Acceptance and use of condoms among school-aged young people in Australia.

BACKGROUND: Despite availability of vaccines or medical prophylaxis for some sexually transmissible infections (STIs), promoting condom use remains an important public health strategy for the prevention of STIs. Recent research shows that regular condom use among young people in Australia has declined over the past decade, while the rate of common STIs has increased. METHOD: In this paper, we report findings from a large survey of school-aged young people in Australia (14-18years old) in which we looked at the association between condom use and positive feelings about sex, beliefs about social acceptability of condoms and confidence talking with partners about sex and condoms. RESULTS: Communication and relational factors supported more consistent condom use. Participants were more likely to regularly use condoms if they discussed condom use with a sexual partner, perceived condom use to be easy (a measure that included perceived ease of discussing condoms with a partner) and perceived condom use to offer social or relational benefits, including perceiving condom use as a demonstration of care for a partner. Young men were more likely to report positive feelings about sex and regular condom use than young women. Young women were less likely than young men or trans and non-binary young people to report regular condom use. CONCLUSIONS: The study shows the importance of supporting young people to build confidence expressing sexual needs and wants with partners. Public health approaches to STI prevention need to consider condom promotion in the context of young people's contemporary sexual, gendered and relationships cultures.

Profiling cognitive workload in an unmanned vehicle control task with cognitive models and physiological metrics.

In the present study, we use Cognitive Metrics Profiling (CMP) to capture variance in cognitive load within a complex unmanned vehicle control task. We aim to demonstrate convergent validity with existing workload measurement methods, and to decompose workload into constituent cognitive resources to aid in diagnosing causes of workload. A cognitive model of the task was developed and examined to determine the extent to which it could predict behavioral performance, subjective workload, and validated physiological workload metrics. We also examined model activity to draw insights regarding loaded cognitive capacities. We found that composite workload from the model predicted physiological metrics, performance, and subjective workload. Moreover, the model indicates that differences in workload were driven largely by procedural, declarative, and temporal memory demands. We have found preliminary evidence of correspondence between workload predictions of a CMP model and physiological measures of workload. This suggests our approach captures interesting aspects of workload in a complex task environment and may provide a theoretical link between behavioral, physiological, and subjective metrics. This approach may provide a means to design effective workload mitigation interventions and improve decision-making about personnel tasking and automation.

"The Wild West:" Nurse Experiences of Responding to the 2017 Las Vegas Mass Shooting.

OBJECTIVE: The purpose of this study was to explore the experiences of nurses who responded to a public mass shooting in 2017. METHODS: This qualitative study was conducted with a sample of nurses who responded to a mass shooting, recruited purposively from a hospital in Las Vegas, Nevada. Intensive interviews were conducted with a total of 7 nurses, audio-recorded and transcribed for thematic analysis. RESULTS: Six themes were developed from interview data: (1) "The worst night of my life": Overrun and overwhelmed; (2) Unexpected altruism and benevolence of patients and staff; (3) "The Wild West": Giving victim care by improvising beyond rules; (4) Experiencing a range of reactions in the immediate aftermath and in the long term; (5) Shifts in nursing practice and evolving team dynamics; and (6) Defining realistic approaches to support staff mental health and mass casualty preparation. CONCLUSION: Nurses who were involved in responding to the public mass shooting described the event as life-altering. Given the critical role of nurses in responding to mass shootings, it is essential to consider how nurses can be supported in the aftermath of these events and how mass disaster preparation can include attention to the needs of nurses.

Homology-independent targeted insertion (HITI) enables guided CAR knock-in and efficient clinical scale CAR-T cell manufacturing.

BACKGROUND: Chimeric Antigen Receptor (CAR) T cells are now standard of care (SOC) for some patients with B cell and plasma cell malignancies and could disrupt the therapeutic landscape of solid tumors. However, access to CAR-T cells is not adequate to meet clinical needs, in part due to high cost and long lead times for manufacturing clinical grade virus. Non-viral site directed CAR integration can be accomplished using CRISPR/Cas9 and double-stranded DNA (dsDNA) or single-stranded DNA (ssDNA) via homology-directed repair (HDR), however yields with this approach have been limiting for clinical application (dsDNA) or access to large yields sufficient to meet the manufacturing demands outside early phase clinical trials is limited (ssDNA). METHODS: We applied homology-independent targeted insertion (HITI) or HDR using CRISPR/Cas9 and nanoplasmid DNA to insert an anti-GD2 CAR into the T cell receptor alpha constant (TRAC) locus and compared both targeted insertion strategies in our system. Next, we optimized post-HITI CRISPR EnrichMENT (CEMENT) to seamlessly integrate it into a 14-day process and compared our knock-in with viral transduced anti-GD2 CAR-T cells. Finally, we explored the off-target genomic toxicity of our genomic engineering approach. RESULTS: Here, we show that site directed CAR integration utilizing nanoplasmid DNA delivered via HITI provides high cell yields and highly functional cells. CEMENT enriched CAR T cells to approximately 80% purity, resulting in therapeutically relevant dose ranges of 5.5 × 108-3.6 × 109 CAR + T cells. CRISPR knock-in CAR-T cells were functionally comparable with viral transduced anti-GD2 CAR-T cells and did not show any evidence of off-target genomic toxicity. CONCLUSIONS: Our work provides a novel platform to perform guided CAR insertion into primary human T-cells using nanoplasmid DNA and holds the potential to increase access to CAR-T cell therapies.

Intratumoral pan-ErbB targeted CAR-T for head and neck squamous cell carcinoma: interim analysis of the T4 immunotherapy study.

BACKGROUND: Locally advanced/recurrent head and neck squamous cell carcinoma (HNSCC) is associated with significant morbidity and mortality. To target upregulated ErbB dimer expression in this cancer, we developed an autologous CD28-based chimeric antigen receptor T-cell (CAR-T) approach named T4 immunotherapy. Patient-derived T-cells are engineered by retroviral transduction to coexpress a panErbB-specific CAR called T1E28ζ and an IL-4-responsive chimeric cytokine receptor, 4αß, which allows IL-4-mediated enrichment of transduced cells during manufacture. These cells elicit preclinical antitumor activity against HNSCC and other carcinomas. In this trial, we used intratumoral delivery to mitigate significant clinical risk of on-target off-tumor toxicity owing to low-level ErbB expression in healthy tissues. METHODS: We undertook a phase 1 dose-escalation 3+3 trial of intratumoral T4 immunotherapy in HNSCC (NCT01818323). CAR T-cell batches were manufactured from 40 to 130 mL of whole blood using a 2-week semiclosed process. A single CAR T-cell treatment, formulated as a fresh product in 1-4 mL of medium, was injected into one or more target lesions. Dose of CAR T-cells was escalated in 5 cohorts from 1×107-1×109 T4+ T-cells, administered without prior lymphodepletion. RESULTS: Despite baseline lymphopenia in most enrolled subjects, the target cell dose was successfully manufactured in all cases, yielding up to 7.5 billion T-cells (67.5±11.8% transduced), without any batch failures. Treatment-related adverse events were all grade 2 or less, with no dose-limiting toxicities (Common Terminology Criteria for Adverse Events V.4.0). Frequent treatment-related adverse events were tumor swelling, pain, pyrexias, chills, and fatigue. There was no evidence of leakage of T4+ T-cells into the circulation following intratumoral delivery, and injection of radiolabeled cells demonstrated intratumoral persistence. Despite rapid progression at trial entry, stabilization of disease (Response Evaluation Criteria in Solid Tumors V.1.1) was observed in 9 of 15 subjects (60%) at 6 weeks post-CAR T-cell administration. Subsequent treatment with pembrolizumab and T-VEC oncolytic virus achieved a rapid complete clinical response in one subject, which was durable for over 3 years. Median overall survival was greater than for historical controls. Disease stabilization was associated with the administration of an immunophenotypically fitter, less exhausted, T4 CAR T-cell product. CONCLUSIONS: These data demonstrate the safe intratumoral administration of T4 immunotherapy in advanced HNSCC.

Direct correction of haemoglobin E ß-thalassaemia using base editors.

Haemoglobin E (HbE) ß-thalassaemia causes approximately 50% of all severe thalassaemia worldwide; equating to around 30,000 births per year. HbE ß-thalassaemia is due to a point mutation in codon 26 of the human HBB gene on one allele (GAG; glutamatic acid → AAG; lysine, E26K), and any mutation causing severe ß-thalassaemia on the other. When inherited together in compound heterozygosity these mutations can cause a severe thalassaemic phenotype. However, if only one allele is mutated individuals are carriers for the respective mutation and have an asymptomatic phenotype (ß-thalassaemia trait). Here we describe a base editing strategy which corrects the HbE mutation either to wildtype (WT) or a normal variant haemoglobin (E26G) known as Hb Aubenas and thereby recreates the asymptomatic trait phenotype. We have achieved editing efficiencies in excess of 90% in primary human CD34 + cells. We demonstrate editing of long-term repopulating haematopoietic stem cells (LT-HSCs) using serial xenotransplantation in NSG mice. We have profiled the off-target effects using a combination of circularization for in vitro reporting of cleavage effects by sequencing (CIRCLE-seq) and deep targeted capture and have developed machine-learning based methods to predict functional effects of candidate off-target mutations.

"Everything's a fight": A qualitative study of the cancer survivorship experiences of transgender and gender diverse Australians.

BACKGROUND: There is widespread recognition of the need to achieve equitable outcomes for all cancer survivors. This requires understanding of the experiences and outcomes of vulnerable groups. People who identify as sexually or gender diverse are known to be at risk of inferior cancer and survivorship outcomes, however, the posttreatment survivorship experiences of transgender and gender diverse (TGD) people have not been well studied. This study explored the survivorship experiences of people who identify as TGD, focusing on the physical and psychological aspects of the posttreatment survivorship phase and their experiences of follow-up cancer care. METHODS: A qualitative study of 10 TGD cancer survivors. Interviews were transcribed verbatim and data were analyzed by thematic analysis. RESULTS: Six themes were generated from the data. TGD people reported (1) anxiety when attending appointments and avoidance of necessary follow-up care, (2) experiences of transphobia and discrimination within oncology care settings and (3) lack of consideration of TGD identity by providers. Themes further describe (4) physical aspects of being both TGD and a cancer survivor, (5) absence of inclusive and diverse supportive care resources, as well as (6) positive growth after cancer. CONCLUSION: Approaches to mitigate these issues are urgently called for. These include training in TGD health for health-care providers, inclusion of TGD health in medical and nursing curricula, processes to collect and use gender identity and preferred pronoun data in clinical settings, and development of TGD-inclusive information and peer-support resources.

Revictimisation of Women in Non-Urban Areas: A Scoping Review.

Literature widely acknowledges that women who experience Child Sex Abuse (CSA) have a higher risk of experiencing sexual revictimisation later in life, yet less is known about experiences of revictimisation in non-urban areas. The aim of this review is to examine what is known internationally regarding revictimisation of non-urban women, and to provide future research, practice and policy recommendations. A total of 2414 articles were identified through a comprehensive search across five broad health sciences and humanities databases; 11 articles met inclusion criteria and were included in this review. This review found a general lack of qualitative revictimisation studies, and limited research focusing on non-urban women. While existing studies included non-urban research samples, few articles (n = 3) explored how non-urban location contextualises revictimisation experiences. Most peer-reviewed articles identified within this paper (n = 7) examined intimate partner violence (IPV) revictimisation, highlighting a significant lack of research on sexual revictimisation within non-urban settings. Findings from the review indicate that experiences of violence in childhood and adulthood are frequent among non-urban women, and that experiencing child abuse is associated with a heightened likelihood of poor mental health and IPV in adulthood. The review also found that non-urban women faced significant structural disadvantage including low levels of employment and income, limited-service sector resources, unsafe family environments and exposure to community violence. Additional qualitative research is needed to better understand the experiences and needs of non-urban revictimised women, particularly within an Australian context.

Sexting and sexual behaviour in Australian adolescents.

BACKGROUND: Previous studies have tried to determine the relationship between sexting and risky behaviour to discover whether sexting fits into a deviance or normalcy discourse. This study investigated the relationship between sexting and sexual risk behaviours, contraception use and gender. METHODS: The design was a cross-sectional analysis of data from the sixth National Survey of Secondary Student and Adolescent Sexual Health, collected in 2018. There were 8263 Australian adolescents (aged 14-18years). Participants were fairly evenly split by gender, and 73% identified as heterosexual. Participants were asked a series of questions about their engagement in sexting, sexual behaviour and sexual health behaviours. RESULTS: A total of 52% of participants had sent a sext in the previous 2months, with most being text-based sexts. Sexters were 3.29times more likely to have engaged in anal or vaginal intercourse, and 2.88times more likely to have gotten pregnant than non-sexters. Sexters (M =2.76) had significantly more partners than non-sexters (M =2.35), t (3763)=-10.99, P X 2 (1)=0.38, P =0.535, or contraceptive use based on sexting status. CONCLUSIONS: Sexters are more likely to have engaged in sexual intercourse and have more partners than non-sexters. Sexting is not strongly associated with other risky behaviours. Evidence for differences between sexters and non-sexters in protecting against STIs and pregnancy was not found, as there were no significant differences in contraceptive use.

Multilobed flaps for intermediate and large nasal defects: Flap characteristics, patient outcomes, and provider experience at two institutions.

BACKGROUND: While bilobed and trilobed transposition flaps are established workhorses for nasal reconstruction, their utility is often limited to defects less than 1.5 cm, subjecting patients to more involved multistage or cosmetically less favorable repairs. We highlight the use of bilobed and trilobed transposition flaps for intermediate (≥ 1.5 cm) and large (≥ 2.0 cm) nasal defects. METHODS: Patients reconstructed with multilobed transposition flaps 2017-2020 were identified at two institutions. Validated scar scale (SCAR) and patient component of the patient and observer scar assessment survey (POSAS) were used to assess patient outcomes at a minimum 5-week follow-up. Statistical analyses were performed. RESULTS: Thirty-four patients were identified with mean defect size 1.8 cm diameter. There were no major postoperative events. Scar revision was performed in 9 patients. The mean provider SCAR score was 3.06 (best possible 0, worst possible 13). The mean patient arm of the SCAR scale and POSAS were 0.07 (best possible 0, worst possible 2) and 10.93 (best possible 6, worst possible 60), respectively. CONCLUSION: Bilobed and trilobed flaps have excellent outcomes for intermediate and large nasal defects.

Descriptive norms about condom use predict odds of using a condom during last sexual experience in a large, national survey of adolescents from Australia.

BACKGROUND: Reducing sexually transmitted infections among adolescents is an important public health goal in Australia and worldwide. This study estimated the association between condom use during last heterosexual sexual experience with two descriptive norms among a large, national sample of secondary school students from Australia. METHODS: A large, national online survey of 14- to 18-year-olds in Australia was conducted in 2018; a sub-analysis of sexually active participants (n =2989) used multivariable logistic regression to estimate the relationships between condom use during last sexual experience and condom use norms. The analysis controlled for the effects of age, sex, sexual orientation, religious affiliation, remoteness and knowledge of sexually transmitted diseases. RESULTS: Overall, 1673 (56.0% [95% confidence interval: 54.2%, 57.8%]) sexually active respondents reported using condoms during their last sexual experience. Perceiving that all same-age peers used condoms conferred higher odds of using condoms during their last heterosexual sexual experience (adjusted odds ratio: 3.06 [95% CI: 1.6, 6.0]). Perceptions about whether the suggestion to initiate condom use came from boys, girls, or both boys and girls was not associated with condom use. Differences in condom use related to socio-demographic characteristics are reported. CONCLUSIONS: As part of a holistic approach to sexuality education, health educators and service providers may emphasise that young people frequently choose to use condoms.

Methodology to streamline flow cytometric-based detection of early stage Plasmodium parasitemia in mice.

Murine infection models are needed to develop therapeutics and vaccines to combat the Plasmodium parasites causing malaria. Herein, we describe an easy to perform flow cytometry-based methodology for detecting green fluorescent protein-expressing Plasmodium berghei in the peripheral red blood cells (RBC) of mice. This methodology uses one-step staining and simplified gating strategies to streamline the process of Plasmodium quantification and can detect parasitemia at an earlier time point after infection compared to traditional light microscopy-based techniques.

Alleviating gender dysphoria: A qualitative study of perspectives of trans and gender diverse people.

OBJECTIVES: This study sought to explore perspectives of trans and gender diverse (TGD) people of ways to alleviate gender dysphoria in service provision and to develop a framework for application in health and other areas that can be used by researchers and service providers to design study protocols, assess organisations and enhance everyday practice in ways that are sensitive to TGD people's experiences. METHODS: Data from a national Australian survey on TGD people conducted in 2018-2019 (n = 340) were used to develop a framework for alleviating dysphoria. Participants were asked an open-ended question on ways that body discomfort could be minimised in clinical encounters. Inductive thematic analysis was used to develop themes true to participant sentiment, which formed the basis for the development of a framework. RESULTS: The sample was overall young, with 60.6% aged 18-24, and a strong representation of gender diverse people (42.6%). The most important theme for participants was the context of the experience, which included the subthemes of the interpersonal qualities of service providers, language and pronouns, and practical aspects. Aspects of systems were also important, with education and awareness being particularly emphasised, followed by inclusive environments. Access to gender affirming medical and surgical procedures was rarely mentioned (2.6%). A minority of participants indicated that there was nothing that could be done to alleviate their gender dysphoria (4.4%). CONCLUSIONS: The study proposes a framework that can help facilitate assessment of a service's current practices, inform a practitioner's daily practice and be used by researchers to appropriately design studies. The most important areas to address centre on the context of the immediate experience, which may be influenced through systems-level characteristics.

Survival and complications in patients with haemoglobin E thalassaemia in Sri Lanka: a prospective, longitudinal cohort study.

BACKGROUND: Worldwide, haemoglobin E ß-thalassaemia is the most common genotype of severe ß-thalassaemia. The paucity of long-term data for this form of thalassaemia makes evidence-based management challenging. We did a long-term observational study to define factors associated with survival and complications in patients with haemoglobin E thalassaemia. METHODS: In this prospective, longitudinal cohort study, we included all patients with haemoglobin E thalassaemia who attended the National Thalassaemia Centre in Kurunegala, Sri Lanka, between Jan 1, 1997, and Dec 31, 2001. Patients were assessed up to three times a year. Approaches to blood transfusions, splenectomy, and chelation therapy shifted during this period. Survival rates between groups were evaluated using Kaplan-Meier survival function estimate curves and Cox proportional hazards models were used to identify risk factors for mortality. FINDINGS: 109 patients (54 [50%] male; 55 [50%] female) were recruited and followed up for a median of 18 years (IQR 14-20). Median age at recruitment was 13 years (range 8-21). 32 (29%) patients died during follow-up. Median survival in all patients was 49 years (95% CI 45-not reached). Median survival was worse among male patients (hazard ratio [HR] 2·51, 95% CI 1·16-5·43), patients with a history of serious infections (adjusted HR 8·49, 2·90-24·84), and those with higher estimated body iron burdens as estimated by serum ferritin concentration (adjusted HR 1·03, 1·01-1·06 per 100 units). Splenectomy, while not associated with statistically significant increases in the risks of death or serious infections, ultimately did not eliminate a requirement for scheduled transfusions in 42 (58%) of 73 patients. Haemoglobin concentration less than or equal to 4·5 g/dL (vs concentration >4·5 g/dL), serum ferritin concentration more than 1300 µg/L (vs concentration ≤1300 µg/L), and liver iron concentration more than 5 mg/g dry weight of liver (vs concentration ≤5 mg/g) were associated with poorer survival. INTERPRETATION: Patients with haemoglobin E thalassaemia often had complications and shortened survival compared with that reported in high-resource countries for thalassaemia major and for thalassaemia intermedia not involving an allele for haemoglobin E. Approaches to management in this disorder remain uncertain and prospective studies should evaluate if altered transfusion regimens, with improved control of body iron, can improve survival. FUNDING: Wellcome Trust, Medical Research Council, US March of Dimes, Anthony Cerami and Ann Dunne Foundation for World Health, and Hemoglobal.

Improving Cervical Screening in Trans and Gender-Diverse People.

BACKGROUND: Trans and gender-diverse people with a cervix experience difficulties accessing cervical cancer screening because of structural, interpersonal, and individual barriers. OBJECTIVE: The aim of this study was to explore issues with cervical cancer screening participation, awareness, and healthcare provider recommendation for trans and gender-diverse people. METHODS: A national Australian survey was conducted in 2018 to 2019. Participants included 196 trans and gender-diverse people with a cervix. Data were analyzed using descriptive and multiple regression analyses. Two awareness items related to cervical cancer screening, healthcare provider recommendation, and cervical cancer screening participation were assessed. Four variables associated with cervical cancer screening were included in the regression: age, healthcare provider recommendation, like for body, and gender. RESULTS: The sample was young; half (52.6%) were aged 20 to 24 years. Almost half (44.6%) had never had a healthcare provider recommend cervical cancer screening to them. Around half (48.0%) had never participated, with 21.9% reporting that they are regular screeners. More than a quarter (27.5%) of people who had screening had an abnormal result. The most common reasons for not participating in screening were that it is emotionally traumatic for them (55.3%) and inability to find a healthcare provider with whom they are comfortable (38.3%). CONCLUSIONS: Trans and gender-diverse Australians with a cervix are unlikely to be regular participants in cervical cancer screening. To continue reducing cervical cancer rates, healthcare providers must address underscreening in this community. IMPLICATIONS FOR PRACTICE: Gender diversity training needs to be provided to healthcare providers. In addition, healthcare providers need to promote participation in cervical screening in this trans and gender-diverse community.

Social impacts of living with chronic hepatitis B in South Sudanese community in Australia.

Introduction: Chronic hepatitis B presents significant issues to public health and clinical practices. The infection requires lifelong clinical management and is a leading risk factor for liver cancer and liver cirrhosis. Limited studies currently exist on the social impacts of living with chronic hepatitis B, especially for people from Sub-Saharan Africa. The current study explored the experiences of South Sudanese people living with chronic hepatitis B in Australia.Methods: A qualitative inquiry using face-to-face interviews with fifteen South Sudanese people with chronic hepatitis B in Australia explored the social impacts of living with chronic hepatitis B. The study purposively sampled participants who self-identified as South Sudanese with hepatitis B and over 18 years of age. Interviews were coded and analysed using thematic analysis.Results: The study identified three themes relating to the experiences of living with chronic hepatitis B, and were grouped into psychological, interpersonal and healthcare system levels. Psychological level experiences related to the uncertainty of living with chronic hepatitis B and liver cancer risk, where participants expressed persistent fear of disease progression and anxiety around death related to the disease. Interpersonal level experiences involved the disclosure of hepatitis B and its social impacts including stigma, social distancing and isolation. Healthcare system level experiences included receiving mixed messages about hepatitis B, challenges with regular testing and difficulty receiving hepatitis B treatment.Conclusion: The findings provide valuable insights into the social impacts of hepatitis B. It underlines the importance of integrated public health interventions at the community level to improve knowledge and awareness of hepatitis B which address stigma in the South Sudanese community in Australia.

Perceptions and Interpretation of Contemporary Masculinities in Western Culture: A Systematic Review.

The social construct of masculinity evolves in response to changes in society and culture. Orthodox masculinity is mostly considered to be hegemonic and is evidenced by the dominance of men over women and other, less powerful men. Contemporary shifts in masculinity have seen an emergence of new masculinities that challenge traditional male stereotypes. This systematic review aims to review and synthesize the existing empirical research on contemporary masculinities and to conceptualize how they are understood and interpreted by men themselves. A literature search was undertaken on 10 databases using terms regularly used to identify various contemporary masculinities. Analysis of the 33 included studies identified four key elements that are evident in men's descriptions of contemporary masculinity. These four elements, (a) Inclusivity, (b) Emotional Intimacy, (c) Physicality, and (d) Resistance, are consistent with the literature describing contemporary masculinities, including Hybrid Masculinities and Inclusive Masculinity Theory. The synthesized findings indicate that young, middle-class, heterosexual men in Western cultures, while still demonstrating some traditional masculinity norms, appear to be adopting some aspects of contemporary masculinities. The theories of hybrid and inclusive masculinity suggest these types of masculinities have several benefits for both men and society in general.

Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells.

Adoptive cancer immunotherapy using chimeric antigen receptor (CAR) engineered T-cells holds great promise, although several obstacles hinder the efficient generation of cell products under good manufacturing practice (GMP). Patients are often immune compromised, rendering it challenging to produce sufficient numbers of gene-modified cells. Manufacturing protocols are labour intensive and frequently involve one or more open processing steps, leading to increased risk of contamination. We set out to develop a simplified process to generate autologous gamma retrovirus-transduced T-cells for clinical evaluation in patients with head and neck cancer. T-cells were engineered to co-express a panErbB-specific CAR (T1E28z) and a chimeric cytokine receptor (4αß) that permits their selective expansion in response to interleukin (IL)-4. Using peripheral blood as starting material, sterile culture procedures were conducted in gas-permeable bags under static conditions. Pre-aliquoted medium and cytokines, bespoke connector devices and sterile welding/sealing were used to maximise the use of closed manufacturing steps. Reproducible IL-4-dependent expansion and enrichment of CAR-engineered T-cells under GMP was achieved, both from patients and healthy donors. We also describe the development and approach taken to validate a panel of monitoring and critical release assays, which provide objective data on cell product quality.