化疗联合G-CSF动员的骨髓及外周血CD34+细胞表达粘附分子的变化

探讨化疗合G－CSF动员前，后患骺髓及外周血CD34^ 细胞表达CD44，CD49d，CD62L及趋化因子CXCR4的动态变化，用免疫荧光直接三角标记和流式细胞术测定G－CSF动员前，后骨髓及外周血CD34^ 细胞的CD44，CD49d，CD62L及趋化因子CXCR4的表达，观察输注各表达亚群细胞数与移植后造血重建的关系。结果发现，动员后骨髓中CD34^ CD44^ 和CD34^＋CD49d^ 细胞的百分比较动员前明显降低, 而外周血中二的比例则显升高；动员前，后骨髓中CD34^＋CD62L^＋和CD34^＋CXCR4^ 细胞的变化并不明显，而外周血中前明显增加，后则显减少。输入CD44^＋，CD49d^ ，CD62L^ 及CXCR4^ 的CD34^＋细胞的量与移植后血中中性粒细胞和血小板恢复的时间未表明有显的相关，结论：G－CSF动员可下调骨髓CD34^＋细胞的CD44，CD49d，CD62L及CXCR4的表达，从而进入外周血循环，输注这些细胞的临床意义有待累积更多病例的分析。     

rhG-CSF动员的外周血采集物与稳态骨髓中CD4^＋CD8^＋初始及记忆T细胞亚群的比较

本研究探讨重组人粒细胞集落刺激因子（rhG—CSF）动员的外周血采集物（G—PB）与稳态骨髓（SS—BM）中CD4^＋、CD8^＋初始、记忆T细胞亚群的差异。依据细胞表面CIM5RA和CD62L的表达将CD4^＋、CD8^＋T细胞划分为初始T细胞（naive，CIM5RA^＋CD62L^＋）、中心记忆T细胞（centralmemory，TCM，CIM5RA—CD62L^＋）、效应记忆T细胞（effeetor memory，TEM，CIM5RA^-CD62L^-）、终末分化效应记忆T细胞（terminally difierentiated effeetor memory，TTTp，CIM5RA^＋CD62L^-）。用流式细胞仪测定G—PB与SS—BM中CD4^＋、CD8^＋初始和记忆T细胞的比例组成，并计算每微升采集物中各细胞亚群的绝对数量。结果表明：G—PB中CD4^＋、CD8^＋T细胞的比例组成及CD4^＋／CD8^＋比值均高于SS—BM（P〈0．05）；G—PB中CD4^＋初始T细胞的比例显著低于SS—BM（P〈0．001），而CD4^＋TEM比例显著高于SS—BM（P〈0．001）；G—PB中CD8^＋初始和记忆T细胞亚群的比例与SS—BM无显著差异（P〉0．05）；与SS—BM相比，G—PB中CD4^＋CD62L^＋T细胞的比例明显降低（P=0．001），每微升G—PB移植物中的各淋巴细胞亚群均明显高于SS—BM（P〈0.001）。结论：G—PB与SS—BM中CD4^＋、CD8^＋初始和记忆T细胞亚群在相对和绝对数量方面存在明显的差异，这种差异可能影响G—PB和SS—BM移植后急性、慢性移植物抗宿主病发生率及其程度的异同。     

应用重组人粒细胞集落刺激因子对不同人群骨髓CD34+细胞和T细胞亚群影响的动态观察

目的　探讨健康人及处于完全缓解的恶性肿瘤患者 ,在应用重组人粒细胞集落刺激因子(G CSF)期间 ,其骨髓CD34 细胞和T细胞亚群的动态变化特点。方法　 12例异基因外周血干细胞移植健康供者和 16例自体外周血干细胞移植患者 ,在接受G CSF(30 0 μg/d ,共 5d)动员外周血干细胞期间 ,应用流式细胞仪测定其骨髓CD34 细胞和T细胞亚群。结果　①恶性肿瘤患者未应用G CSF时 ,骨髓细胞中CD34 细胞的初始值为 (0 .5 2± 0 .31) % ,同等条件下正常供者骨髓细胞中CD34 细胞的测定值为 (1.2 0±0 .6 8) % ,两者有显著差异 (P <0 .0 1)。应用G CSF 4 8h后 ,两者的测定值分别为 (1.2 2± 0 .4 2 ) %和(2 .0 7± 0 .5 2 ) % ,CD34 细胞均较前有明显增加 (P <0 .0 1) ,且两者之间仍存在明显差异 (P <0 .0 1)。CD34 细胞的高峰值出现在应用G CSF 96h后 ,分别为 (2 .2 3± 0 .34) %和 (2 .4 4± 0 .5 4 ) % ,分别较未应用G CSF时差异显著 (P <0 .0 1) ,两者之间无差异 (P >0 .0 5 )。②无论是否应用G CSF ,恶性血液病患者的T淋巴细胞亚群比例均处于倒置状态 ,而正常供者的T淋巴细胞亚群则比例正常。随两者应用G CSF后CD34 细胞的逐渐增加 ,T淋巴细胞亚群变化不明显。结论　完全缓解的恶性血液病患者 ,在应用G CSF4 8     

动员外周血造血干细胞移植物中同时高效去除T／B细胞的体外研究

单倍体相合造血干细胞移植时，去除移植物中的T细胞和B细胞可减少移植物抗宿主病和EBV相关的淋巴细胞增殖性疾病的发生。本研究在国内首次应用CliniMACS系统，通过CD3／CD19磁珠抗体同时有效地去除动员外周血造血干细胞中的T细胞和B细胞。用流式细胞术检测移植物中T细胞和B细胞的去除效率，以集落形成实验评价去除后的干细胞功能。结果表明：T细胞和B细胞去除之前单个核细胞总数为4．88×10^10，去除之后移植物中T细胞比例为0．02％，去除4．4Log；B细胞比例不到0．01％，至少去除3．3Log。移植物中除CD34^＋细胞外。还包括NK细胞，单核细胞和粒细胞。CD34^＋细胞纯度为0．98％，计数1．84×10^8，回收率为69．7％；NK细胞计数2．54×10^9，回收率为71．7％。集落形成实验显示CD3／CD19去除对造血干细胞功能无影响。结论：应用CliniMACS系统可有效地同时去除动员外周血干细胞中的T细胞和B细胞，且不影响造血干细胞的生物学功能，此项研究为去除T／B细胞的单倍体相合造血干细胞移植提供了技术平台。     

G-CSF动员对供者T淋巴细胞亚群功能和急性移植物抗宿主病的影响

为了研究粒细胞集落刺激因子（G—CSF）动员对供者T淋巴细胞亚群功能和急性移植物抗宿主病（aGVHD）的影响，对20例供者G—CSF动员前后的外周血样品，加入佛波醇酯、伊能霉素、莫能霉素体外刺激培养4小时，以3色荧光标记法流式细胞术检测T细胞亚群Ⅰ（分泌INF-γ、INF-α）、Ⅱ型（分泌IL-4、IL-5和IL-10）细胞比例。结果表明：G—CSF动员前外周血CD3^＋IFN-γ^＋、CD4^＋IFN-γ^＋和CD8^＋IFN-γ^＋T细胞的比例分别为3．2％（0％～45．9％）、1．3％（0％～23．8％）和1．5％（0％～22．2％），G—CSF动员后前述细胞比例显著升高，分别为19．2％（0％～53．9％）、9．5％（0％～49．5％）和7.5％（0％～38．1％）。动员前CD3^＋IL-2^＋、CD4^＋IL-2^＋、CD8^＋IL-2^＋T细胞比例分别为1．5％（0％～31％）、0．8％（0％～30.0％）和0％（0％～5．3％），动员后比例亦明显升高，分别为25．7％（0％～51％）、19．8％（0％～39．7％）、4．6％（0％～20．9％）。T细胞各亚群的IL-4阳性细胞比例在动员后升高不明显。移植早期aGVHD发生组供者的Tcl细胞比例显著高于无aGVHD组。动员后高Tc2比例组，其受者中、重度aGVHD的发生率明显低于低Tc2比例组（分别为20．0％和77．8％）。结论：G—CSF动员使供者外周血的Ⅰ型T细胞比例增加，动员后高Tcl细胞比例与移植早期aGVHD发生相关．Tc2比例与中重度aGVHD发生率负相关。     

经粒系集落刺激因子刺激外周血CD34~+细胞动员效果相关因素的考察

目的探讨粒系集落刺激因子(G-CSF)动员健康供者外周血造血干细胞效果的影响因素。方法对24例健康供者皮下注射G-CSF动员造血干细胞,检测外周血T淋巴细胞亚群和血常规数据。结果经G-CSF刺激后,外周血CD3+(%)、CD3+CD4+(%)、白细胞计数、血小板均明显升高(P0.05);而动员第4天、第5天、第6天骨有核细胞密度、CD34+细胞百分比无明显差别。经相关性分析,性别、年龄、体质量与CD34+细胞百分比呈负相关(P0.05),白细胞计数呈正相关(P0.01)。结论在一定范围内男性供者优于女性供者,年龄越小,体质量越轻,白细胞计数越高,经G-CSF动员的外周血造血干细胞CD34+细胞百分比越高。     

女性供者特征对粒细胞集落刺激因子动员的骨髓和外周血混合采集物中造血和免疫细胞组份的影响

目的 探讨女性健康供者特征对粒细胞集落刺激因子（G CSF）预激的骨髓和外周血混合采集物造血细胞和免疫细胞组份的影响。方法 111名健康女性供者应用G CSF动员,用流式细胞仪测定骨髓和外周血混合采集物中的CD34+细胞和T细胞亚群的数量,并分析怀孕、年龄、身高等供者特征对骨髓和外周血采集物中细胞组份的影响。结果 111例女性健康供者骨髓和外周血混合采集物中的CD34+细胞、CD3+T细胞、CD3+CD4+T细胞、CD3+CD8+T细胞和CD3+CD4-CD8-调节性T细胞的数量以每公斤供者体重计算的中位值分别为2.39×106/kg、226.57×106/kg、120.80×106/kg、89.99×106/kg和15.05×106/kg。①年龄对混合移植物中CD3+CD4-CD8-调节性T细胞的数量有影响。②体重指数（BMI）对混合采集物中的CD3+T细胞以及CD3+CD4-CD8-T细胞的数量有影响;③外周血干细胞采集当天的淋巴细胞（LYM）对混合采集物中的CD3+T细胞、CD3+CD4+T细胞和CD3+CD8+T细胞的数量有影响;④骨髓干细胞采集当天的LYM对混合采集物中CD3+T细胞和CD3+CD4-CD8-调节性T细胞的数量有影响;⑤与怀孕供者相比,未怀孕供者混合采集物中含有更高数量的CD3+CD8+ T细胞。结论 供者年龄、BMI、是否怀孕以及骨髓干细胞和外周血干细胞采集当天的LYM是骨髓免疫组份的主要影响因素。     

人重组粒细胞集落刺激因子动员后采集骨髓对健康供者外周血采集物成份的影响

本研究探讨健康供者体内应用人重组粒细胞集落刺激因子（G—CSF）后采集骨髓对外周血采集物成份的影响。62例健康供者皮下注射rhG—CSF5μg/（kg·d），连用5天，其中31例供者在第4、5天分别采集骨髓和外周血采集物（A组）；另31例供者于第4、5天均单采集外周血单个核细胞（B组）。应用流式细胞术检测两组供者外周血采集物中的淋巴细胞、CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD14^＋、CD34^＋细胞以及CD3^＋CD4^-CD8^-T细胞的数量。结果显示，A组供者每微升外周血采集物中单个核细胞中CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD14^＋、CD34^＋细胞以及CD3^＋CD4^-CD8^-T细胞的中位含量分别1．56×10^5μ1、8．56×10^4μl、6．12×10^4μl、3．38×10^4μl、2．27×10^4μl、3．83×10^4μl、744μl及3588μl，与B组的1．40×10^5μl、7．34×10^4μ1、5．32×10^4l、3．06×10^4μl、1．83×10^4μl、3．21×10^4μl、554μl及3120μl的差异无统计学意义（p〉0．05）；A组外周血采集物中CD4^＋细胞与CD8^＋细胞的比值[1．52（0．54—2．87）]、单核细胞与CD3^＋细胞的比值[0．57（0．15—1．64）]以及CD3^＋CD4^-CD8^-T细胞与CD3^＋细胞[0．064（0．018—0．673）]的比值与B组[1．68（0．31—3．35）]、[0．59（0．18—1．25）]、[0．063（0．021—0．136）]的差异均无统计学意义（P〉0．05）。结论：健康供者体内应用rhG—CSF后采集骨髓对外周血采集物成份无影响，对rhG—CSF动员的同一健康供者可单独采集骨髓、外周血采集物或同时采集两种采集物以满足临床移植的需要。     

重组人G-CSF对健康供者外周造血干／祖细胞动员和采集效果的影响因素分析

应用重组人粒系集落刺激因子（rhG—CSF）对健康供者进行动员并采集造血干细胞用于异基因外周血造血干细胞移植已在临床广泛应用，本研究通过对影响外周干细胞动员和采集效果的多因素分析，进一步探讨最佳动员方案及采集时机。采取回顾性方法分析了431例健康供者外周血干细胞动员采集效果，并进一步分析了供者一般特征、rhG—CSF动员天数、每日皮下注射次数、剂量与采集效果的关系。结果表明：rhG—CSF在动员中平均应用剂量为5．7μg／（kg·d），平均采集1．7次，收获单个核细胞数平均为9．57×10^8／kg，CD34^＋细胞平均为4．91×10^6／kg。绝大多数供者不良反应轻微。多因素分析结果显示，采集效率主要与供者体重指数，采集天数相关。rhG—CSF动员第5天采集的供者，其MNC数、CD34^＋细胞数及第一次单采成功率均优于其他时间采集的供者。同时，本组供者应用rhG—CSF剂量较小且剂量范围较窄，rhG—CSF剂量不如采集时间对采集物质量的影响明显。结论：小剂量应用rhG—CSF动员并于第5天开始采集是健康供者造血干细胞动员的较理想方案。     

支原体肺炎患儿淋巴细胞亚群的变化及意义

目的观察支原体肺炎患儿外周血淋巴细胞亚群的变化及临床意义。方法采用流式细胞仪检测20例支原体肺炎患儿和14例健康儿童淋巴细胞亚群（CD3^＋、CD3^＋CD4^＋、CD3^＋CD8^＋、CD4^＋／CD8^＋、CD3^-CD19^＋、CD3^-CD（16＋56）^＋）。两组年龄、性别差异无统计学意义。结果支原体肺炎患儿CD3^＋细胞明显降低（P〈0．01）、CD3^＋CD8^＋细胞降低（P〈0．05）、CD3^-CD19^＋细胞升高（P〈0．01），较对照组差异有统计学意义；CD3^＋CD4^＋、CD4^＋／CD8^＋、CD3^-CD（16＋56）^＋细胞较对照组差异无显著意义（P〉0．05）。结论支原体肺炎患儿存在细胞免疫失调，主要表现为CD3^＋、CD3^＋CD8^＋细胞降低，CD3^-CD19^＋细胞升高。     

福州地区健康儿童外周血淋巴细胞亚群及主要活化相关指标正常参考范围的建立

目的建立福州地区健康儿童外周血淋巴细胞亚群及活化的正常参考范围。方法选取福州地区0~18岁健康儿童593例为研究对象,按年龄分为新生儿组(0~28d)、婴儿组(~12个月)、幼儿组(~3岁)、学龄前组(~7岁)、学龄组(~12岁)和少青组(~18岁),采集外周血以美国BD公司生产的四色荧光抗体进行标记,以FACSCalibur流式细胞仪测定T淋巴细胞亚群CD3+、CD3+CD4+、CD3+CD8+、B淋巴细胞亚群CD3-CD19+、NK细胞CD3-CD16+CD56+及淋巴细胞活化指标CD4+CD38+、CD4+AntiHLA-DR+、CD8+CD38+、CD8+Anti-HLA-DR+的百分率和绝对计数,以Multi SET软件获取数据并分析结果,SPSS 10.0软件进行统计分析。以百分位数法,采用95%可信区间确定正常参考值范围。结果除CD3-CD16+56+绝对计数在各年龄组间无差异外,淋巴细胞亚群及淋巴细胞活化指标在不同年龄组之间的差异均有统计学意义,表现为CD3+绝对计数、CD3+CD4+、CD4+CD38+百分率和绝对计数、CD4+/CD8+比值随着年龄的增长呈降低趋势。CD4+Anti-HLA-DR+的百分率随着年龄的增长呈增高趋势。新生儿组CD3+、CD3+CD4+、CD4+CD38+、CD8+CD38+的百分率和绝对计数及CD4+/CD8+比值明显高于其它年龄组,CD3-CD19+、CD8+AntiHLA-DR+的百分率和绝对计数明显低于其它年龄组。在不同性别之间CD3+CD4+和CD3-CD19+、CD4+CD38+、CD4+Anti-HLA-DR+和CD8+CD38+的百分率和绝对计数的差异有统计学意义,表现为女童明显低于男童。儿童淋巴细胞亚群绝对计数的参考值高于成人。结论健康儿童淋巴细胞免疫表型的分布存在性别和年龄差别,由于这种差别细微,可以在同一种族同一区域建立统一的正常参考值范围。本研究成功建立福州地区健康儿童淋巴细胞亚群及活化的正常参考范围。     

Effect of blood donation on the establishment of normal ranges of lymphocyte subsets

BACKGROUND: Absolute counts of CD4+ T-lymphocytes are used in the management of patients with human immunodeficiency virus infection. Low absolute counts of CD3+CD4+ cells have also been observed in healthy people–a phenomenon called idiopathic CD4 lymphocytopenia. It is common practice for normal ranges for lymphocyte subsets to be derived from samples taken from blood donors. STUDY DESIGN AND METHODS: A sample of EDTA blood was taken through the donation line tubing, after donation from 565 blood donors in Sydney, Australia, who were selected from a range of age groups. An additional 12 donors provided a predonation sample as well as a postdonation sample. Hematologic assays were performed on two analyzers. Samples were stained for CD3, CD4, CD8, CD19, and CD56 and analyzed on a flow cytometer. RESULTS: Three donors were found to have absolute CD3+CD4+ counts < 300 cells per microL. The percentage of CD3+CD4+ cells was found to increase with age. Both the percentage and the absolute count of CD3+CD8+ cells decreased with age, which resulted in an increased CD4:CD8 ratio with age. Men had consistently higher absolute counts of CD3-CD56+ cells than women. The 12 additional donors all had greater percentages of CD3+CD4+ cells and lower absolute counts for CD3+, CD3+CD4+, CD3+CD8+, CD19+ and CD3-CD56+ cells after donation than they had before donation (p < 0.001). CONCLUSION: It is not satisfactory to base normal ranges for lymphocyte subsets on donor blood, from which the blood sample has been obtained after donation.     

African American adult apheresis donors respond to granulocyte-colony-stimulating factor with neutrophil and progenitor cell yields comparable to those of Caucasian and Hispanic donors

BACKGROUND: Granulocyte–colony‐stimulating factor (G‐CSF)‐mobilized peripheral blood progenitor cells (PBPCs) are the most common source of cells used for hematopoietic transplantation. Benign ethnic neutropenia has been found in persons of African descent, affecting circulating white blood cells (WBCs), but not WBC production within marrow. Persons of African descent have reduced neutrophil mobilization after steroid administration, and newborns have fewer nucleated and progenitor cells in their cord blood. STUDY DESIGN AND METHODS: Twenty‐two African American (AA) and 12 Hispanic PBPC donors were age, sex, and weight matched with 34 Caucasian donors. Groups were compared based on WBC and neutrophil counts after mobilization and numbers of CD34+ cells collected on Day 5 of G‐CSF mobilization. RESULTS: AA donors had significantly lower baseline WBC (6.1 ± 1.1 vs. 7.1 ± 1.7, p = 0.04) and neutrophil (3.4 ± 1.1 vs. 4.5 ± 1.3, p = 0.01) counts compared to matched Caucasian donors. G‐CSF–stimulated AAs had a significantly greater increase in WBC and neutrophil counts compared to matched Caucasians (889 ± 293% vs. 665 ± 230% neutrophils, p = 0.02). There was no significant difference in product cell counts when comparing total nucleated, CD3+, CD34+, and mononuclear cells or colony‐forming units (CFUs) between Caucasians and Hispanics or AAs and trends to greater numbers of neutrophils in products from AA donors. CONCLUSION: When stimulated by G‐CSF, AAs are able to increase WBC and neutrophil counts to a higher degree than Caucasians, achieving similar numbers of neutrophil and progenitor cells in apheresis products despite starting from lower baseline blood counts.     

健康供者特征对rhG-CSF预激的骨髓采集物CD34+细胞和T细胞亚群的影响

目的 探讨健康供者特征对rhG-CSF预激的骨髓(G-BM)采集物造血和免疫细胞成分的影响.方法 150名健康供者体内应用rhG-CSF 5 μ·kg-1·d-1连续4～5 d,第4,5天采集骨髓和外周血,用流式细胞术测定G-BM采集物中的CD3+、CD3+CD4+、CD3+CD8+、CD14+、CD34+细胞以及CD3+CD4-CD8-调节性T细胞的数量,并分析供者性别、年龄、体重、是否妊娠等特征对G-BM成分的影响.结果 150名健康供者G-BM所含有核细胞,淋巴细胞,CD3+、CD3+CD4+、CD3+CD8+、CD14+、CD34+细胞以及CD3+CD4-CD8-调节性T细胞的中位值分别为31 050(12 200～58 100)、2122(506～6618)、1344(307～4791)、692(145～3038)、616(112～2575)、986(265～2958)、63(11～505)和83(9～390)/μl.年龄≤38岁的供者G-BM中的有核细胞、CD34+细胞以及CD3+ CD4-CD8-调节性T细胞的数量分别为33800(18600～57100)、76(22～505)和97(11～380)/μl,显著高于年龄＞38岁的供者[分别为28 000(12 200～58 100)、49(11～220)和65(9～389)/μl],P值分别为＜0.05,＜0.001和0.001.外周血单核细胞计数＞0.37×109 /L的供者G-BM中有核细胞的数量[33550(12 200～57 100)/μl]显著高于≤0.37×109 /L的供者[27 150(13 800～58 100)/μl](P=0.005).多因素分析显示年龄和外周血单核细胞是G-BM中有核细胞采集量的影响因素[HR值分别为0.41和2.87;P值分别为0.01和0.003];年龄是G-BM中CD34+细胞采集量的唯一影响因素(HR=0.26;P值=0.001);年龄还是G-BM中CD3+ CD4-CD8-调节性T细胞数量的影响凶素[HR值为0.31;P=0.001].结论 年龄是影响G-BM中有核细胞、CD34+细胞和CD3+ CD4-CD8-调节性T细胞采集数量的因素,年龄≤38岁的供者更易采集满足临床需要的有核细胞和CD34+细胞;外周血单核细胞＞0.37×109 /L的供者采集的G-BM含有较多数量的有核细胞.     

Blood graft lymphocyte subsets after plerixafor injection in non-Hodgkin's lymphoma patients mobilizing poorly with chemotherapy plus granulocyte-colony-stimulating factor

BACKGROUND: A combination of chemotherapy plus granulocyte–colony‐stimulating factor (G‐CSF) (chemomobilization) is commonly used to mobilize CD34+ cells to circulation. Plerixafor, a chemokine CXCR4 antagonist, increases the mobilization of CD34+ cells and may also have effect on graft composition. STUDY DESIGN AND METHODS: We have analyzed lymphocyte subsets in grafts collected on the next morning after plerixafor injection in 13 chemomobilized patients with non‐Hodgkin's lymphoma (NHL) mobilizing poorly. As controls we had the first leukapheresis products from 11 NHL patients mobilized with chemotherapy plus G‐CSF. The analyses were performed from cryopreserved apheresis products. RESULTS: The median counts of both total CD3+ T cells and natural killer (NK) cells (CD3–CD16/56+) in the graft were significantly higher in plerixafor‐treated group compared to the control group. Both helper T‐lymphocytes (CD3+CD4+) and suppressor T‐lymphocytes (CD3+CD8+) were significantly increased in the plerixafor‐treated group so that CD4+/CD8+ ratio in the graft did not differ between the groups. CD19+ cells were evident only at very small amounts in few patients in both groups, and the CD34+ cell content of the graft did not differ between the groups. Engraftment after high‐dose therapy was comparable between the groups. CONCLUSION: Plerixafor added to chemomobilization in poor mobilizers seems to mobilize more T cells and NK cells than chemomobilization. Larger patient numbers and longer follow‐up is needed in regard to evaluate posttransplant complications and risk of relapse in patients receiving plerixafor due to poor mobilization.     

T lymphocyte differentiation in vitro from adult human prethymic CD34+ bone marrow cells

Pluripotent lymphohematopoietic stem cells are probably confined to bone marrow cells expressing CD34 surface molecules. To investigate the capacity of adult human CD34+ bone marrow cells to differentiate along the T lymphoid lineage, we plated purified CD34+ cells from healthy adults in liquid culture on adherent thymic stromal cells prepared from HLA- or blood group-mismatched postnatal thymic tissue. We show that purified CD34+CD3-CD4-CD8- bone marrow cells contained progenitors with the ability to differentiate into CD4+ and CD8+ T lymphocytes expressing surface (s)CD3 and T cell receptor alpha/beta in vitro. These progenitors were found in the CD34+CD2+sCD3-CD4-CD8-, CD34+CD7+sCD3-CD4-CD8-, and CD34+CD2+CD7+sCD3-CD4-CD8-, as well as in the CD34+CD2-sCD3-CD4-CD8-, CD34+CD7-sCD3-CD4-CD8-, and CD34+CD2-CD7- sCD3-CD4-CD8- subsets, indicating that T lymphocyte progenitors sensitive to signals mediated by thymic stroma in vitro are not restricted to CD34+ cells already coexpressing early T lymphocyte- associated markers. Finally, we show that T lymphopoiesis was enhanced by c-kit ligand.     

不同造血生长因子对脐血有核细胞体外扩增作用的实验研究

目的：探讨造血生长因子不同组合方式对脐血有核细胞的扩增作用以及脐血扩增后Ｔ淋巴细胞和ＨＬＡ－ＡＢ、ＤＲ阳性细胞变化情况。方法：在脐血有核细胞体外培养体系中分别加入不同组合的细胞因子，观察细胞数、ＣＤ３４＋细胞、ＣＦＵ－ＧＭ、Ｔ４（ＣＤ４＋ＣＤ８－ＣＤ３＋）细胞、Ｔ８（ＣＤ４－ＣＤ８＋ＣＤ３＋）细胞以及ＨＬＡ－ＡＢ、ＤＲ阳性细胞的变化。结果：脐血有核细胞在体外液体培养体系中，经重组人干细胞因子（ｒｈＳＣＦ）、白细胞介素（ＩＬ）－３、ＩＬ－６、粒细胞集落刺激因子（Ｇ－ＣＳＦ）、粒－巨噬细胞集落刺激因子（ＧＭ－ＣＳＦ）和红细胞生成素（Ｅｐｏ）的作用下扩增效果最佳。结论：脐血有核细胞在各种造血生长因子的协同作用下可以在体外扩增。脐血扩增后Ｔ４、Ｔ８细胞减少，可能会降低脐血移植急性ＧＶＨＤ发生，但ＨＬＡ－ＡＢ和ＨＬＡ－ＤＲ阳性细胞数急剧上升，增加了免疫排斥的可能性。     

Flow cytometry of cerebrospinal fluid (CSF) lymphocytes: alterations of blood/CSF ratios of lymphocyte subsets in inflammation disorders of human central nervous system (CNS).

Flow cytometry was adapted to measure lymphocytes in human cerebrospinal fluid (CSF). The method was sufficiently precise, reproducible and accurate despite low cell counts. In lumbar CSF of controls with 500 to 3500 (10(3)/l) leukocytes, lymphocyte counts correlated with those in corresponding venous blood: blood/CSF ratios of approximately 2000 : 1 were found for total T cells (CD3+) and CD3+ HLA-DR-, CD3+4+, CD3+8+ subsets, ratios were increased for the lymphocyte subsets CD3+ HLA-DR+ < or = CD3+16+56+ < CD16+56+3- < CD8+3- < CD19+; CD8+4+ ratio was half of CD3+ ratio. Data indicate selective barriers (blood-brain and blood-CSF barriers) to blood lymphocyte subsets which favor the transfer of T subsets. Correlation of the subset ratios to the CD3+ ratio indicates distinct barrier properties which changed differently with acute and subacute inflammations and neuroimmunological diseases of central nervous system (CNS) in lumbar or ventricular CSF, but not with simple protein barrier disturbance. HLA DR+ T ratios were higher than HLA DR- T ratios only with controls and some neuroimmunological diseases. Lymphocyte barrier characteristics were related to protein leakage situated at the same barriers, indicating for the lymphocyte subsets selective transfer routes in control subjects and non-selective routes in patients with CNS inflammation where altered ratios revealed a mixture of both routes.