Behçet's disease accompanied by myelodysplastic syndrome with trisomy 8: two case reports and a review of 15 Japanese cases

We describe two cases of Behçet's disease associated with myelodysplastic syndrome (MDS) with trisomy 8. Both cases developed ulceration in the cecum as a gastrointestinal complication of Behçet's disease, after a diagnosis of MDS. We summarized recent case reports of Behçet's disease associated with myelodysplastic syndrome, and studied the clinical manifestations. Most cases showed trisomy 8 as a chromosomal abnormality. Gastrointestinal involvement without eye lesions seems to be characteristic of Behçet's disease associated with MDS.     

Primary acquired sideroblastic anemia, thrombocytosis, and trisomy 8

 Myelodysplastic syndromes are usually associated with pancytopenia. Disorders involving deletion of the long arm of chromosome 5 (5q- syndrome) and, rarely, patients with karyotypic abnormalities involving chromosome 3 associated with abnormal thrombopoiesis may have a normal or even raised platelet count. Other cytogenetic abnormalities in myelodysplasia are invariably associated with cytopenia in one or all cell lineages. We report a patient who initially presented with slight anemia and a raised platelet count. Further investigations suggested a diagnosis of primary acquired sideroblastic anemia. Cytogenetic examination revealed a clone with trisomy 8. We believe this is the first reported case of trisomy 8 with trilineage myelodysplasia and thrombocytosis with primary acquired sideroblastic anemia. Received: 13 August 1996 /Accepted: 12 December 1996     

骨髓增生异常综合征克隆性8号染色体三体和7号染色体缺失的发生及其临床意义

目的：了解＋8和-7／7q-克隆在骨髓增生异常综合征（MDS）中的发生情况，探讨＋8和-7／7q-克隆在MDS发生发展中的可能机制及其临床意义。方法：采用短期培养法和G显带技术，对MDS患者的染色体异常、尤其＋8和-7／7q～核型进行分析。结果：70例原发性MDS患者中，有43例存在染色体异常，异常检出率为61．4％，最常见的染色体异常为＋8（18例）和-7／7q-（8例），其中4例为＋8与-7／7q-并存。＋8克隆多出现在RA阶段，而-7／7q-克隆则出现在疾病进展中。伴有-7／7q-核型的MDS，更容易转化为白血病。结论：＋8和-7／7q-核型仍是MDS最为常见的染色体异常。＋8克隆出现在MDS的早期，可能与Fas介导的细胞凋亡有关；而-7／7q-克隆的形成，导致MDS的进展及向白血病转化，可能与丢失片段的抑癌基因的失活有关。     

Association between trisomy 8 and the immunophenotype of blast cells from acute leukemias secondary to a myelodysplastic syndrome or chronic myeloproliferative disorders

 In the present study we have used FISH to analyze the incidence of trisomy 8 in acute leukemias following either a primary myeloproliferative disorder (MPD) or a myelodysplastic syndrome (MDS) and correlated it with both the immunophenotype and the cell-cycle distribution of the leukemic blast cells. Six of the 21 (28%) acute leukemias studied displayed trisomy 8 by FISH. The number of trisomic cells in these cases ranged from 20 to 84%, with a mean of 46±24%. Trisomy 8 was associated with a homogeneous population of leukemic cells, phenotypically characterized by CD34+ / HLADR+ / CD13+ / CD33+ / CD11b– / CD15– / CD14–. No significant differences were observed on the proliferative rate of cases with trisomy 8, as compared with blast cells from the remaining patients. Overall, our findings suggest that in acute leukemias secondary to MPD or MDS, trisomy 8 is associated with a blockade of myeloid maturation at an early step of the differentiation process. Received: 2 December 1996 / Accepted: 12 February 1997     

Aberrant expression of the major sialoglycoprotein (CD43) on the monocytes of patients with myelodysplastic syndromes

 CD43, a sialylated glycoprotein expressed on the surface of most hematopoietic cells, has been implicated in cell adhesion and signaling. The reduced expression of this antigen in patients with WiscottAldrich syndrome, in which progressive immunodeficiency is a major problem, raised the question whether abnormal expression of this molecule could affect the susceptibility to infections in patients with myelodysplastic syndromes (MDS). We studied the expression of this antigen on the monocytes of ten patients with chronic myelomonocytic leukemia (CMML) and compared the results with 67 patients suffering from other MDS syndromes and with 18 healthy individuals. We chose this series as it plays an important role in MDS patients where in most cases the neutrophils are defective. We also examined the following antigens as indicative of activation and adhesion of the monocytes in these patients: CD11b, CD18, CD35, CD38, CD44, CD69. We found decreased expression of CD43 on the monocytes of the RA, RAS, RAEB, and RAEB-t patients compared with the CMML and controls. The other activation molecules studied were found to be upregulated, suggesting the existence of activated monocytes in these patients. The increased levels of soluble vascular cell adhesion molecule in these patients suggest vascular endothelial activation in the absence of infection. Further experiments are needed to investigate the significance of CD43 downregulation in these patients, its role in cell adherence and tissue migration, and the correlation of the phenomenon to the increased susceptibility to infections observed in these patients. Received: 9 April 1999 / Accepted: 21 September 1999     

Atypical chronic myelogenous leukemia in a patient with trisomy 8 mosaicism syndrome

Summary A 17-year-old woman was admitted for bone marrow transplantation with the diagnosis of atypical Philadelphia-negative chronic myelogenous leukemia (aCML), cytogenetically characterized by trisomy 8 as the sole chromosome aberration. A striking feature was a congenital opacity of the right cornea. Chromosomal analysis of skin fibroblasts were performed and revealed a mosaic for trisomy 8. Commonly, a distinct clinical picture leads to the diagnosis of trisomy 8 mosaicism syndrome (T8ms), but an extreme phenotypic variability has been observed. To our knowledge the development of an aCML in a patient with T8ms has not been reported. A review of the literature revealed that an association to other hematological disorders had been described in two cases. The question of whether our patient's aCML was a random event or not is discussed. The patient is now 24 months post transplant and shows no evidence of disease. Her Karnofsky score is 100%. We conclude that it might be worthwhile to look for an associated constitutional trisomy 8 mosaicism in all patients with trisomy 8 leukemia.     

Molecular and flow cytometric characterization of the CD4 and CD8 T-cell repertoire in patients with myelodysplastic syndrome

We studied 18 patients with myelodysplastic syndrome (MDS), measuring clonality and T-cell receptor Vbeta (TCRBV) expression of CD4 and CD8 T cells by polymerase chain reaction and by flow cytometric analysis of TCRBV families. The CD4 and CD8 T-cell repertoire in most MDS patients is characterized by an abnormal TCRBV-restricted expansion of T cells in CD4 and CD8 cells, and increased expression of the CD8 effector marker CD57 of multiple TCRBV in CD8 cells. Clonality analysis of CD4 and CD8 cells showed that seven of 10 patients analysed had a major clone in the CD8 cells but not in CD4 cells. Furthermore, in one patient we found that both the CD57- and CD57+ fraction contained the clone (which was absent from the TCRBV-negative fraction). These data suggest that, in MDS, multiple T-cell expansions can be found in both helper and cytotoxic T cells, and that, in the CD8 cells, T cells functionally differentiate in vivo from memory to effector T cells. Together, these data support the hypothesis of the involvement of T cells in the pathogenesis of MDS.     

Induction of CD4+CD8+ double positive T cells and increase in CD5+ B cells in efferent lymph in sheep infected with Trypanosoma evansi

The effects of Trypanosoma evansi on efferent lymphocyte phenotypes draining from a lymph node primed with Pasteurella haemolytica vaccine were studied in sheep. The prefemoral efferent lymphatic ducts of the infected sheep along with those of two uninfected sheep were surgically cannulated. Lymph was collected and lymphocytes recovered from it analysed by two-colour indirect immunofluorescence staining and cytofluoremetry in a fluorescence activated cell analyser (FACSCAN). The study showed the appearance and persistence of T. evansi in the efferent lymph for a long period of time and the appearance of CD4⁺CD8⁺ (double positive, DP) T lymphocytes in the efferent lymph of infected animals. The infection also resulted in increases in CD5⁺ B cells in the prefemoral efferent lymph. In addition, there were decreases in the output of conventional B cells, CD5⁺ and CD4⁺ T cell subsets but large increases in CD8⁺ cells followed by terminal depletion of all cell subsets. In contrast, inoculation of sheep with pasteurella vaccine antigen alone produced little alterations in the proportions, but large increases in the numbers of all T cell subsets except that of CD8⁺ cells which also showed little variation; and there was a concurrent increase in the numbers and proportions of efferent B cells. In addition, the abnormal expression of DP and CD5⁺ B cells did not occur in the uninfected vaccinated sheep. It is concluded that these abnormal changes in the kinetics of efferent lymphocyte phenotypes are likely to play a role in the genesis of the generalized immunosuppression seen in trypanosome-infected hosts.     

CD3和CD8免疫反应阳性细胞在人胎回肠的发育

目的: 探讨CD3和CD8免疫反应阳性细胞在人胎回肠淋巴组织发生过程中的分布、定位及发育.方法: 收集因故终止妊娠人胎回肠30例, 采用免疫组织化学SP法染色显示CD3和CD8免疫反应阳性细胞, 用BioMiaspro图象分析软件对免疫反应阳性细胞进行计数, 有关数据作统计学分析.结果: 9 wk, 回肠内可见CD3和CD8阳性反应细胞, 主要定位于肠壁的结缔组织内, 数量较少, 散在分布;11 wk可见CD3和CD8阳性细胞三五成群分布;17 wk, CD3和CD8阳性细胞聚集成群, 主要分布于绒毛中轴以及小肠腺之间固有结缔组织, 回肠黏膜上皮偶见阳性细胞浸润;18 wk, CD3和CD8阳性反应细胞出现在淋巴小结处参与构成淋巴小结;24 wk后, CD3和CD8阳性反应细胞数量增多, 尤以典型的集合淋巴小结处有较多分布, 其中, 在28 wk数量增加最为明显( P<0.05). CD8和CD3阳性细胞数量之间有较强的正相关关系( r = 0.831,P<0.01), CD3阳性细胞在整个发育过程中数量上均较CD8阳性细胞多.结论: 人胎肠相关淋巴组织的形成过程中, 第9周即有T细胞发育. CD8和CD3阳性T细胞的发育时序基本同步.     

Expression of CD22 on peripheral B cells in patients with rheumatoid arthritis: relation to CD5-positive B cells

B cells in patients with rheumatoid arthritis (RA) are hyperactivated. Although B cell receptor signal transduction may be affected by various response regulators, CD22 plays an important role as a response regulator of B cells. Therefore, we investigated and examined CD22 expression on peripheral blood B cells of patients with RA. Thirty-two RA patients and 16 controls were enrolled in this study, and CD22 expressions on B cells were analyzed by flow cytometry. In patients with RA, CD22⁺ B cells significantly decreased in comparison to the controls (ratio: P < 0.05). However, there was no correlation between this decrease and the clinical data. Interestingly, CD5⁺ CD22⁻ B cells significantly increased in RA patients. The decrease in CD22⁺ B cells and increased in CD5⁺CD22⁻ B cells play critical roles in the pathogenesis of RA mediated by the activation of B cells.     

Evaluation of CD5-positive B cells in blood and synovial fluid of patients with rheumatic diseases

Summary CD5 (OKT1, Leu-1) is an antigen originally associated only with T cells. This antigen has recently been detected on a population of B cells that have been implicated in autoimmune diseases, particularly in rheumatoid arthritis (RA). We determined the percentage of these cells in the peripheral blood (PB) and synovial fluid (SF) of patients with RA and other joint diseases (OJD) using flow cytometry and two-colour fluorescence. No significant difference was observed between the percentages of CD5-positive B cells in the PB of these two patient groups or healthy individuals. In comparison with PB, significantly higher percentages of these cells were observed in the SF of patients with RA, but not in SF of OJD patients. Higher percentages of B cells were also noted in RA SF. However, when B-cell percentages were accounted for, no significant difference was observed between the numbers of B cells expressing CD5 in SF. No correlation was observed between the percentages of CD5-positive B cells and the levels of rheumatoid factor (RF) or C-reactive protein (CRP). We conclude that CD5-positive B cells are an integral part of the B-cell pool of PB and that there is increased representation of B cells in SF. The increased percentages of B cells in RA SF, compared with OJD, may reflect the autoimmune phenomenon occurring in the rheumatoid joint.     

Ring chromosome 8 and trisomy 8 in a patient with acute myeloid leukemia

We describe a child with Acute Myeloid Leukemia (AML M7) with trisomy 8 and ring chromosome 8. Ring chromosome 8 associated with AML is uncommon and is reported to have a poor outcome. The combination of trisomy 8 and ring chromosome 8 has not been previously reported. This 15-month-old girl had presented with a history of fever, weight loss of 1 kg, gum bleeds and pallor. Clinical examinations revealed no nodes or organomegaly. Investigations revealed pancytopenia and elevated serum LDH. Bone marrow aspirate confirmed the presence of myeloid blasts positive only for CD 41 and CD 61 on flow cytometry. Chromosomal analysis from the bone marrow showed 46, XX [13]/ 47, XX, +8[2]/ 47, XX, +r (8) [5]. The child was treated as per UK MRC AML protocol (ADE 10+3+5). Bone marrow on day 21 post-induction was in morphological remission. Repeat karyotyping revealed 46,XX suggesting that the patient was in cytogenetic remission. Cytogenetic sub grouping in AML patients provides guidelines for the choice of optimal treatment strategy. There was no HLA matched family donor and hence an unrelated donor search was commenced as she was in the group with unfavourable cytogenetics. She developed acute myelofibrosis soon after the second cycle of chemotherapy with swinging fever and rapidly enlarging spleen. The marrow showed 11% blasts with intense fibrosis. She went through a stormy period during conditioning for unrelated stem cell transplantation. She passed away on day 11 post transplantation of veno-occlusive disease of liver and multiorgan failure. This case illustrates the poor outcome in paediatric AML with trisomy and ring chromosome 8.     

Soluble CD4 and CD8 molecules in patients with systemic sclerosis

Summary The concentrations of sCD4 and sCD8 in 69 patients with systemic sclerosis (SSc) were examined by using a sandwich enzyme-linked immunosorbent assay. The patients with SSc had significantly higher concentrations of sCD8 (mean 249.2 U/ml (SD 155.1), median 224 U/ml) than the normal subjects (mean 149.3 U/ml (SD 42.1), median 148 U/ml). The concentration of sCD4 in patients with SSc were significantly lower (mean 6.2 U/ml (SD 3.8), median 5.0 U/ml) than in the normal subjects (mean 10.9 U/ml (SD 4.1), median 10.3 U/ml). The concentration of sCD8 in patients with diffuse sclerosis tended to be higher than in those with sclerodactyly.     

Cerebellar myeloblastoma formation in CD7-positive, neural cell adhesion molecule (CD56)-positive acute myelogenous leukemia (M1)

 We present a first report of a CD7⁺ acute myelogenous leukemia patient who developed intracranial myeloblastomas. The patient was neurologically normal on physical examination at presentation. The peripheral leukocyte count was extremely high (203.6×10⁹/l). The blasts expressed CD7 and CD56 (neural cell adhesion molecule) in addition to CD13, CD33, CD34, and HLA-DR. The karyotype of bone marrow cells was normal. The patient was diagnosed as having acute myelogenous leukemia (AML, M1). Following a short period of complete remission, bone marrow relapse and meningeal leukemia occurred, and the patient died of respiratory failure. Autopsy revealed that blasts had invaded the subarachnoid space and cerebellum, and two myeloblastomas were found in the cerebellar hemisphere. Both CD7⁺ and CD56⁺ AML have been reported to have a high incidence of central nervous system involvement. CD7⁺ CD56⁺ AML calls for prophylaxis of central nervous system leukemia. Received: 2 May 1997 / Accepted: 17 July 1997     

An aggressive case of Burkitt's lymphoma with t(8;14) and c-myc rearrangement transformed from CD5+ B-cell lymphoma

 We experienced a case of Burkitt's lymphoma showing an unusual surface phenotype, CD5 expression, at an early stage of the disease. Initially, this patient showed massive abdominal para-aortic lymph node swelling which rapidly developed into leukemic change. Based on the clinical course and cytogenetic features of lymphoblasts in the bone marrow, which showed t(8;14) and c-myc gene rearrangement, the patient was diagnosed with Burkitt's lymphoma. Combination chemotherapy induced short-term remission, but central nervous system (CNS) involvement developed, followed by a regrowth of lymphoma cells in the bone marrow. The bone marrow at the end stage showed monotonous expansion of large cells with conspicuous vacuolation in the basophilic cytoplasm. The initial lymphoma cells showed pan-B markers and were CD5 positive but weakly CD10 positive; however, the lymphoma cells obtained from the bone marrow at the terminal stage did not express CD5. The chromosomal t(8;14) was seen, and identical rearrangement of immunoglobulin heavy chain joining gene and c-myc gene were detected by Southern blot analysis in the bone marrow lymphoblasts throughout the clinical course. This case is evidence that remarkable transformation of CD5-positive lymphoblasts to CD5-negative lymphoblasts occurred in an identical clone of Burkitt's lymphoma. Received: 25 April 1997 / Accepted: 19 September 1997     

系统性红斑狼疮患者CD5+B细胞的测定和意义

目的探讨外周血中CD5 B细胞在系统性红斑狼疮(SLE)活动中的作用及相关性。方法利用流式细胞分析法对57例SLE患者和35名正常人群外周血CD5~ B细胞进行检测,并且同时检测抗dsDNA抗体、抗核抗体(ANA)、抗心磷脂抗体(ACL)、补体C3、C4。结果SLE患者CD5~ B细胞水平[(2．1 0．4)%]与正常人[(1．5±0．4)％]比较差异有统计学意义(P＜0．05),活动期SLE患者CD5~ B细胞水平(2．5±0．5)％显著高于稳定期(1．4±0．5)％；CD5 B细胞与dsDNA、ANA、抗心磷脂抗体(ACL)升高呈正相关,与补体c3呈负相关。结论系统性红斑狼疮患者外周血CD5~ B细胞明显升高,与SLE疾病活动有一定关系。     

Role of CXCR5+ CD8+ T cells in human hepatitis B virus infection

The replication of HBV in hepatocytes can be effectively inhibited by lifelong antiviral therapy. Because of the long-term presence of HBV reservoirs, the virus rebound frequently occurs once the treatment is stopped, which poses a considerable obstacle to the complete removal of the virus. In terms of gene composition, regulation of B cell action and function, CXCR5⁺CD8⁺ T cells are similar to CXCR5⁺CD4⁺ T follicular helper cells, while these cells are characterized by elevated programmed cell death 1 and cytotoxic-related proteins. CXCR5⁺CD8⁺T cells are strongly associated with progression in inflammatory and autoimmune diseases. In addition, CXCR5 expression on the surface of CD8⁺ T cells is mostly an indicator of memory stem cell-like failure in progenitor cells in cancer that are more responsive to immune checkpoint blocking therapy. Furthermore, the phenomena have also been demonstrated in some viral infections, highlighting the duality of the cellular immune response of CXCR5⁺CD8⁺ T cells. This mini-review will focus on the function of CXCR5⁺CD8⁺ T cells in HBV infection and discuss the function of these CD8⁺ T cells and the potential of associated co-stimulators or cytokines in HBV therapeutic strategies.     

Immunophenotype and clinical characteristics of CD45-negative and CD45-positive childhood acute lymphoblastic leukemia

 To evaluate the expression pattern of the leukocyte common antigen CD45 in acute leukemias and to investigate whether the lack of CD45 expression in childhood acute lymphoblastic leukemia (ALL) is associated with other immunophenotypic features and a distinct clinical behavior, we have carried out extensive immunophenotypic analyses of bone marrow and peripheral blood samples from 638 patients with childhood B-cell precursor (n=529) or T-lineage ALL (n=109). All 638 patients were enrolled in the German ALL-BFM 90 and ALL-BFM 95 trials. CD45 was detected on the surface of childhood ALL cells (cut-off ≥20% positive cells) in only 88.7% (n=566) of all cases. Among 529 patients with childhood B-cell precursor ALL, 12.9% (n=68) did not express CD45, compared with only 3.7% (n=4) of patients with childhood T-lineage ALL (p<0.001). In the B-cell precursor ALL subtypes, the highest frequency of CD45- cases (15.1%) was observed in common ALL (56/372) compared with only 7.2% in pro-B ALL (3/41) and 7.8% in pre-B ALL (9/116). Assessment of clinical parameters (age, organ enlargement, WBC, etc.) and event-free survival did not reveal significant differences between CD45- and CD45+ patients. Myeloid antigen coexpression was not correlated with CD45 expression. The mean percentage of antigen expression for CD34, CD10, TdT, CD22, and CD24 was significantly higher in children with CD45- B-cell precursor ALL than in those with CD45+ B-cell precursor ALL. In 28 patients with B-cell precursor ALL, cell cycle analyses of freshly isolated leukemic cells were performed with propidium iodide (PI) staining and flow-cytometric analysis. The percentage of cells in S-phase was inversely correlated to the percentage of CD45+ cells (r=-0.48, p<0.05). With two-parameter analysis of CD45-fluorescein isothiocyanate (FITC)- and PI-stained cells in nine patients with a percentage of CD45+ cells between 40 and 60%, two populations were distinguishable in a single patient. It was shown that the CD45- subpopulation had a higher percentage of cells in S-phase than the CD45+ subpopulation (10.7±4.0 vs. 2.7±1.8, p<0.007). We conclude that the lack of CD45 expression contributes to the identification of a distinct functional and immunological subgroup of B-cell precursor ALL, but that it has no significant impact on clinical behavior or on therapy outcome in childhood ALL. Received: February 18, 1998 / Accepted: May 26, 1998     

骨髓增生异常综合征患者JAK2和STAT5基因表达的变化

目的:比较骨髓增生异常综合征(MDS-RA)型与MDS-RAEB型患者外周血JAK2、STAT5基因表达水平,探索JAK2和STAT5信号转导在MDS发病中的作用。方法:建立JAK2、STAT5基因表达的荧光定量FQ-PCR检测方法,检测10例正常人、15例MDS-RA患者和10例MDS-RAEB患者外周血JAK2、STAT5基因表达水平及其自细胞介素(IL)-2、IL-3、γ干扰素(γ-INF)、肿瘤坏死因子α(TNF-α)细胞因子水平。结果:正常人IL-2、IL-3、γ-INF、TNF-α分别为50．28±14．19、29．15±5．47、26．17±5．36、31．12±8．73;MDS-RA患者分别为51．16±13．44、67．72±10．19、43．39±13．08、62．36±12．78;MDS-RAEB患者为19．55±21．86、28．74±15．52、64．34±27．35、82．13±17．79。正常人JAK2、STAT5基因不表达或低表达,拷贝数分别为480．07±609．17、116．05±173．87,MDS-RA患者分别为4 725．63±3 931．59、1 265．36±1 087．80,显著高于正常人(均P<0．01); MDS-RAEB患者分别为23006．11±11 311．10、13 144．55±8 493．36,显著高于MDS-RA患者(均P<0．01)。结论:细胞因子及其相关的JAK2和STAT5基因参与了MDS的发病及其恶性转变,JAK和STAT可能是MDS细胞由低危MDS-RA型向高危MDS-RAEB型恶性转化的信号通路之一。