非呼吸道合胞病毒性肺炎患儿淋巴细胞亚群变化

引起婴幼儿肺炎的病毒病原中,除呼吸道合胞病毒(RSV)外,还与其他许多病毒如腺病毒、副流感病毒、流感病毒等有关。许多研究表明,RSV可引起急性毛细支气管炎,也与儿童哮喘的发生密切相关,其致喘的发病机制与T辅助淋巴细胞亚群的失衡有关,且以Th2增高为特点。为探讨非RSV病毒性肺炎对机体免疫功能产生的影响,我们对33例非RSV感染的病毒性肺炎患儿的细胞免疫功能进行了研究。1对象与方法1.1研究对象2002～2004年期间在我科住院之非RSV病毒性肺炎33例(为病毒组),男21例,女12例;年龄1～72个月,平均(24.21±21.74)个月。除发热、咳嗽等症状…     

静脉应用人血丙种球蛋白对重症感染患儿淋巴细胞亚群免疫调节作用

目的研究静脉应用人血丙种球蛋白(IVIG)对重症感染患儿淋巴细胞亚群的免疫调节作用,在重症感染时使用IVIG的安全性。方法采用单克隆抗体直接免疫荧光法,流式细胞术测定28例重症感染患儿外周血淋巴细胞亚群在应用浓度为0.2%和2.0%IVIG前后的变化。结果低浓度IVIG应用前后CD3、CD25、CD3CD25、CD4、CD28、CD4CD28、CD95、CD152计数均无明显差异(P>0.05);高浓度IVIG应用后仅CD152计数增高(P<0.05),其他CD亚群均无明显差异(P均>0.05)。结论低浓度IVIG对严重全身感染患儿外周血淋巴细胞亚群的影响不明显,高浓度IVIG通过抑制细胞毒性T细胞(Tc)扩增的机制,下调严重全身感染患儿外周血T细胞活化,抑制免疫功能。     

以流式细胞术建立中国汉族健康儿童外周血淋巴细胞亚群相对计数的正常参考值

目的建立中国汉族健康儿童外周血淋巴细胞亚群的正常参考值范围。方法选取首都医科大学附属北京儿童医院入托、入学体检,或术前查体及术后复查(均为对免疫功能影响不大的疾病)的0～18岁汉族儿童为研究对象。按年龄分为婴儿组(28d至12个月),幼儿组(～3岁),学龄前组(～7岁),学龄组(～12岁)和青春期组(～18岁)。采集外周血以双色及四色荧光标记流式细胞术检测淋巴细胞亚群T细胞(CD3+CD19-)、CD4+T细胞(CD3+CD4+)、CD8+T细胞(CD3+CD8+)、B细胞(CD3-CD19+)和NK细胞(CD3-CD16+CD56+)相对计数及CD4+/CD8+比值。比较各年龄组不同性别淋巴细胞亚群分布的差异,建立正常参考值范围。结果由于青春期组所收集的标本数较少,将该研究对象组予以删除。最终纳入28d至12岁儿童592例。①婴儿组T细胞、CD8+T细胞百分比与CD4+/CD8+比值性别差异均有统计学意义,幼儿组CD4+T细胞、CD8+T细胞百分比与CD4+/CD8+比值性别差异均有统计学意义,学龄组CD4+T细胞百分比性别差异有统计学意义;②除了男童T细胞百分比各年龄组间差异无统计学意义外,男女儿童各年龄组间外周各血淋巴细胞亚群分布总体上差异均有统计学意义;进一步行男女儿童淋巴细胞亚群不同年龄组间两两比较,发现多个年龄组间差异有统计学意义;③男女儿童T细胞、CD8+T细胞和NK细胞百分比随年龄增长均呈逐渐升高趋势;CD4+T细胞、B细胞百分比及CD4+/CD8+比值随年龄增长均呈逐渐降低趋势;男女儿童T细胞、B细胞、NK细胞百分比和CD4+/CD8+比值升高或降低的程度略有不同;④中国汉族健康儿童淋巴细胞亚群分布特点与欧、美、非洲国家儿童相比存在相似的升高或降低的趋势,但数值上存在一定的差异。结论儿童淋巴细胞亚群分布存在年龄和性别的差异。本研究成功建立中国汉族28d至12岁健康儿童淋巴细胞亚群相对计数正常参考值范围。     

Lymphocyte subsets in whole blood and isolated mononuclear leucocytes of healthy infants and children

The determination of lymphocyte subsets utilizing monoclonal antibodies and flow cytometry has become essential in the evaluation of immunological status. Using a standardized method it was found that in healthy children the percentage of CD 8+ (Leu 2+) positive cells increases significantly (P<0.01) during infancy, whereas the percentage of CD 4+ (Leu 3+) positive cells decreases with age (P<0.01). The percentage of CD 3+ (Leu 4+) cells remains constant. The ratio of CD 4/CD 8 positive cells is significantly (P<0.001) higher in infants than in older children. Other subpopulations (HLA DR+, Leu 7) were found to be constant in all age groups. For the comparison of data on lymphocyte subsets obtained by flow cytometry a standardized test procedure is important.     

Lymphocyte subset reconstitution in pediatric liver recipients induced with steroid-free rabbit anti-human thymocyte globulin

Multiple measurements of lymphocyte subsets in 91 children treated with steroid-free tacrolimus, and rabbit anti-human thymocyte globulin induction demonstrate early reconstitution of T-cytotoxic cells, and gradual reconstitution of all other subsets, which is complete after one yr. Rejection-prone children demonstrate significantly higher counts of lymphocytes and all subsets prior to liver transplantation, and may exemplify one basis for enhanced baseline immunocompetence.     

Blood T and B lymphocyte subpopulations in healthy infants and children

Monoclonal antibodies and flow cytometry are now used routinely in the diagnosis of many malignant diseases and primary and secondary immunodeficiency states. Technical advances have improved the identification of blood lymphocyte subsets and reliable normal values are now obtainable. Such values have been reported for adults but not for children. We report both absolute and percentage normal values for lymphocytes and their subsets in infants and children of different ages. Our findings show that the absolute and percentage values for most lymphocyte markers differ substantially not only between children and adults, but also between children from different age groups. In infants, erythroid cell contamination of Ficoll gradient-density isolated mononuclear cells must be removed to obtain reliable flow cytometry values.     

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目的了解喘息与非喘息型病毒性肺炎患儿淋巴细胞亚群的改变。方法对2002—2004在北京友谊医院就诊的急性期喘息型病毒性肺炎患儿37例及非喘息型病毒性肺炎患儿32例,采用流式细胞仪对其外周血淋巴细胞亚群进行检测。并以30例健康儿童作为对照组。结果喘息组与非喘息组病毒性肺炎患儿急性期外周血T辅助淋巴细胞亚群Th1细胞百分率分别为(12·61±7·19)%及(17·32±9·92)%,与对照组[(9·16±9·90)%]比较差异均有显著的统计学意义(P<0·001);喘息组与非喘息组患儿Th1/Th2比值均高于对照组(P<0·05,P<0·01)。此外与非喘息组比较,喘息组CD4+/CD8+较高(P<0·05),但其他各项指标差异均无统计学意义。结论病毒性肺炎时不论是否发生喘息,均呈现Th1应答优势。喘息型病毒性肺炎患儿除CD4+/CD8+较非喘息型病毒性肺炎高外,其他各项淋巴细胞亚群差异无明显统计学意义。     

呼吸道合胞病毒及其他病毒性肺炎患儿血淋巴细胞亚群的变化

目的了解不同病原所致病毒性肺炎时血淋巴细胞亚群的改变。方法对2002~2004年确诊为病毒性肺炎的住院患儿,采用流式细胞仪对其急性期血淋巴细胞亚群进行检测。其中呼吸道合胞病毒(RSV)性肺炎组34例,非RSV肺炎组23例。同时与30例健康对照组比较。结果非RSV组Th1细胞百分率为[(15.89±7.88)%],RSV肺炎组为[(11.76±7.31)%],均较对照组显著增高(P=0.011,0);两者亦有显著差异(P=0.026);Th1/Th2在非RSV肺炎组较RSV肺炎组及对照组均明显增高,有非常显著差异(P=0.006,0)。结论病毒性肺炎时呈现Th1应答优势,并以非RSV感染最为明显。与非RSV肺炎相比,RSV感染具有潜在的相对Th2反应倾向。     

Lymphocyte subsets, sIL2-R and sICAM-1 in blood during allergen challenge tests in asthmatic children

In order to study cell activation in peripheral blood on bronchial allergen provocation up to 24 h, we investigated 32 asthmatic children, sensitive to house-dust mites. Six healthy young adult volunteers served as controls. Lymphocyte subsets (CD3, CD19, CD4, CDS) and activation markers (CD25-T, HLADR-T, CD23) in peripheral blood as well as soluble IL2-R and soluble ICAM-1 in scrum were evaluated. In terms of clinical reaction, 23 children exhibited a DAR, 6 an EAR, 6 a LAR and 3 children did not show a bronchoconstrictor response to allergen challenge with house-dust mite extract (NAR). In comparison to controls, asthmatic children showed a significantly higher expression of CD23 on B-lymphocytcs (p < 0.05). Other subsets were in the same range in both groups. After provocation there was a significant increase of CD4/CD8-ratio only in asthmatic children. Serum levels of sIL2-R were significantly higher in asthmatic children compared to controls at baseline as well as at 12 and 24 h after provocation, without variation during observation period, No differences were noted for SICAM-1. Our results confirm the hypothesis that lymphocytes, as important cells in regulation of allergic immune response, are recruited into peripheral blood under allergen challenge conditions in sensitized asthmatic children.     

Lymphocyte subsets may discern treatment effects in children and young adults with post-transplant lymphoproliferative disorder

To identify potential alternatives to Epstein-Barr virus (EBV)-specific cytotoxic T-cell responses, peripheral lymphocyte subsets (PLS) (CD4+, CD8+, CD3+, CD19+, CD56+) were measured by flow cytometry in children with abdominal transplants (n = 22) and heart transplants (n = 2), with (n = 14) and without (n = 10, group C) post-transplant lymphoproliferative disorder (PTLD). PTLD resolved with reduced immunosuppression and antiviral therapy in eight children (group B). Recalcitrant PTLD was observed in six children (group A). Recalcitrant PTLD followed prior antilymphocyte therapy [monoclonal anti-CD3 antibody (OKT3) and thymoglobin (n = 3) and thymoglobin (n = 1)] for refractory rejection in four of these six children, and resolved after treatment with rituximab (anti-CD20 monoclonal antibody). Ten children without PTLD served as a control group (group C). Between group comparisons showed a numeric increase in CD8 + cells and significantly lower CD4:CD8 ratios in both PTLD groups (A and B) compared with group C. Group A children also demonstrated significant depletion of natural killer (NK) cells, and post-rituximab depletion of B-cells compared with group B (no rituximab treatment). We conclude that NK cell depletion with a reversed CD4:CD8 ratio may represent a persistent immunosuppressed state, which may result from prior antilymphocyte therapy and may predispose to recalcitrant EBV-PTLD. Clinical remission with rituximab is accompanied by B-cell depletion. Serial monitoring of PLS from the time of diagnosis of PTLD will be necessary to confirm these observations.     

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??Abstract??Objective To establish the reference ranges for normal values of peripheral blood lymphocyte subsets in healthy Shanxi children of preschool age. Methods Healthy children aged 3-6 in Taiyuan of shanxi province were enrolled in the study. Relative counts and absolute counts of lymphocyte subpopulations of T cell?? CD3+ CD19-????CD4+ T cell ?? CD3+ CD4+????CD8+ T cell?? CD3+ CD8+????B cell?? CD3- CD19+?? and NK cell?? CD3- CD16+ CD56+?? were detected by three-color flow cytometric analysis. Peripheral blood lymphocyte subsets percentage and absolute counts of lymphocyte subsets in peripheral blood of healthy preschoolers and CD4+ /CD8+ were analyzed?? differences of which were compared between male and female. Results The percentages and absolute values of T cell??CD4+ T cell??CD8+ T cell??B cell and NK cell had no statistically significant difference between the genders ??P??0.05?? in 365 healthy preschoolers. The reference ranges for normal values of T cell??CD4+ T cell??CD8+ T cell??B cell and NK cell respectively were 43.8%??80.3% ??1563??3929 cells/μL????18.8%??46.7% ??738??2001 cells /μL????13.9%??36.1% ??5 32??1549 cells/μL????8.5%??24.6% ??261??960 cells/μL ????4.9%??24.6% ??197??786 cells /μL?? and CD4+ /CD8+ was 0.71??2.39. Conclusion It is important and necessary to establish the normal reference values of peripheral blood lymphocyte subsets for healthy children in the same race and district.It's shown here that there is no statistical difference in the peripheral blood lymphocyte subpopulation distribution between male and female in healthy preschool children.     

Lymphocyte subset reconstitution patterns in children with small bowel transplantation induced with steroid-free rabbit anti-human thymocyte globulin

Abstract:  Multiple measurements (n = 212) of lymphocyte subsets in 67 children treated with steroid-free Tacrolimus, and rabbit anti-human thymocyte globulin induction demonstrate early reconstitution of T-cytotoxic and NK cells. Reconstitution of CD4+ cells is complete after the second post-transplant year. During the period at risk for rejection, NK and T-cytotoxic cell counts are significantly higher among Rejectors. During periods at increased risk for EBV viral infection, CD4 counts bear a significant inverse relationship to EBV viral load in a subset of at-risk recipients. Rejection-prone children also demonstrate significantly higher counts of total lymphocytes, Tc and NK cells prior to SBTx, and may illustrate one basis for enhanced baseline immunocompetence.     

毛细支气管炎患儿Th1/Th2的改变

目的了解毛细支气管炎(毛支)患儿淋巴细胞亚群的改变及Th1/Th2平衡状况。方法对2003～2005年确诊为急性期毛细支气管炎的住院患儿34例,采用流式细胞仪对其外周血淋巴细胞亚群进行检测。其中呼吸道合胞病毒(RSV)性毛支23例,非RSV毛细支11例,均与22例健康对照组比较。结果毛细支气管炎组CD3 及CD4 细胞百分率均较对照组高(P<0.05,P<0.01),但CD4/CD8比值毛支组1.95%±0.64%与对照组1.87%±1.18%比较统计学无差异(P>0.05)。Th1/Th2在毛支组明显高于对照组,分别为6.06%±4.15%及2.90%±1.45%,有显著差异(Z=3.121Pa<0.01)。结论毛细支气管炎患儿细胞免疫功能增强,并呈现Th1应答优势。     

Lymphocyte subsets in human tonsils: The effect of age and infection

The aim of the present study was to determine the effect of repeated tonsillitis on the development of lymphocyte subsets in the tonsils and among peripheral blood lymphocytes (PBL) of children. Subsets of T- and B cells were analyzed in the tonsils and in PBL of patients undergoing tonsillecto-my for idiopathic tonsillar hypertrophy, recurrent tonsillitis, or tonsillar hypertrophy and tonsillitis. The majority of the CD4+ cells in the tonsils displayed the CD45RO+ phenotype, while the majority of those in the PBL displayed the CD45RA+ phenotype. Likewise, the proportion of CD45RO+CD8+ cells was higher in the tonsils than among PBL. The proportion of CD4 cells expressing the CD45RO marker increased with age among PBL, but not in the tonsils. B cells, detected by their CD 19, CD20, and CD21 markers, were three times more abundant in the tonsils than in the PBL. The proportion of CD38+ cells showed a negative correlation with age, both in the tonsils and among PBL. Among PBL a striking age-related reduction was seen in the proportion of CD19+, CD21+ and CD38+CD21+ B cells. In contrast, in the tonsils age-related changes could be detected only in the proportion of CD21+CD38+ cells. No difference among patients with various clinical diagnoses was detectable in any of the T- and B cell subsets in the tonsils and PBL. Thus, lymphocyte subsets evolve independently in the tonsils and peripheral blood, with the repeated antigenic challenge of tonsillar lymphocytes not influencing circulating memory cells.     

Infections and sickle cell disease in Eastern Saudi Arabian children

The rate and pattern of infections in 144 Saudi Arabian children with sickle cell disease (SCD) and matched normal control subjects are reported. All diagnoses of SCD were made at birth by means of screening blood from the umbilical cord. The children were prospectively followed up from birth to 4 years of age. Severe bacterial infections occurred in none of the children with SCD; one of the control children developed pneumococcal meningitis. Acute gastroenteritis was significantly more common among patients with SCD. For the first year of life, patients with SCD had significantly more infections than did the control children; but the reverse was true in the group that was 37 to 48 months of age. Considering all types of infections for all age groups, no difference was noted between patients with SCD and control subjects in terms of infection rate or related hospital admission. There were no deaths caused by infection in this series. We conclude that Saudi Arabian infants and young children of oasis origin with SCD are not at increased risk of infections compared with healthy children of the same age.     

地塞米松对病毒性脑炎患儿CD4+、CD8+淋巴细胞水平的影响

目的观察地塞米松(DEX)对病毒性脑炎(病脑)患儿CD4 、CD8 淋巴细胞的影响。方法对82例轻中度病脑患儿应用DEX治疗,用流式细胞仪动态测定其CD4 、CD8 细胞数变化。结果1.治疗前DEX治疗组和无DEX治疗组与正常儿童比较,CD4 淋巴细胞数减少,CD8 淋巴细胞数增加,有显著差异(F=14.3 P<0.05)。2.治疗1周两组与治疗前比较,CD4 淋巴细胞数进一步减少,CD8 淋巴细胞数进一步增加,这些变化DEX治疗组较无DEX治疗组更明显,有显著性差异(P均<0.05)。3.治疗后4周两组CD4 、CD8 、CD4 /CD8 比值均较治疗前和治疗后1周时恢复,但DEX治疗组仍未恢复到对照组水平,差别仍有显著意义(P均<0.05)。结论DEX治疗病脑可抑制CD4 细胞增生,使CD4 细胞数延迟恢复正常,从而促使继发感染的人数增多。对轻中度病脑患儿应用DEX弊大于利,应慎用。     

热性惊厥患儿血淋巴细胞亚群变化的临床意义

目的探讨热性惊厥(FS)患儿外周血淋巴细胞亚群的变化及其临床意义。方法应用流式细胞仪对单纯性FS38例(SFS组)、复杂性FS32例(CFS组)、29例急性上呼吸道感染(上感组)和32例健康儿童(健康对照组)血淋巴细胞亚群CD3 、CD4 、CD8 、CD4 /CD8 、CD1 6CD56 、CD19 进行测定,并进行队列对照研究。结果SFS组血CD3 、CD4 、CD8 、CD4 /CD8 值、CD1 6CD56 、CD19 水平分别为(58.62±13.7)%、(24.54±5.39)%、(29.6±11.39)%、(0.91±0.25)、(12.24±6)%、(18.85±8.22)%;CFS组分别为(55.92±13.43)%、(21.16±5.16)%、(34.24±15.7)%、(0.72±0.26)、(11.94±5.87)%、(21.03±7.85)%;上感组分别为(58.23±12.98)%、(26.72±6.09)%、(23.84±10.56)%、(1.47±0.96)、(14.37±7.39)%、(17.97±8.14)%;健康对照组分别为(60.87±9.59)%、(28.07±6.42)%、(23.62±8.83)%、(1.43±0.85)、(11.94±5.87)%、(21.03±7.85)%。与健康对照、上感组比较,FS组存在CD4 降低,CD8 升高、CD4 /CD8 比值降低,均有统计学意义(Pa<0.05,0.01,0.001);CFS组CD4 、CD4 /CD8 比值较SFS组低,CD8 高,均有统计学意义(P<0.05,0.01)。结论FS患儿体液和细胞免疫均被激活;CFS患儿免疫功能紊乱较SFS更严重。