Background Immune checkpoint inhibitors (ICIs) have transformed the treatment landscape for advanced non-small cell lung cancer (NSCLC), yet primary resistance remains common, with only ~50% of patients responding to first-line chemo-immunotherapy and 20-30% to monotherapy. Existing biomarkers such as PD-L1 expression and Tumor Mutational Burden (TMB) demonstrate limited predictive accuracy, underscoring the need for more comprehensive, integrative approaches. Methods We conducted a prospective, multicenter study involving 439 patients with advanced NSCLC treated with anti-PD-(L)1 ICI across first-line combo with chemotherapy and later-line monotherapy settings. A total of 443 pre-treatment tumor and blood-derived biomarkers-including genomic alterations, immune cell phenotypes, proteic markers, and routine laboratory tests-were profiled. Extensive biostatistics adjusted for PD-L1 expression were conducted. A rigorously benchmarked machine learning (ML) pipeline including 36 feature selection methods embedded into an optimism-correction framework was applied to identify predictors of primary resistance (PrR). Results Single biomarkers showed limited predictive utility, with PD-L1 (AUC 0.62, positive predictive value (PPV) 49.6%), TMB (AUC 0.55, PPV 43.1%), and key gene mutations (e.g., STK11, KEAP1) failing to achieve significance after multiple testing correction. A gradient boosting ML model integrating 18 selected features yielded a corrected AUC of 0.69 and a Positive Predictive Value (PPV) of 60% for PrR, outperforming standard biomarkers. In first-line patients, the model achieved a PPV of 51% and Negative Predictive Value (NPV) of 79% (baseline PrR rate: 29.9%); in subsequent-line patients, PPV reached 64% (PrR rate: 55.1%). Importantly, the signature also stratified Progression-Free Survival (PFS): high-risk patients had a median PFS of 3.9 vs. 14.6 months in low-risk patients (HR 0.307, p < 0.0001). Features from routine blood tests-such as serum chloride, albumin, CRP, and monocyte-to-lymphocyte ratio (MLR)-accounted for half of the final model and demonstrated independent associations with both PrR and PFS (e.g., chloride: OR 0.616, AUC 0.626; HR 0.685, C-index 0.61). SHAP-based individual-level model explainability revealed heterogeneous and nonlinear biomarker contributions, including cases where high CRP, low albumin, or elevated MLR overrode favorable PD-L1 or Treg profiles. A biomarker dashboard including interactive visualizations is available at https://compo.inria.fr/pioneer-website/. Conclusions Multimodal machine learning integration of clinical, genomic, immune, and laboratory data enables improved prediction of ICI resistance in NSCLC beyond current biomarkers. This approach not only captures the multifaceted nature of tumour-host interactions but also highlights the underrecognized predictive value of accessible blood-based markers, offering a path toward individualized immunotherapy decision-making. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement This work benefited from a government grant handled by the French National Research Agency (ANR) as part of the France 2030 investment plan, under the reference ANR-17-RHUS-0007. This work was supported by a partnership of Aix-Marseille Universite (AMU), Assistance Publique Hopitaux de Marseille (APHM), Centre National de La Recherche Scientifique (CNRS), Institut National de la Sante et de la Recherche Medicale (INSERM), Centre Leon Berard (CLB), Institut Paoli Calmettes (IPC), Gustave Roussy (GR), AstraZeneca (AZ), Veracyte (VERA), Innate Pharma (IPH) & ImCheck Therapeutics (ICT), and initiated by Marseille Immunopole. The authors gratefully acknowledge the support of the APHM, which sponsored the PIONeeR clinical studies. Its role was to control the appropriateness of ethical and legal considerations for all centers and to perform the monitoring of the consents signed and the clinical data recorded and coded as part of the study. The authors are grateful to all the patients and their families, as well as all the investigators, for their participation in the study. This work benefited from support from ITMO Cancer AVIESAN and French Institut National du Cancer (grant #19CM148-00) and from the French National Research Agency (ANR), under the France 2030 program, reference ANR-22-PESN-0017. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The study was conducted in accordance with the Helsinki declaration, French laws and regulations and the International Conference on Harmonization (ICH) E6 Guideline for Good Clinical Practice. The study was approved by the French ethics committee (Comite de Protection des Personnes Ouest II Angers, no. 2018/08) and the French drug and device regulation agency (Agence Nationale de Securite du Medicament, no. 2018020500208). Informed consent was obtained from each participant before any study procedure. The study is registered at ClinicalTrials.gov ([NCT03493581][1]). I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors [1]: /lookup/external-ref?link_type=CLINTRIALGOV&access_num=NCT03493581&atom=%2Fmedrxiv%2Fearly%2F2026%2F01%2F11%2F2026.01.09.26343779.atom

Integrating multimodal datasets in clinical oncology is frequently hindered by high dimensionality and blockwise missingness, where entire data sources are unavailable for specific patient subsets. Standard survival models often struggle with these gaps, leading to biased results or patient exclusion. We introduce Multimodality Stacking with Blockwise missing values (MSB), a late-fusion framework for survival analysis that independently models modality-specific features before aggregating predictions via a cross-validated stacking meta-learner. MSB was validated on the PIONeeR study (n=443 patients, 378 biomarkers across eight heterogeneous sources) to predict progression-free survival in advanced non-small cell lung cancer patients receiving immunotherapy. MSB yielded higher predictive performance (C-index) than baseline algorithms. Improvements varied by baseline strength: linear models showed a 15.9 MSB provides a statistically validated framework for multimodal survival prediction with blockwise missingness. By enabling systematic biomarker evaluation without requiring complete data, MSB offers a practical tool for predictive modeling in biomedical research, pending external validation. Implementation is available at https://github.com/MohamedBoussena/MSB under Inria license.

INTRODUCTION:Hemophagocytic lymphohistiocytosis (HLH) is a severe hyperinflammatory syndrome that can lead to early death from multiple organ failure. In secondary HLH, etiological treatment is an emergency, but sometimes impossible until complementary results (microbiology, histology) are available. A "bridge to etiological treatment" with anti-inflammatory drugs (anti-JAK, anti-cytokines or etoposide) is then essential. This study was conducted to determine the right time to start such treatment. Methods: We conducted a retrospective study at the University Hospital of Marseille on ultraselected (HS score greater than 215, probability of HLH = 95%) adult patients treated for secondary HLH in the intensive care unit. Results: Over a 10-year period, we included 23 patients (7 women, 16 men, 49 [37-59] years). The median HS score was 272 (250-294). Fourteen patients had infection-related HLH, four patients had lymphoma-related HLH, and two patients had adult-onset Still disease (AOSD). Seven patients died (30.4%), all of whom received invasive organ support (IOS; invasive mechanical ventilation, noradrenaline, and/or renal replacement therapy). When comparing the 14 patients with IOS with the nine others, we found no differences in terms of age, etiology, clinical-biological characteristics, and HLH-directed therapy. The mortality rate was none in the group without IOS compared with seven deaths (50%) in the IOS group ( P = 0.02). Conclusions: In secondary HLH, symptomatic anti-inflammatory treatment (awaiting etiological treatment) is an absolute emergency to stop macrophage and/or lymphocyte activation before organ failure occurs. A drastic increase in ferritin and a decrease in platelets could be biological warning signs.

Introduction Les lymphocytes T doubles négatifs à TCR αβ (CD3+CD4−CD8−TCRαβ+) sont une population de lymphocytes T présents en faible proportion dans le sang en situation normale. Ils peuvent être anormalement élevés dans diverses situations pathologiques. Observation Une patiente de 64 ans est hospitalisée pour une altération de l’état général. La biologie met en évidence une prolifération monoclonale de lymphocytes T doubles négatifs αβ à 28 % des lymphocytes T (219/mm3). Une tomodensitométrie thoraco-abdomino-pelvienne retrouve deux masses intra hépatiques suspectes, une polyadénomégalie et des nodules péritonéaux. Ces lésions sont hypermétaboliques en tomographie par émission de positon au 18-fluorodésoxyglucose. Une biopsie des masses hépatiques retrouve une histologie de cholangiocarcinome. Conclusion Nous rapportons une observation illustrant comment l’investigation d’une prolifération de lymphocytes T doubles négatifs αβ a mené au diagnostic de cholangiocarcinome. Le lien entre les deux ne peut être prouvé mais nous supposons que la stimulation antigénique tumorale a sélectionné un clone lymphocytaire T.

We describe pulmonary cryptococcosis in a 28-year-old previously healthy man. Exhaustive immunological investigations revealed a primary NK cell deficiency associated with a secondary impaired anti-Cryptococcus CD8 lymphocyte response and the expansion of a CD8Vβ14 + T cell clone. This case illustrates the potential role of NK cells in immunity against Cryptococcus.

Novel Genetic Etiologies of PIDs Preliminary data from this work were accepted and presented as an oral communication at the 21st biennial ESID meeting (October 2024). Epstein-Barr virus (EBV) is an oncogenic virus. Mostly asymptomatic, primary infection may be complicated by nonmalignant, malignant proliferations, and hemophagocytic lymphohistiocytosis. Infection resolution requires an appropriate cytotoxic T cell response, which depends on sustained T cell expansion. Several inherited EBV susceptibilities are associated with a high risk of complicated infection, which are often characterized by defective T cell expansion. Four patients with EBV-related diseases and one with severe CMV were analyzed by whole-exome sequencing, in whom six deleterious biallelic compound heterozygous variants were identified in RECQL4. These mutations are predicted to be highly damaging and absent in exome public databases. RECQL4 is a DNA helicase involved in DNA replication initiation and DNA repair. Patients with RECQL4 variants have been previously reported with several autosomal recessive syndromes associated with poikiloderma and an increased risk of cancer. Few rare cases were also reported with immunodeficiency but no EBV-related diseases. An additional patient carrier of a homozygous null variant in RECQL4 presenting severe combined immunodeficiency and a severe RECQL4-related syndrome was studied. We showed that RECQL4 is upregulated in activated control T cells upon TCR-CD3 activation. In HEK cells, transiently overexpressed RECQL4 variants of the patients result in a reduced or absent RECQL4 expression. In activated T cells from patients, RECQL4 expression is strongly reduced or absent, and T cells exhibit a proliferation defect and an increased activation-induced apoptosis. In Jurkat T cell line, 3 different shRNA-targeting RECQL4 decrease its expression and induce a cell cycle blockade, proliferation arrest, and an increased apoptosis. Similarly in control T cells, silencing RECQL4 results in an expansion disadvantage and an increased apoptosis in the infected cells. RECQL4 deficiency is associated with impaired T cell expansion that may underlie EBV susceptibility in patients.

INTRODUCTION:Double-negative T lymphocytes with αβ TCR (CD3+CD4-CD8-TCRαβ+) are a population of T cells present in low proportions in the blood under normal conditions. They can be abnormally elevated in various pathological situations. CASE REPORT:A 64-year-old female patient was hospitalized due to a general deterioration of health. Laboratory tests revealed a monoclonal proliferation of double-negative αβ T lymphocytes, representing 28% of T cells (219/mm3). A thoraco-abdomino-pelvic CT scan revealed two suspicious intrahepatic masses, multiple enlarged lymph nodes, and peritoneal nodules. These lesions were hypermetabolic on 18FDG PET scan. A biopsy of the hepatic masses showed histological features consistent with cholangiocarcinoma. CONCLUSION:We report a case illustrating how the investigation of a proliferation of double-negative αβ T lymphocytes led to the diagnosis of cholangiocarcinoma. Although a direct link between the two cannot be proven, we hypothesize that tumor antigenic stimulation selected a T lymphocyte clone.

CTLA4 deficiency is an inborn error of immunity (IEI) due to heterozygosity for germline loss-of-function variants of the CTLA4 gene located on chromosome 2q33.2. CTLA4 deficiency underlies pleiotropic immune and lymphoproliferation-mediated features with incomplete penetrance. It has been identified in hundreds of patients but copy number variants (CNVs) have been reported in only 12 kindreds, including nine which displayed large 2q33.1-2q33.2 deletions encompassing CTLA4. We conducted a nationwide study in France to identify patients with 2q33 deletions encompassing CTLA4. We investigated the clinical and immunological phenotypes and genotypes of these patients. We identified 12 patients across six unrelated kindreds with clinical immunodeficiency. Neurological features were recorded in three patients, including one with syndromic neurodevelopmental disorder. Single-nucleotide polymorphism (SNP) or comparative genomic hybridization (CGH) array analysis, and targeted high-throughput sequencing revealed five different heterozygous 2q33 deletions of 26 kilobases to 7.12 megabases in size and encompassing one to 41 genes. We identified a contiguous gene syndrome (CGS) due to associated KLF7 deficiency in a kindred with a neurodevelopmental phenotype. Deletions within the 2q33 region encompassing CTLA4 are rare and not extensively explored, and are probably underdiagnosed in cytogenetic practice. A literature review identified 14 different CGS loci including at least one gene responsible for an IEI. The deletions involved in IEIs should be systematically delimited, to facilitate screening for CGS.

Introduction La maladie associée aux IgG4 (MAG4) est une pathologie fibro-inflammatoire pouvant toucher tous les organes, avec une prédominance d’atteintes pancréato-bilaires, rétro-péritonéales, rénales et salivaire. Ces atteintes présentent des anomalies histologiques communes avec des lésions fibrose dites « storiforme » et un infiltrat inflammatoire lymphoplasmocytaire polyclonal. La fractalkine (FKN), également connue sous le nom de CX3CL1, a des fonctions d’adhésion cellulaire et de chimioattraction. La FKN se lie spécifiquement à son récepteur CX3CR1, exprimé sur des sous-ensembles de cellules immunitaires telles que les monocytes et les lymphocytes. L’axe FKN-CX3CR1 joue un rôle important dans l’initiation de l’inflammation et contribue aux lésions tissulaires dans la polyarthrite rhumatoïde, la sclérodermie systémique, la fibrose pulmonaire et les maladies cardiovasculaires. Son rôle potentiel dans la MAG4 a été peu étudié. Matériels et méthodes Vingt-deux patients atteints d’une MAG4, définie par un score≥20 selon les critères de classification ACR/EULAR, active et non traitée, ont été inclus et comparés à 22 témoins sains. Les concentrations sériques de FKN ont été mesurées par ELISA. L’expression de CX3CR1 a été étudiée par cytométrie en flux sur différents sous-types de cellules mononuclées du sang périphérique (PBMC) : T follicular helper (Tfh) (CD4+CXCR5+PD1+) et T peripheral helper (Tph) -like (CD4+CXCR5-PD1+), les cellules B, les cellules NK et les monocytes. Résultats Les patients atteints de MAG4 étaient principalement des hommes (83 %), avec un âge médian de 60ans et un score ACR/EULAR médian de 38 [20–70]. La FKN sérique était plus élevée chez les patients atteints de MAG4 (médiane de 0,73 ng/mL [0,24–2,24]) que chez les témoins sains (médiane de 0,53 ng/mL [0,13 à 0,70]) (p=0,03). Chez 21 patients dont le sérum était disponible avant (maladie active) et après le traitement (rémission), une diminution significative de la FKN sérique a été observée (p=0,02). Les proportions de cellules circulantes Tph-like (10,3 vs 4,0 % des cellules T CD4+, p<0,0001) et Tfh (3,7 vs 1,7 % des cellules T CD4+, p<0,001) étaient augmentées chez les patients MAG4, mais une surexpression de CX3CR1 n’a été trouvée que sur les cellules Tph-like (9,8 % de cellules CX3CR1+/Tph dans l’IgG4-RD vs 4,0 % chez les contrôles, p<0,001). CX3CR1 n’était pratiquement pas exprimé (<1 %) sur les cellules Tfh, tant chez les patients que chez les témoins. Une corrélation significative a été trouvée entre le pourcentage de cellules Tph-like CX3CR1+ et les taux sériques d’IgG4 (p=0,03), ainsi qu’avec le nombre d’éosinophiles circulants (p=0,01). Une surexpression de CX3CR1 a également été observée sur les monocytes des patients atteints de MAG4 (98,6 contre 95,7 %, p<0,0001). Les monocytes classiques (CD14+CD16-) étaient plus nombreux chez les patients MAG4 (83,1 vs 66,5 % des monocytes totaux, p=0,0003), et une surexpression de CX3CR1 a été observée uniquement dans cette sous-population. CX3CR1 était plus exprimé sur les cellules NK chez les patients MAG4 (97,5 vs 94,7 %, p=0,001). Enfin, CX3CR1 n’était pas exprimé sur les cellules B, définies comme des lymphocytes CD20+, à la fois chez les patients et les témoins sains. Conclusion La FKN sérique et l’expression de son récepteur CX3CR1 sur les cellules Tph-like, les monocytes et les cellules NK sont augmentés dans le sang des patients MAG4. Nos résultats sont en accord avec les données récentes de single-cell RNAseq [1]. Bien que des analyses de tissus devraient compléter ces résultats, l’axe FKN-CX3CR1 pourrait être impliqué dans la physiopathologie de la MAG4, et l’inhibition de l’axe FKN-CX3CR1 pourrait représenter une nouvelle approche thérapeutique dans cette maladie.

The objective was to conduct a comprehensive review of the morbidity and mortality observed in published patients with gastrointestinal defects and immunodeficiency syndrome-1 (GIDID1) related to TTC7A abnormalities. This included phenotypic, genotypic, and therapeutic aspects. Twenty-seven articles were included, which represented a total of 83 patients. Mortality was of 65.8% of the cases with a mean death at 11.8 months. The mortality rate was 197.1 per 1,000 patients-years, which is significantly higher than other enteropathy types caused by defects in epithelial trafficking and polarity (such as MOY5B, STX3, EPCAM, SPINT2, TTC37 and SKIV2L). Prematurity was also significant, with an average gestational age of 34.8 weeks. Antenatal signs were observed in 30 patients, including 14 cases of hydramnios. Three distinct phenotypic associations were identified: immune deficiency and multiple intestinal atresia without enteropathy (ID/MI), immune deficiency and enteropathy without atresia (ID/E), and immune deficiency with multiple intestinal atresia and enteropathy (ID/ MIA/E). The mortality rates for these groups were 91.6%, 47.3% and 55.5%, respectively (p = 0.03), at earlier age of mortality for the ID/MIA phenotype and a later one for the ID/E phenotype. ELA syndrome (Enteropathy, Lymphopenia and Alopecia) was only observed in the ID/E group. Among the three genotypes (double variant Nonsense NS/NS, variant Missense/Nonsense MS/NS, double variant Missense MS/MS), NS/NS was significantly associated with the ID/MIA phenotype (77.8%), while MS/MS was associated with the ID/E phenotype (73.7%). Few therapies have been shown to be effective in treating enteropathy, particularly immunosuppressive therapies and hematopoietic stem cell transplants. The use of Leflunomide in one patient did not yield successful treatment outcomes. In conclusion, we confirm association between mortality and phenotype, which is itself linked to genotype.

Objective: Epicardial adipose tissue (EAT) is a visceral fat that has been associated with coronary artery disease and atrial fibrillation. Previous work has revealed that EAT exhibits beige features. Methods: First, a new pan-genomic microarray analysis was performed on previously collected paired human EAT and thoracic subcutaneous AT (thSAT) from the EPICAR study (n = 31) to decipher a specific immune signature and its link with browning genes. Then, adaptive (T and B cells) and innate lymphoid cell (ILC1, ILC2, and ILC3) immunophenotyping assay panels, including CD127, CD117, and prostaglandin D2 receptor 2, were performed on prospectively collected paired human multiorgan donors (n = 18; INTERFACE study). Results: In the EPICAR study, a positive correlation between the T helper cell subtype Th2 immune pathway and browning genes was found in EAT versus thSAT (r = 0.82; p < 0.0001). In the INTERFACE study, this correlation was also observed (r = 0.31; p = 0.017), and a preponderance of CD4(+)T cells, CD8(+)T cells, and a few B cells was observed in all ATs (p < 0.0001). An increase in ILCs was observed in visceral AT (VAT) (i.e., EAT + VAT; 30 +/- 5 ILCs per gram of AT) compared with subcutaneous counterparts (i.e., thSAT + abdominal SAT; 8 +/- 2 ILCs per gram of AT; p = 0.001), with ILC1 being the most frequent (ILC1 > ILC3 > ILC2). Numbers of ILCs per gram of AT correlated with several Th2 or browning genes (IL-13, TNF receptor superfamily member 9 [TNFRSF9], and alkaline phosphatase, biomineralization associated [ALPL]). Interestingly, a specific increase in EAT-ILC2 compared with other ATs was observed, including a significant proportion expressing CD69 and/or CD25 activation markers (97.9% +/- 1.2%; p < 0.0001). Finally, more natural killer cells were observed in EAT + VAT than in thSAT + abdominal SAT (p = 0.01). Exclusion of patients with coronary artery disease in the EPICAR and INTERFACE studies did not modify the main findings. Gene expression phenotyping confirmed specific upregulation of Th2 pathway and browning genes (IL-33 and uncoupling protein 1 [UCP-1]) in EAT. Conclusions: This is the first study, to our knowledge, to provide a comparison between innate and adaptive lymphoid cells in human EAT. Further studies are ongoing to decipher whether these cells could be involved in EAT beiging. image

Background and Objectives Real-life studies noted that the risk of disease activity in multiple sclerosis (MS) after switching to rituximab (RTX) or ocrelizumab (OCR) may be unequal depending on prior disease-modifying therapy (DMT), with a higher risk associated with fingolimod (FING). Methods We performed a retrospective analysis of a structured prospective data collection including all consecutive patients with relapsing MS who were prescribed RTX/OCR in the MS center of Marseille. Cox proportional hazards models were applied to clinical and MRI outcomes. Results We included 321 patients with a median (interquartile range [IQR]) follow-up of 3.5 years (1.5-5) after RTX/OCR initiation. At the first RTX/OCR infusion, the mean (SD) age of patients was 37 (10) years, and the median (IQR) disease duration was 8 years (3-15): 68 patients did not receive treatment before RTX/OCR and 108 switched from FING, 47 from low efficacy therapy, and 98 from natalizumab. For statistical analysis, the group "FING" was divided into "short-FING" and "long-FING" groups according to the median value of the group's washout period (27 days). On Cox proportional hazards analysis, for only the "long-FING" group, the risk of relapse within the first 6 months of RTX/OCR was increased as compared with patients without previous DMT (hazard ratio [HR]: 8.78; 95% CI 1.72-44.86; p < 0.01). Previous DMT and washout period duration of FING had no effect on B-cell levels at 6 months. Beyond the first 6 months of RTX/OCR, age <40 years was associated with increased risk of relapse (HR: 3.93; 95% CI 1.30-11.89; p = 0.01), male sex with increased risk of new T2 lesions (HR: 2.26; 95% CI 1.08-4.74; p = 0.03), and EDSS >= 2 with increased risk of disability accumulation (HR: 3.01; 95% CI 1.34-6.74; p < 0.01). Previous DMT had no effect on the effectiveness of RTX/OCR beyond 6 months after initiation. Discussion For patients switching from FING to RTX/OCR, the risk of disease reactivation within the first 6 months of treatment was increased as compared with patients with other DMT or no previous DMT only when the washout period exceeded 26 days. Neither FING nor other previous DMT reduced the effectiveness of RTX/OCR beyond the first 6 months of treatment.

Importance Epicardial adipose tissue (EAT) is a biologically active organ surrounding myocardium and coronary arteries that has been associated with coronary artery disease (CAD) and atrial fibrillation. Previous work has shown that EAT exhibits beige features. Objective Our objective was to determine whether the stromal vascular fraction of the human EAT contains innate or adaptive lymphoid cells compared to thoracic subcutaneous (thSAT), visceral abdominal (VAT) and subcutaneous abdominal (abSAT). Participants New pangenomic microarray analysis was performed on previous transcriptomic dataset using significance analysis of microarray and ingenuity pathway analysis (n=41) to identify specific immune signature and its link with browning genes. EAT, thSAT, VAT and abSAT samples from explanted patients with severe cardiomyopathies and multi-organ donor patients (n=17) were used for flow cytometry (FC) immunophenotyping assay. Patients were on average 55±16 years-old; 47% had hypertension and 6% CAD. Phenotypic adaptive and innate immune profiles were performed using a TBNK panel and a specific ILC1-2-3 panel including CD127, CD117, CRTH2 (CD294) and activation markers such as CD25 and CD69. Results Transcriptomic analysis showed a significant positive correlation between the TH2 immune pathway (IL-4, IL-5, IL-13, IL-25, IL-33) and browning genes (UCP-1, PRDM16, TMEM26, CITED1, TBX1) in EAT versus thSAT (R=0.82, P<0.0001). Regarding adaptive immune cells, a preponderance of CD8T cells, a contingent of CD4T cells, and a few B cells were observed in all ATs (P<0.0001). In innate lymphoid cells (ILCs), an increase was observed in visceral ATs (i.e. EAT; VAT 35±8ILCs/g of tissue) compared to their subcutaneous counterpart (i.e. thSAT+abSAT: 8±3 ILCs/g of AT, P=0.002), with a difference in the proportion of the 3 subtypes of ILCs (ILC1>ILC3>ILC2). In addition, we observed an increase in EAT-ILC2 compared to other ATs and almost all these EAT-ILC2 expressed CD69 and/or CD25 activation markers (99.75±0.16%; P<0.0001). We also observed more NKs in EAT and VAT (1520±71 cells/g of AT) than in SATs (562±17 cells/g of AT); P=0.01. Conclusion This is the first study to provide a comparison between innate and adaptive lymphoid cells in human epicardial versus abdominal or thoracic adipose tissues. Further studies are ongoing to decipher whether these cells could be involved in EAT beiging. Trial Registration CODECOH No. DC-2021-4518 The French agency of biomedicine PFS21-005.

Summary of analyzed NK receptors.

Effect of Vemurafenib on BRAF non mutated melanoma cell lines

ERK expression of melanoma cell lines under Vemurafenib treatment.

List of antibodies used for flow cytometry.

Background and Objectives Patients with multiple sclerosis (PwMS) receiving extended dosing of rituximab (RTX) have exhibited no return of disease activity, which suggests that maintenance of deep depletion of circulating B cells is not necessary to maintain the efficacy of RTX in MS. Methods This was a prospective monocentric observational study including all consecutive PwMS who started or continued RTX after 2019, when the medical staff decided to extend the dosing interval up to 24 months for all patients. Circulating B-cell subsets were monitored regularly and systematically in case of relapse. The first extended interval was analyzed. Results We included 236 PwMS (81% with relapsing-remitting MS; mean [SD] age 43 [12] years; median [range] EDSS score 4 [0–8]; mean relapse rate during the year before RTX start 1.09 [0.99]; 41.5% with MRI activity). The median number of RTX infusions before extension was 4 (1–13). At the time of the analysis, the median delay in dosing was 17 months (8–39); the median proportion of circulating CD19+ B cells was 7% (0–25) of total lymphocytes and that of CD27+ memory B cells was 4% (0–16) of total B cells. The mean annual relapse rate did not differ before and after the extension: 0.03 (0.5) and 0.04 (0.15) (p = 0.51). Similarly, annual relapse rates did not differ before and after extension in patients with EDSS score ≤3 (n = 79) or disease duration ≤5 years (n = 71) at RTX onset. During the “extended dosing” period, MRI demonstrated no lesion accrual in 228 of the 236 patients (97%). Five patients experienced clinical relapse, which was confirmed by MRI. In these patients, the level of B-cell subset reconstitution at the time of the relapse did not differ from that for patients with the same extension window. Discussion The efficacy of RTX outlasted substantial reconstitution of circulating B cells in PwMS, which suggests that renewal of the immune system underlies the prolonged effect of RTX in MS. These findings suggest that extended interval dosing of RTX that leads to a significant reconstitution of circulating B cells is safe in PwMS, could reduce the risk of infection, and could improve vaccine efficacy.

Phenotype and degranulation potential of IL-15 activated NK cells

Introduction Anti-MAG neuropathies are associated with an IgM monoclonal gammopathy of undetermined significance (MGUS) or with a malignant haemopathy. Our objective was to determine whether the presence of a haemopathy or somatic mutations of MYD88 and CXCR4 genes influences disease presentation and response to rituximab (RTX). Methods We included 79 patients (mean age 74 years, disease duration 9.68 years) who had a bone marrow aspiration with morphologic and immunophenotypic analysis. MYD88 L265P and CXCR4 mutations were analysed in peripheral B cells. Information collected included: inflammatory neuropathy cause and treatment sensory sum score (ISS), MRC testing, overall neuropathy limitation scale (ONLS), Rash-built Overall Disability Score (RODS), ataxia score, anti-MAG titres, peak IgM dosage, neurofilament light chain levels, motor and sensory amplitudes, motor unit index (MUNIX) and motor unit size index (MUSIX) sum scores. Efficacy of RTX was evaluated at 12 months in 26 patients. Results Malignant haematological disorders were discovered in 17 patients (22%): 13 Waldenstrom macroglobulinemia, 3 marginal zone lymphoma and one mantle cell lymphoma. MYD88 L265P mutation was detected in 29/60 (48%) patients and CXCR4 in 1 single patient. Disease severity, biological and electrophysiological data and response to RTX were comparable in patients with MGUS/lymphoma and patients with/without MYD88 L265P mutation. ISS was lower and MUSIX higher in patients improved by RTX. Conclusions MYD88 L265P mutation and underlying haemopathies are not predictive of a more severe disease. However, in cases of resistant and progressive neuropathy, they provide an opportunity to prescribe newly available drugs such as Bruton tyrosine kinase inhibitors.