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.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:uu-529953 |
| Date | January 2024 |
| Creators | Jang, Inkyung |
| Publisher | Uppsala universitet, Institutionen för biologisk grundutbildning |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/bachelorThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
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