大运动量训练对排球运动员淋巴细胞亚群的影响

本文研究了大运动量训练对上海男子排球运动员ＮＫ细胞活性和淋巴细胞亚群分布的影响。结果指出大运动量训练后即刻ＮＫ细胞活性显著地低于训练前测定值（Ｐ＜０．０１），运动后１小时及１５小时ＮＫ细胞活性恢复到了训练前水平。Ｔ３（ＣＤ３）和Ｔ８（ＣＤ８）淋巴细胞百分数在运动不同时期均无显著改变，但Ｔ４（ＣＤ４）淋巴细胞百分数在运动后１小时及１５小时显著地低于训练前水平，因而ＣＤ４／ＣＤ８细胞比率也相应地显著降低，至于大运动量训练对细胞免疫功能影响的机理有待进一步研究。     

两种自家免疫性甲低患者治疗前后血清TSII、TGII及外周血T淋巴细胞亚群的变化

对９例原发性粘液性水肿和１４例Ｈａｓｈｉｍｏｔｏ甲状腺炎患者测定了血清ＴＳＩＩ、ＴＧＩＩ及外周血Ｔ淋巴细胞亚群，并以３２例正常人作为对照．原发性粘液性水肿和Ｈａｓｈｉｍｏｔｏ甲状腺炎患者血清存在ＴＳＩＩ、ＴＧＩＩ，阳性比例不同，而且外周血ＣＤ８＋百分率降低和ＣＤ４＋／ＣＤ８＋比值增高．ＴＳＩＩ、ＴＧＩＩ活性与ＣＤ４＋／ＣＤ８＋比值无显著相关．经甲状腺激素治疗，两种自家免疫性甲低患者甲状腺功能明显恢复，血清ＴＳＩＩ、ＴＧＩＩ活性无明显变化，原发性粘液性水肿患者外周血Ｔ淋巴细胞亚群未见明显变化，而Ｈａｓｈｉｍｏｔｏ甲状腺炎患者ＣＤ８＋百分率明显回升．     

慢性丙型肝炎外周血T淋巴细胞亚群的实验研究

应用抗人Ｔ淋巴细胞单克隆抗体ＣＤ系列检测３０例慢性丙型肝炎病例以及２０例健康成人的外周血Ｔ淋巴细胞ＣＤ_３、ＣＤ_４、ＣＤ_８、ＣＤ_１６亚群，以ＡＰＡＡＰ免疫酶标染色法测定。结果发现：慢性丙型肝炎组ＣＤ_４细胞数较正常对照组明显降低，ＣＤ_８细胞数较正常对照组明显升高，ＣＤ_３、ＣＤ_１６细胞数与正常对照组无显著差异。认为Ｔ淋巴细胞亚群异常所致的细胞免疫紊乱可能是慢性丙型肝炎发病的重要原因。     

甲肝患者血清SIL－2R及其与T淋巴细胞表面标志的关系

应用双单克隆抗体夹心ＥＬＩＳＡ法对甲肝患者５２例血清可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）水平做了动态观察，同时以ＣＤ系列单克隆抗体ＡＰＡＡＰ法对以上患者外周血Ｔ淋巴细胞表面ＩＬ－２Ｒ（ｍＩＬ－２Ｒ）及Ｔ细胞亚群进行了动态监测，并对其相互关系及其与肝细胞损伤标志ＡＬＴ做了相关分析。发现ｓＩＬ－２Ｒ及ｍＩＬ－２Ｒ在甲肝急性期均显著增高，恢复期则基本恢复正常；两者在急性期、恢复期均存在非常显著的正相关；两者与ＡＬＴ亦显著相关。同时发现甲肝急性期外周血Ｔ淋巴细胞ＣＤ￣＋＿４率显著降低，ＣＤ￣＋＿８率则明显增高，致ＣＤ￣＋＿４／ＣＤ￣＋＿８比值明显下降且倒置；在恢复期以上改变均恢复正常。还发现ＣＤ￣＋＿８率与ｓＩＬ－２Ｒ、ｍＩＬ－２Ｒ及ＡＬＴ之间均存在着程度不同的正相关。此结果在一定程度上提示细胞免疫可能参与了甲肝的发病机制。     

Graves病患者治疗前后血清TSI,TGI及外周血T淋巴细胞亚群的变化

检测了３７例Ｇｒａｖｅｓ病患者血清甲状腺刺激免疫球蛋白（ＴＳＩ）、甲状腺生长免疫球蛋白（ＴＧＩ）和外周血总Ｔ细胞（ＣＤ３）、辅助性／诱导性Ｔ细胞（ＣＤ４）及抑制性／细胞毒性Ｔ细胞（ＣＤ８），并以３２例正常人作对照．发现Ｇｒａｖｅｓ病ＴＳＩ阳性率８３．８％，ＴＧＩ阳性率５８．３０，ＴＳＩ活性与血清ＴＴ４含量呈正相关（Ｐ＜０．０１），ＴＧＩ活性随甲状腺肿大加重而升高．外周血ＣＤ３＋、ＣＤ８＋百分率降低（Ｐ＜０．０１），ＣＤ４＋／ＣＤ８＋比值增高（Ｐ＜０．０５）．其中１８例经他巴唑治疗，甲状腺功能恢复明显，ＴＳＩ活性明显下降，而ＴＧＩ活性、ＣＤ３＋、ＣＤ８＋百分率和ＣＤ４＋／ＣＤ８＋比值无显著变化．对ＴＳＩ、ＴＧＩ及Ｔ细胞亚群在Ｇｒａｖｅｓ病发病中的作用及其临床意义作了讨论．     

两种自家免疫性甲低患者治疗前后血清TSII,TGII及外周血T淋巴…

对９例原发性粘液性水肿和１４例Ｈａｓｈｉｍｏｔｏ甲状腺炎患者测定了血清ＴＳＩＩ、ＴＧＩＩ及外周血Ｔ淋巴细胞亚群，并以３２例正常人作为对照。原发性粘液性水肿和Ｈａｓｈｉｍｏｔｏ甲状腺炎患者血清存在ＴＳＩＩ、ＴＧＩＩ，阳性比例不同，而且外周血ＣＤ＾＋８百分率降低和ＣＤ＾＋４／ＣＤ＾＋８比值增高。ＴＳＩＩ、ＴＧＩＩ活性与ＣＤ＾＋４／ＣＤ＾＋８比值无显著相关。经甲状腺激素治疗，两种自家免疫性甲低患者甲状腺     

冬泳对人体免疫功能的影响

冬泳对人体免疫功能的影响浙江医科大学第二医院肿瘤研究所（杭州３１０００９）王晓稼杭州市冬泳协会本研究通过测定冬泳者和普通正常人（对照）外周血自然杀伤细胞（ＮＫ）活性以及ＮＫ样细胞──大颗粒淋巴细胞（ＬＧＬ）和ＣＤ１６＋抗原表达单个核细胞（ＣＤ１６＋）...     

低剂量辐射诱导小鼠胸腺细胞及其亚组的适应性反应

目的研究低剂量辐射预先照射以及随后大剂量照射后小鼠胸腺细胞Ｔ细胞受体（ＴＣＲ）、ＣＤ３、ＣＤ４和ＣＤ８分子表达的变化。方法ＴＣＲ、ＣＤ３表达采用间接荧光流式细胞术检测,ＣＤ４、ＣＤ８表达采用双参数直接免疫荧光流式细胞术检测。结果实验结果表明：单纯１５ＧｙＸ射线全身照射后ＴＣＲ,ＣＤ３阳性细胞数以及ＣＤ４－Ｄ８－,ＣＤ４＋ＣＤ８＋,ＣＤ４＋ＣＤ８－和ＣＤ４－ＣＤ８＋细胞数明显减少,当１５ＧｙＸ射线全身照射前６小时预先照射００７５Ｇｙ时,可明显减轻其后１５Ｇｙ照射对ＴＣＲ＋,ＣＤ３＋,ＣＤ４＋ＣＤ８＋,ＣＤ４＋ＣＤ８－和ＣＤ４－ＣＤ８＋的损伤作用。表现为各亚组细胞数显著高于单纯１５Ｇｙ照射组。ＣＤ４－ＣＤ８－亚组的细胞数无明显变化。结论００７５ＧｙＸ射线全身照射能够诱导胸腺细胞ＴＣＲ＋,ＣＤ３＋,ＣＤ４＋ＣＤ８＋,ＣＤ４＋ＣＤ８－和ＣＤ４－ＣＤ８＋亚组细胞的适应性反应。     

采用流式细胞仪分析31例优秀运动员T细胞亚群

采用双色荧光法流式细胞仪测定３１例优秀运动员外周血Ｔ淋巴细胞表型的分布特征，并与我国正常成人参考值作比较。结果显示：运动员ＣＤ３＋、ＣＤ４＋、ＣＤ８＋的分布离散度较大，与我国正常成人参考值相比，除ＣＤ３＋无显著性差异外，ＣＤ４＋较正常成人低，ＣＤ８＋较正常成人高，ＣＤ４＋／ＣＤ８＋比值较正常成人低，统计学上均存在显著性差异。     

高海拔地区进展期胃癌患者手术前后T细胞亚群的变化

目的：观察高海拔地区进展期胃癌（ＡＧＣ）患者手术前后Ｔ细胞亚群的变化；方法：采用间接免疫荧光法对３２例进展期胃癌患者手术前后不同时期的Ｔ细胞亚群进行检测；结果：①高海拔地区进展期胃癌患者ＣＤ３、ＣＤ４、ＣＤ８、ＣＤ４／ＣＤ８值均低于正常对照组。②根治性切除术后，患者的ＣＤ３、ＣＤ４细胞值较术前有显著怀差异（Ｐ〈０．０１）；ＣＤ４／ＣＤ８细胞比值亦较术前有显著性差异（Ｐ〈０．０１），但与正常对照线之     

低剂量半身照射对恶性肿瘤患者免疫功能的影响

目的 评价低剂量辐射对恶性肿瘤患者免疫功能的影响。方法 将20例患者(其中非何杰金氏淋巴瘤13例、小细胞肺癌7例)随机分为两组:HBR组和RR组。其中HBR组10例患者采用常规放疗(RR)加低剂量半身照射,低剂量半身照射方法为10 cGy/次,2次/周,总剂量为100 cGy,每次均间隔6～8 h再行常规放疗;RR组10例患者只予以常规放疗。采用流式细胞术(FCM)双标法检测HBR与RR组在放疗前、中、后外周血淋巴细胞CD₄、CD₈、CD₂₅和CD₅₆等指标的变化。结果 照射后RR组患者的CD₄⁺ CD₈⁺降低(P<0.05),HBR组患者的CD₄⁺升高(P<0.05),CD₂₅⁺和CD₅₆⁺分子表达均明显地增加(P<0.05),放疗前和放疗后CD₈⁺均低(P<0.05),放疗中(P<0.05)和放疗后(P<0.01)CD₄⁺ CD₈⁺均高。结论 低剂量半身照射可增强机体的免疫功能。     

Occupational exposure to ionizing radiation has no effect on T‐ and B‐cell total counts or percentages of helper,cytotoxic and activated T‐cell subsets in the peripheral circulation of male radiation workers

Purpose: To investigate changes in immune cell subsets in the peripheral circulation of a male population occupationally exposed to ionizing radiation.

Materials and methods: Peripheral blood samples were taken from 194 male workers with cumulative exposures of >200?mSv (mean exposure 331.5?mSv, mean age 51 years) and from a reference population of 131 male workers with cumulative exposures of <27.5?mSv (mean exposure 13.9?mSv, mean age 47 years). Samples were analysed by flow cytometry for T‐ and B‐cell total counts and for the T‐cell subset percentages of CD4⁺ (helper T‐cells), CD8⁺ (cytotoxic T‐cells) and CD3⁺/HLA‐DR⁺ (activated T‐cells).

Results: Comparison of the >200 and <27.5?mSv exposure groups using linear regression analysis showed no statistically significant differences between the two groups for T‐cell total count, B‐cell total count or for percentages of the T‐cell subsets CD4⁺, CD8⁺ or CD3⁺/HLA‐DR⁺ and CD4⁺:CD8⁺. However, statistically significant increases in both T‐ and B‐cell total counts were observed within the two exposure groups and data pooled from both groups when non‐smokers (never and ex‐smokers) were compared with current smokers. For pooled data T‐cell total count increased in smokers by 35% (p=0.0001) and B‐cell total count increased by 37% (p=0.0004).

Conclusions: No significant immunological effects were observed in male radiation workers with cumulative exposures of >200?mSv when compared with a reference population with cumulative exposures of <27.5?mSv, although highly significant increases in both T‐ and B‐cell total counts were observed in smokers compared with non‐smokers.     

Post-irradiation viability and cytotoxicity of natural killer cells isolated from human peripheral blood using different methods

Purpose We compared the pre- and post-irradiation viability and cytotoxicity of human peripheral natural killer cell (NK) populations obtained using different isolation methods.

Material and methods Three methods were used to enrich total NK cells from buffy coats: (I) a Ficoll-Paque gradient, plastic adherence and a nylon wool column; (II) a discontinuous Percoll gradient; or (III) the Dynal NK cell isolation kit. Subsequently, CD16⁺ and CD56⁺ NK cell subsets were collected using (IV) flow cytometry or (V) magnetic-activated cell sorting (MACS) NK cell isolation kits. The yield, viability, purity and cytotoxicity of the NK cell populations were measured using trypan blue exclusion, flow cytometry using propidium iodide and ⁵¹Cr release assays after enrichments as well as viability and cytotoxicity after a single radiation dose.

Results The purity of the preparations, as measured by the CD16⁺ and CD56⁺ cell content, was equally good between methods I–III (p?=?0.323), but the content of CD16⁺ and CD56⁺ cells using these methods was significantly lower than that using methods IV and V (p?=?0.005). The viability of the cell population enriched via flow cytometry (85.5%) was significantly lower than that enriched via other methods (99.4–98.0%, p?=?0.003). The cytotoxicity of NK cells enriched using methods I–III was significantly higher than that of NK cells enriched using methods IV and V (p?=?0.000). In vitro the NK cells did not recover cytotoxic activity following irradiation. In addition, we detected considerable inter-individual variation in yield, cytotoxicity and radiation sensitivity between the NK cells collected from different human donors.

Conclusions The selection of the appropriate NK cell enrichment method is very important for NK cell irradiation studies. According to our results, the Dynal and MACS NK isolation kits best retained the killing capacity and the viability of irradiated NK cells.     

1.5Gy X射线全身照射对小鼠胸腺细胞CD、CD4、CD8分子表达及

采用单克隆抗体免疫荧光技术,流式细胞术检测,观察了1.5GyX射线单次全身照射(剂量率0.286Gy/min)对小鼠胸腺细胞CD₃分子表达的影响,及胸腺细胞亚群和细胞周期的变化。结果表明,中等剂量照射后,胸腺细胞CD₃分子的百分率明显增高,胸腺细胞CD₃阳性细胞比例为对照组的177.61%.同时,胸腺中CD₄⁺及CD₈⁺细胞比例亦显着增加,分别为对照的202.5%及165.87%.然而,胸腺中CD₄⁺、CD₈⁺细胞的比例显着降低,为对照组的68.28%,与此同时,胸腺细胞周期中S期细胞比例明显降低,为对照组的55.14%.     

Influence of photodynamic therapy on peripheral immune cell populations and cytokine concentrations in head and neck cancer

BackgroundPhotodynamic therapy (PDT) represents a palliative treatment option for a selected group of patients with head and neck squamous cell carcinoma (HNSCC). PDT induces a local inflammatory reaction with the potential to initiate antitumor immune responses. However, the systemic impact on peripheral immune cells has not been described so far.MethodsHNSCC patients (n = 9) were treated with PDT in a palliative setting. All patients had previously undergone several oncologic treatment regimens. Blood samples were taken before, during and after PDT. Age-matched healthy donors served as control group (NC, n = 15). The frequency and absolute number of T- and B-lymphocytes, CD4⁺CD39⁺ regulatory T-cells (Treg) and NK-cells were measured by 10-color flow cytometry. Serum concentrations of T cell related cytokine panel, including HMGB1, IL-6, IL-10 and perforin were measured by bead array and ELISA.ResultsIn heavily pretreated HNSCC patients, the number and frequency of Treg and NK-cells were increased as compared to NC. PDT induced a further increase of the frequency of Treg and NK-cells in the peripheral blood. Additionally, the serum concentrations of HMGB1, IL-6 and IL-10 showed a significant elevation after treatment with simultaneously decreased perforin levels.ConclusionAlthough PDT is a local treatment regimen, a systemic inflammatory response with altered peripheral immune cell populations and cytokine concentrations is visible. The increased Treg and NK cell numbers after PDT support the hypothesis that PDT may successfully be combined with NK cell or T cell activating immune checkpoint modulators in HNSCC patients to improve HNSCC specific immunity.     

不同剂量照射对胸腺细胞四个亚组的影响

本实验采用双荧光流式细胞术研究了不同剂量x射线照射后24小时,昆明小鼠胸腺T细胞四个亚组的变化,结果表明：各亚组对辐射的敏感性有较大的差异。其辐射敏感性依次为CD₄⁺CD₈⁺CD₄^-CD₈^-CD₄^-CD₈⁺CD₄⁺CD₈^-,由于各亚组辐射敏感性的差异,导致了大剂量照射后各亚组之间比例的改变。     

CD34+脐血干细胞转染pIRES2-FL-IL-3后静脉移植对辐射损伤小鼠的影响

目的 观察CD₃₄⁺ 脐血干细胞转染质粒载体pIRES2-FL-IL-3后静脉移植对辐射损伤小鼠的影响并探讨其机制。方法 质粒载体pIRES2-FL-IL-3转染脐血CD₃₄⁺干细胞,将其通过静脉移植到辐射损伤的小鼠体内(5×10⁵个细胞/只,双表达组,12只),观察辐照后2、4和6周动物存活率及外周血象变化。6周时免疫荧光法检测实验小鼠脾脏CD₃₄表达,RT-PCR法和Western blot 法进行IL-3和FL的mRNA和蛋白活性检测,另外3组为单纯CD₃₄⁺细胞组(CD₃₄组)、转染pIRES2-IL-3CD₃₄⁺细胞组(IL-3组)和转染pIRES2-FL CD₃₄⁺细胞组(FL组),各组动物数均为12只。结果 CD₃₄组、IL-3组和FL组动物6周存活率分别为25.0%、50.0%和50.0%,双表达组动物存活率为91.7%,显著高于其余各组。2周时双表达组白细胞数量、红细胞数量及血小板数量显著高于其余3组。双表达组脾脏CD₃₄免疫荧光强度显著高于其余3组,IL-3及FL的mRNA 水平及蛋白活性均显著高于其余3组。结论 真核共表达质粒载体pIRES2-FL-IL-3转染CD₃₄⁺脐血干细胞后对辐射损伤小鼠具有显著的促脾脏造血作用,其机制与移植后小鼠脾脏组织中移植CD₃₄⁺脐血干细胞的聚集、增殖及目的 蛋白的高效表达有关。     

南京“5.7”192Ir源放射事故患者造血系统的改变

目的 通过对南京¹⁹²Ir源放射事故中患者的血液系统进行观察研究,探讨造血系统损伤规律,为核与辐射事故医学应急的救治和随访提供临床资料和积累经验。方法 应用血细胞分析仪动态观察患者的白细胞计数、淋巴细胞计数、血小板计数及血红蛋白值的变化,取其双侧髂骨骨髓涂片观察骨髓象的改变,采用PCR-以测序为基础的分型技术(PCR-SBT)方法检测融合基因突变情况;分别采用流式细胞术和放射免疫分析法测定T淋巴细胞亚群百分率和血清免疫球蛋白(IgA、IgG、IgM)、补体(C3、C4)含量,评价细胞免疫和体液免疫功能。结果 患者骨髓造血轻度障碍,予以粒细胞集落刺激因子(G-CSF)治疗后,短时间内恢复正常,受照后约40 d进入恢复期;受照侧骨髓增生低下;融合基因检测未检测到突变;局部感染导致淋巴细胞计数及免疫功能下降。结论 轻度骨髓型急性放射病造血系统损伤较小,病程分期不明显,造血功能恢复较快,尚未造成免疫抑制,但局部感染可导致免疫功能下降。临床医生需注意全身治疗与局部治疗的统一。     

Modification of X-ray Induced Chromosome Aberration Frequency by Pre- and Postirradiation Hyperthermia of Human Peripheral Lymphocytes

Summary

Human peripheral lymphocytes (HPL) were used to study the synergistic effects of hyperthermia and X-irradiation. The effects on the chromosome aberration frequencies of different combinations of heat and radiation exposure at different cell cycle phases were analysed at the first mitosis after irradiation. When unstimulated HPL were heated after irradiation with 2 and 3 Gy, respectively, the chromosomal aberration frequencies were significantly higher than following radiation exposure alone. Heat treatment during different phases of the cell cycle and irradiation during the G₂ phase led to an increase in the aberration frequencies when the cells were heat-treated not later than 16 h before radiation. Furthermore we show that the number of breaks increased linearly with the duration of hyperthermia when HPL were heated for periods of 1–15 min at 45°C 32 h before irradiation at 48 h after stimulation. Heat treatment alone during any phase of the cell cycle did not induce chromosome aberrations. The increased aberration frequencies are probably the result of inhibited repair of radiation-induced DNA strand breaks.