不同分期的成骨肉瘤患者免疫学的变化

目的：评价成骨肉瘤患者临床分期与免疫学变化的关系及意义。方法：利用流式细胞仪检测120例成骨肉瘤患者淋巴细胞亚群、利用MTT法检测NK细胞活性,应用放射免疫分析法检测IL－2、TNF－α、β2－微球蛋白的变化,应用ELISA法检测SIL－2R的改变。结果：随着Enecking分期的增高,患者NK活性、CD4、CD4／CD8、IL－2逐渐下降,CD8、sIL－2R、β2－MG、TNF－α逐渐升高P＜0．01;Ⅱa期成骨肉瘤患者机体免疫功能活跃,CD3、CD4、NK活性及IL－2、sIL－2R高于对照组P＜0．01,其余指标与对照组相同;Ⅰb及Ⅱb组患者各项指标改变明显;Ⅲ期肿瘤CD4、CD4／CD8、IL－2较Ⅱ期明显降低sIL－2R、TNF－α急剧升高P＜0．01。结论：成骨肉瘤患者免疫功能与疾病分期呈负相关,淋巴细胞亚群、sIL－2R、IL－2可作为监测患者病情改变的指标。     

肝动脉灌注LAK／IL—2及化疗药物治疗晚期肝癌

目的：观察经皮肝动脉插管化疗药物灌注，栓塞术联合LAK／IL－2灌注治疗中晚期肝癌的疗效、T淋巴细胞改变及副反应。方法：96例经确诊的中晚期肝癌，随机分为动脉化疗栓塞联合LAK／IL－2观察组及单纯介入化疗栓塞组各48例，两组均采用经皮穿刺肝动脉插管灌注化疗药物及栓塞。化疗药物5－Fu1000mg＋卡铂300mg 表阿霉素30mg或丝裂霉素10－20mg，栓塞剂为碘化油和明胶海绵，介入治疗3－4周1次，2次1疗程，观察组栓塞后沿导管灌注同种异体LAK细胞1×10^9100ml及IL－240万U。两次介入间隔期间观察组每周3次静滴1×10^9/100mlLAK细胞及每日肌注IL－210万U结果：观察组总有效率（CR＋PR）为72．9％（35／48），对照组总有效率（CR＋PR）淡57．3％（28／48），两组比较有显性差异（P＜0．05）。观察组T淋巴细胞治疗前后变化有统计学意义（P＜0．01），而对照组无统计学意义，结论：同种异体LAK细胞联合化疗及栓塞治疗中晚期肝癌较单纯介入化疗栓塞疗效明显提高，是安全可行的一种治疗方法。     

肝动脉灌注LAK/IL-2及化疗药物治疗晚期肝癌

目的观察经皮肝动脉插管化疗药物灌注、栓塞术联合LAK／IL－2灌注治疗中晚期肝癌的疗效、T淋巴细胞改变及副反应。方法96例经确诊的今晚期肝癌,随机分为动脉化疗栓塞联合LAK/IL－2观察组及单纯介入化疗栓塞组各48例。两组均采用经皮穿刺肝动脉插管灌注化疗药物及栓塞。化疗药物5-Fu1000mg＋卡铂300mg＋表阿霉素30mg或丝裂霉素10～20mg,栓塞剂为碘化油和明胶海绵,介入治疗3～4周1次,2次1疗程。观察组检室后沿导管灌注同种异体LAK细胞1×109/100ml及IL－240万U,两次介入间隔期观察组每周3次静滴1×109／100mlLAK细胞及每日从注IL－210万U。结果观察组总有效率（CR＋PR）为72．9％（35／48）,对照组总有效率（CR＋PR）为58．3％（28／48）,两组比较有显著性差异（P＜0．05）。观察组T淋巴细胞治疗前后变化有统计学意义（P＜0．01）,而对照组无统计学意义。结论同种异体LAK细胞联合化疗及栓塞治疗中晚期肝癌较单纯介入化疗栓塞疗效明显提高,是安全可行的一种治疗方法。     

头颈癌患者T细胞亚群与白细胞介素2及其受体的动态研究

研究对35例头颈癌患者手术前后外周血T细胞亚群、IL－2及sIL－2R表达水平进行了检测，并对其中20例患者进行动态观察。结果显示：头颈癌患者CD；及sIL－2R表达显著增高，比值及IL－2水平明显低于正常人；手术后及sIL－2R表达显菩下降，比位与IL－2水平明显回升；动态观察复发者，比值及IL－2水平再显下降，及sIL－2R表达又见回升。表明：T细胞亚群与IL-2／sIL－2R的检测能反应机体抗肿瘤免疫水平，对监视病情发展、疗效观察及预后判断具有一定的临床价值，动态观察有助于监视肿瘤的复发与转移。     

淋巴细胞亚群比例在原发性肝癌TACE、THPP前后动态变化的临床研究

目的：观察淋巴细胞亚群比例在原发性肝癌患者肝动脉介入＋栓塞（TACE）前后及腹腔热灌注顺铂（THPP）后的动态变化。方法：治疗组（27例）在行肝动脉介入化疗＋栓塞术前后及一周后行腹腔热灌注顺铂后一月，分别应用流式细胞仪（FCM）3次行淋巴细胞亚群测定，对照组（27例）也作介入前后及一月后，作3次淋巴细胞亚群测定。结果：原发性肝癌虱淋巴细胞亚群比例失调。CD4^ 、CD4^ /CD8^ 、CD19^ 、NK^ 明显下降（P＜0.05)，CD3^ 、CD8^ 维持正常或略低下，TACE后更加剧了这种紊乱（P＜0.01)，CD4^ 、CD4^ /CD8^ 、CD19^ 、NK^ 下降更明显（P＜0.05)，CD3^ 、CD8^ 变化不大。THPP后，上述CD4^ 、CD4^ /CD8^ 、CD19^ 、NK^ 上升，CD3^ 、CD8^ 变化不大。表明患者机体免疫功能增强。结论：原发性肝癌患者免疫功能低下，TACE后免疫功能进一步降低。THPP后，淋巴细胞免疫功能明显增强。     

腹部肿瘤病人rIL-2/LAK治疗前后T细胞亚群、淋巴细胞转化试验及NK细胞活性变化

本文对30例晚期腹部肿瘤患者rIL－2／LAK治疗前后的免疫功能进行动态观察。检测指标包括T细胞亚群（CD3、CD4、CD8及CD4／CD8）。淋巴细胞转化试验（LymphocytesBlastogenesisTestLBT）和NK细胞活性。结果表明：肿瘤患者经rIL－2／LAK治疗后，CD3、CD4明显升高．CD8也略有升高．CD4／CD8明显升高；淋巴细胞转化率明显升高；经rIL－2／LAK治疗后，NK细胞活性增强。这反映肿瘤患者经rIL－2／LAK治疗后机体细胞免疫功能得到增强。     

肺癌患者外周血红细胞免疫功能和T淋巴细胞亚群测定的临床观察

本文对62例肺癌患者进行红细胞免疫功能及T淋巴细胞亚群测定，并与20例正常人对照，结果显示：肺癌组红细胞膜C3b受体活性（RBC－C3bRR）；CD3 、CD4 、CD4 ／CD8 比值均低于正常人（P＜0.05～0.01），红细胞膜吸附免疫复合物（RBC－ICR）、CD8 均高于正常人（P＜0.05～0.01），因此认为红细胞免疫功能及T淋巴细胞亚群测定对肺癌的诊断、治疗及预后有一定价值。     

微波固化保肢术对骨肉瘤患者Th1/Th2漂移的逆转

目的：研究微波组织固化保肢技术对患者免疫功能的影响。方法：应用放射免疫分析法以及酶联免疫分析法检测20例行微波组织固化保肢术组骨肉瘤患者及20例其它手术组患者手术前后血清IL－2,IL－4,IL－6,IL－10,IL－12的含量,应用流式细胞仪检测患者淋巴细胞亚群的变化。结果：与正常对照相比,骨肉瘤患者CD4^ T细胞,血清中IL－2,IL－12减少（P＜0．01）,CD8^ T细胞,IL－10,IL－4,IL－6含量升高（P＜0．01）,CD4／CD8比值降低（P＜0．01）,患者免疫功能受到抑制;术后30天截肢组及保肢组CD4^ T细胞,血清中IL－2,IL－12均升高,CD8^ T细胞,IL－10,IL－4,IL－6含量降低（P＜0．01）,CD4／CD8比值升高（P＜0．01）,患者免疫功能较术前改善;与截肢组比较,保肢组患者CD4^ T细胞、IL－2,及CD4／CD8比值30天上升幅度明显高于截肢组（P＜0．01）,CD8^ T细胞,IL－4,IL－6 30天降低幅度明显高于截肢组（P＜0．01）。IL－2,IL－12与CD4^ 细胞成正相关,IL－4,IL－6,IL－10与CD4^ T细胞成负相关。结论：微波固化保肢术可以加速逆转骨肉瘤患者Th1/Th2和Th2漂移,增强患者的免疫功能。     

多西他赛对复发转移性乳腺癌免疫功能的影响

目的：探讨多西他赛化疗前、化疗2周期评价疗效时复发转移性乳腺癌（MBC）患者外周血淋巴细胞亚群比率的变化及影响因素。方法应用流式细胞仪检测34例行多西他赛一线化疗的复发MBC患者化疗前、化疗2周期评价疗效时外周血淋巴细胞亚群[CD3+T淋巴细胞、CD3+/CD4+T淋巴细胞、CD3+/CD8+T淋巴细胞、CD3－/CD16+56+自然杀伤细胞（NK）、CD3+/CD16+56+T淋巴细胞、CD19+B淋巴细胞、CD4+/CD25+调节性T细胞（Treg细胞）、CD8+/CD28－T淋巴细胞和CD8+/CD28+T淋巴细胞]比率,并进一步分析患者临床病理因素对于外周血淋巴细胞亚群变化的影响。结果34例患者化疗后CD3+总T淋巴细胞、CD3+/CD4+T淋巴细胞及CD19+B淋巴细胞比率均较化疗前下降（P=0.002、0.044、0.006）,下降平均比率分别为2.2%、4.7%、3.1%。中位年龄﹥54岁的患者CD19+B淋巴细胞下降比率较中位年龄≤54岁的患者小（P=0.031）;中位OS﹥3.6个月的患者CD3+总T淋巴细胞下降比率较中位OS≤33.6个月的患者小（P=0.038）。结论多西他赛化疗后MBC患者外周血总T、B淋巴细胞比率下降,免疫功能降低,而相对保留较好的T淋巴细胞免疫功能的患者可能有更好的生存获益。     

支气管动脉灌注化疗对肺癌患者血清SIL-2R及T细胞亚群的影响

目的 探讨肺癌患者支气管动脉灌注化疗前后血清可溶性白细胞介素2受体(SIL-2R)、T细胞亚群的变化及临床意义.方法 选择肺癌患者30例,于支气管动脉灌注(BAI)化疗前、化疗后2周、4周,分别采用双抗体夹心(ELISA)法及碱性磷酸酶抗碱性磷酸酶(APAAe)法检测血清SIL-2R、T细胞亚群的变化,并与30例健康者作对比分析.结果 肺癌患者SIL-2R水平、CD8细胞显著高于健康者,同时CD3细胞、CD4细胞、CD4/CD8比值低于健康对照组.按TNM分期,Ⅲ期患者SIL-2R水平高于Ⅱ期(P＜0.01),Ⅳ期高于Ⅲ期(P＜0.05).BAI治疗后2周血清SIL-2R水平、T细胞亚群与治疗前比较未见明显变化(P＜0.01),治疗后4周血清SIL-2R、CD3、CD4、CD4/CD8比值升高,CD8下降(P＜0.01).结论 动态观察肺癌患者BAI治疗前后血清SIL-2R水平、T细胞亚群变化可监测机体的免疫状况,有助于判断病情及预后,指导临床综合治疗;对首次接受BAI治疗的患者在4周后进行第二次治疗较为恰当.     

原发性肝癌肝动脉留置导管灌注LAK细胞与化疗栓塞联合应用的疗效观察

对原发性肝癌肝动脉留置导管灌注同种异体ＬＡＫ细胞与化疗栓塞联合应用的临床疗效进行观察。结果表明：联合治疗组总有效率８４％，明显高于单纯化疗栓塞组（７０％），大多数病人一般症状明显改善，免疫状态增强。治疗副反应为一过性发热。     

IL－2／LAK疗法对肿瘤患者免疫功能的影响

我们观察了２０例肿瘤患者应用ＩＬ－２／ＬＡＫ疗法前后其免疫指标的改善，包括Ｔ淋巴细胞亚群、ＬＡＫ细胞活性、白介素Ⅱ膜受体、白介素Ⅱ分泌细胞及血清白介素Ⅱ受体。通过治疗显示：ＣＤ４／ＣＤ８比值、ＬＡＫ活性、白介素Ⅱ膜受体及白介素Ⅱ分泌细胞的水平均显著升高，而可溶性白介素Ⅱ受体减少，提示ＩＬ－２／ＬＡＫ可以改善机体免疫功能。     

Levels of soluble interleukin-2 receptors are predictive of response in patients treated with interleukin-2 and lymphokine-activated killer cells

Soluble interleukin-2 receptor (sIL-2R) α (CD25) levels were serially determined in the sera of 20 patients who had undergone adoptive immunotherapy with high-dose IL-2 and lymphokine-activated killer (LAK) cells for various types of metastatic solid tumors or Hodgkin’s disease. The treatment course consisted of 5 days of high-dose IL-2 priming followed by the collection of peripheral blood leukocytes by leukapheresis, andin vitro activation of mononuclear cells with IL-2, and the subsequent infusion of such prepared LAK-cells together with IL-2. sIL-2R levels increased in all patients following IL-2 administration, and the ratio of baseline sIL-2R levels to those measured after 5 days of IL-2 was signifıcantly correlated with pre-IL-2 levels (p=0.016) in that higher pre-IL-2 levels resulted in a larger increase upon IL-2 administration. In terms of treatment outcome, the variables analysed included sIL-2R levels, total IL-2 doses administered, the expression of membrane-bound CD25 onin vitro cultured cells (pre- and post-IL-2 exposure), the total number of LAK-cells infused andin vitro cytotoxic activity of LAK-cells against the natural killer cell-resistant cell line Daudi. In a multivariate analysis, low baseline sIL-2R levels (p=0.095) and highin vitro cytotoxic activity of LAK-cells against Daudi cells (p=0.082) were jointly associated with response. Our data suggest that serum sIL-2R levels provide a fast and non-invasive parameter for predicting the response in patients treated with IL-2 and LAK-cells.     

Cellular immune profile of patients with advanced cancer before and after taxane treatment

The purpose of this study is to determine immune recovery and function after treatment with docetaxel or paclitaxel. Peripheral blood mononuclear cells were harvested before chemotherapy and at weekly times afterwards for cycle 1. Leukocyte subsets ICD45hiCD14lo polymorphonuclear neutrophils, CD45hiCD14hi monocytes, CD45hiCD14- lymphocytes, CD3+CD4/CD8+ T cells, CD3-CD19+ B cells, CD3-CD16/CD56+ natural killer (NK) cells], and circulating cytokine levels [tumor necrosis factor-alpha, gamma-interferon (gamma-IFN), and interleukins (IL-2, IL-10, IL-12)] were followed. In addition, T-cell mitogenic function, NK function, and lymphokine activated killer (LAK) function was assessed. Ten patients were entered in the trial. T-cell frequency, B-cell frequency, and CD4/CD8 ratio did not change. IL-10 serum levels significantly decreased in paclitaxel-treated patients (4.4+/-1.3 pg/ml at week 4 versus 7.8+/-2.1 pg/ml at baseline; p < 0.05). IL-2, IL-12, and gamma-IFN levels were not detectable. NK cytotoxic activity decreased in docetaxel-treated patients. LAK cell activity was not altered. Four patients achieved a partial or complete response. They demonstrated higher than normal CD4:CD8 T-cell ratios and an improved phytohemagglutinin stimulation index (SI = 2.5). In conclusion, our findings suggest that immune function was affected more significantly after docetaxel treatment. Investigational approaches, which enhance cellular immunity, may be of greater relevance after treatment with docetaxel. Additional studies monitoring NK function after chemotherapy are recommended.     

Heterogeneity of human lymphokine (IL-2)-activated killer (LAK) precursors and regulation of their LAK induction by blood monocytes

Highly purified lymphocytes (greater than 99%) and monocytes (greater than 90%) were isolated by CCE from peripheral blood of healthy donors. Blood lymphocytes were separated by this CCE into 9 subpopulations. The NK activities of these lymphocyte fractions against NK-sensitive K-562 cells and their LAK activities against NK cell-resistant target (Daudi) cells were assayed promptly or after incubation of the fractions for 4 days with or without an optimal concentration of IL-2. NK and LAK activities were measured by 4-hr 51Cr-release assay. On the basis of their NK and LAK activities, these lymphocyte fractions were classified into 3 subpopulations of LAK precursors: one lacking both NK and LAK activities (Fr.2), one with moderate NK activity but low LAK activity (Fr.5), and one possessing both NK and LAK activities (Fr.8). Addition of autologous fresh monocytes to the lymphocyte cultures resulted in a significant increase in induction of LAK activity in Fr.2 and Fr.5. This up-regulation of lymphocytes in Fr. 2 and Fr.5 by monocytes was confirmed in parallel experiments by measuring the blastogenic response of the lymphocytes to IL-2. Deletion of lymphocytes in Fr. 8 of CD16+ (Leu-11+) NK cells resulted in 74% reduction in LAK induction, whereas depletion of mixtures of monocytes and lymphocytes in Fr. 2 of cells reacting with CD3+ (OKT3+) antibody resulted in a 66% reduction in LAK induction. This up-regulation of LAK cell induction from LAK precursors by monocytes was confirmed using 4 lines of human lung cancer cells as targets for LAK activity. These results clearly indicate that human monocytes may cause up-regulation of the expression of IL-2-induced LAK activity in T cells and in a subpopulation of NK cells.     

磁流体对肝癌细胞的凋亡诱导研究及联合紫杉醇对大鼠肝癌的抑制作用

不同浓度的磁性纳米颗粒作用于肝癌HepG2细胞后,检测其凋亡情况,并电镜观察细胞形态变化。制备抗肿瘤靶向药物紫杉醇白蛋白磁性纳米颗粒（MAG-TAX-NP）,将其联合碘化油治疗大鼠肝癌,观察治疗效果。方法:化学共沉淀法制备磁流体,将其作用于HepG2细胞,流式细胞仪检测细胞凋亡情况。电镜观察磁微球作用于肝癌HepG2细胞后的形态学变化。用乳化-超声-加热固化法制备紫杉醇磁性纳米颗粒。建立大鼠肝癌模型,将荷瘤大鼠30只随机分为5组,除空白对照组外,分别将碘化油﹑碘化油紫杉醇﹑碘化油纳米磁流体、碘化油紫杉醇白蛋白磁性纳米颗粒各0.2 mL注入其余4组大鼠肝固有动脉内。紫杉醇用量为5mg/kg体重,肝肿瘤区外加磁场,14d后处死大鼠,取肝肿瘤组织称重,计算各组抑瘤率。结果:证实了磁流体可诱导肝癌HepG2细胞凋亡。电镜下可见肿瘤细胞吞噬磁流体后细胞内形成凋亡小体,细胞裂解增加。大鼠肝癌模型中除空白对照组外,其余各组的抑瘤率分别为43.2%、51%、57.4%、87.4%。结论:磁微球可诱导肝癌HepG2细胞凋亡。碘化油紫杉醇白蛋白磁性纳米颗粒局部栓塞可抑制肿瘤生长,较其他组具有更强的抑瘤效果。随着深入研究,磁流体有望成为治疗肿瘤的方法,紫杉醇白蛋白磁性纳米颗粒可作为临床紫杉醇药物治疗肝癌的新剂型。      

原发性肝癌肝动脉导管灌注CD_3AK细胞与化疗栓塞联合应用的疗效观察

观察肝动脉导管灌注CD3AK细胞联合TAE治疗晚期HCC的效果。方法：40例晚期HCC分为两组，实验组20例，采用肝动脉导管灌注CD3AK细胞联合TAE治疗；对照组20例，单纯TAE治疗。并检测两组AFP及T细胞亚群的变化。结果：实验组与对照组有效率分别为45％、30％（P＜0．01），治疗后实验组CD4、CD4／CD8比值明显升高（P＜0．01），而对照组则轻度下降。AFP两组均明显下降，以实验组为著（P＜0．005）。结论：肝动脉导管灌注CD3AK细胞联合TAE治疗晚期HCC，是一种最有希望的方法。     

Interleukin-2 Therapy for Myelodysplastic Syndrome: Does It Work?

Recent clinical studies suggested that interleukin-2 (IL-2) has therapeutic potential for some hematologic malignancies, but the therapeutic role of IL-2 for myelodysplastic syndrome (MDS) is still unclear. MDS is a clonal malignant disorder which often involves a variety of immunologic abnormalities. Examination of the effects of IL-2 on MDS in vitro yielded the following results: (1) IL-2 did not induce the proliferation of blasts in most MDS cases. (2) The cytotoxicity of IL-2-induced lymphokine-activated killer (LAK) cells for cell lines and MDS blasts was reduced in the high-risk MDS group (refractory anemia with excess blasts (RAEB), RAEB in transformation and MDS transformed to acute leukemia), but it was still preserved in the low-risk MDS group (refractory anemia (RA) and RA with ringed sideroblasts). However, considerable variation in LAK cell cytotoxicity was noted in each group. (3) The reduced LAK cell cytotoxicity observed in MDS was explained, at least in part, by the presence of a reduced of number of natural killer (NK) cells amongst the LAK cells. (4) MDS patients who have a high blood soluble IL-2 receptor (sIL-2R) level often had defects in NK and CD8+ T cells. These in vitro findings suggest that the response to IL-2 is heterogeneous in MDS patients, and those who have a low-risk MDS subtype and/or a low blood sIL-2R level, may be prone to respond to IL-2 therapy. Clinical trials are mandatory in order to elucidate the efficacy of IL-2 therapy in the treatment of MDS.     

Lymphokine-activated killer-cell function of lymphocytes from peripheral blood, regional lymph nodes and tumor tissues of patients with oral cancer

Lymphokine-activated killer (LAK) cells, generated from peripheral blood lymphocytes (PBL) from patients with oral cancer or oral leukoplakia and from healthy donors showed comparable lysis of 6 target tumor cell lines, including 3 derived from head and neck and oral cancers. The tumor burden of the host did not appear to influence the systemic LAK activity. LAK activity of lymphocytes infiltrating the tumor tissues (TIL) was also comparable to that of the PBL. Both TIL and PBL showed a parallel increase in proportion of HNK-I+ and CD-25+ cells upon activation with IL-2. The lymph-node lymphocytes (LNL) from metastatic (met) and non-metastatic (non-met) draining lymph nodes, however, showed reduced LAK activity and an increase in CD8+ cells, in addition to CD25+ and HNK-I+ cells, when cultured with IL-2. When IL-2-activated LNL were co-cultured with autologous PBL during IL-2 activation of the latter, a strong suppressive effect was exerted by LNL. In contrast, IL-2-activated PBL did not suppress autologous LAK generation in spite of an increase in CD8+ cells seen after activation with IL-2. Frequency distribution of LAK precursors was significantly lower in LNL than in PBL from oral cancer patients. LAK precursor frequency in TIL was comparable to that of PBL. The results show that, in oral cancer, regional lymph nodes may not have adequate IL-2-inducible cytotoxic potential, due to a reduced number of LAK progenitors and possible activation of suppressor cells. Alternatively, TIL can be a potential source for LAK cell function.