Immunomodulatory effects of different Cannabis extracts on human primary lymphocytes

Abstract: Cannabis sativa has been used in traditional medicine for the treatment of different inflammatory diseases and is one of the most ancient herbal remedies. Its use is prohibited in many countries worldwide because of its mind-altering properties. Therefore, new formulations have been developed which contain little or no THC and are rich in other cannabinoids. Cannabis is used for the treatment of inflammatory autoimmune diseases, however, its potential effects and mode of action are not yet understood clearly. In this research work, in vitro cell-based immunomodulatory effects of different cannabis extracts were investigated. Human T lymphocytes were used for our investigations because of their role in autoimmune diseases such as rheumatoid arthritis or multiple sclerosis.

The phytochemical profiling of extracts was done using HPLC-PDA-ELSD-MS analysis. The bioactivity of cannabis extracts was analysed in vitro using flow cytometry-based techniques. Proliferation and apoptosis/necrosis were assessed by CFSE and annexin V/propidium iodide staining, cell cycle distribution by propidium iodide staining of DNA. The viability of the cells was analysed using WST-1 assay, DNA damage was measured using single-cell gel electrophoresis and ROS was measured via EPR spectroscopy, respectively. The effect on T lymphocyte activation was measured via CD25 and CD69 marker expression, degranulation, and altered expression levels of interleukin-2 (IL-2), interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α), as well as autophagy induction using flow cytometry. The influence on the transcription factor activator protein-1 (AP-1), nuclear factor of activated T cells (NFAT), and nuclear factor kappa-light-chain enhancer of activated B cells (NF-кB) was also analysed via flow cytometry, for which Jurkat reporter cell lines were used. Additionally, the effect on the proliferation and IL-2 production was examined using specific CB2 and TRPV1 receptor antagonists.

Santhica (SA) extract contained mostly cannabigerol acid (CBGA) and did not influence the proliferation of activated T lymphocytes but inhibited the activation and functionality by inhibition of CD25 marker expression, degranulation and IL-2 cytokine production in concentrations not inducing apoptosis, necrosis, or affecting viability or causing DNA strand break. Fidora (FI) extract contains cannabidiol acid (CBDA) and CBD. Non-toxic concentrations inhibited T cell signalling via an IL-2 dependent mechanism. Further investigation revealed a reduced expression of CD25 and CD69 activation markers, degranulation as well as reduced IL-2 and TNF-α production of activated T lymphocytes. The Fedora (FE) extract which is rich in CBD was investigated in comparison to pure CBD. FE had, as CBD, stronger immunosuppressive effects than SA and FI. FE extract and CBD both impaired proliferation, activation, and functionality of T lymphocytes. They induced cell cycle arrest and specific impairment of T cell signalling via an IL-2 dependent mechanism as well as via inducing autophagy. AP-1 and NFAT inhibition was found for both FE and the corresponding concentrations of CBD, which could explain the inhibitory effects of FE and CBD on T lymphocyte proliferation, CD25 marker expression, lymphocyte degranulation and IL-2 and IFN-γ production. The specific CB2 and TRPV1 receptor antagonists SR144528 and A78416B reversed FE induced inhibition of lymphocyte proliferation and IL-2 production as well as the CBD induced inhibition of proliferation. Accordingly, most of the immunosuppressive effects mediated by FE can be explained by its CBD content. However, related to equivalent CBD concentrations, FE extract is slightly more effective for the inhibition of proliferation (IC50: FE = 3.8; CBD = 4.7), degranulation (IC50: FE = 2.9; CBD = 9.5), NFAT (IC50: FE = 1.5; CBD = 4.4), indicating that other compounds present in the extract may be relevant for the effects. Altogether, our findings demonstrate that cannabis extracts as well as CBD alone can be potential immune modulators. Further in vivo research is required to corroborate these effects

Object Details

• Location: Deutsche Nationalbibliothek Frankfurt am Main
• Extent: Online-Ressource
• Language: Englisch
• Notes: Universität Freiburg, Dissertation, 2022

Classification and Topics

• Keyword: Signaltransduktion ; Immunsystem ; Immunreaktion ; Autoaggressionskrankheit ; Immunmodulation

Contributors, Places and Time

• Event: Veröffentlichung
• (where): Freiburg
• (who): Universität
• (when): 2022
• Creator: Devi, Seema
• Contributor: Huber, Roman ; Huber, Roman ; Ehl, Stephan ; Albert-Ludwigs-Universität Freiburg. Medizinische Fakultät

Further information

• DOI: 10.6094/UNIFR/231709
• URN: urn:nbn:de:bsz:25-freidok-2317096
• Rights: Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
• Last update: 25.03.2025, 1:42 PM CET