Monoclonal antibody-based immunotherapy has emerged as a promising treatment for B-cell lymphoma, with Rituximab (RTX) IgG1 targeting the CD20 surface protein showing significant clinical success in combination with chemotherapy (Bello & Sotomayor 2007). However, not all patients respond to RTX, whereby further studies are warranted to improve RTX efficacy. In this study, an in-house ELISA method was optimized to analyze cytokines from supernatants from human B cell lymphoma cell lines and monocytic cell cultures. Basal levels of the pro-inflammatory tumor necrosis factor α (TNF-α) and the immunosuppressive interleukin 10(IL-10) cytokines were investigated. The study shows several factors that can affect cytokine measurements in cell lines, such as time of culture, cell passage numbers, and incubation times of standards and samples in the ELISA. Furthermore, the correlation between IL-10 secretion of cell lines and phagocytosis was investigated using supernatant samples from phagocytosis assays of B-cell lymphoma cells treated with RTX and anti-CD47 mAbs. Notably, the pattern of IL-10 production from the samples varied depending on the treated antibodies and not by the intensity of the phagocytosis induced by the mAbs.In summary, this study shows that ELISA methods need to be tailored for analysis of cytokines in cell cultures, and highlights that cytokine levels not necessarily correspond to phagocytosis intensity induced by therapeutic mAbs.

La douleur neuropathique est due à une lésion ou une pathologie du système nerveux somatosensoriel. La prégabaline, un anticonvulsivant, et la duloxétine, un antidépresseur, sont des traitements de référence, efficaces chez un tiers des patients. Mieux comprendre leurs mécanismes d’action est crucial pour améliorer leur tolérance et leur efficacité. En utilisant un modèle murin de douleur neuropathique périphérique, nous montrons que : 1) la prégabaline, dont l’action est indépendante du système opioïdergique, agit sur la composante neuroimmunitaire périphérique de la douleur ; 2) la duloxétine agit via deux mécanismes indépendants, l’un central (contrôles descendants) pour un traitement aigu et l’autre périphérique (ganglion rachidien) pour un traitement chronique. Dans ce cas, l’analyse transcriptomique met en évidence une inhibition de l’inflammation neurogène. La comparaison des taux plasmatiques de duloxétine chez l’homme et chez la souris suggère une action périphérique chez l’homme. / Neuropathic pain is caused by a lesion or a disease of the somatosensory nervous system. Pregabalin, an anticonvulsant, and duloxetine, an antidepressant, are the standard treatments, effective in one-third of patients. A better understanding of their mechanisms of action is a crucial point to improve their tolerance and efficiency. By using a murine model of peripheral neuropathy, we have shown that : 1) pregabalin, whose effect is independent from the opioid system, acts on the peripheral neuroimmune component of pain ; 2) duloxetine acts via two independent mechanisms, one central (descending controls) for an acute treatment and the other peripheral (dorsal root ganglia) for a chronic treatment. In this case, transcriptomic analysis hightlights an inhibition of the neurogenic inflammation. Comparison of duloxetine plasmatic levels in humans and mice suggests a peripheral action in humans.