CHIMERISM AND CYTOTOXIC T LYMPHOCYTE UNRESPONSIVENESS AFTER NEONATAL INJECTION OF SPLEEN CELLS IN MICE EFFECTS OF T CELL DEPLETION AND OF A SEMIALLOGENEIC OR FULLY ALLOGENEIC INOCULUM

ABSTRACT There is an acute dearth of therapeutic interventions against visceral leishmaniasis that is required to restore an established defective cell-mediated immune response. Hence, formulation of effective immunotherapy requires the use of dominant antigen(s) targeted to elicit a specific antiparasitic cellular immune response. We implemented hybrid cell vaccination therapy in Leishmania donovani-infected BALB/c mice by electrofusing dominant Leishmania antigen kinetoplastid membrane protein 11 (KMP-11)-transfected bone marrow-derived macrophages from BALB/c mice with allogeneic bone marrow-derived dendritic cells from C57BL/6 mice. Hybrid cell vaccine (HCV) cleared the splenic and hepatic parasite burden, eliciting KMP-11-specific major histocompatibility complex class I-restricted CD8+ cytotoxic T-lymphocyte (CTL) responses. Moreover, splenic lymphocytes of HCV-treated mice not only showed the enhancement of gamma interferon but also marked an elevated expression of the Th2 cytokines interleukin-4 (IL-4) and IL-13 at both transcriptional and translational levels. On the other hand, IL-10 production from splenic T cells was markedly suppressed as a result of HCV therapy. CD8+ T-cell depletion completely abrogated HCV-mediated immunity and the anti-KMP-11 CTL response. Interestingly, CD8+ T-cell depletion completely abrogated HCV-induced immunity, resulting in a marked increase of IL-10 but not of IL-4 and IL-13. The present study reports the first implementation of HCV immunotherapy in an infectious disease model, establishing strong antigen-specific CTL generation as a correlate of HCV-mediated antileishmanial immunity that is reversed by in vivo CD8+ T-cell depletion of HCV-treated mice. Our findings might be extended to drug-nonresponsive visceral leishmaniasis patients, as well as against multiple infectious diseases with pathogen-specific immunodominant antigens.

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Human bone marrow and peripheral blood T lymphocyte depletion: efficacy and effects of both T cells and monocytes on growth of hematopoietic progenitors

Blood ◽

10.1182/blood.v65.3.663.bloodjournal653663 ◽

1985 ◽

Vol 65 (3) ◽

pp. 663-679

Author(s):

L Levitt ◽

TJ Kipps ◽

EG Engleman ◽

PL Greenberg

Keyword(s):

T Cells ◽

T Cell ◽

T Lymphocyte ◽

Peripheral Blood ◽

Cell Depletion ◽

Target Cells ◽

Facs Analysis ◽

Hematopoietic Progenitors ◽

T Cell Depletion

The efficacy of four separate methods of human bone marrow T lymphocyte depletion was assessed, and the effect of T cells and monocytes on in vitro growth of marrow (CFU-GEMM, BFU-E, and CFU-GM) and peripheral blood (BFU-E) hematopoietic progenitors was determined. Extent of T cell depletion was assessed by multiparameter fluorescent cell sorter (FACS) analysis and by functional studies. Cells staining positively by FACS analysis for one or more of three separate fluorescent pan-T cell monoclonal antibodies (MCAbs) comprised 8.4% to 9.5% of control marrow mononuclear cells (MNCs). T cells constituted 3.2% to 5.1% of marrow following single, sequential, or combination treatment with two different pan-T cell MCAbs (Leu 1 and TM1) plus complement, 1.5% to 2.2% of marrow following solid-phase immunoabsorption (“panning”), 0.2% of marrow after sheep cell rosetting, and only 0.05% of marrow after FACS selective cell sorting and gated separation. T cells made up 59% to 73% of control peripheral blood MNCs and 0.8% to 2.8% of peripheral MNCs following sheep cell rosetting plus treatment with Leu 1 MCAb and complement. Mitogen (PHA, Con A) and allogeneic MLC-induced blastogenic responses (stimulation indices, experimental/control or E/C) revealed a concordant decrement in marrow T cell function after MCAb plus complement (E/C of 3.9 to 9.0), after panning (E/C of 1.6 to 3.5) and after sheep cell rosetting (E/C of 0.7 to 1.3), compared with control marrow (E/C of 5.3 to 15.7). After T cell depletion, marrow BFU-E growth was 95% to 120% of control, CFU-GM growth was 90% to 108% of control, and CFU-GEMM growth was 89% to 111% of control. Marrow T cell and/or monocyte depletion did not alter erythropoietin-dependent BFU-E growth in the absence of Mo-conditioned medium (81% to 95% of control), and the addition of as many as 50 to 100 X 10(3) purified marrow monocytes or T cells to 10(5) autologous nonadherent T cell-depleted marrow target cells had a negligible (P greater than .1) effect on marrow BFU-E growth in vitro. Peripheral blood (PB) BFU-E/10(5) T- depleted target cells were 106% +/- 19% of expected; PB BFU-E growth was significantly diminished after monocyte depletion alone (7% +/- 6% of expected) or after monocyte plus T cell depletion (8% +/- 4% of expected).(ABSTRACT TRUNCATED AT 400 WORDS)

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Induction of neonatal tolerance to H-2k in B6 mice does not allow the emergence of T cells specific for H-2k plus vaccinia virus.

Journal of Experimental Medicine ◽

10.1084/jem.156.3.810 ◽

1982 ◽

Vol 156 (3) ◽

pp. 810-821 ◽

Cited By ~ 3

Author(s):

D H Schwartz ◽

P C Doherty

Keyword(s):

T Cells ◽

T Cell ◽

Vaccinia Virus ◽

Somatic Mutation ◽

T Lymphocyte ◽

Cytotoxic T Lymphocyte ◽

Spleen Cells ◽

Cell Ontogeny ◽

Stimulation Of ◽

Tolerized Mouse

Thymocytes and spleen cells from C57BL/6 mice (H-2b) neonatally tolerized to H-2k alloantigens do not generate an anti-vaccinia response restricted to H-2Kk when adoptively transferred to appropriate irradiated hosts. This is in sharp contrast to the case for negatively selected C57BL/6 spleen cells acutely depleted of alloreactivity. No evidence for suppression was found in cell mixture experiments. We have shown elsewhere that our neonatally tolerized animals have a centrally induced delection-type tolerance in the absence of obvious suppression.2 We now suggest that in the neonatally tolerized mouse, chronic, central delection of anti-H-2k clones during early T cell ontogeny eliminates the major source of cells able to give rise, via somatic mutation and expansion, to anti-H-2Kk + vaccinia specific cytotoxic T lymphocyte precursors (CTL-P) in the adult. A similar mechanism may operate in the (k + b) leads to b chimera; however, the presence of H-2kxb accessory and presenting cells may permit the eventual generation (via cross-stimulation) of an H-2k-restricted vaccinia-specific repertoire. This would account for our observation of such "aberrant recognition" CTL-P emerging in the spleens of older (k x b) leads to b chimeras.

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Indirect allorecognition of platelets by T helper cells during platelet transfusions correlates with anti-major histocompatibility complex antibody and cytotoxic T lymphocyte formation [published erratum appears in Blood 1995 Dec 15;86(12):4710]

Blood ◽

10.1182/blood.v86.2.805.bloodjournal862805 ◽

1995 ◽

Vol 86 (2) ◽

pp. 805-812 ◽

Cited By ~ 40

Author(s):

JW Semple ◽

ER Speck ◽

YP Milev ◽

V Blanchette ◽

J Freedman

Keyword(s):

T Cells ◽

Major Histocompatibility Complex ◽

T Cell ◽

Mhc Class I ◽

T Lymphocyte ◽

Nk Cell ◽

Cytotoxic T Lymphocyte ◽

Spleen Cells ◽

Histocompatibility Complex ◽

Platelet Transfusions

To study the cellular immunology of platelet-induced alloimmunization, a murine transfusion model was developed. BALB/c (H-2d) recipient mice were transfused weekly with 2 x 10(8) platelets or 10(3) leukocytes from C57BL/6 (H-2b) donor mice. Recipient antidonor major histocompatibility complex (MHC) class I alloantibodies could be detected in flow cytometric assays by the fifth platelet transfusion. In contrast, when leukocytes only were transfused, alloantibodies were not detected. In vitro assays demonstrated that murine H-2b platelets were positive for MHC class I expression but lacked MHC class II molecules on their membranes and were unable to stimulate proliferation or cytokine production when incubated with naive H-2d spleen cells. In vivo, however, platelet transfusions induced two distinct patterns of cell-mediated reactivity. First, during the initial transfusions and before alloantibody formation, there was induction of T-cell anergy, characterized by the inability of recipient T cells to respond to Concanavalin A (ConA) or to proliferate in an antidonor mixed lymphocyte reaction (MLR), together with suppressed natural killer (NK) cell activity. This unresponsiveness was associated with a transient increase in nitric oxide (NO)-dependent cytotoxicity and interleukin-1 (IL-1) production. Second, once alloantibodies developed, significantly increased antidonor CD8+ cytotoxic T lymphocyte (CTL) and NK cell responses were observed. At this time, when recipient spleen cells were depleted of CD8+ T cells and incubated with only donor platelets in 7- day antigen-presenting cell (APC) assays, enhanced proliferation and IL- 2 production occurred. These cellular responses were not seen when 10(3) allogeneic leukocytes were transfused. Thus, the results suggest that leukoreduced platelet transfusions induce antidonor MHC antibodies and CD8+ CTL responses in recipient mice. At the same time, the transfusions induced recipient CD4+ T-cell activation when incubated with donor platelets in the presence of syngeneic APCs, an indirect recognition pathway that correlates with the time of alloantibody production.

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Priming of T cells to Fas-mediated proliferative signals by interleukin-7

Blood ◽

10.1182/blood-2007-12-126698 ◽

2008 ◽

Vol 112 (4) ◽

pp. 1195-1204 ◽

Cited By ~ 14

Author(s):

Bence Rethi ◽

Nancy Vivar ◽

Stefano Sammicheli ◽

Caroline Fluur ◽

Nicolas Ruffin ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Lymphocyte ◽

Cell Receptor ◽

Cell Depletion ◽

Activated T Cells ◽

T Cell Depletion ◽

Interleukin 7 ◽

Costimulatory Signals ◽

Opposing Effects

Abstract T-cell depletion associated with HIV infection or cytoreductive therapies triggers potential T-cell regenerative mechanisms such as peripheral T-lymphocyte expansion to weak antigenic stimuli and the increased availability of interleukin-7 (IL-7), a cytokine with potent antiapoptotic and proliferative activities. Deleterious mechanisms also associated with lymphopenia, such as increased Fas expression and apoptosis of T cell, however, may result in opposing effects. In this study, we show that Fas molecules, primarily associated with T-cell depletion in lymphopenic settings, may also contribute to compensatory T-cell expansion through transmitting costimulatory signals to suboptimally activated T cells. Proliferation of T lymphocytes in response to concomitant Fas and T-cell receptor (TCR) triggering was shown to be increased in HIV-infected individuals compared with noninfected controls. As IL-7 levels are often elevated in lymphopenic individuals in association with increased Fas expression, we analyzed whether IL-7 would influence Fas-mediated proliferative signals in T cells. We show that IL-7 is able to increase the efficacy of Fas to induce proliferation of suboptimally activated T cells. Thus, high IL-7 levels associated with lymphopenic conditions may simultaneously induce sensitivity to Fas-mediated apoptosis in nonactivated T cells and increase Fas-induced costimulatory signals in T cells recognizing low-affinity antigens.

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Human bone marrow and peripheral blood T lymphocyte depletion: efficacy and effects of both T cells and monocytes on growth of hematopoietic progenitors

Blood ◽

10.1182/blood.v65.3.663.663 ◽

1985 ◽

Vol 65 (3) ◽

pp. 663-679 ◽

Cited By ~ 18

Author(s):

L Levitt ◽

TJ Kipps ◽

EG Engleman ◽

PL Greenberg

Keyword(s):

T Cells ◽

T Cell ◽

T Lymphocyte ◽

Peripheral Blood ◽

Cell Depletion ◽

Target Cells ◽

Facs Analysis ◽

Hematopoietic Progenitors ◽

T Cell Depletion

Abstract The efficacy of four separate methods of human bone marrow T lymphocyte depletion was assessed, and the effect of T cells and monocytes on in vitro growth of marrow (CFU-GEMM, BFU-E, and CFU-GM) and peripheral blood (BFU-E) hematopoietic progenitors was determined. Extent of T cell depletion was assessed by multiparameter fluorescent cell sorter (FACS) analysis and by functional studies. Cells staining positively by FACS analysis for one or more of three separate fluorescent pan-T cell monoclonal antibodies (MCAbs) comprised 8.4% to 9.5% of control marrow mononuclear cells (MNCs). T cells constituted 3.2% to 5.1% of marrow following single, sequential, or combination treatment with two different pan-T cell MCAbs (Leu 1 and TM1) plus complement, 1.5% to 2.2% of marrow following solid-phase immunoabsorption (“panning”), 0.2% of marrow after sheep cell rosetting, and only 0.05% of marrow after FACS selective cell sorting and gated separation. T cells made up 59% to 73% of control peripheral blood MNCs and 0.8% to 2.8% of peripheral MNCs following sheep cell rosetting plus treatment with Leu 1 MCAb and complement. Mitogen (PHA, Con A) and allogeneic MLC-induced blastogenic responses (stimulation indices, experimental/control or E/C) revealed a concordant decrement in marrow T cell function after MCAb plus complement (E/C of 3.9 to 9.0), after panning (E/C of 1.6 to 3.5) and after sheep cell rosetting (E/C of 0.7 to 1.3), compared with control marrow (E/C of 5.3 to 15.7). After T cell depletion, marrow BFU-E growth was 95% to 120% of control, CFU-GM growth was 90% to 108% of control, and CFU-GEMM growth was 89% to 111% of control. Marrow T cell and/or monocyte depletion did not alter erythropoietin-dependent BFU-E growth in the absence of Mo-conditioned medium (81% to 95% of control), and the addition of as many as 50 to 100 X 10(3) purified marrow monocytes or T cells to 10(5) autologous nonadherent T cell-depleted marrow target cells had a negligible (P greater than .1) effect on marrow BFU-E growth in vitro. Peripheral blood (PB) BFU-E/10(5) T- depleted target cells were 106% +/- 19% of expected; PB BFU-E growth was significantly diminished after monocyte depletion alone (7% +/- 6% of expected) or after monocyte plus T cell depletion (8% +/- 4% of expected).(ABSTRACT TRUNCATED AT 400 WORDS)

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The safety and effctiveness of donor memory T lymphocyte infusions after hematopoietic stem cell transplantation with ab T cell depletion platform in children with acute leukemia

Pediatric Hematology/Oncology and Immunopathology ◽

10.24287/1726-1708-2021-20-2-12-28 ◽

2021 ◽

Vol 20 (2) ◽

pp. 12-28

Author(s):

M. A. Dunaykina ◽

L. N. Shelikhova ◽

Zh. B. Shekhovtsova ◽

S. Yu. Glushkova ◽

R. V. Nikolayev ◽

...

Keyword(s):

T Cell ◽

T Lymphocyte ◽

Conditioning Regimen ◽

Cumulative Risk ◽

Cell Depletion ◽

Control Group ◽

T Cell Depletion ◽

Hematopoietic Stem ◽

Grade Ii ◽

Related Mortality

T-cell ab depletion prevents “graft-versus-host” disease (GVHD), does not impair engraftment, and improves the outcomes of hematopoietic stem cell transplantation (HSCT) from a haploidentical donor. Memory T lymphocyte infusions (CD45RA-depleted) can transfer functional immunity to common pathogens to recipients. In a randomized study, we explored the safety and effctiveness of donor memory T lymphocyte infusions (DLI) in children with leukemia after HSCT with ab T cell depletion platform. The study was approved by the Independent Ethics Committee and the Scientifi Council of the Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology of the Ministry of Healthcare of the Russian Federation. A total of 149 patients were enrolled in the study; 76 patients were randomly assigned to the DLI group and 73 patients were allocated to the control group. Donors were haploidentical related in 91% of cases. The myeloablative conditioning regimen included treosulfan and total body irradiation. Anti-thymocyte globulin (ATG) was excluded from the conditioning regimen, instead, we used a combination of abatacept and tocilizumab. Graft processing involved TCRab-/CD19-depletion. The main parameters of assessment included the cumulative risk of detection of cytomegalovirus (CMV) DNA and the cumulative risk of grade II–IV GVHD. The additional parameters of assessment were the cumulative risk of transplant-related mortality, the cumulative risk of relapse, the overall and event-free survival rates, and the parameters of immune recovery. A historical control group was used to compare the primary outcomes of HSCT with ATG and an alternative immunomodulatory regimen (abatacept and tocilizumab). The cumulative risk of grade II–IV GVHD was 14% in the experimental group and 12% in the control group (p = 0.8). The cumulative risk of CMV viremia was 45% and 55% in the experimental and control groups, respectively (p = 0.4). In the prospective cohort, the rates of transplant-related mortality, the cumulative risk of relapse, and the overall survival were 2%, 25%, and 80%, respectively, without statistical diffrence between the arms. In the experimental group, we noticed a tendency toward an increase in the proportion of patients who developed an immune response to CMV in the early post-HSCT period. The substitution of ATG with tocilizumab and abatacept was not accompanied by a higher incidence of GVHD or graft failure; it was associated with signifiantly lower transplant-related mortality rates (2% vs 13%, p = 0.002) and improved immune recovery in the early post-HSCT period. Prophylactic infusions of donor memory lymphocytes are safe and may be used for further improvement in. the results of HSCT with ab T cell depletion platform.

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