Adoptive immunotherapy with tumor-derived donor lymphocytes after allogeneic hematopoietic stem cell transplantation

Treatment of refractory or recurrent malignancy with donor lymphocyte infusion (DLI) after allogeneic hematopoietic stem cell transplantation (alloHSCT) is limited in efficacy, and graft-vs-tumor (GVT) is often accompanied by graft-versus-host disease (GVHD). After alloHSCT, lymphocytes infiltrating residual tumor are likely of donor origin. Compared with DLI, they may provide enhanced antigen specificity and maintain tumor-specific homing, thus generate better GVT with less GVHD. We are testing this hypothesis through administration of ex-vivo activated tumor-derived lymphocytes (TDL) after alloHSCT. Clinical evaluation of TDL therapy was initiated with a 51 year-old woman for metastatic breast cancer whose disease progressed after matched-sibling alloHSCT and subsequent conventional therapy plus DLI. Metastases were surgically removed two weeks after administration of unmanipulated DLI. Lymphocytes were liberated from 9.4 cm of tumor and expanded for 14 days with anti-CD3/CD28-coated magnetic beads (3:1 bead-to-total nucleated cell ratio) and media containing IL-2 (100 or 1000 IU/mL). The cell products from the two culture conditions were similar. The process yielded 42.5 × 106 cells, 33% expressing CD3, and generated 14.7 × 109 TDL, 85% expressing CD3 (a 3.1-log T cell expansion). There was no tumor contamination of the T cell product by immunohistochemistry. Chimerism analysis revealed the TDL to be of donor origin. Flow cytometry showed an increase in the CD4/CD8 ratio from 1.3 to 1.9 after expansion. 76% of CD8 and 31% of CD4 cells secrete IFN-γ, and none secretes IL-2, IL-4 or IL-10. 76% of CD8 and 57% of CD4 cells expressed CXCR3. Three infusions of TDL were given in a dose-escalating manner (5, 25 and 100 × 106 CD3+ cells/kg). Two additional infusions were given in conjunction with low-dose IL-2, the second of which was preceded by one cycle of paclitaxel and trastuzumab cytoreductive therapy. No infusion-related or delayed toxicities were observed. The patient had no evidence of GVHD, even after the highest dose of 108 allogeneic T cells. Evaluation of the remaining thoracic lesion demonstrated progressive disease after the first two TDL infusions, transient disease stability after the third and fourth infusions, and at present, the patient has stable disease one month after the fifth infusion. This is the first clinical report of the application of TDL and represents a novel approach for adoptive immunotherapy in the setting of alloHSCT. Treatment of refractory or recurrent malignancy with donor lymphocyte infusion (DLI) after allogeneic hematopoietic stem cell transplantation (alloHSCT) is limited in efficacy, and graft-vs-tumor (GVT) is often accompanied by graft-versus-host disease (GVHD). After alloHSCT, lymphocytes infiltrating residual tumor are likely of donor origin. Compared with DLI, they may provide enhanced antigen specificity and maintain tumor-specific homing, thus generate better GVT with less GVHD. We are testing this hypothesis through administration of ex-vivo activated tumor-derived lymphocytes (TDL) after alloHSCT. Clinical evaluation of TDL therapy was initiated with a 51 year-old woman for metastatic breast cancer whose disease progressed after matched-sibling alloHSCT and subsequent conventional therapy plus DLI. Metastases were surgically removed two weeks after administration of unmanipulated DLI. Lymphocytes were liberated from 9.4 cm of tumor and expanded for 14 days with anti-CD3/CD28-coated magnetic beads (3:1 bead-to-total nucleated cell ratio) and media containing IL-2 (100 or 1000 IU/mL). The cell products from the two culture conditions were similar. The process yielded 42.5 × 106 cells, 33% expressing CD3, and generated 14.7 × 109 TDL, 85% expressing CD3 (a 3.1-log T cell expansion). There was no tumor contamination of the T cell product by immunohistochemistry. Chimerism analysis revealed the TDL to be of donor origin. Flow cytometry showed an increase in the CD4/CD8 ratio from 1.3 to 1.9 after expansion. 76% of CD8 and 31% of CD4 cells secrete IFN-γ, and none secretes IL-2, IL-4 or IL-10. 76% of CD8 and 57% of CD4 cells expressed CXCR3. Three infusions of TDL were given in a dose-escalating manner (5, 25 and 100 × 106 CD3+ cells/kg). Two additional infusions were given in conjunction with low-dose IL-2, the second of which was preceded by one cycle of paclitaxel and trastuzumab cytoreductive therapy. No infusion-related or delayed toxicities were observed. The patient had no evidence of GVHD, even after the highest dose of 108 allogeneic T cells. Evaluation of the remaining thoracic lesion demonstrated progressive disease after the first two TDL infusions, transient disease stability after the third and fourth infusions, and at present, the patient has stable disease one month after the fifth infusion. This is the first clinical report of the application of TDL and represents a novel approach for adoptive immunotherapy in the setting of alloHSCT.