An immature B cell population from peripheral blood serves as surrogate marker for monitoring tumor angiogenesis and anti-angiogenic therapy in mouse models

Abstract Females are more susceptible than males to many autoimmune diseases. The processes causing this phenomenon are incompletely understood. Here, we demonstrate that aged female mice acquire a previously uncharacterized population of B cells that we call age-associated B cells (ABCs) and that these cells express integrin αX chain (CD11c). This unexpected population also appears in young lupus-prone mice. On stimulation, CD11c+ B cells, both from autoimmune-prone and healthy strains of mice, secrete autoantibodies, and depletion of these cells in vivo leads to reduction of autoreactive antibodies, suggesting that the cells might have a direct role in the development of autoimmunity. We have explored factors that contribute to appearance of ABCs and demonstrated that signaling through Toll-like receptor 7 is crucial for development of this B cell population. We were able to detect a similar population of B cells in the peripheral blood of some elderly women with autoimmune disease, suggesting that there may be parallels between the creation of ABC-like cells between mice and humans.

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Posttransplant Cyclophosphamide Promote Naïve-Subset Dominant B Cell Recovery Coordinated with Early Treg Expansion; Implication of Reduced Risk of Pathogenesis into Chronic Gvhd

Blood ◽

10.1182/blood-2018-99-120081 ◽

2018 ◽

Vol 132 (Supplement 1) ◽

pp. 4566-4566

Author(s):

Miki Iwamoto ◽

Yusuke Meguri ◽

Takumi Kondo ◽

Hiroyuki Sugiura ◽

Shuntaro Ikegawa ◽

...

Keyword(s):

T Cells ◽

B Cells ◽

B Cell ◽

Cell Population ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Chronic Gvhd ◽

Lymphocyte Subset ◽

Cell Recovery ◽

Haplo Group

Abstract Posttransplant cyclophosphamide (PTCy) is an effective prophylaxis for both acute and chronic graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (HSCT). We recently studied the immune reconstitution dynamics of each lymphocyte subset after PTCy-based transplant using murine haploidentical BMT model and reported that PTCy strongly promoted Treg-dominant T-cell reconstitution and stem cell-derived mature B-cell generation with broad BCR-diversity. We also found that the early reconstitution of Treg could contribute to promote naïve B cell emergence from bone marrow, indicating the T and B cell recovery might be mutually coordinated after PTCy-based transplant (Iwamoto et al, ASH2017). However, the detailed process of immune reconstitution in patients after haploidentical HSCT with PTCy has not been well studied. To address this issue, we here investigated the early dynamics of donor-lymphocyte subset chimerisms in patient after clinical PTCy-based haploidentical HSCT with comparing those in patients after low-dose ATG-based haploidentical HSCT and patients after cord blood transplantation. Laboratory studies were undertaken in 13 adult patients who received HLA-mismatched allogeneic graft; unrelated cord blood (n=5), and haploidentical related peripheral blood after ATG-based conditioning (n=5) and haploidentical related peripheral blood after PTCy-based conditioning (n=5). Blood samples were obtained before and at 1, 2, 4, 6 and 8 weeks after HSCT. Peripheral blood mononuclear cells (PBMCs) were isolated from blood samples by density gradient centrifugation and cryopreserved before being analyzed. After thawing, to analyze the subset-specific chimerism, PBMCs were stained with anti-HLA monoclonal antibodies and other subset-specific antibodies as follows: Pacific Blue conjugated anti-CD4, eFluor450 conjugated anti-CD3, PE-Cy7 conjugated anti-CD25, anti-CD14, APC conjugated anti-CD127, anti-CD56, and APC-eFluor780 conjugated anti-CD8a, anti-CD19. Gated lymphotes (CD4+Tcons, CD4+Tregs, CD8+T cells, B cells, NK cells, Monocytes) were analyzed their chimerism by flowcytometry. To examine the detailed phenotype of B cells, the expression of CD27, CD24, CD38 and IgD were tested. Flowcytometry-based method enables us to analyze the lymphocyte subset chemerism in the very early phase after HSCT. At 2 weeks after HSCT, our analysis revealed that CD4+Tcons, CD4+Tregs and CD8+T cells had already achieved complete donor chimerisms (>95% in all subsets) in patients after ATG-based SCT and had been approaching complete donor chimerisms (85.8%, 75.4% and 87.2%, respectively) in patients after CBT. In contrast, percentage of donor chimerisms of CD4+Tcons, CD4+Tregs and CD8+T cells after PTCy-based haplo-SCT was 73.5%, 59.6% and 59.2%, respectively, and those remained to be in the lower levels than other 2 groups. However, at 4 weeks after HSCT, all examined patients achieved complete donor chimerism of T cells, NK cells and Monocytes (>90%). At 8 weeks after HSCT, the number of B cells in PTCy-based haplo-group was higher than in ATG-based haplo-group (3494 vs 1901/mm3). Of note, B cell population in PTCy-based haplo-group at 8 weeks contained the significantly higher percentage of CD24+CD27-IgD+CD38+ transitional/naïve subset and the significantly lower percentage of CD24+CD27+IgD-CD38neg/dim activated/switched-memory subset when compared to B cell population in ATG-based haplo-group (59.9% vs 10.2%, 2.6% vs 21.5%, P<0.02 respectively), suggesting PTCy treatment might be associated with the favorable B cell reconstitution with naïve-subset dominant composition. Moreover, in patients after PTCy-based haplo-group, the percentage of activated/switched-memory subsets in B cell population at 8 weeks was inversely correlated with percentage of Treg in CD4 T cells at 4 weeks (P<0.05, r2=0.77). Taken together, consistently with our murine study, the current data from clinical samples again suggest that PTCy-based immune-modulation lead to coordinated T and B cell recovery, especially promoting naïve-subset dominant B cell recovery with help of the early expansion of Treg, which might reduce the risk of subsequent chronic GVHD. These data provide the important information for understanding the immunological reconstitution after PTCy-based haploidentical HSCT. Disclosures No relevant conflicts of interest to declare.

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A new CD21low B cell population in the peripheral blood of patients with SLE

Clinical Immunology ◽

10.1016/j.clim.2004.05.010 ◽

2004 ◽

Vol 113 (2) ◽

pp. 161-171 ◽

Cited By ~ 165

Author(s):

Claudia Wehr ◽

Hermann Eibel ◽

Madhan Masilamani ◽

Harald Illges ◽

Michael Schlesier ◽

...

Keyword(s):

B Cell ◽

Cell Population ◽

Peripheral Blood

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A New Memory CD27−IgG+B Cell Population in Peripheral Blood Expressing VHGenes with Low Frequency of Somatic Mutation

The Journal of Immunology ◽

10.4049/jimmunol.177.6.3728 ◽

2006 ◽

Vol 177 (6) ◽

pp. 3728-3736 ◽

Cited By ~ 184

Author(s):

Jessie F. Fecteau ◽

Geneviève Côté ◽

Sonia Néron

Keyword(s):

B Cell ◽

Somatic Mutation ◽

Cell Population ◽

Peripheral Blood ◽

Low Frequency

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Selective expression of CD45 isoforms defines CALLA+ monoclonal B- lineage cells in peripheral blood from myeloma patients as late stage B cells

Blood ◽

10.1182/blood.v78.3.711.711 ◽

1991 ◽

Vol 78 (3) ◽

pp. 711-719 ◽

Cited By ~ 91

Author(s):

GS Jensen ◽

MJ Mant ◽

AJ Belch ◽

JR Berenson ◽

BA Ruether ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Population ◽

Plasma Cell ◽

Late Stage ◽

Peripheral Blood ◽

Plasma Cells ◽

Lower Proportion ◽

Large Subset ◽

B Lineage

Abstract The peripheral blood lymphocytes from 42 patients with multiple myeloma (MM) and 13 patients with monoclonal gammopathy of undetermined significance (MGUS) were studied by three-color immunofluorescence (IF) using antibodies directed to a broad range of B-cell markers (CD19, CD20, CD21, CD24), CALLA (CD10), PCA-1 (a plasma cell marker), and to the high and low molecular weight isoforms of the leukocyte common antigen, CD45RA (p205/220) and CD45RO (p 180). CD45RA is expressed on pre-B and B cells, and a transition from CD45RA to CD45RO defines differentiation towards plasma cells. Peripheral blood mononuclear cells (PBMC) from patients with myeloma included a large subset of B- lineage cells (mean of 39% to 45%) that were CALLA+ and PCA-1+ in all patients studied, including newly diagnosed patients and patients undergoing chemotherapy. Southern blot analysis indicated the presence of monoclonal Ig rearrangements in PBMC and a substantial reduction in the germ-line bands consistent with the presence of a large monoclonal B-cell subset. Avoidance of purification methods involving depletion of adherent cells was essential for detection of the abnormal B cells. Phenotypically, this abnormal B-cell population corresponded to late B or early pre-plasma cells (20% to 80% of PBMC), as defined by the concomitant expression of low densities of CD19 and CD20, moderate densities of CALLA and PCA-1, and strong expression of CD45RO on all B cells, with weakly coexpressed CD45RA on a small proportion. Heterogeneity in the expression of CD45RA and CD45RO within the abnormal B-cell population from any given patient suggested multiple differentiation stages. Abnormal B cells similar to those in MM were also detected in MGUS, although as a lower proportion of PBMC (26%). Abnormal B cells from patients with MGUS expressed predominantly the CD45RO isoform, but had a lower proportion of CALLA+ and PCA-1+ cells than were found on B cells from MM. This work indicates that the large subset of circulating monoclonal B lymphocytes from myeloma patients are at a late stage in B-cell differentiation, continuously progressing towards the plasma cell stage.

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Selective expression of CD45 isoforms defines CALLA+ monoclonal B- lineage cells in peripheral blood from myeloma patients as late stage B cells

Blood ◽

10.1182/blood.v78.3.711.bloodjournal783711 ◽

1991 ◽

Vol 78 (3) ◽

pp. 711-719 ◽

Cited By ~ 3

Author(s):

GS Jensen ◽

MJ Mant ◽

AJ Belch ◽

JR Berenson ◽

BA Ruether ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Population ◽

Plasma Cell ◽

Late Stage ◽

Peripheral Blood ◽

Plasma Cells ◽

Lower Proportion ◽

Large Subset ◽

B Lineage

The peripheral blood lymphocytes from 42 patients with multiple myeloma (MM) and 13 patients with monoclonal gammopathy of undetermined significance (MGUS) were studied by three-color immunofluorescence (IF) using antibodies directed to a broad range of B-cell markers (CD19, CD20, CD21, CD24), CALLA (CD10), PCA-1 (a plasma cell marker), and to the high and low molecular weight isoforms of the leukocyte common antigen, CD45RA (p205/220) and CD45RO (p 180). CD45RA is expressed on pre-B and B cells, and a transition from CD45RA to CD45RO defines differentiation towards plasma cells. Peripheral blood mononuclear cells (PBMC) from patients with myeloma included a large subset of B- lineage cells (mean of 39% to 45%) that were CALLA+ and PCA-1+ in all patients studied, including newly diagnosed patients and patients undergoing chemotherapy. Southern blot analysis indicated the presence of monoclonal Ig rearrangements in PBMC and a substantial reduction in the germ-line bands consistent with the presence of a large monoclonal B-cell subset. Avoidance of purification methods involving depletion of adherent cells was essential for detection of the abnormal B cells. Phenotypically, this abnormal B-cell population corresponded to late B or early pre-plasma cells (20% to 80% of PBMC), as defined by the concomitant expression of low densities of CD19 and CD20, moderate densities of CALLA and PCA-1, and strong expression of CD45RO on all B cells, with weakly coexpressed CD45RA on a small proportion. Heterogeneity in the expression of CD45RA and CD45RO within the abnormal B-cell population from any given patient suggested multiple differentiation stages. Abnormal B cells similar to those in MM were also detected in MGUS, although as a lower proportion of PBMC (26%). Abnormal B cells from patients with MGUS expressed predominantly the CD45RO isoform, but had a lower proportion of CALLA+ and PCA-1+ cells than were found on B cells from MM. This work indicates that the large subset of circulating monoclonal B lymphocytes from myeloma patients are at a late stage in B-cell differentiation, continuously progressing towards the plasma cell stage.

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T and B Cell Populations in Platelet Donors

Tumori Journal ◽

10.1177/030089167806400608 ◽

1978 ◽

Vol 64 (6) ◽

pp. 613-619

Author(s):

B. Lichtiger ◽

J. M. Trujillo ◽

H. E. Fischer ◽

I. S. Cox

Keyword(s):

B Cell ◽

Cell Population ◽

Peripheral Blood ◽

B Lymphocyte ◽

Cell Populations ◽

Appreciable Effect ◽

Lymphocyte Populations ◽

Normal Controls

T and B lymphocyte populations were monitored in the peripheral blood of two long-term plateletpheresis donors. The studies showed minimal fluctuation of both populations compared to the normal controls. The evidence so far accumulated seems to indicate that long-term manual plateletpheresis has no appreciable effect on the level of the T and B cell population of the peripheral blood. Further studies are needed to evaluate larger donor pools.

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Immunophenotypic Analysis Using CD80 and CD86 Staining Improves the Diagnosis of Waldenstrom's Macroglobulinemia by Evidencing a CD80+/CD86- Profile.

Blood ◽

10.1182/blood.v114.22.4389.4389 ◽

2009 ◽

Vol 114 (22) ◽

pp. 4389-4389

Author(s):

Andrea Toma ◽

Stephanie Poulain ◽

Aurore Grelier ◽

Magali Le Garff-Tavernier ◽

Frederic Davi ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Cell Population ◽

Peripheral Blood ◽

Waldenström’S Macroglobulinemia ◽

The Mean ◽

Waldenstrom's Macroglobulinemia ◽

Waldenstrom’S Macroglobulinemia ◽

Waldenström's Macroglobulinemia ◽

Immunophenotypic Analysis

Abstract Abstract 4389 The immunophenotypic characterization is an essential tool in the diagnosis of hematological malignancies but the immunophenotypic profile of Waldenstrom's macroglobulinemia (WM) B cells remains not clearly defined. In order to improve the diagnosis of WM by an immunophenotypic peripheral blood test, we studied within a multicentric protocol the expression of costimulatory molecules CD80 (B7.1) and CD86 (B7.2) on blood B-cells of WM patients diagnosed by monoclonal IgM in the serum and morphological lymphoplasmacytic bone marrow (BM) infiltration. Results were compared to those obtained in healthy controls and other chronic B-cell lymphoproliferative disorders (LPD). Immunophenotypic analysis was performed by flow cytometry using six-colour staining. The CD80 and CD86 staining was concomitant with usual analysis of standard antigens (kappa/lambda immunoglobulin light chains, CD20, CD19, CD79b, CD5, CD10, CD23, FMC7, CD38, CD25, CD43, CD11c, CD1c, CD27, IgM, IgD). We first analyzed 65 cases of WM and compared results to 13 healthy control subjects. Among the WM patients, 45 (69%) showed a monotypic kappa or lambda peripheral blood B-cell population with a Matutes score '3. In these patients, CD80 was always positive (>21% of B-cells) and CD86 negative (<18%). The mean responses obtained in WM compared to controls were: CD80 = 44.2±19.5% versus 13±4.6% (p<0.00001), CD86 = 5.4±5.3% versus 4.5±2.1% (p=0.18). Among the 20 patients without monotypic kappa or lambda B-cell population in peripheral blood, 13 showed a CD80+/CD86- profile (CD80 = 33±7.4%, CD86 = 7.6±4.4%). Six of them were studied by molecular aproach (PCR) showing the presence of a clonal B-cell population in all cases. Overall, 90% of WM patients showed a CD80+/CD86- profile in blood. Seven patients without peripheral monotypic kappa or lambda B-cell population evidenced a CD80-/CD86- profile; among them, one was tested by PCR and showed the lack of a clonal B-cell population in blood. We then studied other chronic B-cell LPD with peripheral blood neoplastic involvement by a monotypic kappa/lambda population, including 15 marginal zone lymphoma (MZL), 4 non-Hodgkin lymphomas (NHL) and 28 chronic lymphocytic leukemia (CLL). The mean expression of CD80 was 37±35.6%, 2.5±1.7%, 12.4±20.6%, and of CD86 30.5±38%, 19±27.5%, 15.4±19.3%, respectively. The expression of these antigens in MZL was heterogeneous, but a WM-type CD80+/CD86- profile was never observed in peripheral blood B cells. No NHL and 4 CLL (14%) exhibited a CD80+/CD86- profile. In conclusion, while immunophenotypic analysis usually performed with standard antigens does not allow to define a typical profile of WM, additional CD80 and CD86 staining in blood samples usefully improves the diagnosis of WM in the context of chronic B-cell LPD with peripheral blood involvement having a Matutes score '3. Furthermore, even in the absence of blood involvement detected by classical imunophenotypical aproach, the CD80 and CD86 staining may prove the presence of a peripheral blood monoclonal population, discriminating WM from other B-LPD with plasmatic IgM and lymphoplasmacytic BM infiltration, and diagnosing a WM in 90% of cases. Disclosures: No relevant conflicts of interest to declare.

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