Management of patients undergoing CAR-T cell therapy in Germany

Introduction: Chimeric antigen receptor positive T cell (CAR-T cell) treatment became standard therapy for relapsed or refractory hematologic malignancies, such as non-Hodgkin’s lymphoma and multiple myeloma. Owing to the rapidly progressing field of CAR-T cell therapy and the lack of generally accepted treatment guidelines, we hypothesized significant differences between centers in the prevention, diagnosis, and management of short- and long-term complications. Methods: To capture the current CAR-T cell management among German centers to determine the medical need and specific areas for future clinical research, the DAG-HSZT (Deutsche Arbeitsgemeinschaft für Hämatopoetische Stammzelltransplantation und Zelluläre Therapie; German Working Group for Hematopoietic Stem Cell Transplantation and Cellular Therapy) performed a survey among 26 German CAR-T cell centers. Results: We received answers from 17 centers (65%). The survey documents the relevance of evidence in the CAR-T cell field with a homogeneity of practice in areas with existing clinical evidence. In contrast, in areas with no – or low quality – clinical evidence, we identified significant variety in management in between the centers: management of cytokine release syndrome, immune effector cell-related neurotoxicity syndrome, IgG substitution, autologous stem cell backups, anti-infective prophylaxis, and vaccinations. Conclusion: The results indicate the urgent need for better harmonization of supportive care in CAR-T cell therapies including clinical research to improve clinical outcome

Ijuhya vitellina sp. nov., a novel source for chaetoglobosin A, is a destructive parasite of the cereal cyst nematode Heterodera filipjevi.

Cyst nematodes are globally important pathogens in agriculture. Their sedentary lifestyle and long-term association with the roots of host plants render cyst nematodes especially good targets for attack by parasitic fungi. In this context fungi were specifically isolated from nematode eggs of the cereal cyst nematode Heterodera filipjevi. Here, Ijuhya vitellina (Ascomycota, Hypocreales, Bionectriaceae), encountered in wheat fields in Turkey, is newly described on the basis of phylogenetic analyses, morphological characters and life-style related inferences. The species destructively parasitises eggs inside cysts of H. filipjevi. The parasitism was reproduced in in vitro studies. Infected eggs were found to harbour microsclerotia produced by I. vitellina that resemble long-term survival structures also known from other ascomycetes. Microsclerotia were also formed by this species in pure cultures obtained from both, solitarily isolated infected eggs obtained from fields and artificially infected eggs. Hyphae penetrating the eggshell colonised the interior of eggs and became transformed into multicellular, chlamydospore-like structures that developed into microsclerotia. When isolated on artificial media, microsclerotia germinated to produce multiple emerging hyphae. The specific nature of morphological structures produced by I. vitellina inside nematode eggs is interpreted as a unique mode of interaction allowing long-term survival of the fungus inside nematode cysts that are known to survive periods of drought or other harsh environmental conditions. Generic classification of the new species is based on molecular phylogenetic inferences using five different gene regions. I. vitellina is the only species of the genus known to parasitise nematodes and produce microsclerotia. Metabolomic analyses revealed that within the Ijuhya species studied here, only I. vitellina produces chaetoglobosin A and its derivate 19-O-acetylchaetoglobosin A. Nematicidal and nematode-inhibiting activities of these compounds have been demonstrated suggesting that the production of these compounds may represent an adaptation to nematode parasitism

Innate immunity mediated by TLR9 modulates pathogenicity in an animal model of multiple sclerosis

Inflammatory diseases of the CNS, such as MS and its animal model EAE, are characterized by infiltration of activated lymphocytes and phagocytes into the CNS. Within the CNS, activation of resident cells initiates an inflammatory cascade, leading to tissue destruction, demyelination, and neurologic deficit. TLRs recognize microbes and are pivotal mediators of innate immunity. Within the CNS, augmented TLR expression during EAE is observed, even in the absence of any apparent microbial involvement. To determine the functional relevance of this phenomenon during sterile autoimmunity, we studied the role of different TLRs as well as their common signaling adaptor MyD88 in the development of EAE. We found that MyD88(–/–) mice were completely EAE resistant. Surprisingly, this protection is partly due to engagement of the CpG receptor TLR9. Restricting the MyD88 or TLR9 mutation to host radio-resistant cells, including the cells within the CNS, revealed that engagement of radio-resistant cells modulated the disease course and histopathological changes. Our data clearly demonstrate that both TLR9 and MyD88 are essential modulators of the autoimmune process during the effector phase of disease and suggest that endogenous “danger signals” modulate the disease pathogenesis

Bayesian inference of infrageneric phylogenetic relationships within <i>Ijuhya</i> based on <i>act</i>, ITS, LSU, <i>rpb1</i>, and <i>ß-tub</i> sequences.

<p>Numbers above nodes are estimates of <i>a posteriori</i> probabilities (≥ 0.9) / NJB and MLB (≥ 70%). The topology was rooted with three distantly related ‘<i>Ijuhya</i>’ species (‘<i>Ijuhya</i>’ <i>antillana</i>, <i>I</i>. <i>dentifera</i>, and ‘<i>Ijuhya</i>’ <i>oenanthicola</i>).</p

Light micrographs of <i>Ijuhya vitellina</i>, formation of microsclerotia.

<p>(A-F) Transformation of hyphae into (A-D) chlamydospore or dictyochlamydospore-like structures, and (E, F) microsclerotia. (G-I) Coiling or coalescence of dictyochlamydospore-like structures. (J) Microsclerotia densely arranged in a chain. (K-N) Pigmentation first observed (K) in cell walls, and later (L-M) intensifying throughout microsclerotia. (O) A single microsclerotium inoculated on agar surface developing hyphae. A-I, K-N: from PDA, J: from CMA, O: from PDA 1/3. Scale bars: (A, C, E-I, K, L, O) = 30 μm; B, J = 50 μm; D = 200 μm; (M, N) = 10 μm.</p

Bayesian inference of phylogenetic relationships of selected taxa of the Bionectriaceae and Nectriaceae (Hypocreales) based on LSU sequences.

<p>Numbers above nodes are estimates of <i>a posteriori</i> probabilities (≥ 0.9) / NJB and MLB values (≥70%). The topology was rooted with <i>Aschersonia placenta</i>, <i>Balansia henningsiana</i>, <i>B</i>. <i>pilulaeformis</i>, and <i>Hypocrella nectrioides</i>, (Hypocreales).</p

Light micrographs of the infection and colonisation process of <i>Ijuhya vitellina</i> in cysts and eggs of <i>Heterodera filipjevi</i>.

<p>(A) Symptomatic cyst, reddish-dotted due to eggs containing reddish, globose microsclerotia. (B-E) Early colonisation of nematode eggs by hyphae becoming chlamydospore- and dictychlamydospore-like to develop microsclerotia inside eggs. (F, G) Dictyochlamydospore-like structures and small microsclerotia. (H) Hyphae penetrating through the eggshell by forming appressorium-like structure (arrows). (I-K) Development of the fungus inside nematodes eggs: (I) Formation of thick-walled hyphal cells, later (J-K) transforming into microsclerotia. The arrow in (J) points at the nematode stylet; in (K) at immature microsclerotium. (L) Egg with mature microsclerotium. (M-N) Near-identical cells of microsclerotium formed in (M) egg and (N) pure culture, forming a <i>textura angulari</i> in optical sections. Material obtained from (B-G, M) infected cysts directly placed and incubated on fungal colony, (H-L) slide cultures, (N) OA. Scale bars: A = 300 μm; (B-N) = 30 μm.</p

Cysts and eggs of <i>Heterodera filipjevi</i> naturally infected with <i>Ijuhya vitellina</i>, and pure cultures obtained from the infected eggs.

<p>(A) Symptomatic, reddish dotted nematode cysts. (B, C) Nematode eggs accommodating reddish microsclerotia. (D, E) Microsclerotial tissue developing inside juveniles. (F) A six-month-old culture that developed from a single infected nematode egg. (G) Surface of colony showing reddish microsclerotia arranged in concentric rings. (H, I) Two-month-old cultures on PDA and CMA. Scale bars: A = 0.5 mm, B = 30 μm, C-E = 50 μm, F = 1 cm (also applying for H, I), G = 400 μm.</p