IL-12hi Rapamycin-Conditioned Dendritic Cells Mediate IFN-γ-Dependent Apoptosis of Alloreactive CD4+ T Cells and Limit Graft-Versus-Host Diseas

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4110-4110
Author(s):  
Elizabeth O. Stenger ◽  
Brian R. Rosborough ◽  
Lisa Mathews ◽  
Huihui Ma ◽  
Markus Mapara ◽  
...  
Keyword(s):  
T Cells ◽  
Stem Cell ◽  
T Cell ◽  
Cell Apoptosis ◽  
Cd4 T Cells ◽  
Annexin V ◽  
Hematopoietic Stem ◽  
Graft Versus Host ◽  
T Cell Apoptosis ◽  
Ifn Γ

Abstract Abstract 4110 Introduction: Rapamycin (RAPA) inhibits the serine/threonine kinase mammalian Target of Rapamycin to profoundly modulate immune cell function. Dendritic cells (DC) exposed to RAPA (RAPA-DC) exhibit tolerogenic properties, including promotion of experimental cardiac allograft survival. RAPA-DC enrich for CD4+FoxP3+ regulatory T cells and induce alloreactive T cell apoptosis and anergy. Yet, paradoxically, RAPA-DC secrete increased IL-12, which is central for the generation of IFN-γ+CD4+ T helper type 1 cells. IFN-γ can be pro-apoptotic, and IL-12-driven IFN-γ inhibits graft-versus-host disease (GVHD) following hematopoietic stem cell transplantation. We hypothesized that IL-12hi RAPA-DC, unlike control (CTR)-DC, would be effective in the prevention of GVHD by supporting IFN-γ-mediated apoptosis of alloreactive T cells. Methods: DC were generated from C57BL/6 (H2-b) or B6.129S7-Ifngr1tm1Agt/J (IFN-γ-R−/− [H2-b]) bone marrow (BM) in 7 day (d) culture in the absence (CTR-DC) or presence of RAPA (RAPA-DC). CTR- and RAPA-DC cultures remained either untreated or were stimulated via TLR4 with LPS (100 ng/ml) for an additional 18 hours. Washed and CD11c+ purified DC from these groups were then used as stimulators of allogeneic CD4+ T cells (BALB/c [H2-d] or CByJ.129S7(B6)-Ifngr1tm1Agt/J [IFN-γ-R−/− {H2-d}]) in 5 d mixed leukocyte reaction (MLR) with or without anti-IFN-γ antibody. T cell apoptosis was quantified using an Annexin V Apoptosis Detection kit. The capacity of DC to prevent GVHD was assessed in irradiated BALB/c mice reconstituted with 5×106 T cell-depleted B6 BM on d0. Recipient mice received 1×106 CD11c+ BALB/c DC (CTR-, RAPA-, or LPS-exposed DC) on d0 and 1×106 B6 T cells on d1. Mice were monitored daily, and moribund mice or those with >20% weight loss were euthanized. Results: Compared to CTR-DC, especially when LPS-stimulated, RAPA-DC induced increased apoptosis (Annexin V+7-AAD+) of alloreactive CD4+ T cells in MLR (13.9±1.6% versus 7.1±0.2%; +LPS: 30.6±3.4% versus 14.7±3.6%; both p<0.05). Neutralization of IFN-γ in co-cultures containing LPS-stimulated RAPA-DC decreased levels of apoptosis to those of LPS-exposed CTR-DC (18.5±1.3% versus 14.7±3.6%; NS). IFN-γ-R−/− CD4+ T cells exposed to LPS-stimulated IL-12hi RAPA-DC exhibited decreased apoptosis when compared to wild-type CD4+ T cells (WT 15.7±3.6% versus IFN-γ-R−/− 4.9±1.1%; p<0.05) (Figure 1). There was no difference in the extent of apoptosis of alloreactive CD4+ T cells induced by IFN-γ-R−/− RAPA-DC compared to WT RAPA-DC (10.6±7.6% versus 12.4±7.6% respectively; +LPS: 30.1±16.3% versus 27.3±12.0%; both p=NS) (Figure 2). The addition of anti-IFN-γ mAb to LPS-stimulated IFN-γ-R−/− RAPA-DC significantly decreased levels of CD4+ T cell apoptosis comparable to LPS-stimulated WT CTR-DC (18.0±10.3% versus 22.0±6.3%, p<0.05). Depletion of IFN-γ did not impact on the Treg-enriching capacity (CD4hiFoxP3+/CD4hiFoxP3−) of RAPA-DC (24.2±0.4% versus 29.2±6.2% with anti-IFN-γ mAb; p=NS). Further, whereas CTR-DC and LPS-exposed CTR-DC did not prolong survival, IL-12hi RAPA-DC significantly prolonged survival from GVHD (median survival in days: GVHD, 17.5 d; RAPA-DC, 27 d, p=NS compared to GVHD; RAPA-DC + LPS, 35 d, p<0.01; syngeneic control, >50 d, p<0.01) (Figure 3). In contrast, CTR-DC had comparable GVHD mortality (21 d, p=NS). Conclusions: Increased apoptosis induced by LPS-stimulated IL-12hi RAPA-DC is mediated directly on CD4+ allogeneic T cells via IFN-γ and not through actions on DC. Thus, increased IL-12 may support IFN-γ-mediated activation-induced cell death while sparing regulatory T cells and may underlie the capacity of LPS-exposed RAPA-DC to prevent GVHD. IL-12hi human RAPA-DC, generated with the aid of endotoxin-free, synthetic TLR4 agonists, may offer a means to harness the demonstrated capacity of both IL-12 and tolerogenic DC to prevent GVHD following hematopoietic stem cell transplant. Disclosures: No relevant conflicts of interest to declare.

Increased Effector CD4+ T-Cells Early Post Hematopoietic Stem Cell Transplantation Using an Alemtuzumab-Based Reduced Intensity Conditioning Regimen Correlate with Subsequent Development of Graft Versus Host Disease.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4999-4999
Author(s):  
Katie Matthews ◽  
ZiYi Lim ◽  
Laurence Pearce ◽  
Khalid Tobal ◽  
Alejandro Madrigal ◽  
...  
Keyword(s):  
T Cells ◽  
Stem Cell ◽  
T Cell ◽  
Hematopoietic Stem Cell Transplantation ◽  
Cd4 T Cells ◽  
Subsequent Development ◽  
Effector T Cells ◽  
Cd4 T Cell ◽  
Hematopoietic Stem ◽  
Graft Versus Host

Abstract Lymphocyte depletion using the anti CD52 monoclonal antibody alemtuzumab reduces the incidence of graft versus host disease (GvHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT), but some patients still develop this potentially life-threatening complication. We previously reported that patients achieving rapid full donor T cell chimerism after fludarabine, busulphan and alemtuzumab (FBC) conditioned allo-HSCT have a significantly increased risk of GvHD compared to patients with prolonged mixed donor chimerism beyond day 100 (Lim et al. Br. J. Haematology 2007). We performed a prospective study of 29 patients who received allo-HSCT with FBC conditioning (median age: 53 years; range: 34 – 69 presenting with AML or MDS) to examine the kinetics of lymphocyte reconstitution in relation to T-cell chimerism patterns and incidence of GvHD. Naïve, memory, effector and terminally differentiated CD4+ and CD8+ T-cells, activated T-cells (CD25+ HLA-DR+); putative regulatory CD4+ CD25high Foxp3+ T-cells, B-cells and NK cells were enumerated in whole peripheral blood of patients at days 30, 60, 90, 180, 270 and 360 after HSCT. Chimerism analysis of purified T-cells was performed by genetic profiling of polymorphic short tandem repeat loci. Ten patients developed GvHD (acute or chronic). Although alemtuzumab induced profound depletion of all T-cell subsets, significantly higher numbers of CD4+ effector (CD45RO+ CD27−) T-cells were detected at day 30 post transplant in patients who later developed GvHD (24 cells/μl; range: 1 – 84 cells/μl) compared to patients without GvHD (5 cells/μl; range: 1 – 40 cells/μl) (p = 0.026). In contrast, there were no significant differences in the numbers or rate of reconstitution of CD8+ T-cell sub-populations, NK cells or B-cells in patients that developed GvHD and those who did not at any time points. T-cells present at day 30 in patients that subsequently developed GvHD were 100% donor whereas the majority of patients that did not develop GvHD exhibited mixed donor and recipient T cell chimerism. Development of GvHD pathology was associated with expansion of these donor effector CD4+ T-cells (at day 60: 35 cells/μl; range: 9 – 154 cells/μl compared to 7 cells/μl; range: 1 – 56 cells/μl for patients without GvHD, p = 0.04). Absolute numbers of CD4+ CD25high Foxp3+ T-cells at day 30 were similar in both groups of patients (p = 0.8). However, of note, a significant deficit of these putative regulatory T-cells in the group that developed GvHD was apparent when numbers were considered relative to CD4+ effector T cells at day 30 (41 CD4+ effector T-cells; range: 28 – 51 /per regulatory CD4+ T cell for the GvHD group compared to 12 CD4+ effector T-cells; range: 2 – 33 /per regulatory CD4+ T-cell for patients without GvHD, p = 0.03). We speculate the higher numbers of effector CD4+ T-cells detected in patients at day 30 post HSCT are donor-derived mature T cells that alemtuzumab fails to deplete. In the solid organ transplant setting, alemtuzumab has been shown to be relatively sparing of effector memory CD4+ T-cells. Our correlation of donor-derived effector CD4+ T-cells with subsequent development of GvHD suggests they are alloreactive and that a deficit of T-regs relative to CD4+ effector T-cells early post HSCT contributes to GvHD.

PD-1/PD-L Pathway Potentially Involved in ITP Immunopathogenesis

2019 ◽  
Vol 119 (05) ◽  
pp. 758-765 ◽  
Author(s):  
Mu Nie ◽  
Yang Liu ◽  
Xiu-xiu Li ◽  
Ya-nan Min ◽  
Dan-dan Yang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
T Cell ◽  
Cd8 T Cells ◽  
Cell Apoptosis ◽  
Cd4 T Cells ◽  
Peripheral Tolerance ◽  
Healthy Controls ◽  
T Cell Apoptosis ◽  
Ifn Γ

AbstractThe binding of programmed death 1 (PD-1) to its ligands PD-L1 and PD-L2 on antigen-presenting cells turns off autoreactive T cells and induces peripheral tolerance. Aberrant PD-1/PD-L signalling could result in a breakdown of peripheral tolerance and lead to autoimmune diseases. In this study, we detected PD-1 and PD-L expression on T cells and dendritic cells (DCs) in immune thrombocytopenia (ITP) patients with active disease by flow cytometry. The effects of PD-L1-Fc fusion protein (PD-L1-Fc) on T cells and on secretion of interferon-γ (IFN-γ) and interleukin-2 (IL-2) were detected by flow cytometry and enzyme-linked immunosorbent assay, respectively. Compared with healthy controls, PD-1 expression was significantly increased in CD4+ T cells and CD8+ T cells from patients with active ITP. However, PD-L1 expression on monocyte-derived DCs was lower in patients with active ITP than in healthy controls. In vitro assays revealed that PD-L1-Fc increased T cell apoptosis, inhibited activation and proliferation of CD4+ T cells and CD8+ T cells and decreased IFN-γ and IL-2 secretion in patients with active ITP. These results suggest that the aberrant PD-1/PD-L negative co-stimulatory pathway may play a role in ITP. Enhancing PD-1/PD-L signalling might be a promising therapeutic approach for ITP patients by enhancing T cell apoptosis, inhibiting T cell activation and proliferation and reducing secretion of inflammatory factors.

Functional Comparison and Longitudinal Assessment of Tri-Functional T-Cells Recognizing CMV pp65 and IE-1 Polypeptides in Hematopoietic Stem Cell and Solid Organ Transplant Recipients.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2936-2936
Author(s):  
Don J. Diamond ◽  
Simon F. Lacey ◽  
Corinna La Rosa ◽  
Wendy Zhou ◽  
Ghislaine Gallez-Hawkins ◽  
...  
Keyword(s):  
T Cells ◽  
Stem Cell ◽  
T Cell ◽  
Solid Organ ◽  
Transplant Recipients ◽  
Hematopoietic Stem ◽  
Specific Ctls ◽  
Tnf Α ◽  
Ifn Γ ◽  
Cmv Disease

Abstract Reconstitution of adaptive T-cell responses to human cytomegalovirus (CMV) is critical to protection from CMV disease following hematopoietic stem cell (HSCT) or solid organ transplantation (SOT). However, there is an incomplete understanding of which CMV antigens and epitopes are most crucial to providing protective responses. The functional status of cytotoxic T-lymphocyte (CTL) populations recognizing cytomegalovirus IE-1 and pp65 polypeptides was investigated in PBMC from either HSCT or SOT recipients. Our previous finding of differing levels of degranulation between CMV IE1 and pp65/pp50 specific T-cells was complicated by the possibility that differences were epitope and/or HLA-specific. We generalized the approach using a combined flow-based CD107a/b degranulation/mobilization and intracellular cytokine (ICC) assays using peptide libraries as antigens. These assays indicated that a significantly higher proportion of pp65-specific CTLs were in a more mature functional state compared to IE-1-specific CTLs. Degranulation/multicytokine ICC assays also indicated that a significantly higher proportion of the pp65-specific versus IE-1-specific CTLs secreted both IFN-γ and TNF-α, in addition to possessing greater cytotoxic potential. These results support our earlier findings of functional differences between CTLs recognizing individual epitopes within the IE-1 and pp65 antigens in HSCT recipients, and extend them to a broader array of HLA-restricted responses to those antigens. A report that a subset of HIV-1 specific CTLs capable of producing both IFN-γ and TNF-α was associated with improved cytotoxic activity prompted us to investigate whether degranulation, a functional correlate of cytotoxicity, was positively associated with dual cytokine production and predicted differences between IE1 and pp65-specific CD8+ T-cells. A higher proportion of pp65-specific compared to IE1-specific T-cells were present in the trifunctional IFN-γ+,TNF-α+, CD107+ population (p=0.008) in HSCT recipients. We have extended these findings to investigate the role of donor CMV status in terms of functional maturity of CMV-specific T cell response in transplant recipients. T cell maturation/function may act as a mechanistic correlate to the survival advantage of recipients receiving a stem-cell graft from CMV sero-positive donors. These principles have also been applied to investigations of a high risk population of sero-negative recipients of a sero-positive liver allograft. Data from this study will also be reviewed in the context of the model of trifunctional T cells being indicative of enhanced protective capacity against CMV disease and associated with survival.

Donor BM Dendritic Cells Expressing IDO Limit Graft-Versus-Host Disease After Allogeneic Bone Marrow Transplant.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 1331-1331
Author(s):  
Ying Lu ◽  
Wayne Harris ◽  
Jian-Ming Li ◽  
Edmund K. Waller
Keyword(s):  
T Cells ◽  
T Cell ◽  
Marrow Transplant ◽  
Allogeneic Bone ◽  
Transplant Recipients ◽  
Hematopoietic Stem ◽  
Graft Versus Host ◽  
Donor T Cells ◽  
Ifn Γ ◽  
Th1 Polarization

Abstract Abstract 1331 Poster Board I-353 Background In contrast to the essential role of host dendritic cells (DC) in the initiation of graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) reactions, less is known about the effects of donor DC on T cells in these processes. We have previously reported that adding donor BM plasmacytoid DC (pDC) progenitors to a murine graft composed of purified hematopoietic stem cells (HSC) and T-cells increased donor activation and Th1 polarization leading to enhanced GVL activity without increasing GVHD (Li et al. 2007 Blood 110:2181), while larger numbers of human donor pDC were associated with less GVL activity following allogeneic bone marrow transplant (BMT) (Waller et al. 2001 Blood 97:2948). To explore the dissociation of GVHD from GVL we tested the hypothesis that activation of donor T-cells by donor pDC leads to reciprocal induction of indoleamine 2,3-dioxygenase (IDO) expression and immune counter-regulatory activity by donor DC that limits donor T-cell allo-reactivity. Methods pDC precursors were purified by high-speed FACS from un-stimulated BM harvested from wild type (WT) and IDO knock-out (IKO) mice. T-cell proliferation and immune polarization in response to indirect antigen presentation by syngenic DC was measured in mixed lymphocyte reaction (MLR) and by recovery of CFSE-labeled donor T-cells from allogeneic transplant recipients. IDO expression in DC was measured by FACS and intracellular staining using pDC from IKO BM as a negative staining control. FACS-purified 5 × 104 pDC either from WT mice or from IKO mice in combination with 3 × 103 c-kit+ Sca-1+ hematopoietic stem cells (HSC) and 3 × 105 T-cells were transplanted in MHC mismatched C57BL/6→B10.BR model following lethal irradiation. Results FACS-purified lineage−CD11cloCD11b− pDC expressed B220 (72%), CD90 (51%), and CD317 (PDCA-1) (93%), had low levels of MHC-II, partial expression of CD4, and lacked expression of CD24, CD80, CD86 and NK cell or granulocytic markers. IDO expression in purified pDC was up regulated by IFN-γ produced by syngenic T-cells in vitro in one-way MLR. In vivo proliferation of CFSE-labeled donor T-cells was enhanced in mice that received pDC from either WT or IKO mice. Co-transplantation of IKO pDC led to higher proliferation rates of CD8+ T-cells but not CD4+ T-cells compared with the proliferation of corresponding donor T-cell subset co-transplanted with WT DC. The incidence and severity of GVHD (weight loss and GVHD score) were markedly increased in recipients receiving pDC from IKO mice as compared with mice receiving WT pDC. The enhanced GVL activity of donor T-cells induced by transplanted donor WT pDC was abolished when IKO pDC were transplanted into tumor-bearing recipients. Transplanting WT donor pDC led to larger numbers of donor-derived CD4+CD25+Foxp3+ T-reg cells in the spleens of transplant recipients compared with mice receiving IKO pDC (p<0.01) in combination with purified HSC and T-cells. Conclusions Taken together, our data suggest IDO expression in pDC as a critical downstream event that inhibits continued T-cell activation and GVHD. We propose a feedback model in which donor pDC initially induce Th1 polarization of activated donor CD8+ T-cells that secret high levels of IFN-γ. IDO expressed by donor pDC in response to local IFN-γ subsequently induces a counter-regulatory effect including the generation of T-reg and down-modulation of CD8+ T-cell allo-reactivity and proliferation, limiting GVHD while preserving the GVL activity of donor T-cells. Disclosures No relevant conflicts of interest to declare.

Increased Functional T Cell Defects in Patients with low CD4 Counts after Allogeneic Hematopoietic Stem Cell Transplantation

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 4588-4588
Author(s):  
Udo Holtick ◽  
Lukas P. Frenzel ◽  
Shimabukuro-Vornhagen Alexander ◽  
Sebastian Theurich ◽  
Julia Claasen ◽  
...  
Keyword(s):  
T Cells ◽  
Stem Cell ◽  
T Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Cd4 T Cells ◽  
Cd4 T Cell ◽  
Hematopoietic Stem ◽  
Cd4 Counts ◽  
Graft Vs Host Disease

Background The recovery of the host immune system after allogeneic hematopoietic stem cell transplantation is pivotal to prevent infections, relapse and secondary malignancies. In particular, numerical CD4 T-cell reconstitution is delayed and CD4-helper cell function considered impaired as consequence of the transplant procedure and concommitant immunosuppressive medication. From HIV/AIDS patients it is known that numerical and functional CD4 defects increase the risk of opportunistic infections. Therefore, even in the absence of immunosuppressants and graft-vs-host disease, anti-infective prophylaxis is usually given for at least six months. We hypothesized that the numerical CD4 defect in patients may be reflected by immunosuppressive RNA fingerprints previously established for certain immuno-inhibitory molecules and tested whether the functional CD4 capacity was different according to the CD4 cell number. Methods RNA was separated from CD4 T-cells of 10 patients with CD4 counts >500/µl, 10 patients with CD4 counts <200/µl and four healthy controls. All patients had to be off immunosuppression and without any clinical signs of graft-vs-host disease. Transcriptional activity was assessed with regard to previously defined fingerprints motives for CTLA-4, IL-10, PD-1, TGF-β and PGE-2. CD4 T-cells from all groups were further tested for their proliferative capacity and cytokine production. Results Hierarchical clustering segregated the three groups. Applying the immunosuppressive fingerprints, patients with CD4 T-cells >500/µl were demonstrated to be under the influence of PGE2, whereas patients with CD4 T-cells <200/µl were demonstrated to be under the influence of PGE2 and CTLA-4. In normal controls, no association was found. The proliferative capacity of patient CD4 T-cells upon CD3-CD28-bead stimulation was not significantly different from healthy controls. The production of IL-2 by stimulated CD4 T-cells was significantly downregulated in patients with CD4 T-cells <200/µl, while there was no difference in IFN-ƴ and TNF-α secretion. Conclusion The severity of the CD4 numerical defect reflects the state of immunosuppression as demonstrated by RNA immuno-inhibitory fingerprint motives. This partially translates into functional differences as measured by decreased IL-2 secretion. In addition to time after transplant, CD4 T-cell numbers should be considered for the decision to stop or maintain anti-microbial prophylaxis in patients after allogeneic stem cell transplantation. (UH, LPF and CW, JMC contributed equally to this work.) Disclosures: No relevant conflicts of interest to declare.

scholarly journals β2- Adrenergic Signaling Regulates Graft Versus Host Disease after Allogenic Transplantation While Preserving Graft Versus Leukemia Effect

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 1915-1915 ◽  
Author(s):  
Hemn Mohammadpour ◽  
Joseph L. Sarow ◽  
George L. Chen ◽  
Cameron R. MacDonald ◽  
Umesh Sharma ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cd4 T Cells ◽  
Adrenergic Receptor ◽  
Graft Versus Host ◽  
Graft Versus Leukemia ◽  
Donor T Cells ◽  
Β2 Adrenergic Receptor ◽  
Ifn Γ

β2 adrenergic receptor signaling is a key regulator of various immune cells, including T cells; however, its role in T cell function in the context of graft versus host disease (GvHD) is poorly understood. We previously showed that housing mice at thermoneutral temperature (TT; 30°C), which reduces systemic adrenergic stress, increased the incidence and severity of GvHD after allogeneic hematopoietic cell transplant (allo-HCT) compared to mice housed at standard temperature (ST; 22°C) which exerts a mild but chronic adrenergic stress (Leigh et al J Immunol 2015). The increased incidence and severity of GvHD in TT mice can be reversed by the administration of a β2-adrenergic receptor (β2-AR) agonist, suggesting an important role of epinephrine and norepinephrine in allo-HCT outcome (Leigh et al., J. Immunol 2015; Mohammadpour et al J Immunol 2018). We investigated the mechanisms and downstream events of β2-AR signaling in donor T cells after allo-HCT by using β2-AR knockout (β2-AR-/-) mice and commercially available β2-AR agonists. The main goal here was to explore whether signaling through β2-AR in donor T cells could control GvHD incidence and severity without minimizing the graft-versus leukemia (GvL) effect. We utilized both a major MHC-mismatch C57B6 (H-2kb) into BALB/c (H-2kd) model and a MHC-matched, multiple minor histocompatibility antigen (miHA) mismatched B6 (H-2kb) into C3H/SW (H-2kb) model. Recipient BALB/c and C3H/SW WT mice were lethally irradiated with 850 and 1100 cGy respectively and injected by tail vein with T cell depleted bone marrow (TCD-BM) alone (3 ×106) or TCD-BM and splenic T cells derived from allogeneic WT or β2-AR-/- B6 donors (0.7 × 106 T cells in B6 → BALB/c and 1.5 × 106 in B6 → C3H/SW). We found that donor T cells express β2-AR after allo-HCT and that β2-AR expression on WT T cells plays an important role in controlling GvHD, as evidenced by less severe weight loss, and increased survival compared to mice receiving β2-AR-/- donor T cells (Figure 1A). Histopathologic examination showed that β2-AR-/- T cells induced more damage in the small and large intestine. To explore further the mechanism(s) by which β2-AR signaling controls the severity of GvHD, we used NanoString analysis and discovered that β2-AR-/- T cells have the Th1 phenotype with an increase in Tbx21, Ifng, Irf8 and Emoes genes, while WT CD4+ T cells had higher levels of Th2 and Treg associated genes, including Foxp3, Ptgs5, Tgfb2, Il10, Il21 and Il22. We also observed a significant increase in the inflammatory cytokines IFN-γ and IL-17 in β2-AR-/- CD4+ T cells from the spleen and liver on days 7 and 14 after allo-HCT as compared to WT T cells (Figure 1B), while the expression of IL-10 was significantly higher in WT T cells compared to β2-AR-/- T cells (P< 0.01). We next sought to determine whether GvL may be affected by use of long acting β2-AR agonist (Bambuterol) to control GvHD. Bambuterol was administered daily at a dose of 1mg/kg from day 0. We observed that Bambuterol controlled the severity and mortality of GvHD after allo-HCT in both major and minor mismatch mouse models, as evidenced by reduced weight loss and an improved clinical score and survival rate in mice receiving Bambuterol compared to vehicle (P<0.001). We showed that treatment increased the expression of IL-10 and decreased the expression of IFN-γ and IL-17 in CD4+ T cells. Interestingly, we found that β2-AR agonist treatment significantly increased the generation of myeloid derived suppressor cells (MDSCs) from WT BM without any effect on β2-AR-/- BM both in vitro and in vivo, suggesting an important role of β2-AR signaling in the generation of MDSCs. To investigate the effect of Bambuterol on GvL, the A20 lymphoma cell line was injected 4 hours before allo-HCT. Using two different doses of T cells (0.5 × 106 and 0.2 × 106) in B6 → BALB/c model, we found that Bambuterol preserved GvL by inducing CD44+ CD62L- NKG2D+ effector cells and CD44+ CD62L+ central memory cells. Since β2-AR agonists can affect cardiac function, we measured heart rate (HR) and blood pressure (BP) using a tail-cuff. There was no difference in BP and HR at day 21 and 28 after allo-HCT between mice receiving Bambuterol compared to mice receiving vehicle. In conclusion, these data reveal how β-AR signaling can influence donor T cell differentiation and function in murine GvHD models without decreasing GvL effect pointing to the feasibility of manipulation of β2-AR signaling to ameliorate clinical GvHD. Disclosures No relevant conflicts of interest to declare.

scholarly journals Prior Use of Mogamulizumab to Allogenic Hematopoietic Stem Cell Transplantation Induces Severe Acute Graft-Versus-Host Disease

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 1940-1940 ◽  
Author(s):  
Takeshi Sugio ◽  
Koji Kato ◽  
Takatoshi Aoki ◽  
Takanori Ota ◽  
Noriyuki Saito ◽  
...  
Keyword(s):  
Bone Marrow ◽  
T Cells ◽  
Stem Cell ◽  
T Cell ◽  
Regulatory T Cells ◽  
Research Funding ◽  
Acute Gvhd ◽  
Cell Lymphoma ◽  
Hematopoietic Stem ◽  
Graft Versus Host

Abstract [Introduction] Adult T-cell leukemia/lymphoma (ATL) is an aggressive peripheral T-cell lymphoma (PTCL) with a dismal prognosis. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the only curative treatment in ATL patients. Mogamulizumab, a humanized anti-CC chemokine receptor 4 (CCR4) monoclonal antibody, is a novel immunotherapeutic agent, effective in treating patients with PTCL such as ATL, PTCL-not specified, and cutaneous T-cell lymphoma. However, in allo-HSCT setting, we should be careful to use mogamulizumab because CCR4 is expressed in regulatory T cells: The mogamulizumab treatment may accelerate GVHD by eradicating regulatory T cells in allo-HSCT patients. Here, we retrospectively analyzed the effect of mogamulizumab on GVHD development in ATL patients treated with mogamulizumab prior to allo-HSCT. [Patients and Methods] Data from the Fukuoka Bone Marrow Transplantation Group were retrospectively analyzed after the approval of mogamulizumab use in Japan. [Results] A total of 24 patients with ATL received mogamulizumab prior to allo-HSCT between April 2012 and April 2015 in our group. The median age at allo-HSCT was 58.5 years (range, 32-72). The median intervals from the last administration of mogamulizumab to allo-HSCT were 25 days (range, 9-126). The median total dose of mogamulizumab was 3 mg/kg (range, 1-8 mg/kg). After treatment with mogamulizumab, 18 patients (75%) had achieved in remission (CR in 4 patients and PR in 14) at allo-HSCT. Ten patients received unrelated bone marrow, 5 received related peripheral blood, and 9 received cord blood as stem cell sources. Eleven patients were treated with full-intensity conditioning and 13 received reduced-intensity conditioning. Graft-versus-host disease (GVHD) prophylaxis consisted of calcineurin inhibitors (cyclosporine or tacrolimus) with short-term methotrexate in 14 patients and mycophenolate mofetil in 9. The cumulative incidence (CI) of acute GVHD at 100 days was 66.6% in grade 2-4 and 33.3% in grade 3-4. The involved organs of acute GVHD were skin in 14 patients, gut in 10, and liver in 4. Among 14 patients who developed grade 2-4 acute GVHD, 5 had severe fluid retention such as pleural effusion or ascites associated with GVHD. Chronic GVHD was observed in 6 patients, and 5 of them were extensive disease. The CI of transplant-related mortality (TRM) and relapse at 1-year were 53.2% (95%CI, 29.3-72.3%) and 29.6% (95%CI, 12.6-48.9%), respectively. The leading cause of death was GVHD (n = 7). The 1-year overall survival and progression-free survival were 19.2% (95%CI, 5.7-38.8%) and 17.2% (95%CI, 4.9-35.7%), respectively. [Discussion] Use of mogamulizumab prior to transplantation in allo-HSCT patients has a merit to decrease the burden of ATL cells. However, it was associated with an increase of TRM due to severe GVHD. Although most of ATL patients achieved better disease status at allo-HSCT through mogamulizumab and the survival rate was expected to be 50% based on the previous data, the survival in the present study was ~20%. These data suggest that mogamulizumab administered before transplantation may have retained until an early phase of post-transplantation, and the donor or host-derived regulatory T cells might be eliminated, allowing the GVHD T-cell clone to expand. Since mogalizumab is a potent anti-ATL agent, we need to develop new treatment protocols integrating mogalizumab at a suitable dose or administration timing, to minimize the unwanted GVHD development in future studies. Disclosures Akashi: Asahi Kasei: Research Funding, Speakers Bureau; Shionogi: Research Funding, Speakers Bureau; Astellas: Research Funding, Speakers Bureau; Celgene: Research Funding, Speakers Bureau; Chugai: Research Funding, Speakers Bureau; Bristol-Myers Squibb: Research Funding, Speakers Bureau; Novartis Pharma K.K.: Consultancy, Research Funding, Speakers Bureau; Kyowa Hakko Kirin Co., Ltd.: Consultancy, Research Funding, Speakers Bureau.

scholarly journals Effects of HCV co-infection on apoptosis of CD4+ T-cells in HIV-positive patients

2009 ◽  
Vol 116 (12) ◽  
pp. 861-870 ◽  
Author(s):  
Christian Körner ◽  
Benjamin Krämer ◽  
Daniela Schulte ◽  
Martin Coenen ◽  
Stefan Mauss ◽  
...  
Keyword(s):  
T Cells ◽  
Hepatitis C ◽  
T Cell ◽  
Antiretroviral Therapy ◽  
Hiv Infection ◽  
Cell Apoptosis ◽  
Cd4 T Cells ◽  
Cd4 T Cell ◽  
Hiv Positive ◽  
T Cell Apoptosis

Apoptosis importantly contributes to loss of CD4+ T-cells in HIV infection, and modification of their apoptosis may explain why HIV/HCV (hepatitis C virus)-co-infected patients are more likely to die from liver-related causes, although the effects of HCV on HIV infection remain unclear. In the present study, we studied in a cross-sectional and serial analysis spontaneous ex vivo CD4+ T-cell apoptosis in HIV/HCV-co-infected and HIV-mono-infected patients before and after HAART (highly active antiretroviral therapy). Apoptosis of peripheral blood CD4+ T-cells was measured by both a PARP [poly(ADP-ribose) polymerase] and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling) assay to detect cells with irreversible apoptosis. Although hepatitis C alone did not increase CD4+ T-cell apoptosis, HCV co-infection disproportionately increased elevated rates of apoptosis in CD4+ T-cells from untreated HIV-positive patients. Increased CD4+ T-cell apoptosis was closely correlated with HIV, but not HCV, viral loads. Under HAART, increased rates of CD4+ T-cell apoptosis rapidly decreased both in HIV-mono-infected and HIV/HCV-co-infected patients, without any significant difference in apoptosis rates between the two patient groups after 4 weeks of therapy. Nevertheless residual CD4+ T-cell apoptosis did not reach the normal levels seen in healthy controls and remained higher in HIV patients receiving protease inhibitors than in patients with other antiretroviral regimens. The results of the present study suggest that HCV co-infection sensitizes CD4+ T-cells towards apoptosis in untreated HIV-positive patients. However, this effect is rapidly lost under effective antiretroviral therapy.

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