Effects of concurrent cisplatinum administration during radiotherapy vs. radiotherapy alone on the immune function of patients with cancer of the uterine cervix

PURPOSE: To compare the effects of concurrent administration of cisplatinum (40 mg/m(2)/weekly) with radiation therapy (C-RT) to those induced by radiation therapy alone (RT) on the immune function of patients with locally advanced cervical cancer. METHODS AND MATERIALS: In 8 prospectively randomized patients (i.e., 4 receiving RT vs. 4 receiving C-RT), lymphocyte populations including CD3+, CD4+ and CD8+ T-cell subsets, B cells (CD19+) and natural killer cells (CD56+, CD16+, CD3-) were studied before, during, and after therapy. Expression of the activation marker CD25 on CD3+ T cells, intracellular levels of perforin in CD8+ and CD56+ cells, and interferon-gamma (IFN-gamma) and IL-2 in CD4+ and CD8+ T cells was also measured. Finally, lymphoblast transformation and natural killer (NK) cytotoxic activity were assessed. RESULTS: Both RT and C-RT significantly decreased the mean absolute number of all lymphocyte subsets compared to pretreatment levels (p > 0.001). However, no differences were detected in the characteristics or the magnitude of the lymphopenia induced by the two treatments. Both RT and C-RT increased similarly the percentages of CD25-positive lymphocytes (p > 0.001), and significantly decreased PHA-induced T-cell lymphoblast transformation (p > 0.001) and NK cytotoxic activity against K562 cells (p > 0.001). The percentage of perforin-positive and CD8+ T cells was not altered during either treatment, whereas the percentage of perforin-positive and CD56+ cells was significantly reduced during both treatments, and correlated with reduced cytotoxicity against K562 cells. The percentages of CD8+ IFN-gamma+ and CD4+ IFN-gamma+ T cells as well as that of CD8+ IL-2+ and CD4+ IL2+ T cells were not significantly altered by C-RT compared to RT alone. Finally, with both regimens, NK cells and B-cell numbers showed a more rapid recovery than T-cell numbers. CONCLUSION: Administration of concurrent cisplatinum to radiation may synergistically increase cytotoxic effects of radiation on tumor cells but does not alter the magnitude and the characteristics of radiation-induced immunosuppression.     

康赛迪胶囊对食管癌放疗患者细胞免疫功能的影响

目的　观察中药复方制剂康赛迪胶囊对食管癌患者放疗前后细胞免疫功能的影响。方法　采用FACSCalibur流式细胞仪 ,SimulSETv3 1分析软件 ,分别检测 2 0例食管癌患者在放疗 6 0Gy前后 (对照组 )和口服康赛迪同时放疗 6 0Gy前后 (实验组 )患者外周血NK细胞活性、T淋巴细胞亚群 (CD3+、CD4 +、CD8+)、CD4 +/CD8+、B淋巴细胞 (CD19+)、NK淋巴细胞 (CD16 +,CD5 6 +)水平。结果　二组患者放疗前外周血NK细胞活性、CD3+、CD4 +、B淋巴细胞百分数、CD4 +/CD8+的平均值均低于健康人参考值 ,其中CD4 +细胞和B淋巴细胞明显下降 ,CD8+细胞明显升高。对照组患者放疗 6 0Gy后 ,除CD8+细胞和NK淋巴细胞百分数稍有升高外 ,其他细胞均略有下降 (P >0 0 5 )。实验组患者在治疗后CD3+、CD4 +、NK淋巴细胞百分数、CD4 +/CD8+明显升高 ,并明显高于对照组 (P <0 0 5或P <0 0 1)。结论　食管癌患者在放疗前后细胞免疫功能持续低下。放疗 6 0Gy后患者细胞免疫功能受到轻度抑制 ,康赛迪可以明显改善食管癌患者放疗后的细胞免疫功能状态。提示食管癌患者在放疗的同时应加强免疫治疗 ,以改善患者的免疫抑制状态。     

Immunological effects of interferon-alpha on chronic myelogenous leukemia

Treatment with interferon-alpha is effective for chronic myelogenous leukemia in the chronic phase (CML-CP), but the immunological mechanisms of the antileukemic effect of this substance are still unclear. The objective of this study was to investigate the immunological effects of interferon-alpha in CML patients. Markers of cellular activation and apoptosis, natural killer (NK) cell cytotoxicity and production of intracellular cytokines (IFN-gamma, IL-2 and IL-4) were determined by flow cytometry in the peripheral blood mononuclear cells (PBMC) of 26 CML-CP patients before and 3, 6 and 9 months after IFN-alpha treatment. The results were correlated with the hematological response. In the whole group of patients, INF-alpha use was followed by a significant increase of lymphocytes producing IL-2 and IFN-gamma, an increase in NK activity and a decrease in the number of CD34+ cells. Out of 26 CML patients, 15 achieved hematological remission and 7 achieved partial cytogenetic remission after 9 months of IFN-alpha treatment. There was an increase in the percentage of CD8/FasL+, DR/CD3+, DQ/CD3+, CD34/Fas+, DR/CD56+, CD56/FasL+ cells and of IFN-gamma- and IL-2-producing lymphocytes and an increase in NK cytotoxicity only in the group of patients who achieved complete hematological remission. Our results indicate that IFN-alpha use in CML-CP reduces the number of CD34+ cells, activates T cells, enhances stem cell apoptotic markers and increases the production of intracellular IFN-gamma and IL-2 by lymphocytes. Taken together, these results indicate that the therapeutic effect of IFN-alpha in CML-CP is mediated at least in part by immunological mechanisms.     

肿瘤患者外周血免疫细胞亚群和有关细胞因子含量的相关性研究

目的:评价血液中CD3+、CD4+、CD8+、CD19+、自然杀伤（NK）细胞含量和细胞因子肿瘤坏死因子（TNF）、白介素-2（IL-2）、IL-6、IL-10的相关性。方法:对80例不同肿瘤患者和20例健康对照者进行细胞亚群和细胞因子检测。使用流式细胞仪测定样本外周血CD3+、CD4+、CD8+、NK、CD19+细胞比例,同时使用酶联免疫吸附试验测定外周血中TNF、IL-2、IL-6、IL-10水平。应用配对资料t检验分析肿瘤患者和正常对照者的细胞亚群和细胞因子有无差异,应用直线相关性t检验分析各细胞亚群和细胞因子之间有无线性相关。结果:肿瘤患者CD8+细胞含量高于正常对照者,NK细胞含量低于正常对照者,血液中CD3+细胞含量和细胞因子TNF、IL-2、IL-6、IL-10均无相关性,CD4+、CD8+细胞含量和IL-2线性相关,NK细胞含量和IL-2、IL-10线性相关,CD19+细胞含量和IL-6线性相关（但差异无统计学意义）。结论:肿瘤患者细胞免疫状况和正常人有统计学差异,细胞因子未发现统计学差异;免疫细胞亚群和细胞因子有一定线性相关。     

A novel natural killer cell line (KHYG-1) from a patient with aggressive natural killer cell leukemia carrying a p53 point mutation.

We present the establishment of a natural killer (NK) leukemia cell line, designated KHYG-1, from the blood of a patient with aggressive NK leukemia, which both possessed the same p53 point mutation. The immunophenotype of the primary leukemia cells was CD2+, surface CD3-, cytoplasmic CD3epsilon+, CD7+, CD8alphaalpha+, CD16+, CD56+, CD57+ and HLA-DR+. A new cell line (KHYG-1) was established by culturing peripheral leukemia cells with 100 units of recombinant interleukin (IL)-2. The KHYG-1 cells showed LGL morphology with a large nucleus, coarse chromatin, conspicuous nucleoli, and abundant basophilic cytoplasm with many azurophilic granules. The immunophenotype of KHYG-1 cells was CD1-, CD2+, surface CD3-, cytoplasmic CD3epsilon+, CD7+, CD8alphaalpha+, CD16-, CD25-, CD33+, CD34-, CD56+, CD57-, CD122+, CD132+, and TdT-. Southern blot analysis of these cells revealed a normal germline configuration for the beta, delta, and gamma chains of the T cell receptor and the immunoglobulin heavy-chain genes. Moreover, the KHYG-1 cells displayed NK cell activity and IL-2-dependent proliferation in vitro, suggesting that they are of NK cell origin. Epstein-Barr virus (EBV) DNA was not detected in KHYG-1 cells by Southern blot analysis with a terminal repeat probe from an EBV genome. A point mutation in exon 7 of the p53 gene was detected in the KHYG-1 cells by PCR/SSCP analysis, and direct sequencing revealed the conversion of C to T at nucleotide 877 in codon 248. The primary leukemia cells also carried the same point mutation. Although the precise role of the p53 point mutation in leukemogenesis remains to be clarified, the establishment of an NK leukemia cell line with a p53 point mutation could be valuable in the study of leukemogenesis.     

Phenotype and function of a CD56+ peripheral blood monocyte.

G-CSF primed CD34 cells cultured for 2-3 weeks in IL-2 and stem cell factor generate CD56(high) cells with phenotypic and morphologic features of NK cells, and a novel adherent CD56(low) CD16- population expressing myeloid markers (CD33 and HLA-DR). We hypothesized that similar cells might also occur in peripheral blood. In 13/13 normal individuals, we found a circulating population of CD56(low), CD33+, FcgammaRI+, FcgammaRII+, HLA-DR+, CD11b(high), CD14+ monocytes closely resembling the cultured CD56(low)CD33+ cells. They may represent a normal counterpart of the CD56+ CD33+ hybrid myeloid/natural killer cell leukemia. Their mean frequency was 1.3+/-1% (standard deviation), range 0.16-3.5%, of total mononuclear cells. CD56(low)CD33+ cells, primed with cytomegalovirus antigen, induced autologous T-lymphocyte proliferation comparably to CD56-, CD14+ peripheral blood monocytes (PBM). Conversely, CD56(low) cells induced greater T-cell proliferation than CD56- PBM when lymphocyte responders were HLA mismatched. Unstimulated CD56(low)CD33+ cells showed a low antiproliferative effect on K562, which was increased upon LPS stimulation. The pattern of cytokine production by CD56(low)CD33+ cells and PBM largely overlapped; however, they produced detectable levels of IL-6 and IL-1beta. These results define a minor monocyte population with distinct phenotypic and functional features.     

DC-CIK联合放化疗对小细胞肺癌患者免疫功能的影响

目的：探讨细胞因子诱导的杀伤细胞（CIK）、树突状细胞（DC）免疫治疗联合放化疗对小细胞肺癌患者外周血淋巴细胞亚群的影响及疗效。方法收集60例小细胞肺癌患者资料,其中32例患者采用DC-CIK联合放化疗（化疗方案为依托泊苷+顺铂/依托泊苷+卡铂/伊立替康+顺铂,放疗方案为适形调强放疗技术）为联合治疗组;28例进行放化疗的患者为对照组。治疗结束2周后采用流式细胞仪检测外周血CD3+、CD3+CD4+、CD3+CD8+、NK细胞、Treg细胞及相关因子IFN-γ、IL-2、IL-10、TGF-β1的变化,并评价疗效。结果联合治疗组IFN-γ治疗后较治疗前升高,IL-10、TGF-β1较治疗前降低（P﹤0.05）;联合治疗组患者外周血CD3+CD8+、CD3+CD4+、NK细胞治疗后水平高于对照组（P﹤0.05）;而Treg细胞、IL-10、TGF-β1治疗后水平低于对照组（P﹤0.05）。对照组患者外周血CD3+、CD3+CD4+、CD3+CD8+、NK细胞治疗后较治疗前下降,差异有统计学意义（P﹤0.05）;细胞因子在治疗前后无变化。虽然联合组疾病控制率较对照组高,但差异无统计学意义（87.50%vs 71.43%,P﹥0.05）。结论自体DC-CIK治疗可以改善小细胞肺癌联合放化疗患者的细胞免疫功能,是否能改善生存需进一步研究。     

Enhanced T-cell response to mosquito extracts by NK cells in hypersensitivity to mosquito bites associated with EBV infection and NK cell lymphocytosis

Hypersensitivity to mosquito bites is characterized by severe systemic as well as local symptoms, and associated with chronic active EBV infection and NK cell lymphocytosis. In this HEN disease, we investigated the response of PBMC to MSG extracts. PBMC were taken from three defined cases of HEN disease, three borderline cases, five individuals with simple exaggerated reactions to mosquito bites without systemic symptoms (simple responders), and eight healthy donors. PBMC, or purified CD4+, CD8+ or CD56+ cells, were cultured with MSG extracts prepared from each of five mosquito species to examine their proliferation and cytokine secretion. The patients with HEN disease had high stimulation indices with variations in responses to the extracts from Aedes albopictus, Aedes aegypti, Anopheles sinensis and Culex pipiens pallens. However, a non-Japan-habitant species Anopheles stephensi did not stimulate the patients' PBMC. Some borderline or simple responders showed moderate proliferation, and healthy donors had no reactive PBMC. In HEN disease, both CD56+ NK cells (producing IFN-gamma) and CD4+ Th0 cells (producing IL-4 and IFN-gamma) were increased in the blood. CD4+ cells, but not CD56+ NK cells or CD8+ cells, propagated in response to MSG extracts. However, this response of CD4+ cells and their IL-4 production were strongly enhanced by coexisting CD56+ cells. We suggest that the CD4+ T cell serving as the primary responder to MSG antigen and the NK cell functioning as the enhancer are both pathogenic in the development of HMB.     

外周血淋巴细胞亚群水平变化在鼻咽癌中的意义

目的 探究外周血淋巴细胞亚群水平变化在鼻咽癌(NPC)中的意义.方法 选取NPC患者60例为试验组,选取健康体检者30例为对照组.采用流式细胞仪检测试验组治疗前后和对照组的外周血中CD3+、CD4+、CD8+细胞亚群及自然杀伤细胞(NK)水平;采用免疫组化法和Western Blot检测试验组治疗前后肿瘤组织中白细胞介素-21(IL-21)蛋白表达水平.结果 治疗前,试验组外周血中CD3+、CD4+、CD4+/CD8+及NK细胞水平明显低于对照组(P﹤0.01),CD8+细胞水平明显高于对照组(P﹤0.01);治疗后,试验组外周血中CD4+、CD4+/CD8+和NK细胞水平均较本组治疗前明显上升(P﹤0.01),CD8+细胞水平较本组治疗前明显下降(P﹤0.01);治疗后,早期NPC患者的总缓解率高于晚期NPC患者(P﹤0.05);免疫组化法和Western Blot检测结果显示,治疗前,NPC患者肿瘤组织中IL-21蛋白的阳性表达率及表达水平均高于治疗后(P﹤0.05).结论 NPC患者外周血中淋巴细胞亚群及NK细胞的检测对鼻咽癌的早期诊断及疗效评判具有重要意义.     

右美托咪定对乳腺癌根治手术患者应激反应及细胞免疫的影响

目的：探讨右美托咪定对乳腺癌根治手术患者围手术期应激反应及术后细胞免疫的影响。方法选择2012年1月至2015年6月在我院择期全身麻醉下行乳腺癌根治术的患者108例,采用随机数字表法分为右美托咪定组（D 组）和对照组（C 组）,每组54例。D 组麻醉前15 min 内静脉滴注右美托咪定1．0μg／kg,麻醉诱导插管后,以0．5μg．kg －1．h －1维持至术毕。C 组给予等容量的生理盐水代替右美托咪定,其余麻醉同 D 组。分别测定麻醉前（T0）、术后4 h（T1）、术后1 d（T2）、术后3 d（T3）和术后7 d（T4）的血浆去甲肾上腺素（NE）、肾上腺素（E）、皮质醇（COR）、血清白细胞介素（IL）-6、T 淋巴细胞亚群（CD3＋、CD4＋、CD8＋、CD4＋／CD8＋）及 NK 细胞的水平。结果与 T0时比较,D 组各点血浆 NE、E、COR 及血清 IL-6水平差异无统计学意义（均 P ＞0．05）。C 组 T1～T3时血浆 NE、E,T2、T3时血浆COR,T2～T4时血清 IL-6水平均显著升高（均 P ＜0．05）。与 C 组比较,D 组 T1～T3时血浆 NE、E,T2和T3时血浆 COR,T2～T4时血清 IL-6水平均明显降低（均 P ＜0．05）。与 T0比较,T1～T3时两组 CD3＋及NK 细胞水平明显降低（均 P ＜0．05）。与 C 组比较,D 组 T1～T3时 CD3＋、NK 细胞明显升高,D 组 T2～T3时 CD4＋及 CD4＋／CD8＋明显升高（P ＜0．05）。结论全身麻醉期间持续静脉滴注右美托咪定可有效减轻乳腺癌根治术患者围手术期的应激反应,改善机体免疫状态。     

化疗对鼻型NK/T细胞淋巴瘤患者细胞免疫功能的影响

目的:通过对鼻型NK/T细胞淋巴瘤患者化疗前、后外周血T 淋巴细胞各亚群及NK细胞检测来探讨化疗对其细胞免疫功能的影响。方法:分组对照研究,鼻型NK/T细胞淋巴瘤患者（实验组）41例,分别在化疗前及化疗2 个周期结束后10d 两次检测外周血T 淋巴细胞各亚群及NK细胞数目。正常健康组35例（对照组）。 对相关实验结果进行统计学分析。结果:鼻型NK/T细胞淋巴瘤患者化疗前与正常健康组相比,T 淋巴细胞亚群比例紊乱,CD3+、CD4+比例和CD4+/CD 8+比值及NK细胞明显下降,CD8+比例及CD4+CD25+调节性T 细胞（Tregulatory cells ,Treg ）明显升高（P<0.05）。 经过2 个周期的化疗治疗,化疗有效组化疗后CD3+、CD4+比例、NK细胞比例、CD4+/CD 8+比化疗前明显升高（P<0.05）,CD8+比例、CD4+CD25+Treg 明显下降（P<0.05）。 化疗有效组化疗后除CD4+、CD8+比例外,其他指标与正常健康组相比差别均无统计学意义（P>0.05）;而无效组患者化疗后CD4+比例、CD8+比例、CD4+/CD 8+比值、NK细胞数目比化疗前则进一步下降（P<0.05）。 结论:鼻型NK/T细胞淋巴瘤患者细胞免疫功能低下,T 淋巴细胞亚群比例紊乱,NK细胞明显下降。有效化疗可通过杀伤、诱导肿瘤细胞凋亡,减轻肿瘤负荷,减少CD4+CD25+Treg ,排除某些免疫抑制因素;改善了患者的细胞免疫功能,化疗无效患者其细胞免疫功能则继续恶化。通过检测患者的T 淋巴细胞各亚群及NK细胞数变化可以反映患者的细胞免疫功能状态,对指导临床治疗、判断预后有重要的意义。      

Expression of signal-transducing zeta chain in peripheral blood T cells and natural killer cells in patients with Hodgkin's disease in different phases of the disease

A number of phenotypic and functional alterations have been described in T cells of cancer patients. These changes are believed to reflect an impaired T-cell mediated immunity, which in turn, may result in a decreased capacity to generate an effective antitumor response. Several mechanisms have been proposed to explain depressed immunity in cancer patients including tumor-derived suppressor factors, abnormal cytokine production, deletion or inactivation of tumor-reactive T-cells. To investigate the mechanism underlying the immunodeficiency in Hodgkin's disease (HD) we studied the expression of T cell receptor zeta chain, which plays a vital role in the cascade of events leading to T and NK cell activation. The expression of the zeta chain of the T cell receptor/CD3 complex was analyzed by dual colour immunofluorescence on peripheral blood T lymphocytes: CD3+, CD4+, CD8+ and NK-cells (CD56+) in patients in different phases of the disease. Zeta chain was significantly reduced on CD3, CD4, CD8, and CD56 positive cells from patients in active phase of the disease compared with normal controls (p=0.05). In patients tested in complete clinical remission the values were normal except for the subpopulation of CD8+ cells in which the expression of zeta chain remained significantly reduced compared with controls. Downregulation of CD3/zeta-chain in PBLs and NK cells in active phase of HD- and to a lesser extent in clinical remission may contribute to immunodeficiency associated with the disease.     

Phenotyping of human tumor-infiltrating lymphocytes before and after exposure to different in vitro stimulation conditions.

Tumor-infiltrating lymphocytes (TiL) and autologous peripheral blood lymphocytes (PBL), mainly from breast and kidney tumor patients were cultivated with high- and low-dose rIL-2 under addition of autologous tumor cells or nonmalignant epithelial cells. Tumor cells added to TIL-cultures induced an additional proliferative response. Before cultivation, CD8+ and CD25+ lymphocytes were more frequent in TIL when compared to PBL, while CD16+, CD19+ and CD56+ cells were rare. After culture, lymphocytes of both origins showed an increase of CD2+, CD25+, CD56+ and HLA-DR+ cells and a relative decrease of CD3+ cells. The CD4+/CD8+ ratios and a number of CD56+ cells were rIL-2 dose dependent.     

The effect of peripheral blood lymphocyte stimulation on zeta chain expression and IL-2 production in Hodgkin's disease

It has been reported that peripheral blood T cells and NK cells express reduced levels of the T-cell receptor signal-transducing zeta chain in Hodgkin's disease (HD). The zeta chain has emerged as a key subunit of the T-cell antigen receptor, which plays a central role in the signal-transducing events leading to T and NK-cell activation. We were interested in determining whether the low zeta chain expression in HD could be corrected by anti-CD3, anti-CD3-rIL-2 ex vivo stimulation. Zeta chain expression was analysed by dual immunofluorescence on permeabilized cells before and after 72 hours of culture. The IL-2 concentration in the culture supernatants was measured by ELISA. Zeta chain was significantly reduced on unstimulated CD4+, CD8+ and CD56+ cells from patients in active disease compared with normal subjects. In patients in complete remission, the values were normal except for CD8+ cells, on which zeta expression remained significantly reduced. Stimulation with anti-CD3 did not change zeta expression. Co-stimulation with rIL-2 increased but did not normalize the proportions of CD4(+)/zeta(+), CD8(+)/zeta(+)and CD56(+)/zeta(+)cells and IL-2 production in active disease. Stimulation of cells from patients in clinical remission with anti-CD3(+)rIL-2 increased the proportion of CD8(+)zeta(+)cells and normalized IL-2 production levels. Considering the pivotal role of CD3-zeta in immune response, our data suggest that successful immunotherapy approaches in active HD should consider inclusion of other potent cytokines, as well as genetically engineered tumour vaccines.     

肺癌患者外周血 T- 淋巴细胞亚群 NK 细胞的基线数值及其与预后的关系*

目的:对比肺癌患者与健康者之间淋巴细胞亚群差异,评估肺癌患者外周血T 淋巴细胞亚群及自然杀伤细胞（NK）之基线值与预后的关系。方法:收集2006年2 月至2013年3 月就诊于天津医科大学肿瘤医院病例资料完整的肺癌患者105 例,其中非小细胞肺癌（NSCLC ）86例、小细胞肺癌（SCLC）19例,另选健康对照35例,对比接受治疗前肺癌患者和健康对照者外周血中的CD3+T 细胞、CD4+T 细胞、CD8+T 细胞及NK细胞所占百分比,并回顾性分析86例NSCLC 患者初治时外周血淋巴细胞亚群与预后的关系。结果:肺癌患者外周血CD3+T 细胞、CD4+T 细胞、NK细胞及CD4/CD 8 比值均明显低于健康对照组（P = 0.011,P = 0.007,P <0.001,P=0.025）,而CD8+T 细胞比例高于健康对照组（P = 0.013）。 当CD8+T ≥ 31.8% 及CD4/CD 8< 1.28时NSCLC 患者可以获得一个更长的OS（中位OS分别为36.2 m vs . 20.0 m ,P = 0.010;30.8 m vs . 20.0 m ,P = 0.035）。 而CD3+T 细胞、CD4+T 细胞及NK细胞百分比对NSCLC 患者预后无显著影响。结论:外周血CD8+T 细胞基线水平较高的NSCLC 患者生存较长,此基线水平可能对NSCLC 患者预后有指示作用。      

不同麻醉及镇痛方法对宫颈癌患者细胞免疫功能和术后恢复的影响

目的:探讨不同麻醉及镇痛方法对宫颈癌患者细胞免疫功能和术后恢复的影响。方法:拟行腹腔镜下宫颈癌根治术患者78例,分为3组（n=26）,Ⅰ组:全凭静脉麻醉+静脉病人自控镇痛（PCIA）,Ⅱ组:全凭静脉麻醉联合硬膜外神经阻滞+硬膜外病人自控镇痛（PCEA）,Ⅲ组:静吸复合麻醉联合硬膜外神经阻滞+PCEA。观察麻醉前（T0）、术毕（T1）、术后24 h（T2）、术后48 h（T3）时点T淋巴细胞亚群及NK细胞百分比的变化。记录患者第一次补救镇痛时间、胃肠道恢复时间、下床时间、术后呕吐及患者满意度。结果:与T0比较,3组患者在T1、T2时点CD3+、CD4+、CD4+/CD8+及NK细胞百分比明显降低,CD8+明显升高（P<0.05）;Ⅱ组和Ⅲ组在T2及T3与Ⅰ组比较,CD3+、CD4+、CD4+/CD8+及NK细胞百分比显著增高,CD8+细胞百分比显著降低（P<0.05）。Ⅱ组和Ⅲ组患者术后恢复明显优于Ⅰ组（P<0.05）,其中术后呕吐和患者满意度Ⅱ组明显优于Ⅲ组（P<0.05）。结论:全凭静脉麻醉联合硬膜外神经阻滞并行硬膜外术后镇痛对宫颈癌患者术后细胞免疫功能的抑制较小,术后恢复快,有益于患者的康复。     

Regeneration of functional and activated NK and T sub-subset cells in the marrow and blood after autologous bone marrow transplantation: a prospective phenotypic study with 2/3-color FACS analysis

The phenotypic reconstitution of lymphoid cells in the bone marrow and peripheral blood was examined prospectively in 27 patients who underwent autologous bone marrow transplantation (ABMT) for leukemia/lymphoma. Patterns of activation within NK and T cell subsets as well as T sub-subsets were studied with monoclonal antibodies in two- and three-color FACS analysis. NK-like cells (CD16+) were found to be increased in the peripheral blood and bone marrow after ABMT. The expression of two activation markers, 4F2 and HLA-DR, was sustainedly increased within this subset. High numbers of CD4+ and CD8+ T cells carrying surface HLA-DR were found early after ABMT both in blood and marrow. The T suppressor inducer cell sub-subset (CD45R+ CD4+) was severely depressed in both the blood and marrow 6-9 months post-ABMT. Using three-color FACS analysis, half of this T sub-subset was shown to express HLA-DR+. The levels of T suppressor effector-like cells (CD11+ CD8+) remained within the normal range in both peripheral blood and bone marrow during follow-up. The HLA-DR expression was elevated and equally distributed between the CD11- CD8+ and CD11+ CD8+ sub-subset cells. There was no major impact of marrow T cell purging or CMV carrier status on the phenotypic NK/T cell reconstitution. The present results provide an immune phenotypic basis for the suggested generation of anti-leukemic NK-like and T suppressor-like activity after ABMT.     

食管癌患者外周血中自然杀伤性T细胞含量的检测及其临床意义

 　目的　研究食管癌患者外周血中自然杀伤性（NK）T细胞在手术前后的表达情况。方法　采用流式细胞术（FCM）分析59例食管癌患者手术前后外周血中CD3、CD56、CD4、CD8抗体的表达,研究NKT细胞及其亚群的表达情况及所占比例。结果　随着CD+3 CD+56 CD+8／CD+3 CD+56 CD+4的比值逐渐升高,Ⅲ～Ⅳ期食管癌患者的比例逐渐降低,ANOVA示组间差异有统计学意义（P＜0.05）;CD+3 CD+56 CD+8／CD+3 CD+56 CD+4的比值与CD+3 CD+56 CD+4 NKT细胞非线性相关;CD+3 CD+56 CD+8 NKT细胞与NK细胞正相关,与CD+3 T细胞负相关。结论　CD+3 CD+56 CD+8／CD+3 CD+56 CD+4 的比值可能与肿瘤负荷有关,并且有助于判断食管癌患者的疾病程度及预后。     

Tumour-associated macrophages are related to progression in patients with metastatic melanoma following interleukin-2 based immunotherapy

The aim of the present study was to analyze whether leukocyte subsets in peripheral blood and tumour biopsies obtained before treatment were able to predict response or survival in patients with metastatic melanoma following Interleukin-2 (IL-2) based immunotherapy. Flow cytometry was performed on peripheral blood for CD4+ T cells, CD8+ T cells and CD56+ natural killer (NK) cells. Immunohistochemical analyses were used to identify CD4+ T cells, CD8+ T cells, CD57+ NK cells and CD64+ (macrophages) cells in tumour biopsies. High numbers of tumour-associated CD64+ macrophages in tumour biopsies were statistically significantly associated with poor response to treatment. Our data suggest that tumour-associated macrophages may correlate negatively with response, which may be of biological importance for IL-2 based immunotherapy of malignant melanoma.     

柔红霉素联合阿糖胞苷治疗小儿急性白血病的临床疗效及对外周血调节性T细胞的影响

目的柔红霉素联合阿糖胞苷治疗小儿急性白血病的临床疗效及对外周血调节性T细胞(Treg)的影响。方法随机数字表法将78例急性白血病患儿分为观察组和对照组,每组39例。对照组患儿接受长春新碱+环磷酰胺+泼尼松治疗,观察组患儿接受柔红霉素+阿糖胞苷(DA方案)治疗。治疗1个月后评估两组患儿的近期疗效及化疗期间不良反应的发生情况;化疗前、化疗后、化疗后随访1个月,比较两组患儿外周血免疫功能指标,包括CD4+CD25+CD127-、NK细胞及T淋巴细胞亚群(CD3+、CD4+、CD8+)水平。结果观察组患儿临床总有效率为82.05%,高于对照组患儿的61.54%,差异有统计学意义(χ2=4.052,P﹤0.05)。化疗后,两组患儿CD3+、CD4+、NK细胞水平均低于本组化疗前,CD8+、CD4+CD25+CD127-水平均高于本组化疗前,差异均有统计学意义(P﹤0.05);观察组患儿CD3+、CD4+、NK细胞水平均高于对照组患儿,CD8+、CD4+CD25+CD127-水平均低于对照组患儿,差异均有统计学意义(P﹤0.05)。随访1个月,观察组患儿CD3+、CD4+、CD8+、NK细胞、CD4+CD25+CD127-水平均逐渐恢复至化疗前水平,与化疗前比较差异均无统计学意义(P﹥0.05)。观察组患儿化疗期间不良反应总发生率为35.90%,与对照组患儿的20.51%比较,差异无统计学意义(P﹥0.05)。结论红霉素联合阿糖胞苷治疗小儿急性白血病的近期疗效较好,可在一定程度上缓解CD4+CD25+Treg细胞介导的肿瘤免疫逃逸,安全性较高。