Autoantibody Production during Chronic Graft-Versus-Host Disease Does Not Associate with Long-Term Persistence of Host B Cells in Humans

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1180-1180
Author(s):  
Simona Piemontese ◽  
Zulma Magnani ◽  
Jacopo Peccatori ◽  
Claudio Bordignon ◽  
Chiara Bonini ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Graft Versus Host Disease ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Autoantibody Production ◽  
Graft Versus Host

Abstract Background. Chronic graft-versus-host disease (cGvHD) is a common complication of allogeneic hemopoietic cell transplantation (allo-HCT). The pathogenesis of cGvHD is poorly understood. In cGvHD, the homeostasis of B lymphocytes is perturbed, as demonstrated by the production of autoantibodies. B-cell depletion with monoclonal antibodies (mAb) interferes with autoantibody production and ameliorates signs and symptoms of cGvHD. In mouse models, cGvHD and autoantibodies associate with the long-term persistence of host B cells after allo-HCT (Sylvain Perruche et al., Transplantation 2006). It has been postulated that host B cells may present alloantigens to donor T cells and, in turn, receive help for autoantibody production. This could be crucial to the pathogenesis of cGvHD. Aim. To investigate whether the long-term persistence of host B lymphocytes is associated with cGvHD and autoantibodies in humans. Patients and methods. We recruited 13 consecutive patients with active cGvHD (4 mild, 5 moderate, 4 severe according to NIH classification) with a median time of onset of 6 months (range 3–36) from HLA-identical sibling (9 patients) and HLA-matched unrelated (4) allo-HCT. As controls, we chose 10 patients that underwent HLAidentical sibling (2), HLA-matched unrelated (5) or haploidentical (3) allo-HCT and never experienced cGvHD. In the two groups, we studied: circulating autoantibodies, including anti-nuclear (ANA), anti-DNA, anti-extractable nuclear antigen, anti-beta2 glycoprotein, anti-neutrophil cytoplasm, anti-thyroid, anti-mytocondria antibodies, rheumatoid factor, absolute numbers of T (CD3+, CD4+, CD8+), conventional B (CD19+), B1 (CD5+/CD19+) and NK cells (CD16+/CD56+) in the graft and in the peripheral blood, microchimerism by short-tandem repeats (STR) on B, T and myeloid cells purified by immunomagnetic cell sorting (sensitivity 0,01%). Results. Patients with cGvHD had high-titer circulating ANA (>1:160) more frequently than controls (54% versus 10%, P<0,05). All other autoantibodies were negative. Peripheral T-cell counts were lower in patients with cGvHD than in controls (for CD8+ cells P<0,05). This was not due to a difference in the absolute numbers of T lymphocytes within the graft between the two groups. Peripheral counts of conventional B and B1 cells in patients with cGvHD were similar to controls. Autoantibodies and cGvHD were not associated with the persistence of host B lymphocytes, since the analysis of STR on purified B cells revealed that they were all of donor origin. T and myeloid cells were also of donor origin. Of interest, in univariate analysis, in vivo B-cell depletion with mAb for the prophylaxis against Epstein-Barr virus-related lymphoproliferative disease showed a trend towards a lower risk of cGvHD (P=0,06). Conclusions. This study indicates that autoantibody production during cGvHD does not associate with long-term persistence of host B cells in humans. Moreover, it suggests that the early depletion of donor B lymphocytes in vivo may be effective for GvHD prophylaxis

AML Relapse after Rituximab Treatment for Graft-Versus-Host Disease: Crucial Role for B Cells in Graft-Versus-Host and Graft-Versus-Leukemia

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 5473-5473
Author(s):  
Marijn Aletta Gillissen ◽  
Sophie E. Levie ◽  
Greta de Jong ◽  
Etsuko Yasuda ◽  
Arjen Q. Bakker ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Cell Lines ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Disease Relapse ◽  
Graft Versus Host ◽  
Graft Versus Leukemia ◽  
Host Tissues

Abstract Patients with hematologic malignancies that develop graft-versus-host disease (GvHD) after allogeneic hematopoietic stem cell transplantation (HSCT) have a reduced risk of disease relapse compared to patients that do not develop GvHD, suggesting that GvHD and graft-versus-leukemia (GvL) responses are co-dependent. T cells are important in these processes, as T cell depletion from the graft reduces the risk of GvHD at the expense of disease relapse. Much less is known about the effect of B cell depletion on GvHD and GvL responses. Small patient series have demonstrated variable efficacy of Rituximab in the treatment of steroid-refractory chronic GvHD but the effect of depleting allo-reactive B cells on disease relapse remains to be determined. We here report on a patient with steroid-refractory GvHD whose AML relapsed after Rituximab treatment. This 39-year old male received an allogeneic HSCT for chemotherapy-related AML (AML-t) that however relapsed 8 weeks after the transplantation. Upon rapid cessation of immunosuppressive therapy (cyclosporine, mycophenolic acid) and without additional chemotherapy he obtained full remission, at the expense of severe GvHD of the skin, liver and intestine that was corticosteroid-refractory. B cell depletion with Rituximab was successful as second-line treatment for GvHD but eliminated the GvL response and the patient died of AML relapse several months after. To evaluate the B cell repertoire of this patient at the moment of maximal GvH and GvL responses, we isolated peripheral blood B lymphocytes that were transduced with Bcl-xL and Bcl-6 to create clonal B cell lines. These B cells were screened for binding to AML and host tissues. One clone was retrieved that specifically bound to AML cell lines and AML blasts freshly isolated from newly diagnosed AML patients. Antibodies from this clone induced antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity of AML cells in vitro. Several other clones were retrieved that were specific for host tissues such as liver (HepG2 and H69 cell lines), skin (primary fibroblasts) and/or colon (CaCo cell line). These data demonstrate the pivotal role of B lymphocytes in anti-leukemia and anti-host immune responses in an allogeneic HSCT recipient with relapsed AML-t. Disclosures No relevant conflicts of interest to declare.

Risk Reduction of Chronic Graft Versus Host Disease by Anti-CD20 Treatment (Rituximab).

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 5567-5567
Author(s):  
Marlies E.H.M. Van Hoef
Keyword(s):  
B Cell ◽  
Risk Reduction ◽  
Graft Versus Host Disease ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Host Disease ◽  
Medline Search ◽  
Graft Versus Host ◽  
Anti Cd20 ◽  
Study Plan

Abstract Introduction: Chronic graft versus host disease (cGVHD) occurs in 30–50% of adults transplanted by non-myeloablative (reduced intensity conditioning) allogeneic transplant, whereas it is rare in children. The syndrome complex manifests itselves as auto-immune disease. Dysregulation of B-cell function has also been reported in auto-immune diseases. This is a rationale for B-cell depletion. We considered that B-cell depletion might have a therapeutic effect in cGVHD and reduce the risks associated with cGVHD. Based on these assumptions we applied risk reduction principles and defined a plan for cure of cGVHD. Methods: To identify the risks associated with B-cell depletion in cGVHD we performed a medline search on anti-CD20 or B-cell depletion or rituximab and cGVHD. The publications were analyzed and those applicable to the topic evaluated. Clinical research methods were applied to define a plan for risk reduction of cGVHD. Results: The medline search revealed that rituximab was used for B-cell depletion in cGVHD and in auto-immune diseases. In advanced cGVHD complete and partial remissions of a variety of manifestations of cGVHD were reported. Complete remissions could also be induced early during the disease course of single manifestations of cGVHD not responsive to immune suppressive treatment or as initial treatment. The dose of rituximab varied from 1 to 4 weekly courses of rituximab 375 mg/m2, with repetitions of this schedule in advanced disease to induce complete response. During reported follow-up responding manifestations did not recur. Rituximab treatment was well tolerated with appropriate anti-allergic prophylaxis. The results support the concept that anti-CD20 treatment might cure cGVHD manifestations at first diagnosis; it is too early to define whether it also serves as prophylactic for development of new manifestations. The observations were discussed with the company marketing rituximab and a study plan was designed. This study plan is being discussed with study centers for risk reduction of cGVHD by rituximab. The risk reduction plan will be presented

scholarly journals Altered B-cell homeostasis and excess BAFF in human chronic graft-versus-host disease

Blood ◽  
2009 ◽  
Vol 113 (16) ◽  
pp. 3865-3874 ◽  
Author(s):  
Stefanie Sarantopoulos ◽  
Kristen E. Stevenson ◽  
Haesook T. Kim ◽  
Corey S. Cutler ◽  
Nazmim S. Bhuiya ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Graft Versus Host Disease ◽  
High Plasma ◽  
Cell Homeostasis ◽  
Hematopoietic Stem ◽  
Host Disease ◽  
Graft Versus Host ◽  
B Cell Homeostasis

Abstract Chronic graft-versus-host disease (cGVHD) causes significant morbidity and mortality in patients otherwise cured of malignancy after hematopoietic stem cell transplantation (HSCT). The presence of alloantibodies and high plasma B cell–activating factor (BAFF) levels in patients with cGVHD suggest that B cells play a role in disease pathogenesis. We performed detailed phenotypic and functional analyses of peripheral B cells in 82 patients after HSCT. Patients with cGVHD had significantly higher BAFF/B-cell ratios compared with patients without cGVHD or healthy donors. In cGVHD, increasing BAFF concentrations correlated with increased numbers of circulating pre–germinal center (GC) B cells and post-GC “plasmablast-like” cells, suggesting in vivo BAFF dependence of these 2 CD27+ B-cell subsets. Circulating CD27+ B cells in cGVHD comprised in vivo activated B cells capable of IgG production without requiring additional antigen stimulation. Serial studies revealed that patients who subsequently developed cGVHD had delayed reconstitution of naive B cells despite persistent BAFF elevation as well as proportional increase in CD27+ B cells in the first year after HSCT. These studies delineate specific abnormalities of B-cell homeostasis in patients with cGVHD and suggest that BAFF targeting agents may be useful in this disease.

Selective Rapid B-Cell Depletion In Vivo by Pharmacologic IKK2 Inhibition.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2481-2481
Author(s):  
Christopher C. Fraser ◽  
Kumiko Nagashima ◽  
Vito Sasseville ◽  
Jim Deeds ◽  
Alice McDonald ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
Oral Dose ◽  
B Cell ◽  
Single Oral Dose ◽  
Colony Growth ◽  
B Cell Development ◽  
Cell Depletion ◽  
B Cell Depletion

Abstract Studies using genetically deficient mice have revealed that members of the NF-kB family play key roles in B-cell development. IKK2 which activates NF-kB by targeting degradation of IkB, is also required for B-cell development. We have studied the role of IKK2 in hematopoiesis using a chemical specific inhibitor (ML120B). Mice given daily oral dosing of ML120B for 4 days had severe B-cell depletion in spleen and bone marrow. B-cells at all stages (pro-B, pre-B, immature and mature B) were depleted 10 fold in the bone marrow while granulocyte numbers were largely unaffected. IKK2 inhibition in vivo showed selective sensitivity of B-cell progenitors (4 fold decrease) in the marrow compared to myeloid progenitors which were unaffected at an equivalent dose. Foci of cells with an apoptotic morphology were visible in bone marrow and spleen within 6 hours of a single oral dose. Apoptotic cells detected by labeling fragmented DNA were increased within splenic follicles (6 fold) and bone marrow. Also an increase in B220+ / annexin V+ cells and a decrease in pre-B (B220+/IgM−) cells in the marrow were observed. RNA expression studies in the marrow 6 hours after a single oral dose revealed a decrease in IL-7 and increased GM-CSF expression. Image analysis of B220 in spleens within 18 hours of a single dose of an IKK inhibitor revealed decreased follicle size. In order to evaluate hematopoietic progenitor sensitivity to NF-kB inhibition, dose responses to ML120B, panepoxydone (PPD) and proteasome inhibitor Lactacystin (Lcyst) were evaluated in B-cell and myeloid bone marrow colony assays. Inhibitors PPD and Lcyst were more effective at inhibiting B-cell colony growth than myeloid colony growth. In summary, pharmacologic inhibition of IKK2 results in a rapid induction of apoptosis with preferential depletion of B-cells and retention of myeloid cells and progenitors within the bone marrow.

scholarly journals Long-term effects of combined B-cell immunomodulation with rituximab and belimumab in severe, refractory systemic lupus erythematosus: 2-year results

2020 ◽  
Author(s):  
Tineke Kraaij ◽  
Eline J Arends ◽  
Laura S van Dam ◽  
Sylvia W A Kamerling ◽  
Paul L A van Daele ◽  
...  
Keyword(s):  
Systemic Lupus Erythematosus ◽  
B Cells ◽  
B Cell ◽  
Lupus Erythematosus ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Systemic Lupus ◽  
Anti Cd20

Abstract Background Anti-CD20 B-cell depletion has not shown superior efficacy to standard immunosuppression in patients with systemic lupus erythematosus (SLE). Besides trial design, potential explanations are incomplete B-cell depletion in relation to substantial surges in B-cell-activating factor (BAFF). To improve B-cell targeting strategies, we conducted the first study in SLE patients aimed at investigating immunological effects and feasibility of combining rituximab (RTX; anti-CD20) and belimumab (BLM; anti-BAFF). Methods Reported is the long-term follow-up of a Phase 2 proof-of-concept study in 15 patients with SLE including 12 (80%) with lupus nephritis (LN). Results In 10/15 (67%) patients, a clinical response was observed by achievement of lupus low disease activity state, of which 8 (53%) continued treatment (BLM + ≤7.5 mg prednisolone) for the complete 2 years of follow-up. Five patients (33%) were referred to as ‘non-responders’ due to persistent LN, major flare or repetitive minor flares. Out of 12 LN patients, 9 (75%) showed a renal response including 8 (67%) complete renal responders. All anti-dsDNA+ patients converted to negative, and both anti-C1q and extractable nuclear antigen autoantibodies showed significant reductions. CD19+ B cells showed a median decrease from baseline of 97% at 24 weeks, with a persistent reduction of 84% up to 104 weeks. When comparing responders with non-responders, CD20+ B cells were depleted significantly less in non-responders and double-negative (DN) B cells repopulated significantly earlier. Conclusions Combined B-cell targeted therapy with RTX and BLM prevented full B-cell repopulation including DN B cells, with concomitant specific reduction of SLE-relevant autoantibodies. The observed immunological and clinical benefits in a therapy-refractory SLE population prompt further studies on RTX + BLM.

scholarly journals Modifying Graft-versus-Host Disease in a Humanized Mouse Model by Targeting Macrophages or B-Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-14 ◽  
Author(s):  
Marieke C. H. Hogenes ◽  
Suzanne van Dorp ◽  
Joyce van Kuik ◽  
Filipa R. P. Monteiro ◽  
Natalie ter Hoeve ◽  
...  
Keyword(s):  
B Cells ◽  
Mouse Model ◽  
B Cell ◽  
Cell Depletion ◽  
Mrna Levels ◽  
B Cell Depletion ◽  
Humanized Mouse ◽  
Humanized Mouse Model ◽  
Graft Versus Host ◽  
Macrophage Depletion

Humanized mouse models can well be modified to study specific aspects of Graft-versus-Host Disease (GvHD). This paper shows the results of both macrophage depletion and (early) B-cell depletion in a humanized mouse model using RAG2-/-γc-/- mice injected with HuPBMCs. Macrophage depletion showed a significant decrease in survival and also lead to a change in the histomorphology of the xenogeneic reaction. Higher levels of infiltrating B-cells were observed in various organs of mice depleted for macrophages. With (early) B-cell depletion using Rituximab, a clear improvement on clinical symptoms was observed, even when probably only inactivated B-cells were deleted. However, the histological examinations only showed a significant morphological effect on liver fibrosis. This may be related to a difference in the mRNA levels of TGF-β. Also, lower mRNA levels of Tregs in some organs were observed after Rituximab treatment, which contradicts that a higher number of Tregs would always be related to less severe GvHD. Our data show that both macrophage depletion and (early) B-cell depletion in a xenogeneic mouse model can influence the clinical, histological, and cytokine production of a GvHD response.

Circulating B-Lymphocyte Subpopulations as Novel Biomarker for Measuring Activity of Chronic Graft-Versus-Host Disease.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2881-2881
Author(s):  
David Pohlreich ◽  
Hildegard T. Greinix ◽  
Karin Feldmann ◽  
Ulrike Koermoeczi ◽  
Imke Lohmann ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Graft Versus Host Disease ◽  
Memory B Cells ◽  
Hematopoietic Stem ◽  
Host Disease ◽  
Graft Versus Host ◽  
Cell Counts ◽  
Severe Infections

Abstract Introduction: Chronic graft-versus-host-disease (cGvHD) is a major complication of allogeneic hematopoietic stem cell transplantation (HSCT) and a leading cause of non-relapse mortality due to profound immunodeficiency. The approach currently used to establish the diagnosis of cGvHD depends almost exclusively on the clinical history, physical examination, and histopathologic confirmation. Biology-based markers for confirmation of diagnosis or monitoring progression of cGvHD are urgently warranted. The pathogenesis of cGvHD is poorly understood. Mounting evidence implies a role of B-cells in this autoimmune-like disorder. In this study we analyzed whether changes in B-cell subpopulations in the peripheral blood (PB) of HSCT recipients could serve as a biomarker for immune-modulatory consequences of cGvHD. Methods: Within six months 70 consecutive patients (45 men, 25 women; median age 47.5 years) with complete donor cell engraftment a median of 45.5 (range, 5–149) months after allogeneic HSCT were enrolled including 21 never having cGvHD and 49 with cGvHD (35 active and/or progressive, 14 resolved at study entry). The latter received standard anti-infective prophylaxis and immunosuppressive therapy with a median duration of 36 (range, 3–104) months. A third of patients with active/progressive cGvHD had more than 2 organs involved. Evaluations in the study consisted of clinical parameters including cGvHD severity and treatment, serum immunoglobulin (Ig) levels, and infections. PB was analyzed by multiparameter flow cytometry to define B-lymphocytes (CD19+), which were further subdivided by staining for surface Ig (IgD+/M+) and the B-cell memory marker CD27+ and more immature B-lymphocytes (CD19+/CD21−). Results: No significant differences in absolute B, T, and NK-cell counts between the groups with and without cGvHD were seen. However, the relative number of both class-switched and non-class-switched memory B-cells was significantly lower (below 3%) in patients with active cGvHD compared to patients never having cGvHD (up to 7%) (p<0.001, c2 =22.0 for class-switched and p=0.002, c2 =9.2 for non-class-switched). Absolute cell counts confirmed these findings (10.3 vs. 27.2 cells/uL, p=0.02 for class-switched and 12.1 vs. 22.1 cells/ul, p=0.047 for non-class-switched). Significantly more patients with active cGvHD had elevated numbers of immature CD19+/CD21− B-cells (>15% cut-off) than the groups with resolved or never having cGvHD (37% vs 21% vs 9%, p=0.024). In patients with >15% immature CD19+/CD21− B-cells significantly more severe infections (according to NCI-CTC Infection module grade III–IV) were seen compared to patients with low CD19+/CD21− B-cell counts (p=0.002, c2 =9.5). In the group of active cGvHD 12/13 (92%) patients with >15% CD19+/CD21− B-cells compared to 4/22 (18%) patients with ≤ 15% CD19+/CD21− B-cells experienced severe infections (p<0.001, c2 =18.1). First results in an ongoing prospective study showed an increase of memory B-cell numbers in patients with decreasing cGvHD activity. Discussion: Monitoring of class-switched and non-class-switched memory B-cells may allow measuring of disease activity of chronic GvHD. In combination with assessment of immature CD19+/CD21− B-cells new insights into immune-modulatory consequences of cGvHD focusing on B-cell defects and activation status are possible.

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