CD38 as a novel therapeutic treatment target for Systemic Lupus Erythematosus

Der Systemische Lupus erythematodes (SLE) ist eine systemische Autoimmunerkrankung mit bisher nur zum Teil geklärter Ätiopathogenese. Ein wichtiger Aspekt der Krankheitsentstehung ist die Entwicklung von antinukleären Autoantikörpern, die von kurz- und langlebigen Plasmazellen sezerniert werden. Langlebige Plasmazellen tragen zur Chronizität der Erkrankung bei und stellen eine therapeutische Herausforderung dar, da sie durch konventionelle immunsuppressive oder B-Zell-gerichtete Therapien nur unzureichend unterdrückt werden können. CD38 ist ein Oberflächenprotein, das auf unterschiedlichen Gruppen von Immunzellen entweder konstitutiv oder nach Stimulation exprimiert wird und insbesondere von Plasmazellen stark exprimiert wird. In dieser Arbeit wurde zunächst die Expression von CD38 auf verschiedenen Immunzellen im Blut von 35 SLE Patienten und 20 gesunden Kontrollen (HC) analysiert. Dabei konnte eine signifikant erhöhte CD38 Expression auf verschiedenen Immunzellsubsets von SLE-Patienten nachgewiesen werden, insbesondere bei plasmazytoiden dendritischen Zellen (pDC), Gedächtnis- B- und T-Zellen und auch zirkulierenden Plasmazellen. Zusätzlich wurde der therapeutische Effekt des gegen CD38 gerichteten monoklonalen Antikörpers Daratumumab bei zwei SLE-Patientinnen mit refraktärem, lebensbedrohlichen SLE untersucht. Dabei erhielten die Patientinnen 4 wöchentliche Infusionen mit Daratumumab und wurden für ein Jahr klinisch und immunologisch nachbeobachtet. Die Daratumumab Behandlung resultierte bei beiden Patientinnen in einer klinisch relevanten Verbesserung der Krankheitsaktivität bei insgesamt guter Verträglichkeit. Insbesondere konnte ein günstiger Effekt auf Lupus-Nephritis und autoimmun-hämolytische Anämie beobachtet werden. Die therapeutischen Effekte waren assoziiert mit einem Abfall von Autoantikörpern und Impftitern, was auf eine relevante Depletion von langlebigen Plasmazellen hindeutet. Zusätzlich war eine Reduktion der Typ-I Interferon-Aktivität und eine modulierte Genexpression von Gedächtnis T-Zellen nachweisbar. Die Depletion von autoreaktiven Plasmazellen ist nicht nur beim SLE ein wichtiges Therapiekonzept – so berichten wir zusätzlich über die Behandlung eines Patienten mit einer therapierefraktären Anti-CASPR2-Antikörper vermittelten Enzephalitis mit Daratumumab. Auch bei diesem Patienten kam es unter der Therapie zu klinischer Besserung der neurologischen Symptomatik und Reduktion der Autoantikörper. Allerdings entwickelte dieser Patient in der Rehabilitation eine Sepsis und verstarb wenig später an der Infektion. Insgesamt weisen diese Daten auf ein hohes Potenzial einer gegen CD38 gerichteten Therapie bei SLE, was möglicherweise auch auf andere Autoantikörper-vermittelte Erkrankungen übertragbar ist. Allerdings muss die Effektivität und Sicherheit von Daratumumab in diesen Erkrankungen in klinischen, kontrollierten Studien untersucht werden.Systemic Lupus erythematosus (SLE) is a rheumatologic autoimmune disease, the pathogenesis and aetiology of which are only partially understood. One important aspect of the mechanisms of disease is the generation and maintenance of short- and long-lived, autoreactive plasma cells, which produce autoantibodies. Long-lived plasma cells with their continuous secretion of autoantibodies contribute to the chronicity of the disease and are not sufficiently targeted by conventional immunosuppressants or B cell targeted therapies. CD38 is a surface protein that is expressed either constitutionally or upon stimulation on a host of different immune cell subsets and that is particularly highly expressed on plasma cells. In this work, we examined the expression of CD38 on different immune cell subsets in the peripheral blood of 35 SLE patients and 20 healthy controls. We were able to show that the expression of CD38 is increased on various subsets of peripheral blood leukocytes of SLE patients, such as plasmacytoid dendritic cells, memory B and T cells, as well as plasma blasts was increased. Additionally, we investigated the therapeutic effects of the anti-CD38 monoclonal antibody Daratumumab in two SLE patients with refractory, life-threatening disease with the anti-CD38 monoclonal antibody Daratumumab. The patients received four weekly infusions of Daratumumab and were followed up clinically and immunologically for one year. The treatment of two SLE patients with Daratumumab was, apart from the expected hypogammaglobulinemia, safe and induced a clinical and immunological improvement of the disease that was sustained after one year of follow-up. Especially, amelioration of lupus nephritis and autoimmune haemolytic anaemia were observed. The therapeutic effects were associated with a decline in the titres of both autoantibodies and vaccine-induced protective antibodies declined, indicating a depletion of (long-lived) antibody-secreting cells. Additionally type 1 interferon activity and single-cell transcription analysis of memory T cells showed a downregulation of T cell transcripts associated with chronic T cell activation. The depletion of autoreactive plasma cells could also be an important framework work the treatment of other autoimmune diseases – here, we additionally report the treatment of a patient with refractory anti-CASPR2 antibody mediated encephalitis with daratumumab. Similar to the SLE patients, the treatment resulted in clinical improvement of the neurological manifestations as well as a reduction in autoantibody titres. However, this patient developed septicemia after discharge into a neurological rehabilitation centre and subsequently died. Our results indicate that a CD38-target therapy is a modality that has large potential for the treatment of SLE and likely also many other autoantibody-mediated diseases. The safety and efficacy of Daratumumab for these diseases, however, will need to be investigated in clinical-controlled Trials

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer‐reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state‐of‐the‐art handbook for basic and clinical researchers.DFG, 389687267, Kompartimentalisierung, Aufrechterhaltung und Reaktivierung humaner Gedächtnis-T-Lymphozyten aus Knochenmark und peripherem BlutDFG, 80750187, SFB 841: Leberentzündungen: Infektion, Immunregulation und KonsequenzenEC/H2020/800924/EU/International Cancer Research Fellowships - 2/iCARE-2DFG, 252623821, Die Rolle von follikulären T-Helferzellen in T-Helferzell-Differenzierung, Funktion und PlastizitätDFG, 390873048, EXC 2151: ImmunoSensation2 - the immune sensory syste

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

Cossarizza A, Chang H‐D, Radbruch A, et al. Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition). European Journal of Immunology. 2021;51(12):2708-3145.The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers.ISSN:0014-2980ISSN:1521-414

Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition)

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples and respective applications of flow cytometry in the context of a variety of autoimmune diseases, cancers as well as acute and chronic infectious diseases such as COVID-19. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer-reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state-of-the-art handbook for basic and clinical researchers