Clinical grade multiparametric cell sorting and gene-marking of regulatory T cells.

BACKGROUND AIMS: Regulatory T cells (Tregs) are the main mediators of peripheral tolerance. Treg-directed therapy has shown promising results in preclinical studies of diverse immunopathologies. At present, the clinical applicability of adoptive Treg transfer is limited by difficulties in generating Tregs at sufficient cell dose and purity. METHODS: We developed a Good Manufacturing Practice (GMP) compliant method based on closed-system multiparametric Fluorescence-Activated Cell Sorting (FACS) to purify Tregs, which are then expanded in vitro and gene-marked with a clinical grade retroviral vector to enable in vivo fate tracking. Following small-scale optimization, we conducted four clinical-scale processing runs. RESULTS: We showed that Tregs could be enriched to 87- 92% purity following FACS-sorting, and expanded and transduced to yield clinically relevant cell dose of 136-732×106 gene-marked cells, sufficient for a cell dose of at least 2 × 106 cells/kg. The expanded Tregs were highly demethylated in the FOXP3 Treg-specific demethylated region (TSDR), consistent with bona fide natural Tregs. They were suppressive in vitro, but a small percentage could secrete proinflammatory cytokines, including interferon-γ and interleukin-17A. CONCLUSIONS: This study demonstrated the feasibility of isolating, expanding and gene-marking Tregs in clinical scale, thus paving the way for future phase I trials that will advance knowledge about the in vivo fate of transferred Tregs and its relationship with concomitant Treg-directed pharmacotherapy and clinical response.

A randomized controlled trial on the effectiveness of the abridged illness management and recovery program for people with schizophrenia spectrum disorders.

Objective: This study aimed to investigate the effectiveness of an abridged version of the Illness Management and Recovery Programme (AIMR) that was modified and developed in Hong Kong through a mult-centre randomized controlled trial for patients with schizophrenia spectrum disorders. Methods: This study was implemented in 10 occupational therapy departments, psychiatric day hospitals of 7 Hospital Authority clusters in Hong Kong. A total of 211 patients with schizophrenia or schizoaffective disorder was recruited and randomized into either the experimental or the control condition. In the control group, the subjects went through conventional occupational therapy programmes. In the experimental group, the subjects went through an additional 10-session programme of AIMR. Participants: were measured at baseline, completion of the AIMR, and 3-month after the AIMR programme. Measures include the expanded version of the Brief Psychiatric Rating Scale (BPRS-E), the client version of the Illness Management and Recovery Scale (IMRS), the Snyder Hope Scale, the Social and Occupational and Occupational Functioning Assessment Scale (SOFAS), the WHO Quality of Life Scale (WHOQOL-BREF), and the Chinese Version of the Short Warwick-Edinburgh Mental Well-Being Scale (C-SWEMWBS). Results: Both the experimental and control cohorts had comparable clinical and socio-demographic characteristics except years of education and duration of illness. These two variables were entered as covariates in the linear mixed model which showed that the experimental group had significantly higher improvement than the control group in terms of illness management (F = 4.82; p = .03; Cohen's d = .45), functional (F = 10.65; p = .001; Cohen's d = .58), and hope (F = 5.52; p = .02; Cohen's d = .08) measures after the completion of treatment. Conclusion: The results supported the effectiveness of the AIMR programm which would be important in the recovery oriented practices in psychiatry.

Brief report: A randomized controlled trial of a compensatory cognitive training to improve prospective memory performance in people with schizophrenia or depression.

People with schizophrenia or depression have prospective memory (PM) deficits, which affect their daily living. Given the paucity of research into training to correct PM deficits, we subjected a group of participants to a Chinese version of the PM module of the Cognitive Compensatory Training (CCT-C-PM) intervention to study its effect on their PM performance. Specifically, we independently randomized two diagnostic cohorts (schizophrenia and depression) into control groups (occupational therapy only) or experimental groups (CCT-C-PM and occupational therapy). The schizophrenia cohort had 17 participants in its control group and 23 participants in its experimental group. The depression cohort had 10 participants in its control group and 12 participants in its experimental group. The sociodemographic information of the participants was collected. Their symptoms and PM performances were measured at baseline and after treatment (after the completion of the CCT-C-PM intervention in the experimental group and the same timeframe in the control group). The treatment effects were examined by a repeated measure analysis of variance/analysis of covariance and a post hoc Scheffé test. The effect sizes (Cohen's d) of treatments against the controls were also calculated. There was no difference between the experimental and control groups in either cohort in terms of sociodemographic data, symptoms, and PM measures at baseline. The sex combination differed across the groups in the depression cohort. We found that the CCT-C-PM improved PM, especially event-based PM, for which large effect sizes were seen. The effect on time-based PM was unclear and requires future study. Our findings suggest that the CCT-C-PM is a viable training method for improving PM.

Nanopore sequencing as a scalable, cost-effective platform for analyzing polyclonal vector integration sites following clinical T cell therapy.

BACKGROUND: Analysis of vector integration sites in gene-modified cells can provide critical information on clonality and potential biological impact on nearby genes. Current short-read next-generation sequencing methods require specialized instruments and large batch runs. METHODS: We used nanopore sequencing to analyze the vector integration sites of T cells transduced by the gammaretroviral vector, SFG.iCasp9.2A.ΔCD19. DNA from oligoclonal cell lines and polyclonal clinical samples were restriction enzyme digested with two 6-cutters, NcoI and BspHI; and the flanking genomic DNA amplified by inverse PCR or cassette ligation PCR. Following nested PCR and barcoding, the amplicons were sequenced on the Oxford Nanopore platform. Reads were filtered for quality, trimmed, and aligned. Custom tool was developed to cluster reads and merge overlapping clusters. RESULTS: Both inverse PCR and cassette ligation PCR could successfully amplify flanking genomic DNA, with cassette ligation PCR showing less bias. The 4.8 million raw reads were grouped into 12,186 clusters and 6410 clones. The 3'long terminal repeat (LTR)-genome junction could be resolved within a 5-nucleotide span for a majority of clusters and within one nucleotide span for clusters with ≥5 reads. The chromosomal distributions of the insertional sites and their predilection for regions proximate to transcription start sites were consistent with previous reports for gammaretroviral vector integrants as analyzed by short-read next-generation sequencing. CONCLUSION: Our study shows that it is feasible to use nanopore sequencing to map polyclonal vector integration sites. The assay is scalable and requires minimum capital, which together enable cost-effective and timely analysis. Further refinement is required to reduce amplification bias and improve single nucleotide resolution.

Phase I Trial of Inducible Caspase 9 T Cells in Adult Stem Cell Transplant Demonstrates Massive Clonotypic Proliferative Potential and Long-term Persistence of Transgenic T Cells.

PURPOSE: Inducible caspase 9 (iCasp9) is a cellular safety switch that can make T-cell therapy safer. The purpose of this phase I trial was to investigate the use of iCasp9-transduced T-cell addback in adult patients undergoing haploidentical stem cell transplantation for high-risk hematologic malignancies. PATIENTS AND METHODS: Patients undergoing myeloablative, CD34-selected haploidentical stem cell transplantation were treated with 0.5-1.0 × 106/kg donor-derived iCasp9-transduced T cells on day +25 or 26 post-transplant, with additional doses allowed for disease relapse, infection, or mixed chimerism. RESULTS: Three patients were enrolled. iCasp9-transduced T cells were readily detectable by 4 weeks post-infusion in all patients and remained at high level (114 cells/µL, 11% of T cells) in 1 patient alive at 3.6 years. One patient developed donor-derived Epstein-Barr virus-associated post-transplant lymphoproliferative disease (EBV-PTLD), which was followed by a marked expansion of iCasp9 T cells and cytokine release syndrome (CRS). These iCasp9-transduced T cells infiltrated the affected lymph nodes and secreted IFNγ and IL-10. They peaked at 1,848 cells/µL and were found to be monoclonal by T-cell receptor (TCR) clonotype and oligoclonal by viral integrant analysis, representing a 6-log in vivo expansion of the dominant T-cell clone. These T cells were not autonomous and contracted with the resolution of EBV-PTLD, which did not recur. CONCLUSIONS: iCasp9-transduced T cells could persist long-term. They retained very high in vivo clonotypic proliferative capacity and function, and could cause CRS in response to de novo lymphoma development.

Prospective Memory Performance in Persons With Schizophrenia and Bipolar Disorder and Healthy Persons.

PURPOSE: There is a paucity of studies comparing prospective memory (PM) impairment between persons with schizophrenia and bipolar disorder. The aim of this study was to directly compare PM performances of these two groups and healthy controls. DESIGN AND METHODS: A total of 44 persons with schizophrenia and 76 with bipolar disorder, and 44 healthy controls formed the study sample. FINDINGS: Patients were found to be impaired in PM relative to controls and the two patient groups showed similar level of PM performance after controlling confounding sociodemographic and clinical variables. PRACTICE IMPLICATIONS: The findings add to the evidence concerning the neurocognitive similarity between cohorts of schizophrenia and bipolar disorder with respect to PM. Rehabilitative effort in PM remedies for both patient groups is warranted.

Prospective memory predicts the level of community living skills in schizophrenia.

Schizophrenia patients are known to have prospective memory (PM) deficits. There is no robust evidence showing that PM deficits have a major impact on community living skills in schizophrenia. The aim of this study was to examine the association between PM and community living skills in schizophrenia. Forty-four individuals with schizophrenia formed the study sample. Participants׳ psychopathology, prospective and retrospective memory, level of intelligence, and community living skills were measured with standardized instruments. In bivariate analyses, community living skills overall but not self-care correlated with PM total and subscales scores. In multivariate analyses, event-based PM was more predictive than time-based PM of the level of community living skills. In conclusion, PM has a significant impact on community living skills in schizophrenia and attention should be paid to this type of memory disturbance in rehabilitation of schizophrenia.

Quality of life of patients with euthymic bipolar disorder and its associations with demographic and clinical characteristics, psychopathology, and cognitive deficits.

PURPOSE: Little is known about quality of life (QOL) in Chinese patients with bipolar disorder (BD) in remission (euthymia). This study examined the QOL of such a cohort of BD patients and its demographic, clinical, and cognitive correlates. DESIGN AND METHODS: Forty-seven euthymic BD patients and 47 matched healthy controls formed the study sample. Socio-demographic characteristics, prospective memory, retrospective memory, intelligence quotient, and executive functioning were measured in all participants together with patients' psychopathology ratings. FINDINGS: Multivariate analyses revealed that compared to controls, euthymic BD patients had significantly lower satisfaction with physical QOL domain. Only subthreshold depressive symptoms independently contributed to reduced satisfaction with physical and environmental QOL domains, whereas no variable predicted its psychological and social domains. PRACTICE IMPLICATIONS: Contrary to findings from Western settings, demographic variables and cognitive deficits had no associations with any QOL domain in euthymic Chinese BD patients. Control of subthreshold depressive symptoms in euthymic BD patients might enhance their QOL.

Predictive value of prospective memory for remission in first-episode schizophrenia.

PURPOSE: The study examined the rate of remission in individuals experiencing a first episode of schizophrenia (FES) in China and explored predictors of remission in the acute phase of the illness. DESIGN AND METHODS: Fifty-five FES patients were randomly treated with risperidone, olanzapine, or aripiprazole at therapeutic doses for 8 weeks, and their clinical profiles and cognition were assessed using standardized assessment instruments at entry and the end of the study. FINDINGS: Of the 55 patients, 30 (54.5%) remitted by the end of the 8-week study. In univariate analyses, shorter duration of untreated psychosis, higher scores on both the time-based prospective memory (TBPM) and event-based prospective memory tasks and the Hopkins Verbal Learning Test-revised, and less severe negative symptoms were significantly associated with remission. In stepwise multiple logistic regression analyses, only higher scores on the TBPM significantly predicted remission. Individuals having higher scores reflecting better TBPM at baseline were more likely to achieve remission after 8 weeks of optimized antipsychotic treatment. PRACTICE IMPLICATIONS: TPBM may be useful in helping clinicians identify those FES patients most likely to achieve a favorable treatment response.

Assessment of prospective memory in schizophrenia using the Chinese version of the Cambridge Prospective Memory Test: a controlled study.

INTRODUCTION: Data on the psychometric assessment of prospective memory (PM) are limited. The Cambridge Prospective Memory Test (CAMPROMPT) and its Chinese version (CAMPROMPT-C) have been applied to a variety of clinical conditions except for chronic schizophrenia. This controlled study explored the clinical utility of the CAMPROMPT-C in patients with schizophrenia by comparing their PM performance with that of normal controls. METHODS: Forty-four schizophrenia patients and 44 normal controls formed the study sample. Sociodemographic characteristics, PM, retrospective memory, and intelligence were measured in all subjects. Patients' psychopathology was rated with a standardized instrument. RESULTS: Patients performed worse than normal controls on both the sum and subscale scores of the CAMPROMPT-C. Patients had comparable performances in PM subtypes. Bivariate analyses revealed that education level, intelligence, and retrospective memory were associated with PM functions. DISCUSSION: The study supports the clinical utility of the CAMPROMPT-C in chronic schizophrenia and corroborated the significant relationship between PM and education, intelligence, and retrospective memory.

Prospective memory impairment and its implications for community living skills in bipolar disorder.

OBJECTIVE: Prospective memory (PM) refers to the ability to undertake intended actions in the future. The functional significance of PM in bipolar disorder (BD) has not yet been investigated. This study examined PM impairment and its role in community living skills in clinically stable individuals with BD. METHODS: Seventy-six individuals with BD and 44 healthy individuals were assessed with the Chinese version of the Cambridge Prospective Memory Test. Socio-demographic characteristics, retrospective memory (RM; the ability to recall or recognize past information), and intelligence were also measured in all participants. The clinical condition and community living skills of patients with BD were rated independently with standardized instruments. RESULTS: Patients with BD showed PM impairment and their PM total scores independently predicted the level of community living skills in regression analyses. In follow-up analyses, the contribution was found to be accounted for by the time-based PM scores. CONCLUSIONS: Given the expanding body of research on the utility of PM in predicting functioning, these findings further support the role of PM in community living skills in individuals with BD. The results suggest that PM training might be an integral part of clinical rehabilitation devised for individuals with BD.

Prospective memory deficits in euthymic bipolar disorder patients: a preliminary study.

INTRODUCTION: Prospective memory refers to the ability to remember to do something in the future. To date, little is known about prospective memory deficits in bipolar disorder (BD) in remission (euthymia). This study examined the nature and correlates of prospective memory in these patients. METHODS: Forty-seven euthymic BD patients and 47 matched healthy controls formed the study sample. Socio-demographic and basic clinical characteristics, prospective memory (Cambridge Prospective Memory Test [CAMPROMPT]), retrospective memory (immediate Logical Memory subtests of the Wechsler Memory Scales-Revised [WMS-R]), IQ (Raven's Progressive Matrices) and executive functioning (Wisconsin Card Sorting Test) were measured in all participants; patients' symptoms were rated with the 17-item Hamilton Depression Rating Scale and the 11-item Young Mania Rating Scale. RESULTS: Patients performed significantly worse on time-based prospective memory compared to controls. Multivariate analyses revealed that patients' lower score on Raven's Progressive Matrices significantly contributed to poor performance on time-based prospective memory, whereas lower scores on WMS-R Logical Memory subtest contributed to poor performance on event-based prospective memory; in controls, lower education level and older age significantly contributed to poor performance on time-based and event-based prospective memory, respectively. DISCUSSION: Prospective memory deficits persist in remitted BD patients suggesting that prospective memory impairment constitutes a trait deficit in BD.

Characteristics and clinical correlates of prospective memory performance in first-episode schizophrenia.

OBJECTIVE: The aim of this study was to examine prospective memory (PM) and its socio-demographic, clinical, and neurocognitive correlates in first episode schizophrenia (FES). METHODS: Fifty-one FES patients and 42 healthy controls formed the study sample. Time- and event-based PM (TBPM and EBPM) performance were measured with the Chinese version of the Cambridge Prospective Memory Test (C-CAMPROMPT). A battery of neuropsychological tests was also administered. Patients' clinical symptoms were evaluated with the Positive and Negative Symptom Scale (PANSS). RESULTS: Patients performed significantly worse in both TBPM (8.7 ± 5.3 vs. 14.8 ± 3.5) and EBPM (11.3 ± 4.7 vs. 15.7 ± 2.7) than the controls. After controlling for age, gender, education level and neurocognitive test score, the difference in performance on the two types of PM tasks between patients and controls was no longer present. In multiple linear regression analyses, longer duration of untreated psychosis (DUP), lower scores of the Hopkins Verbal Learning Test-Revised (HVLT-R) and the categories completed of the Wisconsin Card Sorting Test (WCST-CC) and higher score of the Color Trails Test-2 (CTT-2) contributed to poorer TBPM performance, while lower score of HVLT-R, higher score of the perseverative errors of the Wisconsin Card Sorting Test (WCST-PE) and longer DUP contributed to worse performance on EBPM. CONCLUSIONS: Both subtypes of PM are impaired in first-episode schizophrenia suggesting that PM deficits are an integral part of the cognitive dysfunction in the disease process.

The effect of added fullness and ventilation holes in T-shirt design on thermal comfort.

This paper reports on an experimental investigation on the effect of added fullness and ventilation holes in T-shirt design on clothing comfort measured in terms of thermal insulation and moisture vapour resistance. Four T-shirts in four different sizes (S, M, L, XL) were cut under the traditional sizing method while another (F-1) was cut with specially added fullness to create a 'flared' drape. A thermal manikin 'Walter' was used to measure the thermal insulation and moisture vapour resistance of the T-shirts in a chamber with controlled temperature, relative humidity and air velocity. The tests included four conditions: manikin standing still in the no-wind and windy conditions and walking in the no-wind and windy condition. It was found that adding fullness in the T-shirt design (F-1) to create the 'flared' drape can significantly reduce the T-shirt's thermal insulation and moisture vapour resistance under walking or windy conditions. Heat and moisture transmission through the T-shirt can be further enhanced by creating small apertures on the front and back of the T-shirt with specially added fullness. STATEMENT OF RELEVANCE: The thermal comfort of the human body is one of the key issues in the study of ergonomics. When doing exercise, a human body will generate heat, which will eventually result in sweating. If heat and moisture are not released effectively from the body, heat stress may occur and the person's performance will be negatively affected. Therefore, contemporary athletic T-shirts are designed to improve the heat and moisture transfer from the wearer. Through special cutting, such athletic T-shirts can be designed to improve the ventilation of the wearer.

Prospective memory deficits in patients with bipolar disorder: a preliminary study.

Prospective memory (PM) is the ability to remember to do something in the future without explicit prompts. To date, little has been known about PM deficits in bipolar disorder (BD). This study examined the nature and correlates of PM in patients with BD. Forty clinically stable BD patients and 40 matched healthy controls formed the study sample. Socio-demographic characteristics, PM, psychosocial functioning, retrospective memory (RM), and IQ were measured in all participants, whereas clinical condition was measured in patients with standardized assessment instruments. Patients performed significantly more poorly on the time-based PM task than controls (10.6 ± 5.0 vs. 14.6 ± 3.0, p < .001). In correlation analyses, older age, lower education, more severe depressive and manic symptoms, poor psychosocial functioning, poor RM, and lower scores in IQ were significantly associated with poor performance in the time-based PM task, whereas poor RM and lower scores in IQ associated with poorer performance in the event-based PM task in patients. In multivariate analyses, severity of depression and older age significantly contributed to poor performance in the time-based PM task, whereas poor RM contributed to poor performance in the event-based PM task in patients. The time-based PM is impaired in BD patients. Depressive symptoms, age, and RM were determinants of certain aspects of impaired PM performance in BD patients.

Cross-cultural validation of the St. Louis Inventory of Community Living Skills for Chinese patients with schizophrenia in Hong Kong.

AIM: The study validated a culturally sensitive community living skills rating scale for Chinese patients by adapting the St. Louis Inventory of Community Living Skills (SLICLS). METHOD: The Chinese version (SLICLS-C) was produced by forward and backward translation. An expert panel evaluated its content validity. Its internal consistency, inter-rater reliability, construct and concurrent validity were tested on 80 DSM-IV schizophrenia inpatients in a long-term facility. For predictive validity, the above sample was extended to ensure at least 20 subjects discharged to each of three levels of community care were included in the study sample. RESULTS AND CONCLUSION: The SLICLS-C was psychometrically sound and could be used for predicting level of community care, program evaluation and measuring outcome.