Skin biopsies for early diagnosis and prognosis of graft-versus-host disease in recipients of allogeneic stem cells from blood or bone marrow

We have investigated the effects of the in vitro depletion of LFA1 positive cytolytic T lymphocytes, natural killer (NK) cells, and monocytes on the afferent phase of graft-versus-host disease (GVHD). Lethal GVHD was induced across the murine major histocompatibility complex by injecting C57BL/6 (H-2b) bone marrow (BM) cells (a source of stem cells) and splenocytes (S) (a source of T cells) into lethally irradiated B10.BR (H-2k) recipients. Because anti-LFA1 does not bind complement (C′) effectively, we conjugated anti-LFA1 alpha chain monoclonal antibody (MoAb) to ricin toxin A chain (RTA) as a means of facilitating target cell elimination. A 2-hour preincubation of C57BL/6 bone marrow/spleen (BMS) with anti-LFA1-RTA in the presence of ammonium chloride (a potentiator of immunotoxin toxicity), but not a control immunotoxin (IT), reduced CTL activity by greater than 2 logs, significantly reduced NK cell activity, and prevented B10.BR mice from developing GVHD. Depletion of target cells by toxin-labeled-MoAb and not the blockade of the LFA1 molecule by the anti-LFA1 MoAb accounted for our results, because incubating cells with IT in the absence of a potentiator had no effect on GVHD prevention. In contrast, C57BL/6 recipients of C3H BMS grafts only partially benefited from anti-LFA1- RTA preincubation, demonstrating that in this system, different cells not expressing LFA1 were involved in GVHD generation. The same findings observed with anti-LFA1-RTA preincubation were observed with preincubation with L-leucyl-L-leucine methyl ester, a chemical compound eliminating cytolytic cells, providing further support that GVHD induction in the C3H/HeJ into C57BL/6 system is not entirely mediated by classical cytolytic T cells. We next tested anti-LFA1-RTA in a model devised to measure its effect on alloengraftment (B10.BR recipients given lower doses of irradiation). Anti-LFA1-RTA BM preincubation selectively reduced alloengraftment in the model. This observation, combined with experiments showing that LFA1-RTA preincubation, but not anti-Thy 1.2 + C' or control IT preincubation, reduced colony-forming unit-spleen formation, indicates that anti-LFA1 alpha chain IT may remove accessory cells or stem cells critical to engraftment. Still, anti-LFA1-RTA may be useful for clinical GVHD prevention when combined with positive selection techniques designed to enrich for stem cells.

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Ex Vivo Gene Therapy: Graft-versus-host Disease (GVHD) in NSG™ (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) Mice Transplanted with CD34+ Human Hematopoietic Stem Cells

Toxicologic Pathology ◽

10.1177/0192623319844484 ◽

2019 ◽

Vol 47 (5) ◽

pp. 656-660 ◽

Cited By ~ 3

Author(s):

Sundeep Chandra ◽

Patrizia Cristofori ◽

Carlos Fonck ◽

Charles A. O’Neill

Keyword(s):

Stem Cells ◽

Gene Therapy ◽

Bone Marrow ◽

Hematopoietic Stem Cells ◽

Graft Versus Host Disease ◽

Ex Vivo ◽

Hematopoietic Stem ◽

Host Disease ◽

Graft Versus Host ◽

Human Cd34

A therapeutic option for monogenic disorders is gene therapy with ex vivo-transduced autologous hematopoietic stem cells (HSCs). Safety or efficacy studies of ex vivo-modified HSCs are conducted in humanized mouse models after ablation of the murine bone marrow and transfer of human CD34+ HSCs. Engrafted human CD34+ cells migrate to bone marrow and differentiate into various human hematopoietic lineages. A 12-week study was conducted in NSG™ mice to evaluate engraftment, differentiation, and safety of human CD34+ cells that were transduced ( ex vivo) with a proprietary lentiviral vector encoding a human gene (BMRN-1) or a mock (green fluorescent protein) vector. Several mice intravenously injected with naive CD34+ cells or transduced CD34+ cells had variable lymphohistiocytic inflammatory cell infiltrates and microgranulomas in the liver and lungs consistent with graft-versus-host disease (GVHD). Spleen, bone marrow, stomach, reproductive tract, but not the skin had similar inflammatory changes. Ex vivo viral transduction of CD34+ cells did not impact engraftment or predispose to xenogeneic GVHD.

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Rat Bone Marrow Mesenchymal Stem Cells Cotransplanted with BM Improve Hematopietic Reconstitution and Immunoreconstitution with Alleviating Graft-versus-Host Disease.

Blood ◽

10.1182/blood.v104.11.4248.4248 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4248-4248

Author(s):

Jun-xia Lei ◽

Shu-nong Li ◽

Xiu-ming Zhang ◽

Xin Du ◽

Peng Xiang

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Graft Versus Host Disease ◽

Histological Analysis ◽

Host Disease ◽

Graft Versus Host ◽

Hematopoietic Reconstitution ◽

Marrow Mesenchymal Stem Cells ◽

Rat Bone

Abstract Prolonged immunodeficiency after allogeneic bone marrow transplantation (BMT) causes significant morbidity and mortality from infection, while graft versus host disease (GVHD) after allogeneic BMT and immunosuppression therapies against GVHD further deteriorate this process. Adult bone marrow mesenchymal stem cells (MSC) have recently been shown to inhibit T-cell proliferation and reduce GVHD after allo-BMT. In this study, we characterized the effect of MSC on immunoreconstitution and hematopoietic-reconstitution after bone marrow transplantation. BMT model from Fischer344 rats (RT-1Al) to WF rats (RT-1Au) was established for this experiment. Effects of MSCs on hematopoietic reconstitution, immunoreconstitution and GVHD were studied by survival rate, peripheral blood counts, histological analysis and FACS at day +30 after transplantation. Immune function recovery were assessed by lymphocyte proliferation reaction stimulated with ConA and LPS and allogeneic mixed-lymphocyte reaction. At day 30 postransplant, compared with BMT groups, we observed that cotransplantation of MSC and bone marrow promoted the recovery of peripheral blood white blood cells (5.47±1.11x109/L vs7.12±1.70x109/L, p<0.05), lymphocytes and platelets. Accordingly, it was noticed that cotransplantation of MSC with BM not only improved recovery of bone marrow cellularity, but also enhanced B lymphopoiesis (4.66±1.03x109/L vs 6.05±1.39x109/L, p<0.05) and megakaryocytopoiesis (402.50±63.70 x109/L vs 594.33±121.09x109/L, p<0.05). MSC was also shown to improve thymic and spenic architecture reconstitution. Histological analysis showed that near normal thymus architecture and normal spenic architecture, with well-developed red and white pulp and intact lymphoid follicles in the cotransplantation group, while loss of demarcation between cortex and medulla in the thymus and lymphocytic depletion of spenic arteriolar sheaths in BM transplantation rats.The total number of thymus cells and spleen cells were also increased in cotransplantation group compared to only BM transplantation group. Most notable, the higher percentage of CD3+CD4+ was observed in the spleens of cotransplantation group (11.47±3.68% vs 19.14±4.03%, respectively), while no difference in the percentage of CD3+CD8+ between two groups (10.61±3.37% vs 12.13±2.27%, respectively). So the ratio of CD4+/CD8+ cells increased and inclined to normal in cotransplantation groups compared to only BM transplantation group.There are no difference in the percentage of natural killer (NK) cells and monocytes between BMT group and BMT with MSC group. The immuno-inhibitory effect of MSC on reducing GVHD in recipients of an MHC-mismatched BMT was also evident in our study, nonetheless, splenic cells from recipients of MSC cotransplantation group displayed improved non-specific proliferation capability (against ConA LPS stimulation) and alloreactivity (against the third party splenocytes) compared to BM transplantation group. In conclusion, rat bone marrow mesenchymal stem cells cotransplanted with BM improves hematopietic reconstitution and immunoreconstitution with alleviating graft-versus-host disease.

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Phenotypical and Functional Characterization of Bone Marrow Mesenchymal Stem Cells in Patients with Chronic Graft-versus-Host Disease

Biology of Blood and Marrow Transplantation ◽

10.1016/j.bbmt.2015.02.013 ◽

2015 ◽

Vol 21 (6) ◽

pp. 1020-1028 ◽

Cited By ~ 7

Author(s):

Binsheng Wang ◽

Yongxian Hu ◽

Lizhen Liu ◽

Kaimin Hu ◽

Ruxiu Tie ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Graft Versus Host Disease ◽

Functional Characterization ◽

Host Disease ◽

Graft Versus Host ◽

Marrow Mesenchymal Stem Cells

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Wnt/β-catenin signaling mediates the abnormal osteogenic and adipogenic capabilities of bone marrow mesenchymal stem cells from chronic graft-versus-host disease patients

Cell Death and Disease ◽

10.1038/s41419-021-03570-6 ◽

2021 ◽

Vol 12 (4) ◽

Author(s):

Han-zhou Qi ◽

Yi-ling Ye ◽

Yuan Suo ◽

Hong Qu ◽

Hai-yan Zhang ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Graft Versus Host Disease ◽

Underlying Mechanism ◽

Hematopoietic Stem ◽

Host Disease ◽

Graft Versus Host ◽

Ppar Γ ◽

Marrow Mesenchymal Stem Cells

AbstractChronic graft-versus-host disease (cGVHD) is the main cause of non-relapse mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Mesenchymal stem cells (MSCs) in bone marrow (BM) remain unclear in the pathophysiology of cGVHD. In this study, we analyzed BM-MSCs from 66 patients after allo-HSCT, including 33 with active cGVHD and 33 without cGVHD. BM-MSCs showed similar morphology, frequency, phenotype, and proliferation in patients with or without cGVHD. MSCs from the active cGVHD group showed a decreased apoptosis rate (P < 0.01). Osteogenic capacity was increased while adipogenic capacity was decreased in the active cGVHD MSCs compared with no-cGVHD MSCs. The expressions of osteogenic gene RUNX2 and COL1A1 were higher (P < 0.001) while adipogenic gene PPAR-γ and FABP4 were lower (P < 0.001) in the active cGVHD MSCs than no-cGVHD MSCs. These changes were associated with the severity of cGVHD (P < 0.0001; r = 0.534, r = 0.476, r = −0.796, and r = −0.747, respectively in RUNX2, COL1A1, PPAR-γ, and FABP4). The expression of Wnt/β-catenin pathway ligand Wnt3a was increased in cGVHD-MSCs. The dysfunction of cGVHD-MSCs could be reversed by Dickkopf related protein 1(DKK1) to inhibit the binding of Wnt3a. In summary, the differentiation of BM-MSCs was abnormal in active cGVHD, and its underlying mechanism is the upregulated of Wnt3a through Wnt/β-catenin signaling pathway of MSCs.

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Treatment of Intractable Graft-Versus-Host Disease with Bone Marrow Derived Mesenchymal Stem Cells - A Pilot Clinical Trial.

Blood ◽

10.1182/blood.v110.11.4974.4974 ◽

2007 ◽

Vol 110 (11) ◽

pp. 4974-4974 ◽

Cited By ~ 1

Author(s):

Je-Hwan Lee ◽

Seong-Jun Choi ◽

Jung-Hee Lee ◽

Miee Seol ◽

Young-Shin Lee ◽

...

Keyword(s):

Clinical Trial ◽

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Graft Versus Host Disease ◽

Ex Vivo ◽

Performance Status ◽

Hematopoietic Stem ◽

Host Disease ◽

Graft Versus Host

Abstract Recent studies have shown that mesenchymal stem cells (MSCs) have profound immunomodulatory function, both in vitro and in vivo. There are several reports to treat effectively therapy-resistant graft-versus-host disease (GVHD) using ex vivo expanded MSCs. We performed a pilot clinical trial to treat intractable GVHD with bone marrow derived MSCs. All of 5 patients, 3 males and 2 females, with steroid-refractory GVHD were included in this study. Age range was 27 to 48 years old. Diagnosis of underlying disease was CML in 2, AML in 1, ALL in 1, and MDS in 1. Hematopoietic stem cell (HSC) donor was a sibling in 2 and an unrelated volunteer in 3. The donor of MSCs was a sibling in all patients: two donors were the same to HSC donors, but other three were not. About 20 mL of bone marrow was aspirated from donors and MSCs were cultured ex vivo. After about 3 weeks, MSCs were harvested for the first infusion, and 4 more weeks’ culture was done for the planned second infusion. The infused doses of MSCs were 5.3 to 6.9 x 106/kg for the first infusion and 1.8 to 7.0 x 106/kg for the second infusion. The onset of GVHD was post-transplant day 24 to 191 and the times from the onset of GVHD to the infusion of MSCs were 73 to 2469 days. There were no adverse events related to the infusion of MSCs. Three patients did not show any response to the treatment of MSCs for GVHD and two showed minimal response: transient improvement of jaundice and diarrhea without improvement of skin GVHD lesions in one patient, and improvement of performance status without significant increase of pulmonary function test parameters in another patient with lung involvement of GVHD. There were no significant changes in hemoglobin, and peripheral blood counts of platelets, leukocytes, lymphocytes, CD3+ cells, CD4+ cells, CD8+ cells, NK cells and B cells over 6 months’ period after the infusion of MSCs. Results of our pilot study suggest that the treatment effects of MSCs may be limited in patients with chronic established GVHD. Further studies on MSCs for GVHD should be focused on acute or exacerbated GVHD.

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