Abstract: EP911
Type: e-Poster
Background
The hallmark of both activated and dysfunctional T cells is the upregulation of inhibitory signal (“checkpoint”) receptors (PD-1, TIM-3, LAG-3 etc). The counts of T cells expressing checkpoint molecules are increased in various hematologic malignancies, including multiple myeloma (MM). While high-dose chemotherapy (HDC) with autologous hematopoietic stem cell transplantation (AHSCT) is a milestone of the treatment for eligible MM patients, there is a lack of data regarding the post-transplant reconstitution of PD-1- and TIM-3-positive T cells.
Aims
The purpose of the study was to evaluate the recovery of PD-1+ and TIM-3+ T cells and their functional activity following AHSCT in MM patients.
Methods
Peripheral blood (PB) samples of 25 healthy donors, 26 MM patients in complete remission/partial response before conditioning, 28 patients at the day of engraftment and 15 individuals following 6 post-transplant months were obtained during routine diagnostic procedures. Simultaneously, bone marrow aspirates were collected from the same pre-conditioning patients (n=18) and 15 individuals following 6 months post-AHSCT. The expression of PD-1 and TIM-3, intracellular production of IFNγ and Granzyme B (GrB), intracellular expression of Ki-67 by CD4+ and CD8+ T cells were evaluated using flow cytometry.
Results
Relative counts of PD-1+ and TIM-3+ subsets of CD4+ and CD8+ T cells were higher in PB of MM patients before HDC and following AHSCT (both at the engraftment day and 6 month follow-up) compared with the healthy donors. Absolute amounts of PD-1+ and TIM-3+ subsets did not differ between patients and donors, except CD4+TIM3+ and PD-1+TIM-3+ subsets of CD4+ and CD8+ T cells. Percentages of the evaluated T cell subsets restored rapidly following AHSCT; the majority of them (except PD-1+ subset of CD8+ T cells) were significantly higher at the day of engraftment compared with the pre-transplant levels. Absolute counts of studied subsets following AHSCT were nearly equal or lower compared with the values before HDC. Relative counts of PD-1+ subset of CD8+ T cells and both PD-1+ and TIM-3+ subsets of CD4+ T cells were higher in BM compared with PB. BM samples also contained higher amounts of double-positive PD-1+TIM-3+ subsets of CD8+ and CD4+ T cells compared with PB (Figure).
PD-1+ T cells before and after AHSCT had a pronounced GrB and IFNγ production, the cytotoxic and cytokine-producing activity of TIM3+ (especially PD-1+TIM-3+) T cells was significantly reduced, compared with PD-1- and TIM-3-negative subsets, respectively. Before conditioning, the percentages of Ki-67+ cells were the highest in CD4+PD-1+ cells and the lowest in CD8+PD-1+TIM-3+ T cells. At the engraftment day the proportion of Ki-67+ cells dramatically increased: the lowest value was 28.9 % (14.9—64.1 %) for CD4+TIM-3+ T cells, and the highest was 62.4 % (49.1—66.8 %) for CD8+PD-1+ subset. Following six months, proliferative capacity of studied subsets decreased, but remained higher compared with PD-1- and TIM-3-negative subsets.
Conclusion
T cells expressing PD-1 and/or TIM-3 recovered quickly following HDC with AHSCT, and reached or exceeded pre-conditioning counts at the engraftment. PD-1-positive T cells possessed preserved or increased functional properties before and after AHSCT in MM patients; “more dysfunctional” T cells were associated with TIM-3+ subsets. Proliferative ability at early post-transplant period increased even in dysfunctional TIM-3+ T cell subsets.
The Russian Science Foundation under grant № 18-75-00050 supports this work.
Session topic: 13. Myeloma and other monoclonal gammopathies - Biology & Translational Research
Keyword(s): Autologous hematopoietic stem cell transplantation, Multiple myeloma, T cell activation
Abstract: EP911
Type: e-Poster
Background
The hallmark of both activated and dysfunctional T cells is the upregulation of inhibitory signal (“checkpoint”) receptors (PD-1, TIM-3, LAG-3 etc). The counts of T cells expressing checkpoint molecules are increased in various hematologic malignancies, including multiple myeloma (MM). While high-dose chemotherapy (HDC) with autologous hematopoietic stem cell transplantation (AHSCT) is a milestone of the treatment for eligible MM patients, there is a lack of data regarding the post-transplant reconstitution of PD-1- and TIM-3-positive T cells.
Aims
The purpose of the study was to evaluate the recovery of PD-1+ and TIM-3+ T cells and their functional activity following AHSCT in MM patients.
Methods
Peripheral blood (PB) samples of 25 healthy donors, 26 MM patients in complete remission/partial response before conditioning, 28 patients at the day of engraftment and 15 individuals following 6 post-transplant months were obtained during routine diagnostic procedures. Simultaneously, bone marrow aspirates were collected from the same pre-conditioning patients (n=18) and 15 individuals following 6 months post-AHSCT. The expression of PD-1 and TIM-3, intracellular production of IFNγ and Granzyme B (GrB), intracellular expression of Ki-67 by CD4+ and CD8+ T cells were evaluated using flow cytometry.
Results
Relative counts of PD-1+ and TIM-3+ subsets of CD4+ and CD8+ T cells were higher in PB of MM patients before HDC and following AHSCT (both at the engraftment day and 6 month follow-up) compared with the healthy donors. Absolute amounts of PD-1+ and TIM-3+ subsets did not differ between patients and donors, except CD4+TIM3+ and PD-1+TIM-3+ subsets of CD4+ and CD8+ T cells. Percentages of the evaluated T cell subsets restored rapidly following AHSCT; the majority of them (except PD-1+ subset of CD8+ T cells) were significantly higher at the day of engraftment compared with the pre-transplant levels. Absolute counts of studied subsets following AHSCT were nearly equal or lower compared with the values before HDC. Relative counts of PD-1+ subset of CD8+ T cells and both PD-1+ and TIM-3+ subsets of CD4+ T cells were higher in BM compared with PB. BM samples also contained higher amounts of double-positive PD-1+TIM-3+ subsets of CD8+ and CD4+ T cells compared with PB (Figure).
PD-1+ T cells before and after AHSCT had a pronounced GrB and IFNγ production, the cytotoxic and cytokine-producing activity of TIM3+ (especially PD-1+TIM-3+) T cells was significantly reduced, compared with PD-1- and TIM-3-negative subsets, respectively. Before conditioning, the percentages of Ki-67+ cells were the highest in CD4+PD-1+ cells and the lowest in CD8+PD-1+TIM-3+ T cells. At the engraftment day the proportion of Ki-67+ cells dramatically increased: the lowest value was 28.9 % (14.9—64.1 %) for CD4+TIM-3+ T cells, and the highest was 62.4 % (49.1—66.8 %) for CD8+PD-1+ subset. Following six months, proliferative capacity of studied subsets decreased, but remained higher compared with PD-1- and TIM-3-negative subsets.
Conclusion
T cells expressing PD-1 and/or TIM-3 recovered quickly following HDC with AHSCT, and reached or exceeded pre-conditioning counts at the engraftment. PD-1-positive T cells possessed preserved or increased functional properties before and after AHSCT in MM patients; “more dysfunctional” T cells were associated with TIM-3+ subsets. Proliferative ability at early post-transplant period increased even in dysfunctional TIM-3+ T cell subsets.
The Russian Science Foundation under grant № 18-75-00050 supports this work.
Session topic: 13. Myeloma and other monoclonal gammopathies - Biology & Translational Research
Keyword(s): Autologous hematopoietic stem cell transplantation, Multiple myeloma, T cell activation
Abstract: EP911
Type: e-Poster
Background
The hallmark of both activated and dysfunctional T cells is the upregulation of inhibitory signal (“checkpoint”) receptors (PD-1, TIM-3, LAG-3 etc). The counts of T cells expressing checkpoint molecules are increased in various hematologic malignancies, including multiple myeloma (MM). While high-dose chemotherapy (HDC) with autologous hematopoietic stem cell transplantation (AHSCT) is a milestone of the treatment for eligible MM patients, there is a lack of data regarding the post-transplant reconstitution of PD-1- and TIM-3-positive T cells.
Aims
The purpose of the study was to evaluate the recovery of PD-1+ and TIM-3+ T cells and their functional activity following AHSCT in MM patients.
Methods
Peripheral blood (PB) samples of 25 healthy donors, 26 MM patients in complete remission/partial response before conditioning, 28 patients at the day of engraftment and 15 individuals following 6 post-transplant months were obtained during routine diagnostic procedures. Simultaneously, bone marrow aspirates were collected from the same pre-conditioning patients (n=18) and 15 individuals following 6 months post-AHSCT. The expression of PD-1 and TIM-3, intracellular production of IFNγ and Granzyme B (GrB), intracellular expression of Ki-67 by CD4+ and CD8+ T cells were evaluated using flow cytometry.
Results
Relative counts of PD-1+ and TIM-3+ subsets of CD4+ and CD8+ T cells were higher in PB of MM patients before HDC and following AHSCT (both at the engraftment day and 6 month follow-up) compared with the healthy donors. Absolute amounts of PD-1+ and TIM-3+ subsets did not differ between patients and donors, except CD4+TIM3+ and PD-1+TIM-3+ subsets of CD4+ and CD8+ T cells. Percentages of the evaluated T cell subsets restored rapidly following AHSCT; the majority of them (except PD-1+ subset of CD8+ T cells) were significantly higher at the day of engraftment compared with the pre-transplant levels. Absolute counts of studied subsets following AHSCT were nearly equal or lower compared with the values before HDC. Relative counts of PD-1+ subset of CD8+ T cells and both PD-1+ and TIM-3+ subsets of CD4+ T cells were higher in BM compared with PB. BM samples also contained higher amounts of double-positive PD-1+TIM-3+ subsets of CD8+ and CD4+ T cells compared with PB (Figure).
PD-1+ T cells before and after AHSCT had a pronounced GrB and IFNγ production, the cytotoxic and cytokine-producing activity of TIM3+ (especially PD-1+TIM-3+) T cells was significantly reduced, compared with PD-1- and TIM-3-negative subsets, respectively. Before conditioning, the percentages of Ki-67+ cells were the highest in CD4+PD-1+ cells and the lowest in CD8+PD-1+TIM-3+ T cells. At the engraftment day the proportion of Ki-67+ cells dramatically increased: the lowest value was 28.9 % (14.9—64.1 %) for CD4+TIM-3+ T cells, and the highest was 62.4 % (49.1—66.8 %) for CD8+PD-1+ subset. Following six months, proliferative capacity of studied subsets decreased, but remained higher compared with PD-1- and TIM-3-negative subsets.
Conclusion
T cells expressing PD-1 and/or TIM-3 recovered quickly following HDC with AHSCT, and reached or exceeded pre-conditioning counts at the engraftment. PD-1-positive T cells possessed preserved or increased functional properties before and after AHSCT in MM patients; “more dysfunctional” T cells were associated with TIM-3+ subsets. Proliferative ability at early post-transplant period increased even in dysfunctional TIM-3+ T cell subsets.
The Russian Science Foundation under grant № 18-75-00050 supports this work.
Session topic: 13. Myeloma and other monoclonal gammopathies - Biology & Translational Research
Keyword(s): Autologous hematopoietic stem cell transplantation, Multiple myeloma, T cell activation
Abstract: EP911
Type: e-Poster
Background
The hallmark of both activated and dysfunctional T cells is the upregulation of inhibitory signal (“checkpoint”) receptors (PD-1, TIM-3, LAG-3 etc). The counts of T cells expressing checkpoint molecules are increased in various hematologic malignancies, including multiple myeloma (MM). While high-dose chemotherapy (HDC) with autologous hematopoietic stem cell transplantation (AHSCT) is a milestone of the treatment for eligible MM patients, there is a lack of data regarding the post-transplant reconstitution of PD-1- and TIM-3-positive T cells.
Aims
The purpose of the study was to evaluate the recovery of PD-1+ and TIM-3+ T cells and their functional activity following AHSCT in MM patients.
Methods
Peripheral blood (PB) samples of 25 healthy donors, 26 MM patients in complete remission/partial response before conditioning, 28 patients at the day of engraftment and 15 individuals following 6 post-transplant months were obtained during routine diagnostic procedures. Simultaneously, bone marrow aspirates were collected from the same pre-conditioning patients (n=18) and 15 individuals following 6 months post-AHSCT. The expression of PD-1 and TIM-3, intracellular production of IFNγ and Granzyme B (GrB), intracellular expression of Ki-67 by CD4+ and CD8+ T cells were evaluated using flow cytometry.
Results
Relative counts of PD-1+ and TIM-3+ subsets of CD4+ and CD8+ T cells were higher in PB of MM patients before HDC and following AHSCT (both at the engraftment day and 6 month follow-up) compared with the healthy donors. Absolute amounts of PD-1+ and TIM-3+ subsets did not differ between patients and donors, except CD4+TIM3+ and PD-1+TIM-3+ subsets of CD4+ and CD8+ T cells. Percentages of the evaluated T cell subsets restored rapidly following AHSCT; the majority of them (except PD-1+ subset of CD8+ T cells) were significantly higher at the day of engraftment compared with the pre-transplant levels. Absolute counts of studied subsets following AHSCT were nearly equal or lower compared with the values before HDC. Relative counts of PD-1+ subset of CD8+ T cells and both PD-1+ and TIM-3+ subsets of CD4+ T cells were higher in BM compared with PB. BM samples also contained higher amounts of double-positive PD-1+TIM-3+ subsets of CD8+ and CD4+ T cells compared with PB (Figure).
PD-1+ T cells before and after AHSCT had a pronounced GrB and IFNγ production, the cytotoxic and cytokine-producing activity of TIM3+ (especially PD-1+TIM-3+) T cells was significantly reduced, compared with PD-1- and TIM-3-negative subsets, respectively. Before conditioning, the percentages of Ki-67+ cells were the highest in CD4+PD-1+ cells and the lowest in CD8+PD-1+TIM-3+ T cells. At the engraftment day the proportion of Ki-67+ cells dramatically increased: the lowest value was 28.9 % (14.9—64.1 %) for CD4+TIM-3+ T cells, and the highest was 62.4 % (49.1—66.8 %) for CD8+PD-1+ subset. Following six months, proliferative capacity of studied subsets decreased, but remained higher compared with PD-1- and TIM-3-negative subsets.
Conclusion
T cells expressing PD-1 and/or TIM-3 recovered quickly following HDC with AHSCT, and reached or exceeded pre-conditioning counts at the engraftment. PD-1-positive T cells possessed preserved or increased functional properties before and after AHSCT in MM patients; “more dysfunctional” T cells were associated with TIM-3+ subsets. Proliferative ability at early post-transplant period increased even in dysfunctional TIM-3+ T cell subsets.
The Russian Science Foundation under grant № 18-75-00050 supports this work.
Session topic: 13. Myeloma and other monoclonal gammopathies - Biology & Translational Research
Keyword(s): Autologous hematopoietic stem cell transplantation, Multiple myeloma, T cell activation