Bone Marrow Histology in Monoclonal B-Cell Lymphocytosis Shows Various B-Cell Infiltration Patterns

Key Points Donor T-cell infiltration of the bone marrow is associated with impaired B-cell immunity after allogeneic HSCT. Quantification of κ-deleting recombination excision circles as a biomarker for bone marrow B-cell output in different clinical episodes.

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Impaired B Lymphopoiesis Concurrent with Marrow Infiltration By Allogeneic Donor T-Cells: Further Evidence for Bone Marrow Graft-Versus-Host Disease

Blood ◽

10.1182/blood.v126.23.3148.3148 ◽

2015 ◽

Vol 126 (23) ◽

pp. 3148-3148

Author(s):

Armin Rashidi ◽

Tandon Bevan ◽

Amanda F. Cashen

Keyword(s):

Bone Marrow ◽

T Cells ◽

T Cell ◽

B Cell ◽

Cell Count ◽

Cell Infiltration ◽

B Lymphopoiesis ◽

Cell Recovery ◽

Marrow Graft ◽

T Cell Infiltration

Abstract Background: The recently introduced concept of bone marrow graft-versus-host disease (BM-GvHD) representing destruction of the host hematopoietic niche in the marrow by allogeneic donor T-cells is loosely defined. Otherwise unexplained B lymphocytopenia and other cytopenias of unclear etiology that frequently occur in the early post-transplant period are often attributed to BM-GvHD. B lymphocytopenia can co-exist with systemic GvHD, supporting the theory of concurrent donor T-cell-induced damage to the marrow causing suppressed B lymphopoiesis. However, demonstration of such correlations has been difficult in humans due to the lymphotoxic effect of steroids which are the frontline therapy for GvHD. The best evidence comes from a recent study showing a correlation between delayed recovery of B lymphopoiesis and donor T-cell infiltration in the marrow 3-4 weeks post-transplant. The potential confounding effect of steroids was not evaluated. The purpose of the present study was to assess whether there is significant donor T-cell infiltration in the marrow at the time of B lymphocytopenia in carefully selected, fully chimeric allo-SCT recipients on minimal or no steroids and with minimal or no systemic GvHD. Methods: A total of 11 patients who underwent allo-SCT for myeloid malignancies were retrospectively studied. Inclusion criteria were: (i) bone marrow biopsy available on days 90-100 or 170-190 post-SCT concurrent with peripheral blood B-cell count using flow cytometry, (ii) full donor chimerism at the time of bone marrow biopsy, (iii) no B lymphodepleting therapy post-SCT, (iv) not on more than 15 mg/d of prednisone on the day of measurement, (v) no GvHD other than acute stage I skin GvHD, and (vi) delayed B-cell recovery defined as <10 CD19+ B-cells/µl on days 90-100 or <100 cells/µl on days 180-200. Peripheral B-cell count measurements were not due to specific clinical indications and were either based on the treating physician's routine practice or the protocols patients were enrolled to. Similar to previous studies, increased T-cell infiltration was defined as ≥5% of total nucleated cells in the core determined by anti-CD3 antibody labeling in a fully chimeric recipient. We determined the frequency of increased marrow T-cell infiltration (as a marker of acute BM-GvHD) on the same day when delayed B-cell recovery (as a marker of impaired B lymphopoiesis) was diagnosed. Results: 11 patients (10 males) with a median (range) age of 60 (32-67) years were studied. Measurements were made between days 90-100 and 170-190 post-SCT in 4 and 7 patients, respectively. The underlying diagnosis was acute myeloid leukemia (n = 7) or myelodysplastic syndrome (n = 4). The donor was a matched sibling (n = 1), matched unrelated donor (n = 4), or haploidentical donor (n = 6). Conditioning was ablative in 5 patients. Leukopenia, anemia, and thrombocytopenia were present in 3, 9, and 11 patients, respectively. The median (range) B-cell count on days 90-100 and 180-200 was 7 (0-9) and 19 (0-65) cells/µl, respectively. All patients had ≥5% T-cells/µl in the concurrent core biopsy with one exception. This patient had zero B-cells on day 180 but no evidence of concurrent BM-GvHD while on 12 mg/d of prednisone for appetite stimulation. Conclusions: Using a carefully selected cohort of fully chimeric allo-SCT recipients with delayed B lymphopoiesis, on no or minimal amounts of steroids, and with minimal or no systemic GvHD, we demonstrated a high frequency of concurrent increased marrow T-cell infiltration. These results support the recently introduced concept of BM-GvHD and highlight its negative effect on B lymphopoiesis. We show that bone marrow damage by allogeneic T-cells can occur even in the absence of systemic GvHD. Given the difficulties in quantification of marrow T-cells on the core, a more reproducible definition for BM-GvHD is needed. Disclosures No relevant conflicts of interest to declare.

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Acute Bone Marrow GvHD Is Associated With Delayed B Cell Neogenesis and Impaired Natural Antibody Response After Allogeneic Hematopoietic Stem Cell Transplantation

Blood ◽

10.1182/blood.v122.21.4605.4605 ◽

2013 ◽

Vol 122 (21) ◽

pp. 4605-4605

Author(s):

Angela Mensen ◽

Korinna Jöhrens ◽

Ioannis Anagnostopoulos ◽

Sonya Demski ◽

Christoph Ochs ◽

...

Keyword(s):

Bone Marrow ◽

T Cells ◽

T Cell ◽

B Cells ◽

B Cell ◽

Cell Infiltration ◽

Strong Increase ◽

Cell Recovery ◽

T Cell Infiltration ◽

Transitional B Cells

Graft-versus-host disease (GvHD) and severe infections are main complications limiting the success of allogeneic hematopoietic stem cell transplantion (alloHSCT). Delayed B cell reconstitution followed by B cell immune dysfunction considerably contributes to an increased risk for life-threatening infections. Several studies have shown that B cell regeneration is impaired in patients with systemic GvHD. Bone marrow (BM) suppression is often observed in parallel as GvHD symptoms appear suggesting the BM as a target of GvHD. Thus far, little is known about mechanisms of BM dysfunction during GvHD in alloHSCT patients. In this study, we investigated the reconstitution kinetics of peripheral blood B cell subsets in adult acute leukemic patients (n=52) before and within six months after alloHSCT by flow cytometry and correlated the data with RT-PCR quantified numbers of kappa-deleting-recombination-excision-circles (KREC), which are stable episomal plasmids generated during BM B cell development. Furthermore, we determined specific B cell antibody responses after in vitrostimulation with CpG, CD40L and T cell cytokines by EliSPOT analysis. To investigate BM as a direct target of allo-reactive T cells we performed histopathological stainings of BM biopsy samples obtained 3-4 weeks after alloHSCT. T cells were detected by specific anti-CD3 antibody staining and osteoblasts were morphologically evaluated. We observed in all patients a profound B cell immune deficiency already pre-transplant that proceeded within the first months post alloHSCT (mean B cells/ml blood±SEM: 11±3 pretransplant, 3±1 day14, 3±1 day28 post alloHSCT; 83±13 healthy control). Onset of B cell reconstitution is characterized by transitional B cell recovery representing the first B cell subset which emigrates from the BM. B cell reconstitution occurred either early (37% of patients) with a strong increase of transitional B cells between days 60-90 (mean transitional B cells/ml blood±SEM: Day 60, 36±10) or late (33% of patients) with delayed recovering transitional B cells (Day 180, 5±2). KREC copy numbers correlated highly positive and significantly with transitional B cell numbers (Spearman 0.94, p=0.017). Less correlation was obtained with naïve and CD27+ memory B cell recovery. Delayed onset of B cell reconstitution was significantly associated with both presence of systemic acute GvHD and full-intensity conditioning therapy (GvHD 71% vs non-GvHD 32%, Fisher´s exact p=0.044; full-intensity 41% vs reduced-intensity 5%, p=0.016). Supporting the hypothesis of bone marrow GvHD we could show a stronger infiltration of CD3+ T cells in the BM in late than in early recovering patients (≥5% T cell infiltration: 64% vs 17%, p=0.010). This increased T-cell infiltration was associated with reduced numbers of osteoblasts, known in mice to support B cell lymphopoiesis (no/few osteoblasts: 65% vs 17%, p=0.011). Impaired B cell lymphopoiesis further resulted in a delayed naïve and IgM memory B cell recovery compared to early recovering patients. No recovery of switched-memory B cells was seen for both patient groups within the analyzed time-period. Functionally, ex vivoactivation of patient B cells revealed higher numbers of IgM producing B cells specific for pneumococcal polysaccharide (PnP) at day 180 post alloHSCT in early than in late recovering patients. Polyclonal IgG producing B cells were significantly diminished in all patients. We conclude from these data, that early onset of B cell reconstitution is characterized by strong increase in regenerating transitional B cells within three months after alloHSCT. Herein, KREC appears as a suitable biomarker to monitor BM B cell output post-transplant. B cell regeneration is significantly delayed in patients showing increased occurrence of systemic acute GvHD and stronger T cell infiltration with loss of osteoblasts in the BM. Thus, delayed onset of B cell reconstitution might result from acute BM GvHD in which alloreactive T cells lead to an osteoblast niche destruction. Increased PnP specific IgM antibody responses are most likely result of higher numbers of early reconstituted transitional and IgM memory B cells but not naïve B cells that were shown not to produce IgM upon CpG stimulation (Capolunghi F et al. 2008). Thus, early B cell reconstitution might provide a first natural antibody immunity after alloHSCT, emphasizing the importance of a functional bone marrow niche. Disclosures: No relevant conflicts of interest to declare.

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Faculty Opinions recommendation of Bone marrow microenvironmental changes underlie reduced RAG-mediated recombination and B cell generation in aged mice.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1020680.232720 ◽

2004 ◽

Author(s):

Cornelis Murre

Keyword(s):

Bone Marrow ◽

B Cell ◽

Cell Generation ◽

Aged Mice

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Faculty Opinions recommendation of Effects of CD20+ B-cell infiltration into allografts on kidney transplantation outcomes: a systematic review and meta-analysis.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.727697810.793533406 ◽

2017 ◽

Author(s):

Minnie Sarwal

Keyword(s):

Systematic Review ◽

Kidney Transplantation ◽

B Cell ◽

Meta Analysis ◽

Cell Infiltration ◽

Transplantation Outcomes

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A Challenging Case of A Myeloid Sarcoma Misdiagnosed as High Grade B-Cell Lymphoma

American Journal of Clinical Pathology ◽

10.1093/ajcp/aqaa161.212 ◽

2020 ◽

Vol 154 (Supplement_1) ◽

pp. S97-S97

Author(s):

A Herrmann ◽

B Mai ◽

S Elzamly ◽

A Wahed ◽

A Nguyen ◽

...

Keyword(s):

Bone Marrow ◽

B Cell ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Myeloid Sarcoma ◽

Proliferation Index ◽

High Grade ◽

Cell Markers ◽

Thoracolumbar Region ◽

The Right

Abstract Introduction/Objective A 46-year-old female presented with severe back pain associated with progressive bilateral lower extremity weakness and paresthesia, urinary retention, and constipation. Computed tomography revealed a retroperitoneal mass encasing the right psoas muscle, obstructing the right kidney, and extending to the thoracolumbar region resulting in severe spinal compression. An epidural tumor resection was subsequently performed at an outside hospital. Methods Histological sections showed sheets of blastoid neoplastic cells with intermediate to large nuclei, irregular membranes, fine chromatin, and prominent nucleoli. Immunohistochemical stains showed that these cells were positive for CD43, CD79a (weak, focal), BCL2, C-MYC, and PAX5 (weak, focal) and negative for CD10, CD20, CD30, ALK1, BCL6, MUM1, and Tdt. The Ki-67 proliferation index was 75-80%. With this immunophenotype, this patient was diagnosed with a high grade B-cell lymphoma and transferred to our institution for further work-up. On review of the slides, further immunohistochemical testing was requested which revealed positivity for CD117 and myeloperoxidase (MPO). Results The overall morphological and immunophenotypical features are most compatible with myeloid sarcoma (MS) with aberrant expression of B-cell markers and this patient’s diagnosis was amended. Interestingly, the patient’s bone marrow examination only showed 2% myeloblasts with left shifted granulocytosis and concurrent fluorescence in situ hybridization (FISH) studies were negative. Conclusion A literature review showed that 40-50% of MS are misdiagnosed as lymphoma. MS can frequently stain with B-cell or T-cell markers, as seen in this case, which makes it challenging for an accurate diagnosis and sub- classification. In addition, our case is interesting in that there was only extramedullary presentation without bone marrow involvement. Typically, MS develops after the diagnosis of acute myeloid leukemia (AML) with an incidence of 3–5% after AML. It can also manifest de novo in healthy patients, who then go on to develop AML months to years later. Therefore, this patient will require close follow-up.

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BONE MARROW INFILTRATION ASSESSMENT BY FDG 18 ‐PET: CAN THIS IMAGING TEST REPLACE BONE MARROW TREPHINE BIOPSY IN DIFFUSE LARGE B CELL LYMPHOMA STAGING?

Hematological Oncology ◽

10.1002/hon.84_2880 ◽

2021 ◽

Vol 39 (S2) ◽

Author(s):

S. Duarte ◽

C. Afonso ◽

B. Marques ◽

C. Barros Lima ◽

D. Neves ◽

...

Keyword(s):

Bone Marrow ◽

B Cell ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Bone Marrow Infiltration ◽

Trephine Biopsy ◽

Bone Marrow Trephine ◽

Bone Marrow Trephine Biopsy ◽

Large B Cell Lymphoma ◽

Large B Cell

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Tumor staging in a Beagle dog with concomitant large B-cell lymphoma and T-cell acute lymphoblastic leukemia

Journal of Veterinary Diagnostic Investigation ◽

10.1177/10406387211011024 ◽

2021 ◽

pp. 104063872110110

Author(s):

Alessandro Ferrari ◽

Marzia Cozzi ◽

Luca Aresu ◽

Valeria Martini

Keyword(s):

Bone Marrow ◽

Lymph Node ◽

T Cell ◽

B Cell ◽

Cell Lymphoma ◽

Tumor Staging ◽

Lymphoblastic Leukemia ◽

B Cell Lymphoma ◽

Large B Cell Lymphoma ◽

Cell Acute Lymphoblastic Leukemia

An 8-y-old spayed female Beagle dog was presented with peripheral lymphadenomegaly. Lymph node cytology and flow cytometry led to the diagnosis of large B-cell lymphoma (LBCL). We detected minimal percentages of LBCL cells in peripheral blood and bone marrow samples. However, a monomorphic population of neoplastic cells different from those found in the lymph node was found in the bone marrow. T-cell acute lymphoblastic leukemia was suspected based on flow cytometric immunophenotyping. PCR for antigen receptor rearrangement (PARR) revealed clonal rearrangement of both B-cell and T-cell receptors, and the presence of both neoplastic clones in the lymph node, peripheral blood, and bone marrow. The dog was treated with multi-agent chemotherapy but died 46 d following diagnosis. Tumor staging and patient classification are needed to accurately establish a prognosis and select the most appropriate therapeutic protocol.

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