Search

Chronic granulomatous disease is an inborn error of immunity due to disrupted function of the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase complex. This results in impaired respiratory burst of phagocytes and insufficient killing of bacteria and fungi. Patients with chronic granulomatous disease are at increased risk for infections, autoinflammation and autoimmunity. Allogeneic hematopoietic stem cell transplantation (HSCT) is the only widely available curative therapy. While HSCT from human leukocyte antigen (HLA) matched siblings or unrelated donors are standard of care, transplantation from HLA-haploidentical donors or gene therapy are considered alternative options. We describe a 14-month-old male with X-linked chronic granulomatous disease who underwent a paternal HLA-haploidentical HSCT using T-cell receptor (TCR) alpha/beta+/CD19+ depleted peripheral blood stem cells followed by mycophenolate graft versus host disease prophylaxis. Decreasing donor fraction of CD3+ T cells was overcome by repeated infusions of donor lymphocytes from the paternal HLA-haploidentical donor. The patient achieved normalized respiratory burst and full donor chimerism. He remained disease-free off any antibiotic prophylaxis for more than three years after HLA-haploidentical HSCT. In patients with x-linked chronic granulomatous disease without a matched donor paternal HLA-haploidentical HSCT is a treatment option worth to consider. Administration of donor lymphocytes can prevent imminent graft failure.

International audience; Patients with inherited CARMIL2 or CD28 deficiency have defective T cell CD28 signaling, but their immunological and clinical phenotypes remain largely unknown. We show that only one of three CARMIL2 isoforms is produced and functional across leukocyte subsets. Tested mutant CARMIL2 alleles from 89 patients and 52 families impair canonical NF-κB but not AP-1 and NFAT activation in T cells stimulated via CD28. Like CD28-deficient patients, CARMIL2-deficient patients display recalcitrant warts and low blood counts of CD4 + and CD8 + memory T cells and CD4 + T REG s. Unlike CD28-deficient patients, they have low counts of NK cells and memory B cells, and their antibody responses are weak. CARMIL2 deficiency is fully penetrant by the age of 10 yr and is characterized by numerous infections, EBV + smooth muscle tumors, and mucocutaneous inflammation, including inflammatory bowel disease. Patients with somatic reversions of a mutant allele in CD4 + T cells have milder phenotypes. Our study suggests that CARMIL2 governs immunological pathways beyond CD28.

Additional file 2. Questionnaire of the online survey among health-care staff and extract of the questionnaire for caregivers (English translation).

Additional file 1: Figure S1: ALK inhibitor treatment of ALK-mutated neuroblastoma cell lines for optimized screeningconditions (related to Figure 1). Figure S2: Quality control of CRISPR/Cas9 knockout screen (related to Figure 1). Figure S3: Lorlatinib- and Ceritinib- resistant NBLW-R neuroblastoma cells grow as aggressive tumors in the kidney capsule of nude mice (related to Figure 3). Figure S4: Tetracycline induced NRASQ61K expression (related to Figure 4). Figure S5: Computational modeling of ALK downstream signaling using STASNet (related to Figure 6). Figure S6: MEK inhibitor treatment of ectopic NRASQ61K expression models (related to Figure 7). Figure S7: High-throughput drug screening of LAN-5 and LAN-5 NF1 KO#2 clone (related to Figure 7). Table S1. Full clinical data on neuroblastoma patients.

Additional file 1. SARS-CoV-2 diagnostic test procedures.

Additional file 6. Supplementary Materials and Methods.

Additional file 3. Results of survey on the perception of infection control measures by hospital staff, stratified by profession.

Background Long-term outcomes of hematopoietic stem cell transplantation (HSCT) in children with juvenile metachromatic leukodystrophy (MLD) have been investigated systematically, while short-term effects of HSCT on the course of the disease remain to be elucidated. Results In this study, the clinical course was evaluated over the first 24 months following HSCT, conducted at our center in 12 children with juvenile MLD (mean follow-up 6.75 years, range 3–13.5) and compared with 35 non-transplanted children with juvenile MLD. Motor function (GMFM-88 and GMFC-MLD), cognitive function (FSIQ), peripheral neuropathy (tibial nerve conduction velocity), and cerebral changes (MLD-MR severity score) were tested prospectively. Seven children remained neurologically stable over a long period, five exhibited rapid disease progression over the first 12 to 18 months after transplantation. In the latter, time from first gross motor symptoms to loss of independent walking was significantly shorter compared with non-transplanted patients at the same stage of disease (p < 0.02). Positive prognostic factors were good motor function (GMFM = 100%, GMFC-MLD = 0) and a low MR severity score (≤ 17) at the time of HSCT. Conclusions Our results show that if disease progression occurs, this happens early on after HSCT and proceeds faster than in non-transplanted children with juvenile MLD, indicating that HSCT may trigger disease progression.

BZgA-Forschungsbericht

The resurgence of haploidentical stem cell transplantation (HaploSCT) over the last decade is one of the most important advances in the field of hematopoietic stem cell transplantation (HSCT). The modified platforms of T cell depletion either ex vivo (CD34+ cell selection, "megadoses" of purified CD34+ cells, or selective depletion of T cells) or newer platforms of in vivo depletion of T cells, with either post-transplantation high-dose cyclophosphamide or intensified immune suppression, have contributed to better outcomes, with survival similar to that in HLA-matched donor transplantation. Further efforts are underway to control viral reactivation using modified T cells, improve immunologic reconstitution, and decrease the relapse rate post-transplantation using donor-derived cellular therapy products, such as genetically modified donor lymphocytes and natural killer cells. Improvements in treatment-related mortality have allowed the extension of haploidentical donor transplants to patients with hemoglobinopathies, such as thalassemia and sickle cell disease, and the possible development of platforms for immunotherapy in solid tumors. Moreover, combining HSCT from a related donor with solid organ transplantation could allow early tapering of immunosuppression in recipients of solid organ transplants and hopefully prevent organ rejection in this setting. This symposium summarizes some of the most important recent advances in HaploSCT and provides a glimpse in the future of fast growing field.

Based on an unbalanced panel of all Bavarian cooperative banks for the years of 1989-97 which includes information on 283 mergers, we analyze motives and cost effects of small-scale mergers in German banking. Estimating a frontier cost function with a time-variable stochastic efficiency term we show that positive scale and scope effects from a merger arise only if the merged unit closes part of the former branch network. When we compare actual mergers to a simulation of hypothetical mergers, size effects of observed mergers turn out to be slightly more favorable than for all possible mergers. Banks taken over by others are less efficient than the average bank in the same size class, but exhibit on average the same efficiency as the acquiring firms. For the post-merger phase, our empirical results provide no evidence for efficiency gains from merging, but point instead to a leveling off of differences among the merging units.

Purpose: Age dependent radiation sensitivity for DNA damage after in vitro blood exposure by computer tomography (CT) was investigated. Materials and methods: Radiation biomarkers (dicentrics and gammaH2AX) in blood samples of newborns, children under five years and adults after sham exposure (0 mGy), low-dose (41 mGy) and high-dose (978 mGy) in vitro CT exposure were analyzed. Results: Significantly higher levels of dicentric induction were found for the single and combined newborns/children group compared to adults, by a factor of 1.48 (95% CI 1.30–1.68), after exposure to 978 mGy. Although a significant dose response for damage induction and dose-dependent repair was found, the gammaH2AX assay did not show an age-dependent increase in DNA damage in newborns/children compared to adults. This was the case for the gammaH2AX levels after repair time intervals of 30 minutes and 24 hours, after correcting for the underlying background damage. For the low dose of 41 mGy, the power of the dicentric assay was also not sufficient to detect an age-dependent effect in the sample size investigated. Conclusion: A 1.5-fold increased level of dicentric aberrations is detected in newborns and children under five years after 1 Gy radiation exposure.

Purpose: Age dependent radiation sensitivity for DNA damage after in vitro blood exposure by computer tomography (CT) was investigated. Materials and methods: Radiation biomarkers (dicentrics and gammaH2AX) in blood samples of newborns, children under 5 years and adults after sham exposure (0 mGy), low dose (41 mGy) and high dose (978 mGy) in vitro CT exposure were analysed. Results: Significantly higher levels of dicentric induction were found for the single and combined newborns/children group compared to adults, by a factor of 1.48 (95% CI 1.30-1.68), after exposure to 978 mGy. Although a significant dose response for damage induction and dose dependent repair was found, the gammaH2AX assay did not show an age dependent increase in DNA damage in newborns/children compared to adults. This was the case for the gammaH2AX levels after repair time intervals of 30 minutes and 24 hours, after correcting for the underlying background damage. For the low dose of 41 mGy, the power of the dicentric assay was also not sufficient to detect an age dependent effect in the sample size investigated. Conclusions: A 1.5-fold increased level of dicentric aberrations is detected in newborns and children under five years after 1 Gy radiation exposure.

Purpose: Age dependent radiation sensitivity for DNA damage after in vitro blood exposure by computer tomography (CT) was investigated. Materials and methods: Radiation biomarkers (dicentrics and gammaH2AX) in blood samples of newborns, children under five years and adults after sham exposure (0 mGy), low-dose (41 mGy) and high-dose (978 mGy) in vitro CT exposure were analyzed. Results: Significantly higher levels of dicentric induction were found for the single and combined newborns/children group compared to adults, by a factor of 1.48 (95% CI 1.30–1.68), after exposure to 978 mGy. Although a significant dose response for damage induction and dose-dependent repair was found, the gammaH2AX assay did not show an age-dependent increase in DNA damage in newborns/children compared to adults. This was the case for the gammaH2AX levels after repair time intervals of 30 minutes and 24 hours, after correcting for the underlying background damage. For the low dose of 41 mGy, the power of the dicentric assay was also not sufficient to detect an age-dependent effect in the sample size investigated. Conclusion: A 1.5-fold increased level of dicentric aberrations is detected in newborns and children under five years after 1 Gy radiation exposure.

In dieser Arbeit wurde die Flexibilität, Steuerbarkeit, Effektivität und Selektivität der AdCAR-T-Zell-Therapie exemplarisch bei fluiden (BCP-ALL, B-NHL) und soliden Tumoren (Neuroblastom und Mammakarzinom) in vitro gezeigt. Als zentrales Element wurde die Surface Activation Matrix als neues immuntherapeutisches Prinzip bei kombinatorischem Targeting untersucht. AdCAR-T-Zellen sind in der Lage, Antigen und LLE-mAb-spezifisch Target-Zellen zu lysieren. Die Aktivität der AdCAR-T-Zellen ist durch die Zugabe der LLE-mAb an- und ausschaltbar. Entsprechend des SAM-Modells ist zur Induktion der Zielantigen-abhängigen Lyse das Überschreiten einer Schwellenwertkonzentration von LLE-mAb nötig. Unterhalb dieser Konzentration wurde beim einzelnen Targeting keine spezifische Lyse erzielt. Durch multiples, synchrones Targeting mit jeweils subtherapeutischen LLE-mAb-Konzentrationen konnte bei allen Tumorzelllinien eine hohe additive Tumorlyse erreicht werden. Mit JeKo-1 Wildtyp- und Knockout-Varianten wurde die Situation eines Antigenverlustes von Tumorzellen simuliert. AdCAR-T-Zellen lysierten spezifisch und zeitlich flexibel Zielantigen-positive Target-Zellen. Die AM können jederzeit flexibel angepasst werden. Durch zeitlich versetztes, sequenzielles Targeting kann Therapieversagen durch Immunevasion des Tumors vermieden werden. Das Prinzip des differenziellen Targetings wurde mit Nalm-6 Tumorzellen und gesunden B-Zellen in Kokultur untersucht. Entsprechend des SAM-Modells führte nur die Expression der Gesamtheit der Zielantigene beim Targeting mit subtherapeutischen LLE-mAb-Konzentrationen zu einer Aktivierung der AdCAR-T-Zellen. Es ist daher irrelevant, dass andere gesunde Körperzellen und lebenswichtige Gewebe einzelne der Zielantigene exprimieren, wenn diese jeweils lediglich mit subtherapeutischen LLE-mAb-Konzentrationen adressiert werden. Die Anwendung der SAM sowie das sequenzielle und differenzielle kombinatorische Targeting von tumorspezifischen Antigenen ermöglichen eine AdCAR-T-Zellen- induzierte zielgerichtete Tumorlyse bei gleichzeitig verringerter oder zumindest zeitlich begrenzter on target, off tumor-Toxizität. Dadurch können potenziell lebensbedrohliche akute und chronische Nebenwirkungen der Tumortherapie kontrolliert und reduziert werden. Zusammenfassend stellt das AdCAR-T-Zell-System eine effiziente, sichere und universelle Therapiestrategie bei heterogenen soliden und fluiden Tumorzelllinien dar.

The resurgence of haploidentical stem cell transplantation (HaploSCT) over the last decade is one of the most important advances in the field of hematopoietic stem cell transplantation (HSCT). The modified platforms of T cell depletion either ex vivo (CD34+ cell selection, "megadoses" of purified CD34+ cells, or selective depletion of T cells) or newer platforms of in vivo depletion of T cells, with either post-transplantation high-dose cyclophosphamide or intensified immune suppression, have contributed to better outcomes, with survival similar to that in HLA-matched donor transplantation. Further efforts are underway to control viral reactivation using modified T cells, improve immunologic reconstitution, and decrease the relapse rate post-transplantation using donor-derived cellular therapy products, such as genetically modified donor lymphocytes and natural killer cells. Improvements in treatment-related mortality have allowed the extension of haploidentical donor transplants to patients with hemoglobinopathies, such as thalassemia and sickle cell disease, and the possible development of platforms for immunotherapy in solid tumors. Moreover, combining HSCT from a related donor with solid organ transplantation could allow early tapering of immunosuppression in recipients of solid organ transplants and hopefully prevent organ rejection in this setting. This symposium summarizes some of the most important recent advances in HaploSCT and provides a glimpse in the future of fast growing field.