Gene expression profile of human T cells following a single stimulation of peripheral blood mononuclear cells with anti-CD3 antibodies

Abstract

Background Anti-CD3 immunotherapy was initially approved for clinical use for renal transplantation rejection prevention. Subsequently, new generations of anti-CD3 antibodies have entered clinical trials for a broader spectrum of therapeutic applications, including cancer and autoimmune diseases. Despite their extensive use, little is known about the exact mechanism of these molecules, except that they are able to activate T cells, inducing an overall immunoregulatory and tolerogenic behavior. To better understand the effects of anti-CD3 antibodies on human T cells, PBMCs were stimulated, and then, we performed RNA-seq assays of enriched T cells to assess changes in their gene expression profiles. In this study, three different anti-CD3 antibodies were used for the stimulation: two recombinant antibody fragments, namely, a humanized and a chimeric FvFc molecule, and the prototype mouse mAb OKT3. Results Gene Ontology categories and individual immunoregulatory markers were compared, suggesting a similarity in modulated gene sets, mainly those for immunoregulatory and inflammatory terms. Upregulation of interleukin receptors, such as IL2RA, IL1R, IL12RB2, IL18R1, IL21R and IL23R, and of inhibitory molecules, such as FOXP3, CTLA4, TNFRSF18, LAG3 and PDCD1, were also observed, suggesting an inhibitory and exhausted phenotype. Conclusions We used a deep transcriptome sequencing method for comparing three anti-CD3 antibodies in terms of Gene Ontology enrichment and immunological marker expression. The present data showed that both recombinant antibodies induced a compatible expression profile, suggesting that they might be candidates for a closer evaluation with respect to their therapeutic value. Moreover, the proposed methodology is amenable to be more generally applied for molecular comparison of cell receptor dependent antibody therapy. Electronic supplementary material The online version of this article (10.1186/s12864-019-5967-8) contains supplementary material, which is available to authorized users.

Dietary Supplementation with Transgenic Camelina sativa Oil Containing 20:5n-3 and 22:6n-3 or Fish Oil Induces Differential Changes in the Transcriptome of CD3+ T Lymphocytes

“…The most altered pathways were associated with cell proliferation and immune response in stimulated compared to unstimulated cells. A previous RNAseq analysis of PBMC cultures stimulated with anti-CD3 antibodies also showed increased expression of pathways involved in immune function and cell division [51], although differences between this previous report and the present findings may be due to analysis of a mixture of cell types in the previous study. Both FO and tCSO increased the number of up-regulated transcripts in pathways involved in cell division [52] and immune-regulatory pathways such as interleukin-17 and interleukin-10 signalling [20,53].…”

Suppressing Kv1.3 Ion Channel Activity with a Novel Small Molecule Inhibitor Ameliorates Inflammation in a Humanised Mouse Model of Ulcerative Colitis

“… 39 In addition, the activation of T cells by anti-CD3 antibodies results in increased expression of various cytokines, including IFNy. 30 As shown in this study, DES1 and ShK are potent inhibitors of all three outcomes, as expected based on ~1000-fold differences in potencies against Kv1.3 obtained from patch clamp measurements. DES1 required ~10x–100x higher concentrations than ShK to elicit its ex vivo efficacy [IC50 values of ~100 nM vs ~50–500 pM for DES1 and ShK, respectively, against Kv1.3 obtained from electrophysiology with the channel expressed in mammalian cells].…”

“…DES1 inhibited the expression of these surface markers in a concentration-dependent manner, and the inhibition reached significance at 1000 nM [CD25+: p = 0.001; CD69+: p = 0.0005]. As T-cell activation by anti-CD3 antibodies results in cytokine release, 30 supernatants were additionally analysed by Luminex assays for IFNy, IL-4, and IL-10 levels. As shown in Figure 2C , anti-CD3 significantly increased the secretion of cytokines [IFNy: p = 0.0006; IL-4: p = 0.002; IL-10 p = 2 E-05].…”

Insights From Single Cell RNA Sequencing Into the Immunology of Type 1 Diabetes- Cell Phenotypes and Antigen Specificity

“…scRNAseq of T cells from the clinical trials of teplizumab and other immunotherapies in T1D could offer an amazing opportunity to identify all biomarkers predictive of successful treatment. For example, scRNAseq studies have shown a variety of phenotypic markers induced in vitro with anti-CD3 antibodies in human PBMC, including a variety of interleukin receptors and markers of regulation and exhaustion including FOXP3, CTLA4, TNFRSF18, LAG3 and PDCD1 (102). In contrast, anti-CD3/ CD28 stimulation of PBMC analysed with scRNAseq and CITEseq, showed phenotypes strongly associated with activation (although memory subsets also upregulated senescence) (103).…”