Myeloma Cells Have the Ability to Suppress Th1-Inducing Capacity but Enhance Th2-Mediated Response of Dendritic Cells By Producing Factors Other Than Soluble SLAMF7
Authors: Azuma Yoshiko , Ito Tomoki , Inaba Muneo , Imai Kai , Yoshimura Hideaki , Hotta Masaaki , Nakanishi Takahisa , Fujita Shinya , Nakaya Aya , Satake Atsushi , Ishii Kazuyoshi , Nomura Shosaku 
Published: October 2019

Abstract
Background: Lenalidomide (LEN), as a backbone drug for multiple myeloma (MM), has both direct tumoricidal and immunomodulatory effects in MM. LEN displays immunopotentiating activity, including augmentation of T and natural killer cell function and to induce selective reduction of regulatory T cells. Thus, the immune-based cancer treatments are currently important in MM therapy. Since myeloma cells together with immune cells are immersed in the pathogenic micro-environment, understanding the influence of micro-environment against myeloma cells and immune system may lead to the development of novel treatment strategies for MM. Series of analyses have clarified a functional plasticity of dendritic cells (DCs) to induce Th1 or Th2 response and DCs are pivotal in orchestrating both innate and acquire immune responses also in the anti-tumor immunity. Here, we focused on the effects of myeloma environment to modulate the human myeloid DCs (mDCs), which are the major regulators to induce Th1 or Th2 responses. Soluble form of SLAMF7 (sSLAMF7) is a pivotal environmental component of MM and functions as a self-ligand of myeloma cells. sSLAMF7 has been detected in the serum of patients with MM at higher levels than healthy individuals and therefore may presumably play some roles in pathophysiological condition of MM. To evaluate the functions of soluble factors/components derived from myeloma cells, we analyzed whether supernatants of myeloma cell lines could modulate mDC functions. Methods: Using cell sorting, blood human CD11+ mDCs were isolated from healthy adult volunteers. The DC functions were analyzed after culture for 24 h with supernatant of myeloma cell lines, NCI-H929 (H929) or MM.1R. This study was approved by the Institutional Review Board of Kansai Medical University.Results: We here found that both supernatants of myeloma cell lines, H929 and MM.1R did not affect CD86 expression on mDCs in response to toll-like receptor (TLR)-ligand (R848), but upregulated their CD86 expression in response to Th2-inducing cytokine, thymic stromal lymphopoietin (TSLP). Of note, the supernatants inhibited mDC-derived IL-12 production in response to R848 (p < 0.05: H929; 25-30% of cont. and MM1.R; less than 50 % of cont.), while in response to TSLP, dose-dependently enhanced the Th2-recruiting chemokine CCL17/TARC production (p < 0.05: H929; 130- 210 % of cont. and MM1.R; 120-185 % of cont.), which functions as chemoattractant for memory Th2 cells and contribute to promote humoral response. In addition, we revealed that these capacities were further enhanced by the addition of LEN at clinical in vivo plasma concentration of 1 µM. Thus, MM micro-environment could have the potential to shift DC-mediated response from Th1 to Th2. To clarify whether sSLAMF7 in the supernatant of myeloma cell lines play a role in the Th1/Th2-modulating effect, we calculated the sSLAMF7 concentration in the supernatants and analyzed an inhibitory effect of elotuzumab for the DC function induced by the supernatants. sSLAMF7 could not be detected at all in the supernatants of both myeloma cell lines, even in any concentration of cells. Furthermore, addition of 30 to 100 ug/ml (clinical trough serum concentration) elotuzumab into the culture of mDCs and the supernatants did not affect the CD86 expression and IL-12/CCL17 production. Hence, factors other than sSLAMF7 from myeloma cells can trigger DCs to suppress Th1 response and to promote Th2 response.Conclusion: Our data suggest that the MM micro-environment, while suppress the Th1-inducing capacity, rather lead to enhance Th2 response at DC phase. As sSLAMF7 is detected in the serum of patients with MM but not in the supernatants of MM cell lines in the experimental setting, there is a limitation that our data does not consider the action of sSLAMF7. However, the present result provides an important clue for understanding the biological basis for the MM micro-environment against host immune defense at onset.

Disclosures
Ito: Bristol-Myers Squibb: Honoraria, Research Funding; Takeda: Honoraria; Novartis Pharma: Honoraria; Pfizer: Honoraria; Mundipharma: Honoraria; Celgene: Honoraria.