Anti-Granzyme A Antibody [GA29] (A269845)

The pleiotropic cytokine interleukin (IL)-10 is best characterized by its ability to downregulate inflammation and promote peripheral tolerance. On the other hand, IL-10 was also found to maintain the effector response of CD8⁺ T cells and promote the expansion of tumor-resident CD8⁺ T cells. In diffuse large B cell lymphoma (DLBCL), the role of IL-10 has been characterized in tumor cells but not in CD8⁺ T cells. We found that CD8⁺ T cells in DLBCL presented robust interferon (IFN)-? expression early during TCR-activation but could not maintain this response later on, which was characterized by significantly lower CD8⁺ T cell degranulation and higher apoptosis. These observations were associated with higher PD-1 expression in DLBCL CD8⁺ T cells. Furthermore, the PD-1⁺ cells were strongly enriched in the IFN-?⁺, but not the IFN-?^-, fraction. Interestingly, exogenous IL-10 significantly improved the survival of DLBCL CD8⁺ T cells, and resulted in significantly higher IFN-?, ganzyme A and granzyme B expression in the absence of CD19⁺ tumor cells, and significantly improved CD8⁺ T cell-mediated specific lysis of CD19⁺ tumor cells. IL-10 did not alter the expression of PD-1 in DLBCL CD8⁺ T cells, but curiously, IL-10-treated DLBCL CD8⁺ T cells were less susceptible to PD-L1-mediated apoptosis. We then demonstrated that IL-10 treatment significantly elevated the expression of pro-survival factor Bcl-2. Blocking IL-10 resulted in higher apoptosis, fewer IFN-?⁺ CD8⁺ T cells, and lower Bcl-2 expression. IL-10 also significantly increased STAT3, but not STAT1, phosphorylation in CD8⁺ T cells. Together, these results suggested that IL-10 could enhance CD8⁺ T cell inflammation in DLBCL patients.

Diffuse large B-cell lymphoma (DLBCL) is a common and aggressive subtype of non-Hodgkin's lymphomas, with limited treatment options in refractory and relapsed patients. Growing evidence supports the notion that CD8⁺ T cell immunity could be utilized to eliminate B cell lymphomas. CXCR5⁺CD8⁺ T cell is a novel cell subtype and share CXCR5 expression with CD19⁺ tumor cells. In this study, we investigated the frequency and function of existing CXCR5⁺CD8⁺ T cells in DLBCL patients. We found that DLBCL patients as a group demonstrated significantly higher level of CXCR5⁺CD8⁺ T cells than healthy individuals, with huge variability in each patient. Using anti-CD3/CD28-stimulated CD8⁺ T cells as effector (E) cells and autologous CD19⁺ tumor cells as target (T) cells, at high E:T ratio, no difference between the intensities of CXCR5⁺CD8⁺ T cell- and CXCR5^-CD8⁺ T cell-mediated cytotoxicity were observed. However, at intermediate and low E:T ratios, the CXCR5⁺CD8⁺ T cells presented stronger cytotoxicity than CXCR5^-CD8⁺ T cells. The expressions of granzyme A, granzyme B, and perforin were significantly higher in CXCR5⁺CD8⁺ T cells than in CXCR5^-CD8⁺ T cells, with no significant difference in the level of degranulation. Tumor cells in DLBCL were known to secrete high level of interleukin 10 (IL-10). We therefore blocked the IL-10/IL-10R pathway, and found that the expressions of granzyme A, granzyme B, and perforin by CXCR5⁺CD8⁺ T cells were significantly elevated. Together, these results suggest that CXCR5⁺CD8⁺ T cells are potential candidates of CD8⁺ T cell-based immunotherapies, could mediate elimination of autologous tumor cells in DLBCL patients, but are also susceptible to IL-10-mediated suppression.