Immunotherapy: FAMILIAL CD45RA DEPLETED T CELLS TO TREAT SEVERE REFRACTORY INFECTIONS IN IMMUNOCOMPROMISED PATIENTS

Background & Aim: Immunocompromised patients are susceptible to high-risk opportunistic infections and malignant diseases. If available, most antiviral and antifungal drugs are quite toxic, relatively ineffective, and induce resistance in the long term. Methods, Results & Conclusion(s): We have previously demonstrated the safety of adoptive cell therapy for COVID-19 patients with CD45RA negative cells containing SARS-CoV-2-specific T cells from a donor, chosen based on HLA compatibility and cellular response to SARS-CoV-2 peptide pools. After finishing a Phase 2 randomized multicenter clinical trial (RELEASE, NCT04578210), we concluded that the infusion is safe, effective, accelerates lymphocyte recovery and shows hallmarks of an immune response. To use adoptive cell therapy to treat COVID-19 it would be necessary to develop a biobank of living drugs. For that, we examined the immune evolution performing a longitudinal analysis from previously SARS-CoV-2 infected and infection- naive individuals covering 21 months from infection. Cellular responses were maintained over time while humoral responses increased after vaccination but were gradually lost. Therefore, the best donors would be recovered individuals and two months after vaccination. We also evaluated the effect of dexamethasone (current standard of care treatment for COVID-19 and other infections involving lymphopenia) and Interleukin-15 (cytokine involved in T-cell maintenance and survival) on CD45RA negative. Dexamethasone did not alter cell functionality, proliferation or phenotype at a clinical-practice concentration, while interleukin-15 increased the memory T-cell and T-regulatory cell activation state, and interferon gamma release. Furthermore, we applied the adoptive passive transfer of CD45RA negative cells containing pathogen-specific memory T-cells to other infectious diseases characterized by sustained lymphopenia. We infused six immunocompromised patients with Cytomegalovirus, BK virus, Aspergillus, and Epstein-Barr virus lymphoproliferative disease. Patients experienced pathogen clearance, resolution of symptoms and lymphocyte increase. Transient microchimerism was detected in three patients. The use of CD45RA negative cells containing specific memory T cells of a third-party donor for treating severe pathogenic diseases in immunocompromised patients is feasible, safe, and effective, and has an advantage over other cell therapies such as lower costs and a less complex regulatory environment.Copyright © 2023 International Society for Cell & Gene Therapy

Immunotherapy: TIMING OF DONOR SELECTION ON CD45RA-MEMORY T CELLS AS ADOPTIVE CELL THERAPY FOR COVID-19

Background & Aim: The COVID-19 pandemic has resulted in significant morbidity and mortality worldwide. The vaccines had dramatically decreased infection rates, number of deaths, and hospitalizations, but they are not 100% effective and immunity is lost gradually over time. We have previously shown how we are able to detect, isolate and produce at clinical scale SARS-CoV-2-specific T cells within CD45RA-memory T cells from COVID-19 convalescent donors. In a phase I clinical trial we have proved that treatment with these cells of hospitalized patients with moderate/severe COVID-19 is safe and feasible. Understanding the durability and the level of cellular immunity within the CD45RA- memory T cells and how changes with immunization are critical for the development of a biobank of living drugs to treat future COVID-19 patients. We performed a longitudinal exploratory analysis of the SARS-CoV-2 specific humoral and cellular immunity within the memory CD45RA- T cells in naive and previously infected individuals at different time points after two doses of BNT162b2 BioNTech/Pfizer vaccine Methods, Results & Conclusion: We studied the cellular and humoral response of SARS-CoV-2 specific memory T cells from recovered patients and controls at different time points: 2 weeks after recovering from COVID-19, 9 months after the infection/just before mRNA immunization, 10 and 65 days after full immunization. Detection of SARSCoV- 2- Specific Memory T Cells was performed by IFNg assay after exposure of cells to the M, N, and S SARS-CoV-2 peptides. Our data shows that memory T cell responses within the CD45RA- memory T cell subpopulation and most of the subsets tend to be higher in recovered individuals at all time points. The cellular response produced by control individuals to the S peptide is like the one from recovered patients at the same time point. Humoral responses were higher in recovered individuals after full immunization. Antibodies titer was not boosted after the late vaccine time point. An exploratory analysis of non-parametric Spearman’s rho correlation of humoral and cellular responses shows a positive correlation after infection with the 3 peptides and 65 days after immunization. In conclusion: We have analyzed the SARS-COV-2 specific T cells within the CD45RA- memory T cell subpopulation and the different subsets at different time points after (Figure Presented) (Figure Presented) infection and fully vaccinated. We claim that the best donors would be immunized individuals recovered from COVID-19 ideally in a time frame not higher than 6 months.

A Phase I/II Dose-Escalation Multi-Center Study to Evaluate the Safety of Infusion of Natural Killer Cells or Memory T Cells As Adoptive Therapy in Coronavirus Pneumonia and/or Lymphopenia: (RELEASE NCT04578210)

Background: Adoptive cell immunotherapies for opportunistic virus in immunocompromised patients using haploidentical memory T cells have shown to be safe and effective. Since severe cases of COVID-19 present a dysregulated immune system with T cell lymphopenia and a hyper-inflammatory state we have proposed that a similar strategy could be proven to be efficient for COVID-19 patients. This is a study protocol of an open-label, multicenter, double-arm, randomized, dose-finding phase I/II clinical trial to evaluate the feasibility, safety, tolerability, and efficacy of the administration of a single dose of allogenic SARS-CoV-2 specific memory CD45RA - T cells and Natural Killer (NK) cells in COVID-19 patients with lymphopenia and pneumonia. The aim of the study is to find efficient treatments for patients with moderate/severe COVID-19. Identification of Specific memory T cells and NK cells: i)Memory T Cells: we first determined the existence of SARS-CoV-2 specific T cells within the CD45RA - T memory cells of the blood of convalescent donors. Memory T cells can respond quickly to the infection and provide long-term immune protection to reduce the severity of the COVID-19 symptoms without inducing classically T cell alloreactivity. Also, CD45RA - memory T cells confer protection for other pathogens the donors encountered in their life. ii)NK cells: we determined the phenotype of NK cells after COVID-19 and the main characteristic of SARS-CoV-2 specific NK population in the blood of convalescent donors, as it has been shown for cytomegalovirus infections. Also, NK cells confer protection for other pathogens the donors encountered in their life. Pilot Phase I- Safety, feasibility, and dose escalation: Between September and November 2020 a phase 1, dose-escalation, single-center clinical trial was conducted to evaluate the safety and feasibility of the infusion of CD45RA - memory T cells containing SARS-CoV-2 specific T cells as adoptive cell therapy against moderate/severe cases of COVID-19. Nine participants with pneumonia and/or lymphopenia and with at least one human leukocyte antigen (HLA) match with the donor were infused. The first three subjects received the lowest dose (1x10 5 cells/kg), the next three received the intermediate dose (5x10 5 cells/kg) and the last three received the highest dose (1x10 6 cells/kg) of CD45RA - memory T cells. Clinicaltrials.gov registration: NCT04578210. Findings: All participants' clinical status measured by National Early Warning Score (NEWS) and 7-category point ordinal scales showed improvement six days after infusion. No serious adverse events were reported. Inflammatory parameters were stabilized post-infusion and the participants showed lymphocyte recovery two weeks after the procedure. Donor microchimerism was observed at least for three weeks after infusion in all patients. Interpretation: This study provides preliminary evidence supporting the idea that treatment of COVID-19 patients with moderate/severe symptoms using convalescent SARS-CoV-2 specific CD45RA - memory T cells is feasible and safe. We did not find dose-liming toxicity. The Recommended Phase 2 dose was 1x10 6 CD45RA - T cells. Phase II- Efficacy: Between January 2021 and July 2021 patients have been enrolled based on the matched with the HLA genotype of the convalescent donors and following the protocol inclusion/exclusion criteria. The primary outcome is the incidence of patient recovery at day 14, defined as normalization of fever and oxygen saturation or lymphopenia recovery. Secondary outcomes are the time to normal level of lymphocytes, the proportion of patients showing clinical improvement at day 7, time to first negative SARS-CoV-2 PCR, the incidence of treatment-related adverse events, duration of hospitalization, time to discharge, time to improvement by one category a 7-point ordinal scale or NEWS score, the proportion of patients requiring intensive care unit, and all-cause mortality. In addition, lymphocyte recovery by multiparametric flow cytometry and donor chimerism by real-time PCR in the e perimental arm was monitored weekly during the first month. This study provides preliminary evidence supporting the idea that treatment of COVID-19 patients with moderate/severe symptoms using convalescent CD45RA - memory T cells is safe and feasible. The phase II clinical trial is ongoing to demonstrate efficacy. [Formula presented] Disclosures: Soria: Celgene: Other: Fees;Gilead: Other: Fees;AbbVie: Other: Fees.

Phase I dose-escalation single centre clinical trial to evaluate the safety of infusion of memory T cells as adoptive therapy in COVID-19 (RELEASE).

BACKGROUND: Effective treatments are still needed to reduce the severity of symptoms, time of hospitalization, and mortality of COVID-19. SARS-CoV-2 specific memory T-lymphocytes obtained from convalescent donors recovered can be used as passive cell immunotherapy. METHODS: Between September and November 2020 a phase 1, dose-escalation, single centre clinical trial was conducted to evaluate the safety and feasibility of the infusion of CD45RA- memory T cells containing SARS-CoV-2 specific T cells as adoptive cell therapy against moderate/severe cases of COVID-19. Nine participants with pneumonia and/or lymphopenia and with at least one human leukocyte antigen (HLA) match with the donor were infused. The first three subjects received the lowest dose (1 × 105 cells/kg), the next three received the intermediate dose (5 × 105 cells/kg) and the last three received the highest dose (1 × 106 cells/kg) of CD45RA- memory T cells. Clinicaltrials.gov registration: NCT04578210. FINDINGS: All participants' clinical status measured by National Early Warning Score (NEWS) and 7-category point ordinal scales showed improvement six days after infusion. No serious adverse events were reported. Inflammatory parameters were stabilised post-infusion and the participants showed lymphocyte recovery two weeks after the procedure. Donor microchimerism was observed at least for three weeks after infusion in all patients. INTERPRETATION: This study provides preliminary evidence supporting the idea that treatment of COVID-19 patients with moderate/severe symptoms using convalescent CD45RA- memory T cells is feasible and safe. FUNDING: Clinical Trial supported by Spanish Clinical Research Network PT17/0017/0013. Co-funded by European Regional Development Fund/European Social Fund. CRIS CANCER Foundation Grant to AP-M and Agencia Valenciana de Innovación Grant AVI-GVA COVID-19-68 to BS.

A phase I/II dose-escalation multi-center study to evaluate the safety of infusion of natural killer cells or memory T cells as adoptive therapy in coronavirus pneumonia and/or lymphopenia: RELEASE study protocol.

BACKGROUND: Moderate/severe cases of COVID-19 present a dysregulated immune system with T cell lymphopenia and a hyper-inflammatory state. This is a study protocol of an open-label, multi-center, double-arm, randomized, dose-finding phase I/II clinical trial to evaluate the safety, tolerability, alloreactivity, and efficacy of the administration of allogeneic memory T cells and natural killer (NK) cells in COVID-19 patients with lymphopenia and/or pneumonia. The aim of the study is to determine the safety and the efficacy of the recommended phase 2 dose (RP2D) of this treatment for patients with moderate/severe COVID-19. METHODS: In the phase I trial, 18 patients with COVID-19-related pneumonia and/or lymphopenia with no oxygen requirement or with an oxygen need of ≤ 2.5 liters per minute (lpm) in nasal cannula will be assigned to two arms, based on the biology of the donor and the patient. Treatment of arm A consists of the administration of escalating doses of memory T cells, plus standard of care (SoC). Treatment of arm B consists of the administration of escalating doses of NK cells, plus SoC. In the phase II trial, a total of 182 patients with COVID-19-related pneumonia and/or lymphopenia requiring or not oxygen supplementation but without mechanical ventilation will be allocated to arm A or B, considering HLA typing. Within each arm, they will be randomized in a 1:1 ratio. In arm A, patients will receive SoC or RP2D for memory T cells plus the SoC. In arm B, patients will receive SoC or RP2D for NK cells plus the SoC. DISCUSSION: We hypothesized that SARS-CoV-2-specific memory T-lymphocytes obtained from convalescent donors recovered from COVID-19 can be used as a passive cell immunotherapy to treat pneumonia and lymphopenia in moderate/severe patients. The lymphopenia induced by COVID-19 constitutes a therapeutic window that may facilitate donor engraftment and viral protection until recovery. TRIAL REGISTRATION: ClinicalTrials.gov NCT04578210 . First Posted : October 8, 2020.

SARS-CoV-2-Specific Memory T Lymphocytes From COVID-19 Convalescent Donors: Identification, Biobanking, and Large-Scale Production for Adoptive Cell Therapy.

Syndrome coronavirus 2 (SARS-CoV-2) pandemic is causing a second outbreak significantly delaying the hope for the virus' complete eradication. In the absence of effective vaccines, we need effective treatments with low adverse effects that can treat hospitalized patients with COVID-19 disease. In this study, we determined the existence of SARS-CoV-2-specific T cells within CD45RA- memory T cells in the blood of convalescent donors. Memory T cells can respond quickly to infection and provide long-term immune protection to reduce the severity of COVID-19 symptoms. Also, CD45RA- memory T cells confer protection from other pathogens encountered by the donors throughout their life. It is of vital importance to resolve other secondary infections that usually develop in patients hospitalized with COVID-19. We found SARS-CoV-2-specific memory T cells in all of the CD45RA- subsets (CD3+, CD4+, and CD8+) and in the central memory and effector memory subpopulations. The procedure for obtaining these cells is feasible, easy to implement for small-scale manufacture, quick and cost-effective, involves minimal manipulation, and has no GMP requirements. This biobank of specific SARS-CoV-2 memory T cells would be immediately available "off-the-shelf" to treat moderate/severe cases of COVID-19, thereby increasing the therapeutic options available for these patients.

Use of standard formulations in pediatric home parenteral nutrition – the experience of a portuguese center during the COVID-19 pandemic

Rationale: Standard solutions for parenteral nutrition (PN) are safe and can meet the nutritional requirements of a significant number of pediatric patients. However, they may not always meet the individual requirements of those on long term PN, particularly those with significant gastrointestinal fluid losses. We evaluated the possibility of replacing individually tailored prescription for standard formulations in children on home PN. Methods: Retrospective analysis of the individual prescriptions of metabolically stable pediatric patients on home PN over a one-year period (march 2019 to march 2020). These were compared with 13 available standard solutions with electrolytes, and replacement was considered adequate if three successive criteria were met: non protein calorie to volume ratio (maximum variation 15%);non protein calorie to nitrogen ratio (NPC:N) (maximum variation either 20% for possible long term use or 35% for short term use);electrolyte concentration (maximum increase 20%). Results: A total 24 patients were included (67% male;median age 7.5 years). The most common diagnosis was short bowel syndrome (58%). Allowing a variation of 35% in NPC:N, replacement with a standard formulation was considered appropriate for 16 (67%) patients. For a variation of 20% in NPC:N, the number of patients decreased to 10 (42%). For those with stomas (8 patients), a standard formulation was only appropriate for two (25%). The maximum number of possible standard formulations per patient was two. These patients required between one and five standard bags per day. Conclusion: Standard PN solutions can be adequate for a significant proportion of pediatric patients on home PN. Their use in the short term may also be appropriate in settings of limited resources or restricted access to hospital facilities, such as those imposed by the Covid-19 pandemic. Disclosure of Interest: None declared

The impact of COVID-19 on children with autism spectrum disorder. / Impacto de la COVID-19 en niños con trastorno del espectro autista.

INTRODUCTION: Children with autism spectrum disorder (ASD) often experience changing routines as a major challenge. For that reason, the need for adaptation during COVID-19 pandemic may have brought major problems to families with children with this pathology. AIM: To explore how children with ASD and their parents experienced the social isolation during COVID-19 outbreak period. SUBJECTS AND METHODS: We conducted an observational, cross-sectional and analytical study. We applied an anonymous questionnaire that included children's demographic and clinical characteristics, along with the impact of the COVID-19 outbreak in different aspects of family's daily life. RESULTS: Out of 99 questionnaires obtained, 43 were related to children with ASD and 56 to control group. Children with ASD predominantly had changes in behavior, while children from control group mostly found no changes. The majority of parents of ASD children reported a negative impact in emotion management against those in control group reporting mostly positive or no impact. Caregivers reported higher mean scores of anxiety levels in themselves than in their children. ASD children and their parents had higher levels of anxiety than healthy ones. In the group with ASD, children that did not maintain routines had higher mean levels of anxiety than children that maintained routines. CONCLUSION: Our results show a potential important psychological impact of the COVID-19 pandemic not only in children with neurodevelopmental disorders but in their caregivers as well. Physicians must be prepared for the post-pandemic surveillance of mental disorders among families.

TITLE: Impacto de la COVID-19 en niños con trastorno del espectro autista.Introducción. Los niños con trastorno del espectro autista (TEA) a menudo experimentan el cambio de rutinas como un desafío importante. La necesidad de adaptación durante la pandemia por la COVID-19 puede haber causado problemas a estos niños. Objetivo. Conocer cómo vivieron los niños con TEA y sus familias el aislamiento social durante la cuarentena. Sujetos y métodos. Se realizó un estudio observacional, transversal y analítico. Se aplicó un cuestionario anónimo que incluía las características demográficas y clínicas de los niños, junto con el impacto que tuvo la pandemia en diferentes aspectos de la vida diaria de las familias. Resultados. De los 99 cuestionarios obtenidos, 43 eran niños con TEA y 56 niños del grupo control. Los niños con TEA tuvieron predominantemente cambios en el comportamiento, a diferencia de los del grupo control. La mayoría de los niños con TEA tuvo un impacto negativo en el manejo de las emociones frente a los del grupo control, que expresaron un impacto mayoritariamente positivo/nulo. Los cuidadores puntuaron niveles de ansiedad más altos en ellos mismos que en sus hijos. Los niños con TEA y sus padres tenían niveles más altos de ansiedad que los controles. En el grupo con TEA, los niños que no mantuvieron las rutinas tuvieron niveles de ansiedad más altos. Conclusión. Los resultados muestran un potencial impacto psicológico de la pandemia de la COVID-19 no sólo en los niños con trastornos del neurodesarrollo, sino también en sus cuidadores. Debe estarse preparado para la vigilancia de los trastornos mentales en las familias tras la pandemia de la COVID-19.