulcers. MDS with trisomy 8 has been observed in adult patients with Behçet syndrome with some cases developing prior to the clinical manifestations of the latter. We present a female with a similar association and explain the importance of identifying the coexisting conditions. The immunological abnormalities, which may be observed in MDS and their possible mechanisms, are also discussed.

Trends and regional variation in the incidence of head and neck cancers in England: 2002 to 2011.

Recent studies show an increased incidence of head and neck cancers worldwide. The present study evaluated the trend in the incidence of head and neck cancers in England during 2002-2011. Data were extracted from the database of Office for National Statistics. The study population was categorised according to age, residential area, gender and cancer sub-types. Overall trend in incidence of head and neck cancer and some subtypes were examined using Poisson regression models. In total, 71,457 head and neck cancers were registered in England between 2002 and 2011 and 68% of patients were males. Statistically significant increases in incidence of 27.0 and 32.4% were documented in males and females, respectively (p<0.001) with the largest increase in the 60+ age category. Potentially HPV-associated cancers, oral cavity cancers and laryngeal cancers increased by 47.1, 24.1 and 1.7% in males and 37.5, 25.5 and 7.7% in females, respectively (p<0.001). Regional differences were also noted with the highest incidence (18.0 and 17.0 per 100,000, respectively) in the North East and North West of England. Our results for England showed an increase in the incidence of both oral cavity and oropharyngeal cancer in both genders, whilst laryngeal cancer incidence remained stable.

Knowledge of oral cancer, distress and screening intentions: longer term effects of a patient information leaflet.

Study aim was to determine the influence of a patient information leaflet (PIL) on mouth cancer to improve knowledge, reduce distress and increase intention to accept a mouth screen over a 2-month period. The design was a randomised controlled trial. Two dental practices in the northwest of England participated. Standardised multi-item scales of the three outcome measures were employed. The PIL was given to a randomised intervention group of patients in waiting room. Single sheet questionnaire was completed by both groups of patients at baseline in waiting room (immediately following leaflet administration in intervention arm of study). Repeat questionnaire completion at 8 weeks by all patients through postal system. Mann-Whitney U-tests comparing outcome variables between patients with and without access to the leaflet at baseline and 8 weeks were performed. Multiple logistic regression was used to predict re-reading of the leaflet at home. Useable replies were received from 317 patients (60% response rate). All measures showed some benefit of immediate exposure to the leaflet at follow up. Older patients, less initial knowledge, and self-reported smoking positively predicted the re-reading of the leaflet. The introduction of a mouth cancer PIL into dental practice may help to inform patients about oral cancer, moderate distress and encourage acceptance of an oral health screen.

Mental nerve anaesthesia; a complication of sickle cell crisis during childbirth.

This report describes the case of an Afro-Caribbean lady diagnosed with sickle cell anaemia, who presented with permanent mental nerve anaesthesia as a result of a sickle cell crisis during childbirth.

Comparison of lentiviral and sleeping beauty mediated αß T cell receptor gene transfer.

Transfer of tumour antigen-specific receptors to T cells requires efficient delivery and integration of transgenes, and currently most clinical studies are using gamma retroviral or lentiviral systems. Whilst important proof-of-principle data has been generated for both chimeric antigen receptors and αß T cell receptors, the current platforms are costly, time-consuming and relatively inflexible. Alternative, more cost-effective, Sleeping Beauty transposon-based plasmid systems could offer a pathway to accelerated clinical testing of a more diverse repertoire of recombinant high affinity T cell receptors. Nucleofection of hyperactive SB100X transposase-mediated stable transposition of an optimised murine-human chimeric T cell receptor specific for Wilm's tumour antigen from a Sleeping Beauty transposon plasmid. Whilst transfer efficiency was lower than that mediated by lentiviral transduction, cells could be readily enriched and expanded, and mediated effective target cells lysis in vitro and in vivo. Integration sites of transposed TCR genes in primary T cells were almost randomly distributed, contrasting the predilection of lentiviral vectors for transcriptionally active sites. The results support exploitation of the Sleeping Beauty plasmid based system as a flexible and adaptable platform for accelerated, early-phase assessment of T cell receptor gene therapies.

Immune responses elicited in tertiary lymphoid tissues display distinctive features.

During chronic inflammation, immune effectors progressively organize themselves into a functional tertiary lymphoid tissue (TLT) within the targeted organ. TLT has been observed in a wide range of chronic inflammatory conditions but its pathophysiological significance remains unknown. We used the rat aortic interposition model in which a TLT has been evidenced in the adventitia of chronically rejected allografts one month after transplantation. The immune responses elicited in adventitial TLT and those taking place in spleen and draining lymph nodes (LN) were compared in terms of antibody production, T cell activation and repertoire perturbations. The anti-MHC humoral response was more intense and more diverse in TLT. This difference was associated with an increased percentage of activated CD4+ T cells and a symmetric reduction of regulatory T cell subsets. Moreover, TCR repertoire perturbations in TLT were not only increased and different from the common pattern observed in spleen and LN but also "stochastic," since each recipient displayed a specific pattern. We propose that the abnormal activation of CD4+ T cells promotes the development of an exaggerated pathogenic immune humoral response in TLT. Preliminary findings suggest that this phenomenon i) is due to a defective immune regulation in this non-professional inflammatory-induced lymphoid tissue, and ii) also occurs in human chronically rejected grafts.

Itch-/- alphabeta and gammadelta T cells independently contribute to autoimmunity in Itchy mice.

E3 ubiquitin ligases determine which intracellular proteins are targets of the ubiquitin conjugation pathway and thus play a key role in determining the half-life, subcellular localization and/or activation status of their target proteins. Itchy mice lack the E3 ligase, Itch, and show dysregulation of T lymphocytes and the induction of a lethal autoimmune inflammatory condition. Itch is widely expressed in hematopoietic and nonhematopoietic cells, and we demonstrate that disease is transferred exclusively by hematopoietic cells. Moreover, distinct manifestations of the autoimmune inflammatory phenotype are contributed by discrete populations of lymphocytes. The presence of Itch-deficient alphabeta T cells drives expansion of peritoneal B1b cells and elevated IgM levels, which correlate with itching and pathology. In contrast, Itch(-/-) interleukin-4-producing gammadelta T cells, even in the absence of alphabeta T cells, are associated with elevated levels of IgE and an inflammatory condition. These data indicate that disruption of an E3 ubiquitin ligase in alphabeta T cells can subvert a B-cell subpopulation, which normally functions to control particular microbial pathogens in a T-independent manner, to contribute to autoimmunity. In addition, disruption of Itch in innate gammadelta T cells can influence autoimmune pathology and might therefore require distinct therapeutic intervention.

Lymphoid neogenesis in chronic rejection: evidence for a local humoral alloimmune response.

Recent advances indicate that, in various chronic inflammatory disorders, the activation of the immune system is triggered locally rather than in lymphoid organs. In this study, we have evaluated whether the humoral alloimmune response involved in chronic rejection is elicited within the graft. We used the rat aortic interposition model and microdissected the adventitia of the graft. Over time, the T cell infiltrate shifted toward a B helper phenotype. B lymphocyte clusters were detected and were the site of intense proliferation and apoptosis. Simultaneously, adventitial vascular endothelium acquired a high endothelial venule phenotype. Similar features were evidenced in the interstitium of chronically allografts (hearts and kidneys). Strikingly, ganocultured graft interstitial tissue was found to be the site of production of antibodies directed against donor MHC-I molecules. These findings, therefore, document the appearance of germinal centers in chronically rejected tissues. This lymphoid neogenesis implies that the graft is not only the target of the alloimmune response but also a site where this response actually develops, so as to optimize the communication between the targeted tissue and the immune effectors.

Neonatal tolerance to a Th2-mediated autoimmune disease generates CD8+ Tc1 regulatory cells.

Immunological tolerance can be achieved in animals by exposure of newborn to a foreign antigen. Depending on the dose and timing of the antigenic challenge, tolerance has been reported to result in clonal deletion, anergy or active suppression. In this latter case, regulatory T cells prevent autoimmunity by suppressing the reactivity of pathogenic self-reactive T cells. We have previously reported the generation of a neonatal, mercury-specific, and dominant tolerance to autoimmunity induced by mercury salts in rats. Chronic exposure to mercury salts can lead to SLE-like autoimmune responses, mediated by autoreactive CD4+ Th2 cells, that regulate and are followed by a resistant state mediated by protective CD8+ T cells. The aim of the study was to compare the resistance to the neonatal tolerance to mercury disease, and to further characterize the CD8+ T cells endowed with regulatory capacity in the neonatal tolerance model. We report here that resistance to mercury disease is long lasting and not mercury-specific, suggesting that different CD8+ T cells are involved in resistance and neonatal tolerance, and that regulatory CD8+ Tc1 cells generated in tolerance are required to control the CD8- cell population from developing Th2-mediated autoimmunity. Upon mercury recall, CD8+ CD45RC(high) T cells, that represent the Tc1 subset in the rat, expanded and were polarized towards IFNgamma production. Interestingly, identical results were obtained with the CD8+ CD25+T cell population. Substantial amounts of FasL gene expression were detected in CD8+ T lymphocytes upon recall with the tolerogen. AICD may be one of the regulatory mechanisms used by these regulatory CD8+ Tc1 cells that control neonatal tolerance to a Th2-mediated autoimmune disorder.

Regulatory CD8+ T cells control neonatal tolerance to a Th2-mediated autoimmunity.

Exposure of newborn animals to a foreign Ag may result in immunological tolerance to that specific Ag, a phenomenon called neonatal tolerance. We have previously reported that neonatal administration to Brown-Norway rats of mercury, a heavy metal toxicant, induces a dominant tolerance, specific for the chemical otherwise responsible for Th2 cell-mediated autoimmune responses in this susceptible strain of rats. Neonatal exposure to Ags can prime immunity, rather than inactivate or delete responses, and sustain regulatory functions effective against autoreactive T cells. Here, we address whether such a tolerant response is due to the generation of regulatory cells. The results suggest that the CD8(+) T cell subset is involved in neonatal tolerance to mercuric salt-induced Th2 autoimmune disease. Thus, we demonstrate that in vivo CD8 depletion breaks tolerance following mercury recall in animals under a neonatal tolerance protocol. Furthermore, adoptive cotransfer of splenocytes from naive and tolerant rats as well as transfer of CD8(+) T cells from tolerant animals prevent naive syngeneic rats from developing pathologic Th2 immune responses. These observations indicate that CD8(+) T cells are endowed with regulatory functions in neonatal tolerance and mediate active suppression. Moreover, neonatal tolerance induced the expansion of CD8(+)CD45RC(high) T cells and the emergence of a high percentage of IFN-gamma-synthesizing CD8(+) T cells, which probably reflects the implication of regulatory Tc1 cells. Thus, in vivo induction of neonatal tolerance suppresses Th2 autoimmune responses via generation of a CD8(+) cell-mediated regulatory response.

Validation of the Sicca Symptoms Inventory for clinical studies of Sjögren's syndrome.

OBJECTIVE: Oral, ocular, and other dryness are the hallmark features of Sjögren's syndrome (SS). We constructed a new measure of sicca symptoms, the Sicca Symptoms Inventory, for the evaluation of patients with primary SS. METHODS: Female Caucasian groups of patients with primary SS, systemic lupus erythematosus, and rheumatoid arthritis and healthy controls were assessed for tear and saliva production and also completed a symptoms-profiling inventory construct-validated from primary SS patients' own vocabulary, augmented with sicca items from publications and participating clinicians. Multi-item facets of sicca and other discomfort were validated by factor analysis. RESULTS: Primary SS and other "sicca" conditions were highly discriminated from other rheumatic disorders and healthy controls on each dryness-related facet of oral and ocular discomfort. Selected symptom scores were as sensitive and specific to primary SS as the scores for saliva and tears, respectively, although the severity scores of symptoms and signs were only moderately correlated. CONCLUSION: These multiple-question scales distinguish patients with primary SS from controls more precisely than previously used measures. Future studies will test if change in these symptom scores can serve as an outcome measure for clinical trials in SS.