A case of acute lymphoblastic leukemia presented with multiple liver tumors.
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症例は17歳, 女性. 既往歴, 家族歴に特記すべきことなし. 1997年1月下旬より咽頭痛, 38℃台の発熱が続き, 肝障害, 腹部エコーにて肝及び脾に多発性の腫瘤性病変を認めたため精査目的にて入院. 入院時に著明な肝脾腫を認めた. 末梢血像は正常であったが, 肝腫瘍生検にて著明な小円形細胞の腫瘍性増殖を認めた. 骨髄穿刺にてリンパ球表面マーカーはCD10, CD19, CD20, CD34陽性でありALL (L1) に合致した所見であった. その後末梢血中に腫瘍細胞が出現し, 化学療法にて肝腫瘤像の著明な改善を認めた. 白血病細胞は高頻度に肝に浸潤するが, 重症肝障害を来すことは稀とされる. 本例は入院時に肝障害を認め, 診断時において末梢血像に腫瘍細胞の出現なく, 多発性の肝腫瘤を契機にALLの診断に至った. 白血病においては一般的に肝生検などによる診断は禁忌な場合が多いとされるが, 早期に肝生検を施行し診断できた貴重な症例と思われ報告した.B cells from systemic lupus erythematosus (SLE) patients display signalling defects that may underlie disease pathogenesis activity.CD19 and CD22 play a major role as regulators of B-cell response. The aim of this study was to clarify the relationship between B cell surface markers namely CD19, CD20 and CD22 expression and clinical and laboratory indices of SLE activity. The study included 33 SLE patients and 20 healthy children and adolescents as controls. Flowcytometric assay of dual markers, CD19/CD20, and CD20/CD22 was done. SLE disease activity was assessed by SLEDAI score. CD22% was significantly higher while CD20% was significantly lower in the study compared to the control group. No significant difference was observed in both groups with respect to CD19% or CD19/CD22% ratio. The level of CD22 expression was significantly lower in high and very high active cases than in mild and moderate cases and negatively correlated with SLDEAI score and ESR. Results obtained showed that, B cell surface receptors CD20 and CD22 are significantly affected in patients with SLE, pointing to their possible involvement in the aetiopathogenesis of the disease and in the regulatory mechanisms in response to the immune disturbance.
CD22
Pathogenesis
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Abstract CD20 is a B cell specific membrane protein and a target of therapeutic antibodies such as rituximab (RTX) 1 . In spite of the prominent usage of anti-CD20 antibodies in the clinic little is known about the biological function of CD20 2 . Here we show that CD20 controls the nanoscale organization of receptors on the surface of resting B lymphocytes. A CRISPR/Cas-based ablation of CD20 in Ramos B cells results in a relocalisation of the IgM B cell antigen receptor (IgM-BCR) and the co-receptor CD19. The resulting IgM-BCR/CD19 signaling synapse leads to transient B cell activation followed by plasma cell differentiation. Similarly to CD20-deficient Ramos cells, naïve human B cells treated with rituximab in vitro or isolated from patients during rituximab administration display hallmarks of transient activation characterized by the formation of the IgM-BCR/CD19 signaling synapse, followed by CD19 and IgM-BCR downregulation. Moreover, increased expression of specific plasma cell genes can be observed after rituximab treatment in relapsed CLL patients. In summary we identify CD20 as a gatekeeper of the resting state on human B cells and demonstrate that a disruption of the nanoscale organization of the B cell surface via CD20 deletion or anti-CD20 treatment profoundly alters B cell fate.
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Objective To detect the expressions of CD19 and CD20 of B-lymphocytes (CD19+B, CD20+B) and natural killer (NK) cells in the peripheral blood of epileptic children and explore their significances. Methods Four hundred and fifty-eight epileptic children, admitted to our hospital from January 2008 to December 2010, were chosen as patient group; another 52 healthy subjects were chosen as controls. The expressions of CD19+B and CD20+B and NK cells were detected by flow cytometry; their results were compared. Ninety-two epileptic children were treated with intravenous immunoglobulin (IVIG) and the effect of IVIG therapy was studied. Results The CD19+B level ([22.35±6.54]％) and CD20+B level ([21.50±8.41]％) in the epileptic children were obviously higher than those in the healthy controls ([16.86±4.02)％,[16.13±4.19]％, P＜0.05); and the level of NK cells ([9.1 1±4.90]％) in the epileptic children was significantly decreased as compared with that in the healthy controls ([14.72±4.15]％, P＜0.05). The CD19+B level ([18.26±5.03]％) and CD20+B level ([16.74±5.12]％) 6months after MG treatment were decreased significantly as compared with those before treatment ([22.74±6.25]％,[21.61±8.03]％, P＜0.05]; while the level of NK cells ([14.65±4.58]％) 6 months after IVIG treatment was increased significantly as compared with that before treatment ([9.07±4.76]％,P＜0.05). Among the 92 patients treated with IVIG, 70 enjoyed good results and 22 had non-effective resutls; The changesofCD19+Blevel ([7.99±5.34]％) and CD20+B ([8.21±5.21]％) before and after treatment in effectively treated patients by IVIG were significantly different as compared with those in ineffectively treated patients by MG ([3.78±2.76]％,[3.66±2.48]％, P＜0.05); however, the changes of level of NK cells before and after treatment showed no significant difference between effectively treated patients and ineffectively treated patients ([5.28±4.55]％,[4.53％±4.43]％, P＞0.05). Conclusion Dysfunctions of B-lymphocytes and NK cells exist in epileptic children, and MG treatment shows good effect on immune dysfunction of them. The levels of CD19-B and CD20-B can be used as monitoring indicators in the IVIG treatment of epileptic children.
Key words:
Epilepsy; Immunoglobulins; Natural killer cell; CD19; CD20
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CD80
CD86
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Abstract T cells contribute to both acute and chronic allograft rejection, but the roles of B cells and alloantibody are less clear. Therefore, CD20 and CD19 mAbs were used to deplete mature or total B cells, respectively, in mice before allografting. Both CD20 and CD19 mAbs effectively depleted mature B cells, but CD19 mAb also depleted >70% of CD138+ plasma cells in flow cytometry assays, as well as >66% of Ab-secreting cells in ELISPOT assays. Consequently, CD19 mAb treatment depleted serum IgG levels by 80% in naïve mice, while CD20 mAb did not. Neither CD20 nor CD19 mAb treatment affected acute cardiac allograft rejection, with all grafts rejected within 7-9 days. However, CD19 mAb treatment prevented the generation of allo-IgG. In a second model of rejection, treatment with CD19 mAb before renal transplant significantly delayed and prevented chronic renal allograft rejection, reduced kidney pathology, and abrogated the development of allo-IgG, while CD20 mAb did not. In additional experiments, CD19 mAb treatment also depleted pre-existing allo-IgG levels in serum. In a third allograft model, pretreatment with CD20 mAb exacerbated acute skin allograft rejection and augmented the proliferation of adoptively transferred alloantigen-specific CD4+ T cells by 2-fold, indicating that B cells negatively regulate acute skin graft rejection and CD4+ T cell responses. Thus, depending on the nature of the allograft, B cells can either positively or negatively regulate allograft rejection.
ELISPOT
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In this study we describe a fast and sensitive method using three‐colour immunofluorescence for the detection of cells with phenotypes that are rare in normal bone marrow (BM) but occur frequently in children with precursor B acute lymphoblastic leukaemia. We show that, in the first year after initiation of therapy, in 17/18 patients (10 patients were analysed after first diagnosis and nine patients after first BM relapse; one patient was analysed on both occasions) the percentage of CD10 + CD19 + cells and CD20 − CD22 + cells in the CD34 + cell population indicated the likelihood of relapse. A suppression of cells expressing these phenotypes after initiation of therapy was followed by an outgrowth of normal precursor B cells after 12 months. Therefore this early test for impending relapse (which occurred 10–28 months after starting chemotherapy) was only applicable in the first year after beginning the treatment. However, despite this predictive value, comparison of fluorescence data with PCR results obtained from the same BM samples indicated that only a subpopulation of the CD34 + CD10 + CD19 + and CD34 + CD20 − CD22 + cells above the determined threshold value represented malignant cells. A large prospective study to confirm the predictive value of this three‐colour immunofluorescence assay is warranted.
Immunofluorescence
Immunophenotyping
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To assess the presence and phenotype of B-lineage cells in the bone marrow (BM) of rheumatoid arthritis (RA) patients after rituximab therapy.Six patients were studied. BM aspirates were collected 3 months after the treatment and analysed using the four-colour flow cytometry.CD19+ (B-lineage) cells in BM samples varied from 0.1 to 3.25% in the lymphoid gate. CD34+ cells varied from 1.23 to 4.86%. The proportion of CD34+ cells committed to the B-lineage varied between 0 and 42.19%. Pro-B-cells were undetectable in one case. The majority of B-cell precursors were pro-B-cells in Patients 5 and 6 (50 and 62% of CD19+ cells, respectively), pre-B-cells in Patients 3 and 4 (64 and 70%) and immature B-cells in Patient 1 (44%). Detectable CD20 expression on CD19+ cells was either low or absent. Plasma cells varied from 0.01 to 0.36% of the total nucleated cells. There was a trend towards longer duration of clinical response in patients with evidence of more complete depletion in BM.In this small cohort of RA patients treated with rituximab, differences in proportion and phenotype of CD19+ BM cells were detected. These differences suggest variation in the degree of depletion achieved and correlate with time to relapse. Although pro-B-cells are not targeted directly by rituximab as they do not express CD20, the levels were unexpectedly low.
Immunophenotyping
CD5
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Normal B-cell differentiation has been characterized extensively, but discrepancies persist regarding the exact sequence of antigen expression. Few systematic studies focusing on identification of the minor or undetectable B-cell subsets in normal human bone marrow (BM) which are frequently found in leukemic cells have been performed. Such studies could help to monitor minimal residual disease (MRD) in precursor-B-acute lymphoblastic leukemia (precursor-B-ALL). The aim of the present study was to analyze the sequence of antigen expression among normal human CD19+ B cells from adult BM. Our major goal was to identify infrequent and undetectable B-cell phenotypes that could be used for the detection of MRD in patients with precursor-B-ALL.Adult BM samples from a total of 33 healthy volunteers were analyzed using triple stainings, and measured by flow cytometry. A sensitive method based on the two-step acquisition procedure was used for the identification and characterization of cells present at very low frequencies.Five different subsets of CD19+ cells were identified in normal BM samples according to their degree of maturation: 1) CD19+/CD34+/CD10-/CD20-/CD22dlm+ (0.5 +/- 0.4% B cells); 2) CD19+/CD34-/CD10++/CD20-/CD22dlm+ (3.4 +/- 2.7%); 3) CD19+/CD34-/CD10+/CD20-/CD22dlm+ (3.5 +/- 2.2%); 4) CD19+/CD34-/CD10+/CD20+,++/CD22dlm+ (21 +/- 11%), and 5) CD19+/CD34-/CD10-/CD20++/CD22+ (73 +/- 19%). We observed that several B-cell phenotypes are frequent among precursor-B-ALL, but are infrequent or undetectable in normal human B cell differentiation. Accordingly, in all normal BM samples analyzed, less than 4 x 10(-5) cells co-expressed CD19 and CD117; CD20strong+/CD34+ and CD22strong+/CD34+ events were found at frequencies less than 5 x 10(-4), while CD20+/CD34+ phenotypes were found in less than 1 x 10(-3) BM cells. Although both CD19+/CD13+ and CD19+/CD33+ events were found at frequencies of up to 3 x 10(-3), they never formed a well-defined population of cells and therefore these latter phenotypic patterns could also be of use for MRD investigation in CD13+ and/or CD33+ precursor-B-ALL cases.Our results show that in adult BM normal B-cells display constant patterns of maturation as regards both their phenotypic characteristics and their relative distribution. Abnormalities in these patterns provide a potentially useful tool for monitoring MRD in precursor-B-ALL patients who achieve cytomorphologic complete remission.
Minimal Residual Disease
Immunophenotyping
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Refractory (planetary science)
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<p>T cell populations become CD8-dominant post CART19/20 cell infusion. % CD8+ among (A) CAR-expressing T cells and (B) all CD3+ T cells was quantified by flow cytometry. Data are shown for CAR-T cells only up to day 90 post infusion as values post day 90 become unreliable due to low CAR+ cell count detected by flow. FP: final product (i.e., cryopreserved CART19/20 cells).</p>
Refractory (planetary science)
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