Takayasu arteritis is characterised by disturbances of B cell homeostasis and responds to B cell depletion therapy with rituximab

2011 ◽  
Vol 71 (1) ◽  
pp. 75-79 ◽  
Author(s):  
Bimba F Hoyer ◽  
Imtiaz M Mumtaz ◽  
Konstanze Loddenkemper ◽  
Anne Bruns ◽  
Claudia Sengler ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Takayasu Arteritis ◽  
Lupus Erythematosus ◽  
Peripheral Blood ◽  
Plasma Cells ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Cell Homeostasis ◽  
B Cell Homeostasis

IntroductionTakayasu arteritis (TA) is a large vessel vasculitis involving the aorta and its major branches. T cell-mediated autoimmunity is thought to play a major role in its pathogenesis, while the role of B cells is still unclear.MethodsB cell subsets in the peripheral blood of 17 patients with TA were analysed and compared with nine patients with active systemic lupus erythematosus (SLE) and nine healthy controls by flow cytometry. Based on these findings, three patients with active refractory TA were treated with B cell depletion therapy (BCDT) using monoclonal anti-CD20 antibodies (rituximab).ResultsThe absolute number and frequency of peripheral blood CD19+/CD20−/CD27high antibody-secreting cells in patients with active TA was significantly higher than in healthy donors. As in active SLE, the majority of these cells are newly generated plasmablasts which significantly correlated with TA activity. Three patients with active refractory TA and expansion of plasmablasts were successfully treated with BCDT, which resulted in remission.ConclusionDisturbances of B cell homeostasis may be critical in TA. Circulating plasmablasts could be a useful biomarker of disease activity and a tool for selecting appropriate candidates for BCDT. B cells and plasmablasts/plasma cells may therefore represent novel targets for effective therapies for TA.

Steady-state generation of mucosal IgA+ plasmablasts is not abrogated by B-cell depletion therapy with rituximab

Blood ◽  
2010 ◽  
Vol 116 (24) ◽  
pp. 5181-5190 ◽  
Author(s):  
Henrik E. Mei ◽  
Daniela Frölich ◽  
Claudia Giesecke ◽  
Christoph Loddenkemper ◽  
Karin Reiter ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Steady State ◽  
B Cells ◽  
B Cell ◽  
Peripheral Blood ◽  
Plasma Cells ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Ki 67

AbstractThe anti-CD20 antibody rituximab depletes human B cells from peripheral blood, but it remains controversial to what extent tissue-resident B cells are affected. In representative patients with rheumatoid arthritis, we here demonstrate that recently activated presumably short-lived plasmablasts expressing HLA-DRhigh and Ki-67 continuously circulate in peripheral blood after B-cell depletion by rituximab at 26%-119% of their initial numbers. They circulate independent of splenectomy, express immunoglobulin A (IgA), β7 integrin, and C-C motif receptor 10 (CCR10) and migrate along CCL28 gradients in vitro, suggesting their mucosal origin. These plasmablasts express somatically hypermutated VH gene rearrangements and spontaneously secrete IgA, exhibiting binding to microbial antigens. Notably, IgA+ plasmablasts and plasma cells were identified in the lamina propria of patients treated with rituximab during peripheral B-cell depletion. Although a relation of these “steady state”–like plasmablasts with rheumatoid arthritis activity could not be found, their persistence during B-cell depletion indicates that their precursors, that is, B cells resident in the mucosa are not deleted by this treatment. These data suggest that a population of mucosal B cells is self-sufficient in adult humans and not replenished by CD20+ B cells immigrating from blood, lymphoid tissue, or bone marrow, that is, B cells depleted by rituximab.

scholarly journals Takayasu's arteritis characterised by disturbances of B cell homeostasis responds to B cell depletion with rituximab

2010 ◽  
Vol 69 (Suppl 2) ◽  
pp. A11-A11
Author(s):  
B F Hoyer ◽  
K Loddenkemper ◽  
I M Mumtaz ◽  
A Bruns ◽  
C Sengler ◽  
...  
Keyword(s):  
B Cell ◽  
Takayasu’S Arteritis ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Takayasu's Arteritis ◽  
Cell Homeostasis ◽  
B Cell Homeostasis

MO251HIGHLY SENSITIVE FLOW CYTOMETRIC DETECTION OF RESIDUAL B-CELLS AFTER RITUXIMAB IN ANTI-NEUTROPHIL CYTOPLASMIC ANTIBODY-ASSOCIATED VASCULITIS PATIENTS*

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y K O Teng ◽  
L Van Dam ◽  
Jelle Oskam ◽  
S W A Kamerling ◽  
E J Arends ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
B Cells ◽  
B Cell ◽  
Mononuclear Cells ◽  
Plasma Cells ◽  
Memory B Cells ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Peripheral Blood Mononuclear

Abstract Background and Aims B-cell depletion with rituximab (RTX) is an effective treatment for anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis (AAV) patients. Nevertheless, relapses are frequent after RTX, often preceded by B-cell repopulation suggesting that residual autoreactive B-cells persist despite therapy. Therefore, this study aimed to identify minimal residual autoimmunity (MRA) in the B-cell compartment of AAV patients treated with RTX. Method EuroFlow-based highly-sensitive flow cytometry (HSFC) was employed to study B-cell and plasma cell (PC) subsets in-depth in AAV patients before and after RTX treatment. Additionally, peripheral blood mononuclear cells (PBMCs) of these RTX-treated AAV patients were cultured and in vitro stimulated with CpG, IL-2, and IL-21 to induce antibody-secreting cells (ASC). (ANCA)-IgG was measured in these supernatants by ELISA. Results By employing EuroFlow-based HSFC, we detected circulating CD19+ B-cells at all timepoints after RTX treatment, in contrast to conventional low-sensitive flow cytometry. Pre-germinal center (Pre-GC) B-cells, memory B-cells and CD20+CD138− plasmablasts (PBs) were rapidly and strongly reduced, while CD20−CD138− PrePC and CD20-CD138+ mature (m)PCs were reduced slower and remained detectable. Both memory B-cells and CD20− PCs remained detectable after RTX. Serum ANCA-IgG decreased significantly upon RTX. Changes in ANCA levels strongly correlated with changes in naive, switched CD27+ and CD27− (double-negative) memory B-cells, but not with plasma cells. Lastly, we demonstrated in vitro ANCA production by AAV PBMCs, 24 and 48 weeks after RTX treatment reflecting MRA in the memory compartment of AAV patients. Conclusion We demonstrated that RTX induced strong reductions in circulating B-cells, but never resulted in complete B-cell depletion. Despite strongly reduced B-cell numbers after RTX, ANCA-specific memory B-cells were still detectable in AAV patients. Thus, MRA is identifiable in AAV and can provide a potential novel approach in personalizing RTX treatment in AAV patients.

HuMax-CD20 Fully Human Monoclonal Antibody in Follicular Lymphoma. First Human Exposure: Early Results of an Ongoing Phase I/II Trial.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1400-1400 ◽  
Author(s):  
Anton Hagenbeek ◽  
Torben Plesner ◽  
Jan Walewski ◽  
Andrzej Hellmann ◽  
Brian K. Link ◽  
...  
Keyword(s):  
B Cells ◽  
Adverse Events ◽  
B Cell ◽  
Cell Count ◽  
Peripheral Blood ◽  
Tumor Response ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Cell Counts ◽  
Grade 3

Abstract HuMax-CD20 is a fully human monoclonal IgG1 antibody targeting a unique extracellular epitope of the CD20 molecule on B-cells. HuMax-CD20 stops growth of engrafted B-cell tumors in SCID mouse tumor models more efficiently than Rituximab®, and i.v. infusion of HuMax-CD20 in cynomolgus monkeys has led to profound, long lasting, dose-dependent B-cell depletion. A total of 40 patients with CD20+ relapsed or refractory follicular non-Hodgkin’s lymphoma grade I-II will be enrolled in this open-label, dose-escalating, international, multi-center clinical trial. Cohorts of 10 patients will receive i.v. infusions at doses of either 300, 500, 700 or 1000 mg once weekly for 4 weeks. The patients are followed for 12 months. Patients receive oral acetaminophen and i.v. antihistamin before infusion. In case of adverse events of CTC grade 3 or higher, i.v. glucocorticosteroids are given. The endpoints are CT scan verified tumor response according to the Cheson criteria, B-cell depletion in peripheral blood and lymph nodes, time to next anti-lymphoma treatment, duration of response, BCL2 conversion, pharmacokinetics, and adverse events. Tumor and bone marrow biopsies and CT scans are assessed centrally. The first 17 patients treated with HuMax-CD20 are the subject of this report. Mean age is 60 years. In the 300 mg group all 10 patients have received all 4 infusions. Seven patients have been enrolled in the 500 mg group; three of them have received 4 infusions, two have received 3 infusions, and two patients have received 2 infusions. Baseline B-cell count was in the range of 11-382 x 106 cells per L with a median of 114 x 106. One week after the first infusion the median B-cell count available in 16 patients was 8 x 106 cells per L with a range of 0–19 x 106. In six of the 16 patients no B-cells were detected. B-cell counts measured one week after the 4th infusion are available for 10 patients. Eight patients had no detectable B-cells, one patient had 11 x 106 and one had 34 x 106 cells per L. B-cell counts eight weeks after the 4th infusion are available for two patients. No B-cells were detectable in these two patients. No dose limiting toxicity has been reported with administration of 300 or 500 mg. One serious adverse event assessed as not related to HuMax-CD20 has been reported in the 300 mg group. Infusion related adverse events have primarily been seen during the first infusion of HuMax-CD20. The events have, as expected, predominantly been signs and symptoms of cytokine release, e.g. pruritus, dyspnoea, rigors/chills, nausea, hypotension, urticaria, fatigue, fever and rash. In 15 of the 17 patients, 51 adverse events have been reported. Nine adverse events were CTC grade 3, 16 were grade 2, and 26 events were grade 1. In conclusion, this analysis based on preliminary data for the first 17 patients treated with HuMax-CD20 demonstrated significant depletion of peripheral blood B-cells and a favorable safety profile. An updated report of results for all 40 patients including preliminary tumor response data will be presented.

scholarly journals Expansion of Activated Peripheral Blood Memory B Cells in Rheumatoid Arthritis, Impact of B Cell Depletion Therapy, and Biomarkers of Response

PLoS ONE ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. e0128269 ◽  
Author(s):  
Diana G. Adlowitz ◽  
Jennifer Barnard ◽  
Jamie N. Biear ◽  
Christopher Cistrone ◽  
Teresa Owen ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
B Cells ◽  
B Cell ◽  
Peripheral Blood ◽  
Memory B Cells ◽  
Cell Depletion ◽  
B Cell Depletion

Emergence of Long-Lived Autoreactive Plasma Cells in the Spleen of Primary Warm Auto-Immune Hemolytic Anemia Patients Treated with Rituximab

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 569-569
Author(s):  
Matthieu Mahevas ◽  
Marc Michel ◽  
Benoit Vingert ◽  
Julien Moroch ◽  
David Boutboul ◽  
...  
Keyword(s):  
Bone Marrow ◽  
B Cells ◽  
B Cell ◽  
Blood Cells ◽  
Plasma Cells ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Bone Marrow Plasma ◽  
Previously Treated

Abstract Introduction: We recently proposed that B-cell depletion in immune thrombocytopenia (ITP) promotes the generation of long-lived plasma cells in the spleen, some of them being auto-reactive; but it remained possible that this observation was related to ITP itself rather than to B-cell depletion. Primary warm autoimmune hemolytic anemia (wAIHA) is a rare disease characterized by IgG auto-antibodies directed against antigens at the surface of red blood cells (RBCs) antigens, leading to their accelerated destruction. The use of B-cell depletion in wAIHA leads to 60 % to 70% of overall response at one-year and beyond. Nevertheless, 30-40 % of patients may resist to rituximab and then require a splenectomy. Nothing is known about antibody secreting cells (ASC) in the spleen of wAIHA patients, who have previously been treated or not with rituximab. In this study, we analyzed at the single cell level the splenic ASC from patients with chronic and active wAIHA, previously treated or not with rituximab (RTX), and we compared them with splenic ASC from ITP patients, and with splenic and bone marrow plasma cells from healthy donors (HD). Methods: We took advantage of the different therapeutic outcomes to analyze the splenic B-cell compartment of wAIHA patients, not previously treated with RTX (n=6), or after failure RTX treatment (n=3).Splenic tissues from organ donors and bone marrow from cardiovascular thoracotomy were used as controls. Blood samples from wAIHA (n=19), and (HD) (n=8) enrolled in this study were obtained after giving written informed consent in accordance with the Declaration of Helsinki. Results: We observed by flow cytometry and microscopy that the spleen from wAIHA patients who received less than 3 months of steroid therapy was the site of a B-cell response characterized by the presence of Bcl6+ germinal-center (GC) B-cells. Furthemore, splenic ASC secreted anti-red blood cell IgG in vitro. In line with this observation, we observed in the peripheral blood from patients with a newly diagnosed wAIHA (n=11), that short-lived IgG plasmablasts were increased compared with HD (n=8) (Mean 4.2 ± 0.84 % vs 0.99 ± 0.19% of CD19+ cells, p< 0.01). Moreover, for patients receiving long term steroid therapy (> 6 months) the plasmablast response was suppressed in the peripheral blood (Mean: 0.68 ± 0.2 % of CD19+ cells, n=8) and the splenic GC B-cell reaction was impaired (n=3). We conclude that short-lived IgG ASC result from an over-activity of GC reactions in wAIHA. We then analyzed the spleen of 3 patients who failed to respond to RTX, and observed a residual population of CD19+B-cells (median: 0.9% of lymphoid cells), including non-proliferative memory B-cells and plasma cells (PC). A fraction of these residual PC secreted anti-red blood cells IgG in vitro, thus accounting for the faillure of the B-cell depletion therapy. By using a single cell multiplex quantitative RT-PCR (Fluidigm dynamic arrays), we showed that such RTX-resistant plasma cells display a long-lived transcriptional program, which differs from PC from untreated wAIHA patients or HD, as well as from plasmablasts. Interestingly, the gene expression profile of wAIHA long-lived plasma cells segregated with long-lived PC previously observed in the spleen of ITP patients treated with rituximab. By a principal component analysis, we observed a gradient of maturation from plasmablasts to bone marrow plasma cells in which PC from RTX-treated spleens segregated close to bone marrow PC. We also observed that the cytokine BAFF was increased in the supernatants of spleen cell cultures from wAIHA patients treated with rituximab compared with controls (p< 0.05), suggesting, in keeping with our previous report in ITP, a role for BAFF in the differentiation of short-lived plasma cells into long-lived plasma cells. Conclusion: The presence of splenic long-lived autoreactive PC in wAIHA may explain why some patients cannot achieve a response after RTX. Our results show that, the B-cell depletion induced by rituximab itself, as opposed to the underlying auto-immune condition, promotes a suitable environment for the maturation of auto-immune long-lived plasma cells in the spleen. Targeting specifically some factors such as BAFF right after rituximab injection could be an interesting therapeutic option in the future. Disclosures No relevant conflicts of interest to declare.

ZAP-70+ B Cell Subset Influences Response to B Cell Depletion Therapy and Early Repopulation in Rheumatoid Arthritis

2012 ◽  
Vol 39 (12) ◽  
pp. 2276-2285 ◽  
Author(s):  
ELISA GREMESE ◽  
BARBARA TOLUSSO ◽  
ANNA LAURA FEDELE ◽  
SILVIA CANESTRI ◽  
STEFANO ALIVERNINI ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
B Cells ◽  
Clinical Outcome ◽  
B Cell ◽  
Peripheral Blood ◽  
Cell Subset ◽  
Cell Depletion ◽  
Poor Responders ◽  
B Cell Depletion ◽  
Switch Memory

Objective.To define the role of ZAP-70+ B cells (CD19+/ZAP-70+) as a biomarker of response to B cell depletion therapy (BCDT), their relationship with clinical outcome, and their behavior during repopulation of peripheral blood in patients with rheumatoid arthritis (RA).Methods.Thirty-one patients with RA underwent BCDT and were followed for 12 months. Disease activity was assessed with the European League Against Rheumatism (EULAR) criteria. Cytofluorimetric analysis of peripheral blood B cell subsets at baseline and at 6- and 12-month intervals after BCDT was performed using surface markers (CD45, CD3, CD56, CD19, IgD, CD38, CD27) and intracellular ZAP-70.Results.A moderate/good EULAR response was achieved in 66.6% of the RA cohort. The baseline percentage of CD19+/ZAP-70+ cells was lower in good responder patients (1.8% ± 1.7%) compared to poor responders (5.6% ± 4.9%; p = 0.02). A decrease of plasmablasts (IgD-CD27+CD38+) and pre-switch memory (IgD+CD27+) B cells occurred after BCDT. Recovery of B cells in peripheral blood after the first course of BCDT was characterized by the reappearance of B cell subtypes that showed a naive, activated phenotype, coupled with a decrease in memory cells. B cells carrying intracytoplasmic ZAP-70 increased significantly from the baseline value of 4.4% ± 4.5% to 12.4% ± 9.2% (p = 0.001) at the 6-month and to 9.4% ± 6.4% (p = 0.002) at the 12-month followup.Conclusion.Baseline percentage of CD19+/ZAP-70+ cells is associated with the clinical outcome after BCDT in patients with RA. Depletion of plasmablasts and pre-switch memory B cells and increase of CD19+/ZAP-70+ cells are features of the recovery of the B cell pool after BCDT.

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.

Preclinical Pharmacology and Toxicology of Humanized Anti-B-Cell Antibodies (Anti-CD22 and Anti-CD20) in Cynomolgus Monkeys (CM).

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1471-1471
Author(s):  
Puja Sapra ◽  
Gary Kikuchi ◽  
Ashwini Venkatasamy ◽  
Marianne K. Hayes ◽  
Christina M. Satterwhite ◽  
...  
Keyword(s):  
B Cells ◽  
Autoimmune Diseases ◽  
B Cell ◽  
Plasma Cells ◽  
Clinical Signs ◽  
T Cell Subsets ◽  
Cell Depletion ◽  
B Cell Depletion ◽  
Significant Difference ◽  
Hodgkin's Lymphomas

Abstract hLL2 (epratuzumab) and hA20 are humanized IgG1 monoclonal antibodies (MAbs) recognizing B-cell antigens, CD22 and CD20, respectively. CD22 and CD20 are expressed at all stages of B-cell development, except progenitor stem cells and plasma cells. Both hLL2 and hA20 have shown promising clinical activity in patients with non-Hodgkin’s lymphomas (NHL). Further, hLL2 has demonstrated clinical efficacy in patients with autoimmune diseases, such as systemic lupus erythematosus and Sjögren’s syndrome. Here, we discuss preclinical toxicology studies and compare the pharmacodynamics (PD) and pharmacokinetics (PK) of hA20 and hLL2 in CD20/CD22-crossrecative CM. For hLL2, we performed two toxicology studies. In the first 14-Day tolerability study, CM received hLL2 as two intravenous (IV) infusions at 0 (vehicle), 10, 60 and 160 mg/kg given one week apart, followed by necropsy on Day 14. In a second 6-month PK/PD study, hLL2 was administered in 2 cycles of 4-weekly infusions, with a 1-month treatment-free period between cycles followed by necropsy at the end of the second cycle. For hA20, CM received 4-weekly infusions of hA20 at 0 (vehicle), 8, 24, 80, and 240 mg/kg-doses and were either sacrificed 1 week after the last infusion or followed for recovery. Both hA20 and hLL2 were well tolerated. There were no drug-related clinical signs or changes in body weight, food intake, ophthalmic condition, EKG, blood pressure, and standard hematology, serum chemistry, coagulation or urinalysis parameters. Also, there were no changes in organ weights, gross necropsy, or histopathology findings. For hLL2, PD indicated a mean reduction from baseline for all B-lymphocytes (CD20+B-cells, or activated CD20+HLA-DR+ and CD20+CD27+ B-cells) of approximately 30–50%, starting on Day 3 in both studies, with no apparent dose-response. In the cyclical regimen study, this effect continued through Day 113 (day after the end of 2nd cycle). In contrast, for hA20, almost 85–95% B cell depletion was observed in all treatment groups on Day 6 and there was a trend towards dose-related depletion. Both hLL2 and hA20 had little discernable effect on T cell subsets, monocytes, or plasma cells. hLL2 and hA20 had a t1/2 of 3–5 days after the first infusion, and hLL2 had a t1/2 of 6–7 days after the fourth infusion. There was no significant difference between PK of hLL2 after the first and second cycles. With regard to immunogenicity, primate antibodies to either hA20 or hLL2 were not detected. In conclusion, hA20 demonstrates significantly higher B-cell depletion, similar to Rituximab, compared to hLL2 in CM. Clinical results obtained with hLL2 as well as studies of in vitro mechanism of action suggest that partial B-cell modulation by hLL2 in autoimmune diseases may provide benefit, and may function by different mechanisms than B-cell depletion related to hA20. In NHL, combined therapy of CD20 and CD22 may be more effective than single-MAb treatment, as has been suggested clinically.

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