Dynamics of Donor T Cell Activation During Early Phase of Graft Versus Host Disease (GVHD): Donor Antigen Presenting Cells Can Activate Allogeneic T Cells

Abstract Growth arrest-specific gene 6 (Gas6) is expressed in antigen-presenting cells and endothelial cells (ECs) but not in T cells. When wild-type (WT) or Gas6−/− mice received allogeneic non–T cell–depleted bone marrow cells, hepatic graft-versus-host disease (GVHD) was alleviated in Gas6−/− recipients regardless of donor genotype, but not in WT recipients. T-cell infiltration was more prominent and diffuse in WT than in Gas6−/− recipients' liver. When mice received 0.5 × 106 allogeneic T cells with T cell–depleted allogeneic bone marrow, clinical signs indicated that GVHD was less severe in Gas6−/− than in WT recipients, as shown by a significant improvement of the survival and reduced liver GVHD. These data demonstrate that donor cells were not involved in the protection mechanism. In addition, lack of Gas6 in antigen-presenting cells did not affect WT or Gas6−/− T-cell proliferation. We therefore assessed the response of WT or Gas6−/− ECs to tumor necrosis factor-α. Lymphocyte transmigration was less extensive through Gas6−/− than WT ECs and was not accompanied by increases in adhesion molecule levels. Thus, the lack of Gas6 in ECs impaired donor T-cell transmigration into the liver, providing a rationale for considering Gas6 pathway as a potential nonimmunosuppressive target to minimize GVHD in patients receiving allogeneic hematopoietic stem cell transplantation.

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111Selective depletion of human GVH-reactive T-cells by anti-CD95 mediated activation-induced cell death or via the T-cell activation antigen CD69 for preventing graft-versus-host disease in allo-BMT

Biology of Blood and Marrow Transplantation ◽

10.1016/s1083-8791(03)80112-x ◽

2003 ◽

Vol 9 (2) ◽

pp. 98

Author(s):

M. Nonn ◽

I. Tubbe ◽

C. Huber ◽

U.F. Hartwig

Keyword(s):

T Cells ◽

Cell Death ◽

T Cell ◽

T Cell Activation ◽

Graft Versus Host Disease ◽

Cell Activation ◽

Host Disease ◽

Graft Versus Host ◽

Activation Induced Cell Death ◽

Activation Antigen

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Plasmacytoid dendritic cells prime alloreactive T cells to mediate graft-versus-host disease as antigen-presenting cells

Blood ◽

10.1182/blood-2008-07-168609 ◽

2009 ◽

Vol 113 (9) ◽

pp. 2088-2095 ◽

Cited By ~ 77

Author(s):

Motoko Koyama ◽

Daigo Hashimoto ◽

Kazutoshi Aoyama ◽

Ken-ichi Matsuoka ◽

Kennosuke Karube ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

T Cell ◽

Graft Versus Host Disease ◽

Antigen Presenting Cells ◽

Host Disease ◽

Graft Versus Host ◽

Alloreactive T Cells ◽

Antigen Presenting

Dendritic cells (DCs) can be classified into 2 distinct subsets: conventional DCs (cDCs) and plasmacytoid DCs (pDCs). cDCs can prime antigen-specific T-cell immunity, whereas in vivo function of pDCs as antigen-presenting cells remains controversial. We evaluated the contribution of pDCs to allogeneic T-cell responses in vivo in mouse models of graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation by an add-back study of MHC-expressing pDCs into major histocompatibility complex-deficient mice that were resistant to GVHD. Alloantigen expression on pDCs alone was sufficient to prime alloreactive T cells and cause GVHD. An inflammatory environment created by host irradiation has the decisive role in maturing pDCs for T-cell priming but this process does not require Toll-like receptor signaling. Thus, functional outcomes of pDC–T-cell interactions depend on the immunologic context of encounter. To our knowledge, these results are the first to directly demonstrate an in vivo pathogenic role of pDCs as antigen-presenting cells in an antigen-specific T cell–mediated disease in the absence of other DC subsets and to provide important insight into developing strategies for tolerance induction in transplantation.

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Immunomodulatory Effects of Vitamin D: Implications for the Prevention and Treatment of Graft Versus Host Disease

Blood ◽

10.1182/blood.v112.11.1251.1251 ◽

2008 ◽

Vol 112 (11) ◽

pp. 1251-1251

Author(s):

Jacalyn Rosenblatt ◽

Adam Bissonnette ◽

Zekui Wu ◽

Baldev Vasir ◽

Corrine Zarwan ◽

...

Keyword(s):

Vitamin D ◽

T Cells ◽

T Cell ◽

Graft Versus Host Disease ◽

Tritiated Thymidine ◽

Antigen Presenting Cells ◽

Host Disease ◽

Graft Versus Host ◽

Post Transplant ◽

Antigen Presenting

Abstract Allogeneic bone marrow transplantation is a uniquely curative therapy for a subset of patients with hematologic malignancies. However, morbidity and mortality related to graft versus host disease remain substantial. Dendritic cells (DCs) are highly potent antigen presenting cells that play a crucial role in maintaining the balance of immune activation and tolerance as well as the nature of immune reconstitution post-transplant. Persistence of host DCs in the early post transplant period has been shown to play an important role in the activation and expansion of alloreactive lymphocytes and the concomitant risk of GVHD. A major focus of research to enhance outcomes following allogeneic transplantation involves the manipulation of DC recovery post-transplant to minimize activation of alloreactive lymphocytes while preserving anti-tumor immunity responsible for the graft versus disease effect. Vitamin D is a hormone involved in bone metabolism and calcium homeostasis. More recently, vitamin D has been shown to have potent immunomodulatory effects. We have evaluated the effect of vitamin D on the phenotypic and functional characteristics of DC and T cell populations. We have demonstrated that vitamin D hinders the maturation of DCs such that cells that are differentiated in its presence exhibit decreased expression of costimulatory molecules. Peripheral blood mononuclear cells (PBMCs) were isolated from leukopaks obtained from normal donors by ficoll density centrifugation. DCs were generated by culturing the monocyte enriched adherent fraction with GM-CSF (1000 U/ml) and IL-4 (1000 U/ml) for 5 days, followed by maturation in the presence of TNFa (25 ng/ml) for 48 hours. DCs were generated in the presence and absence of 10nM of 1,25 hydroxyvitamin D. Mean expression of the costimulatory molecule CD80 and the maturation marker CD83 decreased from 60% to 37% and 53% to 27% respectively in the presence of vitamin D (N=3). To assess the effect of vitamin D on the functional potency of DCs as antigen presenting cells, the capacity of mature DCs to stimulate allogeneic T cell proliferation in the presence of vitamin D was determined. Mature DCs were generated from peripheral blood precursors and then cultured with allogeneic T cells isolated by T cell separation column at a ratio of 1:10. After 5 days, cocultures were pulsed with tritiated thymidine overnight and proliferation was determined by measuring the stimulation index (SI) as defined by uptake of tritiated thymidine of cocultured cells/uptake of tritiated thymidine of T cells alone. Results of 10 serial studies demonstrated the addition of vitamin D resulted in a blunted T cell proliferative response, with mean SI that decreased from 13 to 5. Similarly, the addition of vitamin D to a coculture of DCs and autologous T cells resulted in a 50% reduction in the T cell proliferative response to tetanus toxoid, a recall antigen. In addition, T cells stimulated by allogeneic DCs in the presence of vitamin D were polarized to secrete Th2 cytokines. In three experiments, the mean percentage of T cells secreting IL-10 increased from 2.7% to 4.4%, while expression of IFNγ decreased from a mean of 2.6% to 1.9% in the presence of vitamin D. The presence of vitamin D did not induce FOXP3 expressing regulatory T cell populations, which accounted for 8% and 7% of the T cell population following stimulation by allogeneic DCs in the presence and absence of vitamin D respectively. These data suggest that exposure to vitamin D exerts a tolerizing influence on T cells mediated by its impact on antigen presenting cells. Vitamin D may therefore have a role in the prevention and treatment of graft versus host disease. A clinical trial evaluating the use of vitamin D in the early post-transplant period for the prevention of GVHD is planned.

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Abortive Proliferation of Rare T Cells Induced by Direct or Indirect Antigen Presentation by Rare B Cells In Vivo

Journal of Experimental Medicine ◽

10.1084/jem.187.10.1611 ◽

1998 ◽

Vol 187 (10) ◽

pp. 1611-1621 ◽

Cited By ~ 67

Author(s):

Sarah E. Townsend ◽

Christopher C. Goodnow

Keyword(s):

T Cells ◽

T Cell ◽

B Cells ◽

Antigen Presentation ◽

Cell Fate ◽

T Cell Activation ◽

Cell Activation ◽

Antigen Presenting Cells ◽

Antigen Presenting

Antigen-specific B cells are implicated as antigen-presenting cells in memory and tolerance responses because they capture antigens efficiently and localize to T cell zones after antigen capture. It has not been possible, however, to visualize the effect of specific B cells on specific CD4+ helper T cells under physiological conditions. We demonstrate here that rare T cells are activated in vivo by minute quantities of antigen captured by antigen-specific B cells. Antigen-activated B cells are helped under these conditions, whereas antigen-tolerant B cells are killed. The T cells proliferate and then disappear regardless of whether the B cells are activated or tolerant. We show genetically that T cell activation, proliferation, and disappearance can be mediated either by transfer of antigen from antigen-specific B cells to endogenous antigen-presenting cells or by direct B–T cell interactions. These results identify a novel antigen presentation route, and demonstrate that B cell presentation of antigen has profound effects on T cell fate that could not be predicted from in vitro studies.

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Expression of the T-Cell Activation Antigen, OX-40, Identifies Alloreactive T Cells in Acute Graft-Versus-Host Disease

Blood ◽

10.1182/blood.v89.12.4652 ◽

1997 ◽

Vol 89 (12) ◽

pp. 4652-4658 ◽

Cited By ~ 66

Author(s):

Thomas V. Tittle ◽

Andrew D. Weinberg ◽

Cara N. Steinkeler ◽

Richard T. Maziarz

Keyword(s):

T Cells ◽

T Cell Activation ◽

Graft Versus Host Disease ◽

Peripheral Blood ◽

Cd4 T Cells ◽

Cell Activation ◽

Peripheral Blood Lymphocytes ◽

Host Disease ◽

Graft Versus Host ◽

Acute Graft

Abstract The OX-40 molecule is expressed on the surface of recently activated T lymphocytes. The presence of OX-40 on CD4+ T cells was analyzed in a rat haplo-identical (parental → F1) bone marrow transplant model of acute graft-versus-host disease (aGVHD). Increased numbers of activated CD4+ T cells that expressed the OX-40 antigen were detected in peripheral blood soon after transplantation before the earliest sign of disease. The peak of OX-40 expression occurred 12 days posttransplantation with a range of 18% to 36% of circulating T cells and remained 10-fold above background, never returning to baseline. A slight increase in OX-40 expression (range, 1% to 6%) was also detected on peripheral blood lymphocytes from control syngeneic F1 → F1 recipients. OX-40+ T cells were isolated from spleen, skin, lymph node, and liver tissue of rats undergoing aGVHD, but not in syngeneic transplants. OX-40+ T cells isolated from these tissues were of donor origin and were shown to be allo-reactive. These data raise the possibility of using the OX-40 antibody to detect and deplete selectively the T cells that cause aGVHD.

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Inhibition of inositol kinase B controls acute and chronic graft-versus-host disease

Blood ◽

10.1182/blood.2019000032 ◽

2020 ◽

Vol 135 (1) ◽

pp. 28-40 ◽

Cited By ~ 2

Author(s):

Govindarajan Thangavelu ◽

Jing Du ◽

Katelyn G. Paz ◽

Michael Loschi ◽

Michael C. Zaiken ◽

...

Keyword(s):

T Cell ◽

T Cell Activation ◽

Graft Versus Host Disease ◽

Cell Activation ◽

Cytoplasmic Calcium ◽

Host Disease ◽

Graft Versus Host ◽

T Cell Apoptosis ◽

Graft Versus Leukemia ◽

Intracellular Ca

T-cell activation leads to regulated increases in cytoplasmic calcium through inositol 1,4,5-triphosphate (IP3), a process balanced by phosphorylation and inactivation of IP3 by inositol 1,4,5-trisphosphate 3-kinase B (Itpkb). The investigators demonstrate that inhibition of Itpkb sustains increased intracellular Ca, leads to T-cell apoptosis, and inhibits graft-versus-host disease without impairing graft-versus-leukemia effects.

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Immunologic Effects of CCR5 Blockade in Graft-Versus-Host Disease Prophylaxis

Blood ◽

10.1182/blood.v126.23.920.920 ◽

2015 ◽

Vol 126 (23) ◽

pp. 920-920 ◽

Cited By ~ 1

Author(s):

Ryan H Moy ◽

Austin P Huffman ◽

Lee P Richman ◽

Lisa Crisalli ◽

James A Hoxie ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Activation ◽

Graft Versus Host Disease ◽

Cell Activation ◽

Ccr5 Expression ◽

Allogeneic Hsct ◽

Graft Versus Host ◽

And Control ◽

Ccr5 Blockade

Abstract Background: Acute graft-versus-host disease (GVHD) is a major cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT). Experimental models and clinical studies suggest that blocking lymphocyte trafficking with the chemokine receptor CCR5 antagonist maraviroc (MVC) ameliorates visceral GVHD. However, the effects of CCR5 blockade on immune reconstitution and activation in allogeneic HSCT recipients remain unknown. Methods: We previously reported a phase I/II study of 38 high-risk patients undergoing reduced-intensity allogeneic HSCT with standard GVHD prophylaxis (tacrolimus, methotrexate) combined with brief (33-day) MVC treatment. We compared the clinical and immunologic outcomes of these patients to a contemporary control cohort of 115 consecutive patients undergoing allogeneic HSCT with standard GVHD prophylaxis alone. Multivariate analysis was used to assess the effect of MVC treatment on the incidence of GVHD. Flow cytometry on day 30 and day 60 PBMC samples was used to monitor immune cell subsets, T cell activation markers and chemokine receptor expression. We also used a multiplex Luminex assay to quantify serum cytokine levels at day 30. Results: Since our initial report, longer follow up (median 37 months) now shows that patients who received MVC have reduced incidence rates of acute grade II-IV GVHD (adjusted hazard ratio [aHR]=0.42, 95% CI 0.21-0.84, p=0.015) and grade III-IV GVHD (aHR=0.43, 95% CI 0.17-1.09, p=0.075) compared to our control cohort (Figure 1). There was no difference in chronic GVHD between the groups. Early lymphocyte recovery was similar between MVC and control cohorts, with no significant difference in absolute lymphocyte, CD4 T cell and CD8 T cell counts at day 30. We observed increased CCR5 expression on CD4 T cells, CD8 T cells and B cells at day 30 in response to MVC treatment (Figure 2A-B). However, CCR5 expression on these cell types equalized between MVC and control patients by day 60. Day 30 MVC samples also demonstrated decreased percentages of CD38+ CD4 T cells and HLA-DR+ CD8 T cells (Figure 2C-D), an effect that also faded by day 60. These data suggest that inhibition of lymphocyte trafficking with CCR5 blockade dampens peripheral T cell activation. Notably, some patients developed acute GVHD despite MVC prophylaxis, and therefore we investigated differences between MVC responders and non-responders. Patients who developed GVHD had a higher percentage of activated CD38+ CD4 T cells in peripheral blood (Figure 3A) and an increased proportion of naive CD4 (36.5% vs 18.1% p=0.005) and CD8 (35.5% vs 11.9% p=0.0008) T cells, which has been associated with the development of GVHD. We also observed higher CCR5 expression on CD8 T cells from MVC non-responders (CCR5 MFI 8800 vs 6595, p=0.02). Intriguingly, while MVC responders and non-responders had similar serum concentrations of CCR5 ligands (MIP-1a, MIP-1b, Rantes), MVC non-responders had elevated levels of CXCR3 ligands (IP-10, MIG) (Figure 3B-C). Taken together, these data suggest that lymphocyte migration via CXCR3 may be a potential escape mechanism for GVHD initiation and T cell activation in the setting of CCR5 blockade. Conclusion: Brief CCR5 blockade in high-risk patients inhibits T cell activation and reduces the incidence of GVHD. MVC does not inhibit early lymphocyte recovery, which was similar in treated and control patients. This supports the hypothesis that MVC treatment would not impair graft-versus-leukemia activity. Our analysis of immunologic outcomes of MVC reveals a distinct immunologic effect for CCR5 blockade in allogeneic HSCT recipients and suggests a novel resistance mechanism. These studies bolster CCR5 antagonism as an effective strategy for GVHD prevention and provide support for extended MVC treatment duration and investigation into CXCR3 as a therapeutic target. Figure 1. Incidence of GVHD in control and MVC cohorts Figure 1. Incidence of GVHD in control and MVC cohorts Figure 2. MVC increases CCR5 expression and dampens T cell activation at day 30 *p<0.05; **p<0.01; ***p<0.001 Figure 2. MVC increases CCR5 expression and dampens T cell activation at day 30 *p<0.05; **p<0.01; ***p<0.001 Figure 3. Increased T cell activation and serum CXCR3 ligands in MVC non-responders *p<0.05; ***p<0.001 Figure 3. Increased T cell activation and serum CXCR3 ligands in MVC non-responders *p<0.05; ***p<0.001 Disclosures Off Label Use: Off label use of CCR5 antagonist maraviroc for the prevention of graft-versus-host disease. Vonderheide:Pfizer: Research Funding. Porter:Pfizer: Research Funding.

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