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Wijkhuisen, A.

Publications and source records attributed to Wijkhuisen, A..

2 recordsLinked to original sources

Dual blockade of LILRB1 and LILRB2 enhances antiviral immune responses in SIV infection

Restoring effective antiviral immunity remains a major challenge in HIV infection. Among emerging immune checkpoint molecules, the inhibitory receptors LILRB1 and LILRB2 have been proposed as therapeutic targets, yet their in vivo function remains undefined due to the lack of cross-reactive blocking antibodies for relevant preclinical models. To address this, we developed a dual-specific blocking monoclonal antibody, mac20G10, targeting cynomolgus macaque LILRB1 and LILRB2 and assessed its immunomodulatory activity in an SIV model of infection. Pharmacodynamics analyses demonstrated that mac20G10 persisted in circulation and engaged target myeloid cells for up to 14 days without detectable adverse effects. A single administration prior to SIVmac251 infection enhanced early myeloid immune activation, characterized by increased frequencies of CD80+ pDC and CD80+ monocyte/macrophage subsets in blood and lymphoid tissues. These changes were accompanied by increased plasma levels of IFN-{lambda}, IL8, and IL-1RA during acute infection. Although viral replication remained unchanged, mac20G10 treatment promoted the development of SIV-specific memory CD8 T-cell responses. Together, these findings provide in vivo evidence that LILRB1 and LILRB2 function as myeloid immune checkpoints restraining antiviral priming, supporting this pathway as a rational target for combination immunotherapeutic strategies aimed at achieving durable HIV remission during analytic treatment interruption.

immunology↗

Targeted fusion of Antibody-Secreting Cells: unlocking monoclonal antibody production with hybridoma technology

Monoclonal antibodies (mAbs) produced by hybridoma technology have extensively proved their value for therapeutic, diagnostic, and biomedical research applications, despite the reported low fusion yields between short-lived B cells and immortal myeloma cells. To improve the efficiency of this process and accelerate the development of new mAbs, we characterized and isolated antibody-secreting cells (ASCs) from the spleen of immunized mice before cell fusion. This approach resulted in a high yield of hybridoma generation by increasing the probability of successive pairing between the most suitable cell fusion partners. Specifically, we developed an optimized workflow combining Fluorescence-Activated Cell Sorting (FACS) with antibody secretion assays, using a panel of five cell-surface markers (CD3, TACI, CD138, MHC-II, and B220) that allowed us to identify a particular ASC subset with key characteristics. Such ASCs exhibited a plasmablast phenotype with high MHC-II expression and secreted high levels of Ag-specific antibodies in immunized mice. These features were also found in hybridomas, suggesting a preferential fusion of myeloma cells with this ASCs subset. Finally, the targeted electrofusion of TACIhighCD138high sorted ASCs led to a 100% fusion yield compared to a non-targeted approach. In particular, over 60% of these generated hybridomas secreted Ag-specific mAbs. Collectively, these results pave the way for a highly efficient method to produce new mAbs by cell fusion, which could facilitate hybridoma generation and expand therapeutic applications of mAbs.

immunology↗