Sinus Histiocytosis with Massive Lymphadenopathy: A Spectrum of Disease Associated with Immune Dysfunction

A detailed immunologic study of three cases of sinus histiocytosis with massive lymphadenopathy (SHML) was performed to better characterize this rare disorder. One patient had prominent cervical lymphadenopathy that regressed spontaneously, whereas the other two patients had persistent cervical lymphadenopathy and recurrent infections. The first patient was otherwise healthy and had normal immunologic studies. One of the latter patients had a relative increase in blood B cells, a decreased level of serum immunoglobulin A (IgA), decreased blood lymphocyte mitogenic responses to multiple mitogens (37-42% of controls), and cutaneous anergy. The other patient with persistent disease also had a relative increase in blood B cells, polyclonal hypergammaglobulinemia, and circulating immune complexes, as well as decreased blood T cells and markedly decreased blood lymphocyte responses to mitogens (12-37% of controls). Immunohistochemical stains of the lymph nodes of the three patients revealed a characteristic phenotype for the sinus histiocytes: S-100 protein, 3/3; CD14 (Leu M3), 3/3; CD11c (Leu M5), 1/1; CD71 (OKT9), 3/3; CD4 (Leu 3a), 2/3; CD1a (OKT6), 1/3; alpha-1-antitrypsin, 3/3; alpha-1-antichymotrypsin, 3/3; CD35 (C3b), 1/1; CD11b (Mo1), 0/3; CD15 (Leu M1), 0/3; HLA-DR, 0/3; and lysozyme, 0/3. This phenotype suggests that the cells of SHML have features of both the Langerhans/interdigitating cell and mononuclear phagocyte lineages. Emperipolesis by the histiocytes of B cells, T cells, and natural killer cells was demonstrated by a double-staining technique. Our findings indicate that patients with SHML may have a variably expressed immunodeficiency that predisposes them to recurrent infections.     

Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease): clinico-pathological study of three cases

Introduction We report three cases of sinus histiocytosis, a rare disease of unknown aetiology with massive lymphadenopathy (SHML), also known as Rosai-Dorfman (RD) disease, in a paediatric population. This proliferative histiocytic disorder is defined by histological and immunohistochemical (IHC) characteristics and can manifest as nodal involvement with variable enlargement of the lymph nodes (two cases) and extranodal manifestations involving skin and larynx involvement (one case). One patient had hypergammaglobulinemia. The morphological investigation revealed that all lymph nodes showed hyperplasia of sinuses with abundant histiocytic cell with intracytoplasmic lymphocytes. Skin and larynx biopsies showed a histiocyte and lymphocyte infiltrate with similar characteristics. An ultrastructural study was carried out on material from one patient. In the IHC study, SHML cells expressed phagocytic markers such as CD68 and S100, but markers for Langerhan’s (CD1a) or dendritic cells (DRC, CD23 and CNA42) were absent. Two patients had a complete remission after surgical excision and no other treatment, but the third patient was treated with radiotherapy after a relapse with obstruction of the upper airway.Conclusion This disorder must be considered in the differential diagnosis of young patients who exhibit massive or multiple lymphadenopathies, especially when involvement of the cervical area occurs. Due to the good outcome of the disease, a conservative approach is justified.     

Bone Marrow Transplantation for Chronic Granulomatous Disease Associated with Cytocrome B Deficiency

The diagnosis of acute leukemia consists of morphological FAB classification, genetic abnormality assay, cytochemical analysis, and immunophenotype based on surface and cytoplasmic expression of antigens of blastic cells.^1-6 The commonly used immunophenotypic classification of acute lymphoblastic leukemia (ALL) includes null ALL (only HLA-DR and/or CD19 antigens are present on blast surface), common ALL (HLA-DR, CD19, and CD10 expression on blasts), and pre-B subtype with cytoplasmic μ chain in CD10, HLA-DR, and CD19 positive cell.^1,2,4,7 The heterogeneity of early B-cell differentiation antigens expression (atypical patterns) was described in a few cases, more often in null ALL subtype.^3,5 It suggested that the earliest stages of B-cell development showed less stable antigen expression. The clinical meaning of this heterogeneity was unknown.^6,8     

慢性扁桃腺炎患儿T淋巴细胞表型及功能分析

目的　探讨慢性扁桃腺炎患儿T淋巴细胞表型和功能障碍情况,为分析其病因和发病机制提供临 床和理论依据。方法　采用免疫荧光标记和流式细胞仪技术检测了27例慢性扁桃腺炎患儿和21例健康儿童外 周血T细胞亚群、B细胞及NK细胞的表面标记和T细胞活化后表面分子(CD3+/HLA DR+T和CD3+/CD25+)的 表达;同时采用ELISA方法检测了患儿和对照组血清中Th1型细胞因子IL 2和IFN γ的水平。结果　与对照组比 较,慢性扁桃腺炎患儿CD4+T细胞和CD3+/HLA DR+T活化细胞的阳性率显著降低(28.6%±4.4%vs25.4% ±4.5%,P<0.05;5.7%±1.9%vs3.9%±1.6%,P<0.01),CD4+/CD8+比值降低(1.17±0.30vs0.92± 0.18,P<0.01);但患儿CD3+T细胞、CD8+T细胞和CD3+/CD25+阳性的T细胞与对照组比较差异均无显著意 义。患儿CD19+B细胞阳性率、与Th1细胞功能有关的IL 2和IFN γ表达水平也较对照组显著降低(P<0.05或 P<0.05);而两组CD3 /CD(16+56)+NK细胞的阳性率无显著差异。结论　慢性扁桃腺炎患儿存在CD4+T细 胞减少,T细胞活化障碍,Th1细胞功能异常及B细胞减少,这可能是患儿反复感染和病程迁延的主要原因之一。     

高效抗逆转录病毒治疗对艾滋病患儿CD8+T细胞活化分子CD38和人类白细胞抗原DR表达水平的影响及其与病毒载量的关系

目的 观察高效抗逆转录病毒治疗(HAART)对我国艾滋病患儿CD8+T细胞活化标志分子CD38和人类白细胞抗原DR(HLA-DR)表达水平的影响及其与病毒载量的关系.方法 对194例接受HAART的艾滋病患儿进行横断面研究,用流式细胞术检测CD4+、CD8+T细胞数,以及CD8+/CD38+和CD8+/HLA-DR+T细胞比例,RT-PCR检测血浆HIV RNA载量.并检测52名健康儿童CD8+/CD38+和CD8+/HLA-DR+T细胞比例作为正常对照.结果 本组中,135例病毒载量＜400 copies/ml,59例病毒载量≥400 copies/ml.病毒载量≥400 copies/ml组CD8+/CD38+T和CD8+/HLA-DR+T细胞水平显著高于病毒载量＜400 copies/ml组,差异有统计学意义(29.6±10.1 vs19.9±9.8;17.7±6.4 vs 9.6±6.1,P＜0.05);病毒载量＜400 copies/ml组,CD8+/CD38+T细胞接近正常水平,而CD8+/HLA-DR+T细胞仍高于正常水平,差异有统计学意义(19.9±9.8 vs 15.6±9.0;9.6±6.1 vs 5.8±3.3,P＜0.05).CD8+/CD38+和CD8+/HLA-DR+T细胞百分比均与病毒载量成正相关(前者相关系数R=0.403,P=0.03,后者相关系数R=0.569,P=0.09).结论 艾滋病患儿CD8+T细胞活化程度与病毒载量成正比,有效的HAART治疗能够显著地降低HIV感染者免疫活化程度,CD8+/CD38+和CD8+/HLA-DR+T细胞百分比可能是资源有限地区替代病毒载量检测的潜在指标.

Abstract：

Objective To study the expression of CD38 and HLA-DR on CD8 + T cells in pediatric AIDS patients receiving highly active antiretroviral therapy (HAART) and the relationship of immune activation and disease progression. Methods A cross-section study of 194 pediatric AIDS patients receiving HAART was carried out and 52 age-matched healthy children were recruited as control. The percentage of CD4+ , CD8+ , CD8+/CD38 + and CD8+/HLA-DR+ T cells was tested using flow cytometry, and HIVRNA in plasma was detected by quantitative RT-PCR. Results One hundred and ninety-four pediatric AIDS patients were divided into two groups according to the viral load: 59 patients with VL≥400 copies/ml and 135 patients with VL＜400 copies/ml. The percentage of CD8 +/CD38+ and CD8 +/HLA-DR+ T cells of patients with VL≥400 copies/ml was significantly higher than that of patients with VL ＜ 400 copies/ml (P ＜ 0. 05 ). Of patients with VL ＜ 400 copies/ml, the percentage of CD8 +/CD38 + T cells was nearly normal, and the percentage of CD8 +/HLA-DR+ T cells was higher than normal level ( P ＜ 0. 05 ). There was a positive correlation between percentage of CD8+/CD38+ and of CD8 +/HLA-DR+ T cells and viral load ( R = 0. 403, P = 0. 03 for the former and R = 0. 569, P = 0. 09 for the later). Conclusions Effective HAART could decrease immune activation of HIV-infected children significantly. And there was a positive correlation between percentage of CD8 +/CD38 + and of CD8 +/HLA-DR + T cells and viral load, suggesting that the two indicators might be used as the substitution of viral load in resource-limited areas.     

A Basis for Distinguishing Cultured Dendritic Cells and Macrophages in Cytospins and Fixed Sections

There is a burgeoning literature on the contrasting role of intratumoral dendritic cells (DCs) and tumor-associated macrophages, making reliable identification of both cell types in clinical and experimental tissue sections important. However, because these cell types are closely related and share several differentiation antigens, their absolute distinction in tissue sections is difficult. We differentiated DCs and macrophages from monocytes in vitro, prepared cytospins and paraffin-embedded sections of the various cell populations, and tested a variety of antibodies that purportedly recognize monocytes and DCs for their capacity to react and distinguish cells after conventional formalin fixation. Cultured DCs but not macrophages were detected by fascin, DC-LAMP, and CD83 with a predictable increase in staining that paralleled their maturation. Staining by CD1a was found on immature DCs but was weak and absent on mature DCs and macrophages, respectively. CD14 and CD163 were characteristic for macrophages and absent on DCs. CD68, HLA-DR, and S100 did not discriminate between DCs and macrophages. We conclude that antigens such as HLA-DR and S100 are not in themselves sufficient for identification of DCs in formalin-fixed tissue sections, but that additional macrophage-specific (CD14, CD163) and DC-specific (CD1a, CD83, fascin, DC-LAMP) antigens should be used to distinguish cell types from each other and to provide information on their state of maturation.     

T-lymphocyte and monocyte activation in the course of infection in children with neoplastic disease

One of the side effects of antineoplastic treatment is immunosuppression. A consequence of this are frequent infections, sometimes with fatal outcome. The immunological system "answers" to infections with changes in number and quality of cells participating in the inflammatory process. The aim of the study was to investigate the subpopulations of mononuclear cells in patients with neoplastic (haematologic) diseases during infections. We studied 16 children with acute lymphoblastic leukaemia and with lymphomas (Hodgkin and non-Hodgkin). Among our patients we note 25 episodes of infections requiring hospitalization. Mean percent values of mononuclear antibodies: CD14-PE/CD45-FITC, CD3-FITC/CD19-PE, CD4-FITC/CD8-PE, CD3-FITC/CD16+CD56-PE, CD14-PerCP/CD54-PE, CD3-PerCP/CD54RA-FITC/CD45RO-PE, CD3-PerCP/HLA-DR, CD14-PerCP/HLA-DR. In the examination carried out during acute infection, we found higher mean percent values of CD3+ HLDAR+, CD14+ CD54+ and CD3+ CD45RO+ cells than in analogous values in the remaining examinations. After the infection we found higher percent of CD3 CD45RA+/CD3+CD45RO+ cells. In a patient with cytomegalovirus pneumonitis our examination showed a depression of HLA-DR antigens on monocytes during acute infections (10th day of treatment) and return to normal values after 20 days of treatment. Conclusions: (1) The most sensitive expression of infection in children with neoplastic disease was the variable percent of HDLA-DR molecules on T lymphocytes. (2) Presence of activation indicators on T lymphocytes and monocytes can testify that there is persistent function of the immunological system in spite of immunosuppression in patients with neoplastic disease. (3) Depression in percent value of monocytes with HLA-DR antigens in the course of cytomegalovirus pneumonitis confirms its immunosuppressive influence on the human organism.     

Systemic lupus erythematosus of childhood onset: correlation between T cells expressing early and late activation antigens and disease activity

Correlation between T cell phenotypes, especially activated T cells expressing early (EA1) and late (HLA-DR) activation antigens and clinical features were investigated in 22 patients with systemic lupus erythematosus (SLE) of childhood onset. Percentages of T cells expressing EA1 and HLA-DR in 22 patients with SLE were significantly higher than those in controls. Comparison of T cell phenotypes between patients with active and inactive SLE showed that eight patients with active disease had significantly increased percentages of HLA-DR positive T cells than 14 with inactive disease (P<0.01). Serial examinations showed that the percentages of HLA-DR positive T cells were decreased after therapy in seven with active non-renal or active non-renal and renal diseases but not in one with only active renal disease. A possible significance of T cells expressing EA1 and HLA-DR in the management of patients with SLE is discussed.Abbreviations HLA-DR human leukocyte antigens-DR - SLE systemic lupus erythematosus     

Changes in lymphocyte subsets in children with newly diagnosed type 1 diabetes mellitus

The aim of this study was to assess changes in selected peripheral blood lymphocyte subsets in children and adolescents with newly diagnosed type 1 diabetes mellitus (DM) and determine the correlation between these changes and other immunological markers. The study involved a group of 39 patients aged 2-14 years and a control group. The number of T- and B-lymphocytes and the number of CD4, CD8, CD4/HLA-DR, CD8/HLA-DR, CD5/CD20 subsets were measured by flow-cytometry using monoclonal antibodies. Islet cell antibodies (ICA) and antibodies to glutamic acid decarboxylase (GADA) were assessed. In both the diabetic and control groups the number of T-and B-lymphocytes were within normal limits. In patients with DM, the percentage of CD5+/CD20+ cells was significantly increased compared with the control group (p < 0.0001). ICA were positive in 80% of patients and GADA in nearly 65%. A positive correlation between the CD5/CD20 subset and ICA and GADA was found. In patients with a high percentage of CD5+/CD20+ lymphocytes, a higher percentage of activated subsets (CD4/HLA DR and CD8/ HLA DR) was detected. In conclusion: CD5/ CD20 lymphocyte subsets are a good additional marker of autoimmunological processes in DM.     

维生素A对脐血树突状细胞分化与成熟及其功能的影响

目的　研究维生素A在体内的代谢活性产物视黄酸 (retinoicacid ,RA)对树突状细胞(dendriticcells ,DC)分化、成熟及功能的影响 ,进一步探索维生素A对免疫功能调节的机理。方法　取健康足月儿脐血 9份分离单个核细胞后 ,进行体外培养诱导DC的同时加入生理浓度的RA ,采用流式细胞仪检测DC的表面分子CD1a、CD83、HLR DR以判断RA对DC分化、成熟的影响 ;混合淋巴反应的强度观测RA对DC抗原递呈功能的作用 ;荧光定量PCR法检测细胞因子IL 12p35、IL 12p4 0、IFN γ(Th1)、IL 4、IL 10 (Th2 )mRNA转录水平 ,分析RA对DC诱导Th细胞分化的影响。结果　培养第 6天RA使DC数量显著下降 (P <0 0 0 1) ,培养第 9天 ,DC总数相仿 ,但RA处理后不成熟DC百分比增高 ,而成熟DC百分比降低 ,差异有显著意义 (P <0 0 0 1) ;DC混合淋巴反应的cpm值降低2 9 4 % (P <0 0 0 1)。细胞因子IL 12、IFN γmRNA下降 ,而IL 4、IL 10的mRNA水平升高 ,差异均具有显著意义。结论　维生素A延迟体外培养的脐血单个核细胞向DC的分化和成熟 ,且降低其混合淋巴细胞反应的能力 ,减少Th1细胞因子的产生而增加Th2细胞因子的产生 ,使免疫反应向Th2方向偏移。     

儿童急性淋巴细胞白血病免疫分型特征及其临床意义

目的：探讨急性淋巴细胞白血病（ALL）患儿的免疫分型及其与临床特征的关系。方法：139例ALL患儿为研究对象,取肝素抗凝的骨髓或静脉血2~3 mL,利用流式细胞仪进行免疫分型分析。结果：139例ALL患儿中,B-ALL 103例（74.1 %）,T-ALL 24例（17.3 %）,T/B双表型ALL 12例（8.6 %）。B-ALL主要表达的抗原有CD19（90.3%）、CD10（83.5%）、CD20（27.2%)。T-ALL主要表达的抗原有CD3（79.2%）、CD7（66.7%）、CD5（33.3%）。B/T-ALL主要表达的T系抗原有CD7（50.0%）、CD5（41.7%）;B系抗原有CD19（50.0%）、CD10（33.3%）。139例ALL患儿中,32例伴髓系抗原表达（My+）,主要表达的抗原有CD13、CD33、CD14、MPO等。139例ALL患儿中,31例表达CD34;而My+ ALL中CD34阳性表达（15.6%）明显低于My- ALL（24.3%）。139例ALL患儿中,82例表达HLA-DR。CD10、CD34、HLA-DR在标危、中危、高危型ALL中的表达差异有统计学意义。My+ ALL组性别、出血发生率与My- ALL组比较差异有统计学意义（P<0.05）。结论：免疫分型可正确区分儿童ALL的来源。CD10、CD34、HLA-DR抗原表达与ALL的临床分型有关。     

Strong response of T cells in infants with dual infection by Epstein–Barr virus and cytomegalovirus

BACKGROUND: Although a reversed CD4/CD8 ratio and increased proportion of CD8+ HLA-DR+ T cells are well known as the characteristic immune response in infectious mononucleosis (IM), it has not been elucidated whether these immune responses are affected by patient age and pathogenetic viruses. METHODS: T cell subsets were analyzed by two-color flow cytometry using fluorescein isothiocyanate- and phycoerythrin-conjugated monoclonal antibodies in 115 infants and children aged from 4 months to 10 years with IM due to Epstein-Barr virus (EBV), cytomegalovirus (CMV) and dual infection with both viruses. RESULTS: A reversed CD4/CD8 ratio and increased proportions of CD4+/HLA-DR+ T cells, CD8+ T cells and CD8+/HLA-DR+ T cells became more prominent as the age of the patients became older. No differences were observed in proportions of T cell subsets between EBV- and CMV-infection among patients aged from 6 to 17 months. Although the responses of these T cells were weak in infants with single virus infection by EBV and CMV, markedly strong T cell responses comparable with those in older children were observed in infants with EBV/CMV dual infection. Clinical symptoms were more severe in patients with EBV/CMV dual infection than those with EBV or CMV alone. CONCLUSION: The manner of these T cell responses in the acute phase of IM was considered to be age dependent, although strong T cell responses and severe disease were observed in EBV/CMV dual infection irrespective of patient age.     

101例儿童急性髓细胞性白血病免疫表型与预后相关分析

目的探讨免疫表型对儿童急性髓细胞性白血病的预后价值。方法采用流式细胞术检测101例儿童急性髓细胞性白血病(AML)患儿相关免疫表型,分析免疫表型对完全缓解(CR)及无疾病生存期(DFS)的影响。结果CD34阴性组及CD34、HLA-DR同时阴性组一疗程CR率均明显高于非阴性组,差异有统计学意义(P=0.008,0.000);DFS的CD34阴性组,HLA-DR阴性组以及CD34和HLA-DR同时阴性组均明显高于非阴性组,差异有统计学意义(P=0.004,0.006,0.040),而CD14,CD15,CD7,CD19差异对CR和DFS均无统计学意义。结论免疫表型对评估AML患儿的预后有一定意义。     

Phenotypic analysis of circulating dendritic cells during the second half of human gestation

Dendritic cells (DCs) have been characterized as having an immature phenotype in infants when compared with adults; but it is unclear whether the phenotype or function of these populations changes during human intrauterine development. Three-colour flow cytometry was used to phenotype fetal/neonatal circulating DCs during the second half (>20-wk gestation) of pregnancy, (n = 34) and adults (n = 9). DCs were identified from peripheral blood mononuclear cells (PBMCs) or cord blood mononuclear cells (CBMCs) as staining brightly for HLA-DR but negative for T cell, B cell, monocyte, and NK cell lineage markers. The surface molecule of interest was detected in a third colour. During gestation CD34, a marker of immaturity was significantly higher, and CD4, a differentiation marker, was significantly lower than adult levels. The percentage of CD11c+ cells did not differ significantly at any age, although a trend to reduced intensity of expression at earlier stages of gestation was observed. Significantly fewer DCs expressed the IgG receptors CD32 and CD64 at all gestations. The percentage of HLA-DR+/lin- cells expressing CD40 was lowest at 20–23 wks and was always significantly lower on DCs from cord blood vs. adult blood. Similarly, the percentage of CD86+ and CD54+ DCs was significantly lower than adults throughout gestation. Thus, immaturity of cord blood DCs is likely to arise as a consequence of decreased ability to take up antigen (at least via IgG-mediated mechanisms) and reduced provision of co-stimulation.     

Characterization of Inflammation in Syphilitic Villitis and in Villitis of Unknown Etiology

Chronic villitis is a histologic diagnosis that may be either associated with infection, or termed villitis of undetermined etiology (VUE). The lymphocytic infiltrate in VUE has been reported to consist of maternal lymphocytes, but the origin of the lymphocytic infiltrate in infectious villitis has not been identified. The purpose of our study was to compare the maternal vs. fetal origin of the infiltrating lymphocytes in VUE and syphilitic villitis, and to expand the immunophenotypic data provided by previous studies. Paraffin-embedded placentas from four males with VUE and two males with syphilitic villitis were subjected to fluorescence in situ hybridization (FISH) for the X and Y chromosomes. Serial sections were stained with antibodies to CD3, CD4, CD8, CD68, HLA-DR, and CD20. Quantitation of the relative number of cells marking with each antibody was done for four villi in each slide. CD3 lymphocytes predominated in both VUE and syphilitic villitis, with slightly more CD8 cells compared to CD4 cells. CD68 and HLA-DR positive cells were as frequent as CD3 cells, and B-lymphocytes were rare. Maternal cells were the predominant intravillous population in both VUE and syphilitic villitis, and neutrophils in syphilitic villitis were also maternal. These data indicate that the immune response in both syphilitic villitis and VUE is similar, raising the possibility of a similar immunopathogenetic pathway.     

DOR-1, A novel CD10+ stromal cell line derived from progressive Langerhans cell histiocytosis of bone

BACKGROUND: Langerhans cell histiocytosis (LCH) is granulomatous proliferative disorder characterized by the presence of activated Langerhans cells admixed with macrophages, lymphocytes, and eosinophils. In an effort to obtain an LCH ex vivo model, we succeeded in establishing the DOR-1 cell line from an LCH lesion of bone in a 3-year-old girl. PROCEDURE: The DOR-1 cell line was established from a CD1a immunoreactive LCH lesion of bone maintained in long-term cell culture. The phenotypic characteristics were assessed by immuno-cytochemistry and fluorescence activated cell sorter (FACS) analysis. Cytogenetic analysis was performed by RHG-banding that was supplemented by fluorescence in situ hybridization (FISH). RESULTS: The DOR-1 cells grew in vitro as a poorly differentiated mesenchymal-like cells with a doubling time between 72 and 96 hr. The cells exhibited pleomorphism and consistent immuno-reactivity for CD10 (50%), CD13 (55%), CD68 (65%), and CD117 (70%) while CD1a, Langerin and HLA-DR were not detected. By RHG-banding, several aberrant chromosomes were detected including the t (9; 17) (p23; p13) translocation and a pair of long dicentric marker chromosomes indicating clonal abnormality. Functionally, exposure to 33 nM 12-O-tetradecanoyl phorbol mirystate-13-acetate (TPA) induced DOR-1 cell differentiation with appearance of cytoplasmic extensions. CONCLUSIONS: The DOR-1 cell line exhibits distinct immuno-cytochemical features and carries the t (9; 17) (p23; p13) translocation suggesting involvement of stromal-like cell lineage in LCH initiation and progression.     

肠道病毒71型感染患儿免疫功能探讨

目的 探讨肠道病毒71型(EV71)感染患儿免疫功能变化与病情程度的关系.方法 患儿46例,健康同龄儿童12例,根据病情由轻到重将患儿分为4组:手足口病组11例、中枢神经系统病变组20例、中枢神经系统病变伴自主神经功能失调组10例、神经源性肺水肿组(pulmonaryedema,PE组)5例.进行下述检测:CD14~+单核细胞人类白细胞DR抗原(HLA-DR)表达率、淋巴细胞免疫分型、CD4~+CD25~+Foxp3~(high)调节性T细胞(regulatory T cells,Treg cells)及TH17细胞比例;白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、白细胞介素10(IL-10)、转录生长因子β(TGF-β)、白细胞介素6(IL-6)、白细胞介素17A(IL-17A)血浓度;CD~4T细胞Foxp3、ROR-γt基因表达;血清免疫球蛋白及补体等.结果 (1)前炎症细胞因子TNF-α及IL-1β在轻症患儿中增高,随病情加重而下降,PE组明显降低(P<0.05);抗炎细胞因子IL-10及IL-10/TNF-α比值随病情加重增高,PE组增高明显(P<0.05).(2)HLA-DR、CD3~+T细胞、CD4~+T细胞、CD8~+T细胞、NK细胞随病情加重呈现逐步下降趋势,PE组下降最为明显(P<0.05).各组间B淋巴细胞及抗体差异无统计学意义.(3)Treg细胞比例、转录因子Foxp3 mRNA及诱导因子TGF-β血浓度随病情加重降低,而TH17细胞比例、IL-17A血浓度、转录因子ROR-γt mRNA及诱导因子IL-6血浓度随病情加重升高.结论 EV71感染患儿机体免疫功能随病情程度而变化,轻症患儿处于全身炎症反应状态,重症或危重症病例处于代偿性抗炎症反应或混合性拮抗反应状态,对EV71感染的免疫调控治疗应强调分阶段、个体化.     

Depressed levels of interferon-gamma and HLA-DR+CD3+ T cells in infants with transient hyperferritinemia

Familial hemophagocytic lymphohistiocytosis (FHL), which typically has its onset during infancy, is uniformly fatal if not treated. It therefore requires prompt therapeutic intervention. Although hyperferritinemia has been emphasized as a useful marker for FHL, some nonfatal cases in infants with spontaneous remission also manifest with hyperferritinemia. However, distinguishing them is difficult because initial clinical features of these infants are similar. The authors encountered 14 infants with hyperferritinemia (serum ferritin >674 ng/mL), which normalized within 3 weeks following a benign clinical course. The authors compared the levels of HLA-DR+CD3+ T-cell subsets and interferon-gamma (IFN-γ) in the peripheral blood between these infants and FHL cases: one of the authors' own patients and others from the literature. Serum IFN-γ was not detected in infants with hyperferritinemia. Moreover, levels of HLA-DR+CD3+ T cells were extremely depressed. In contrast, serum IFN-γ was elevated and HLA-DR+CD3+ T cells were not depressed in FHL. Measurement of activated T cells and serum IFN-γ might help differentiate FHL in febrile infants with transient hyperferritinemia.     

CD45+和CD45-儿童急性B淋巴细胞白血病的临床特征分析

目的探讨CD45抗原表达与儿童急性B淋巴细胞白血病(B-ALL)的临床相关性。方法利用多参数流式细胞术(FCM)以CD45/SSC设门对61例B-ALL患儿进行免疫分型检测;采用染色体核型分析和多重巢式RT-PCR技术进行染色体、融合基因分析。结果免疫表型分析显示CD45+组CD13、HLA-DR抗原表达明显高于CD45-组(P<0.05);CD45-组的融合基因主要为HOX11和E2A/PBX1,而CD45+组主要为TEL/AML1、MLL/AF9、HOX11;相关临床参数、染色体核型及疗效比较两组间差异均无显著性(P>0.05)。结论 CD45抗原的表达与否和B-ALL患儿的临床特征、染色体畸变、治疗效果无统计学相关性;CD45+组较CD45-组高表达CD13和HLA-DR。     

婴幼儿脓毒症不同免疫状态细胞因子变化探讨

目的 检测不同免疫状态脓毒症婴幼儿细胞因子浓度的变化,探讨婴幼儿脓毒症免疫功能紊乱的可能机制.方法 深圳市儿童医院重症监护室2007年5月至11月收治的脓毒症患儿36例为研究组,16例正常婴幼儿为对照组.根据人类白细胞抗原(HLA)-DR的测定值,将患儿分为免疫激活组(DR-H组)和免疫抑制组(DR-L组).用流式细胞术检测CD14+单核细胞HLA-DR表达率;实时荧光定量PCR检测CD4+T细胞IL-10、IL-6 mRNA表达;ELISA检测IL-1β、IL-6、TNF-α、IL-10血浆浓度.结果 (1)小儿危重病例评分DR-H组高于DR-L组(90.15±4.43 vs 81.91±7.45,P＜0.05).(2)细胞因子变化:两组前炎症细胞因子IL-1β、IL-6血浓度均明显高于对照组(P＜0.05),DR-H组TNF-α高于对照组.两组问比较,IL-1β及IL-6差异无显著性,DR-H组TNF-α高于DR-L组(P＜0.05).两组IL-6基因表达均高于对照组(P＜0.05),以DR-L组增高尤为显著.两组IL-10均高于对照组(P＜0.05),DR-L组IL-10基因表达高于对照组及DR-H组.结论 CD14+单核细胞HLA-DR表达检测对婴幼儿脓毒症危重程度及预后有一定判断价值.婴幼儿脓毒症发病过程中,前炎症细胞因子始终处于高表达状态,导致免疫抑制的主要原因可能是抗炎因子IL-10过表达.