Lactoferrin-induced up-regulation of zeta (zeta) chain expression in peripheral blood T lymphocytes from cervical cancer patients

Alteration of T-cell-associated signal transduction molecules has recently been implicated in immune suppression in tumour-bearing hosts. Here we report the immunoregulatory effects of human lactoferrin (LF) on zeta-chain expression in peripheral blood T lymphocytes from cervical cancer patients and healthy donors. By quantitative flow cytometry analysis, we demonstrated that the mean zeta-chain expression was significantly higher in freshly-isolated T lymphocytes from healthy donors (69%), compared with the patients (38%). Following 3-day culture under standard conditions, zeta-chain expression in T lymphocytes from the patients increased significantly, whereas it dropped in the cells from healthy donors. Anti-CD3 MoAb as well as LF, significantly increased expression of zeta-chain in T cells both from patients and control subjects. The addition of LF to the anti-CD3 MoAb cell cultures resulted in an even higher stimulation of the zeta-chain expression. The results suggest that, in patients with cervical cancer, zeta-chain defects could be corrected by the therapeutic application of LF.     

Spontaneous ex vivo apoptosis of peripheral blood mononuclear cells in patients with head and neck cancer.

Proportions of apoptotic (TUNEL+) peripheral blood mononuclear cells (PBMCs) were measured by flow cytometry in patients with head and neck cancer and normal controls at the time of blood draws (0 time) and after 24-h incubation. PBMCs were incubated at 37 degrees C in medium (spontaneous apoptosis) and in the presence of CH-11 antibody (anti-Fas) or tumor necrosis factor (TNF)-alpha, both capable of inducing DNA fragmentation in activated T cells expressing the TNF family of receptors. PBMCs obtained from the patients had significantly higher (P < 0.0001) proportion of apoptotic cells than PBMCs of controls at 0 time as well as after 24-h incubation. Ex vivo apoptosis included all subsets of PBMCs: CD3+ T cells, CD16+ CD56+ natural killer cells, CD19+ B cells, and CD14+ monocytes, as determined by two-color flow cytometry. However, T cells represented the largest PBMC subset undergoing apoptosis, and lymphocytes rather than monocytes were the major TUNEL+ PBMC population. Among T cells, the level of spontaneous ex vivo apoptosis was nearly as high as that of CH-11 antibody-induced or TNF-alpha-induced apoptosis, indicating that activated Fas+ and TNFR1+ T cells were preprogrammed in vivo to die. Also, elevated levels of spontaneous apoptosis at time 0 in patients with head and neck cancer (P < 0.0001) indicated that a higher fraction of PBMCs was undergoing apoptosis in vivo in patients than controls. Together, the data suggest that an increased rate of turnover of lymphocytes is associated with cancer and may be responsible for functional lymphocyte imbalance, even in treated patients who have no evident disease.     

乳腺癌患者外周血Th1／Th2细胞因子的漂移及其与病理和临床病程的关系

 目的 检测乳腺癌患者外周血CD+3 T细胞分泌细胞因子的水平，分析Th1和Th2细胞的细胞因子免疫变化，为肿瘤的免疫治疗提供实验依据。方法 用刺激物刺激细胞，增加细胞内细胞因子的表达，加入荧光标记的特异性抗细胞因子单克隆抗体，特异性抗原抗体结合，应用流式细胞仪分析特异性细胞因子表达水平。同时用酶联免疫吸附法（ELISA）检测血清中相应的细胞因子。结果 CD+3 T细胞分泌的细胞因子水平及血清中的细胞因子皆表现Th1型细胞因子干扰素-γ（IFN-γ）、白细胞介素-2（IL-2）、白细胞介素-12（IL-12）表达水平较正常对照组显著降低，Th2型细胞因子白细胞介素-4（IL-4）和白细胞介素-10（IL-10）表达水平较正常对照组升高，差异有统计学意义。肿瘤坏死因子-α （TNF-α）表达水平较正常对照组升高，差异有统计学意义。结论 乳腺癌患者体内Th2型细胞因子模式占优势状态，这可能是肿瘤细胞发生免疫逃逸，从而导致肿瘤的发生或者转移的原因之一。     

Effects of ERCC1 expression in peripheral blood on the risk of head and neck cancer.

Squamous cell carcinoma of the head and neck (SCCHN) accounted for 3.5% of all cancers registered in Korea during 2001. Although tobacco smoking and alcohol consumption are known risk factors of SCCHN, only a small fraction of the populations exposed to tobacco or alcohol develop SCCHN. Therefore, differences in the susceptibility of SCCHN with respect to DNA repair and xenobiotic metabolisms have focused upon attempts to determine the causes of SCCHN. Here, we investigated factors that affect ERCC1 mRNA expression in peripheral blood. The study subjects were all Koreans (67 patients and 73 control subjects); 72.9% of all subjects were male; 68.4% were current or former smokers; and 62.4% were current or former alcohol drinkers. We also studied the association between ERCC1 mRNA expression and the C8092A polymorphism of ERCC1, and found lower ERCC1 mRNA expression in SCCHN patients than in controls (P<0.01). In the present study, we found that ERCC1 C8092A polymorphism is not related to the risk of SCCHN or expression of ERCC1 mRNA. In addition, we found a positive association between ERCC1 mRNA expression in peripheral blood and tumour tissues and inverse associations between ERCC1 mRNA expression and age or the number of cigarettes smoked. Therefore, our study suggests that ERCC1 mRNA expression is reduced by age and smoking and has a weak effect on SCCHN risk as compared with the effects of age and tobacco-smoking expression.     

大肠癌患者外周血前列腺素E_2水平的临床意义

研究测定大肠癌患者外周血PGE2 及其临床意义。方法 :1997年 11月— 1999年 3月 ,共 31例大肠癌患者 ,男 14例、女 17例 ,平均年龄 5 5 6± 11 6岁 ;按有否远处转移分为两组 ,其中同时性远处 (肝、肺等 )转移组15例 ,无远处转移组 16例 ,对照组为健康体检者 ;行根治性切除 16例、姑息性切除 13例 ,剖腹探查 2例。病理诊断DukesB期 13例 ,DukesC期 3例 ,DukesD期 15例。用PGE2 酶免药盒分别测定患者手术前后的外周血PGE2 、及术中肿瘤组织和灌流血中的PGE2 。结果 :大肠癌患者外周血PGE2 明显升高 ,外周血PGE2 与肿瘤大小呈正相关 ,DukesD期者显著高于B、C期者。两组肿瘤静脉血PGE2 均高于伴行动脉血 ,其V A值有显著性差异。肿瘤组织的PGE2 含量较正常肠粘膜高 2 8倍。手术切除原发肿瘤两周后外周血PGE2 显著下降 ,其中根治性切除术组与对照组相比无显著差异 ,姑息性切除术组仍显著高于对照组。结论 :外周血PGE2 有可能视为大肠癌根治术后有否复发或转移的一项辅助性监测指标     

大肠癌患者外周血前列腺素E2水平的临床意义

目的研究测定大肠癌患者外周血PGE2及其临床意义.方法1997年11月-1999年3月,共31例大肠癌患者,男14例、女17例,平均年龄55.6±11.6岁;按有否远处转移分为两组,其中同时性远处(肝、肺等)转移组15例,无远处转移组16例,对照组为健康体检者;行根治性切除16例、姑息性切除13例,剖腹探查2例.病理诊断DukesB期13例,DukesC期3例,DukesD期15例.用PGE2酶免药盒分别测定患者手术前后的外周血PGE2、及术中肿瘤组织和灌流血中的PGE2.结果大肠癌患者外周血PGE2明显升高,外周血PGE2与肿瘤大小呈正相关,DukesD期者显著高于B、C期者.两组肿瘤静脉血PGE2均高于伴行动脉血,其V/A值有显著性差异.肿瘤组织的PGE2含量较正常肠粘膜高2.8倍.手术切除原发肿瘤两周后外周血PGE2显著下降,其中根治性切除术组与对照组相比无显著差异,姑息性切除术组仍显著高于对照组.结论外周血PGE2有可能视为大肠癌根治术后有否复发或转移的一项辅助性监测指标.     

肺癌患者外周血和骨髓中MUC1 mRNA表达的临床意义

目的:检测非小细胞肺癌(non smal celllungcancer,NSCLC)外周血和骨髓中粘蛋 白1(mucin1,MUC1)mRNA的表达。方法:应用 RT PCR技术,对31例肺癌患者、10例肺良性病 变患者和8名健康人外周血及骨髓中MUC1基 因mRNA表达进行检测。结果:31例肺癌患者 中10例检测到外周血,7例检测到骨髓中有 MUC1mRNA表达,10例肺良性病变患者和8名 健康人中未检测到MUC1mRNA表达。肺癌患 者外周血及骨髓中MUC1mRNA表达分别为 32.3%(10/31)和22.6%(7/31),两者之间存在 正相关关系,P<0.05;外周血和骨髓中MUC1 mRNA的表达与肺癌组织学类型、细胞分化程度 及pTNM分期均密切相关,P<0.05。结论:肺癌 患者外周血和骨髓中存在常规方法检测不出的 肿瘤细胞;应用巢式RT PCR法可检测到肺癌患 者外周血和骨髓中MUC1mRNA表达,可为制 定治疗方案和评估预后提供重要参考依据。     

大肠癌患者外周血与肿瘤组织中浸润淋巴细胞Fas及FasL表达的意义

目的 :探讨Fas/FasL在大肠癌患者外周血淋巴细胞及肿瘤浸润淋巴细胞的表达变化的意义。方法 :用流式细胞仪对 3 6例大肠癌患者外周血淋巴细胞与肿瘤浸润淋巴细胞(tumorinfiltratinglymphocyte ,TIL)的Fas及FasL进行定量分析。结果 :大肠癌患者外周血淋巴细胞上Fas/FasL表达明显高于正常对照 ,TIL上Fas/FasL表达高于外周血淋巴细胞上表达。结论 :Fas/FasL在大肠癌患者外周血淋巴细胞与TIL的不同分布说明 ,Fas/FasL不仅在肿瘤浸润区域而且在全身参与了免疫系统与肿瘤的相互斗争。     

Clinical significance of poor CD3 response in head and neck cancer.

PURPOSE: The objective of our investigation was to prospectively study what the implications of an unresponsive CD3 receptor are on clinical outcome in advanced-stage head and neck cancer patients. EXPERIMENTAL DESIGN: Lymph node mononuclear cells were purified from cancer patients and stimulated with immobilized anti-CD3 in vitro for 8 days. Two populations were identified, nonresponders (NRs) with [(3)H]thymidine-counts per min (cpm) <3500 and responders (Rs) with cpm >or=3500. NRs and Rs were prospectively followed for a minimum of 24 months, and clinical outcomes were compared. Postoperative complications, length of hospitalization, toxicities associated with chemotherapy or radiation therapy, survival, and disease-free status were measured. RESULTS: Twenty-six patients were followed, of which 19 Rs [[(3)H](X) = 37,819 +/- 24,979 cpm (mean proliferative count +/- SD)] and 7 NRs ([(3)H](X) = 1,375 +/- 1,102 cpm) were identified. There were no phenotypic differences in lymph node T-cell subpopulations (CD3, CD4, CD8, CD28, CD45RO) between groups. There was a 71% (5/7) incidence of recurrent cancer in NRs compared with 16% (3/19) in Rs; the median disease-free interval was significantly less in NRs (P = 0.03). The risk ratio of Rs to develop a recurrent cancer was 0.237 (95% confidence interval, 0.057-0.994), much less than for NRs. CONCLUSIONS: Patients with an unresponsive CD3 receptor as measured by in vitro response to anti-CD3 monoclonal antibodies had a significantly higher incidence of recurrent cancer. Analyses using Cox proportion hazards models demonstrated that CD3 response was the single greatest predictor of reduced disease-free interval. This is the first prospective study to confirm the importance of regional lymph node mononuclear cell CD3 receptor function in head and neck squamous cell carcinoma patients for tumor control.     

外周血癌胚抗原mRNA在大肠癌组织中的表达及其临床意义

目的:应用RTPCR技术检测大肠癌患者外周血CEAmRNA,探讨与不同临床病理指标之间的关系,评估复发转移的早期预测意义。方法:应用RTPCR技术,选择特异性CEAmRNA引物,检测29例大肠癌患者外周血CEAmRNA,同期定量检测外周血CEA和CA199糖蛋白。另抽取12例健康志愿者外周血标本作为对照。结果:29例大肠癌患者外周血中CEAmRNA、CEA、CA199阳性率分别为24.14%(7/29)、17.24%(5/29)、13.79%(4/29)。12例对照组外周血中CEAmRNA、CEA、CA199均阴性。有淋巴结转移者外周血CEAmRNA阳性率(35.29%,6/17)高于无淋巴结转移者(8.33%,1/12),但χ2=2.79,P=0.095。TNM分期Ⅳ期中外周血CEAmRNA阳性率66.67%(4/6),Ⅲ期16.67%(2/12),Ⅰ、Ⅱ期9.09%(1/11),差异有统计学意义,χ2=7.65,P=0.022。外周血CEAmRNA表达在患者的年龄、性别、病理类型等差异无统计学意义,P>0.05。结论:CEAmRNA、CEA和CA199在大肠癌患者中的阳性率,以CEAmRNA阳性率最高,随TNM分期进展,阳性率更高,提示预后不良,应加强治疗。     

外周血癌胚抗原mRNA在大肠癌组织中的表达及其临床意义

目的：应用RT—PCR技术检测大肠癌患者外周血CEAmRNA，探讨与不同临床病理指标之间的关系，评估复发转移的早期预测意义。方法：应用RT—PCR技术，选择特异性CEAmRNA引物，检测29例大肠癌患者外周血CEAmRNA，同期定量检测外周血CEA和CA19-9糖蛋白。另抽取12例健康志愿者外周血标本作为对照。结果：29例大肠癌患者外周血中CEAmRNA、CEA、CAl9-9阳性率分别为24．14％（7／29）、17．24％（5／29）、13．79％（4／29）。12例对照组外周血中CEAmR—NA、CEA、CA19-9均阴性。有淋巴结转移者外周血CEAmRNA阳性率（35．29％，6／17）高于无淋巴结转移者（8．33％。1／12），但x^2=2．79，P=0．095。TNM分期Ⅳ期中外周血CEAmRNA阳性率66．67％（4／6），Ⅲ期16．67％（2／12）。Ⅰ、Ⅱ期9．09％（1／11），差异有统计学意义。x^2=7．65，P=0．022。外周血CEAmRNA表达在患者的年龄、性别、病理类型等差异无统计学意义，P〉0．05。结论：CEAmRNA、CEA和CA19-9在大肠癌患者中的阳性率，以CEA mRNA阳性率最高，随TNM分期进展，阳性率更高，提示预后不良，应加强治疗。     

N-acetyl-beta-glucosaminidase of peripheral blood lymphocytes in patients with cancer of the larynx.

In 20 men, age 35 to 55 years, with cancer of the larynx of N-acetyl-beta-glucosaminidase in peripheral blood lymphocytes was determined cytochemically according to Hayashi's method. Numbers of enzyme-positive cells were counted with regard to the type of cytochemical reaction (granular, granular-diffuse, or diffuse), and to the number of enzyme-positive lysosomal granules within a single cell. In comparison to 20 healthy men, age 20 to 30 years, the following changes were noted in the patients: 1) the decrease of the total count of enzyme-negative lymphocytes; 2) the decrease of the total count of lymphocytes with granular type of reaction; 3) the increase of the counts of lymphocytes with granular-diffuse and diffuse type of reaction. The authors suggest that these changes could be related to the immune reaction of specific tumor antigens.     

T-cell receptor gene expression in tumour-infiltrating lymphocytes and peripheral blood lymphocytes of patients with nasopharyngeal carcinoma.

The T-cell receptor (TCR) repertoire expression of tumour-infiltrating lymphocytes (TILs) from 19 nasopharyngeal carcinoma (NPC) biopsies was compared with those of lymphocytes from 18 control nasopharyngeal biopsies. mRNA was extracted from these lymphocytes and the cDNA transcribed. A panel of 18 V alpha- and 21 V beta-specific primers was used to detect the TCR gene use from cDNA. The use of V alpha and V beta genes was restricted in TILs compared with lymphocytes from biopsies. The frequencies of V alpha 2, V alpha 3, V alpha 9, V alpha 10, V alpha 11, V alpha 13, V alpha 14, V alpha 15, V beta 11, V beta 15 and V beta 20 were decreased and the frequencies of V alpha 10 [Pc = 0.04; relative risk (RR) = 0.05], V alpha 11 (Pc = 0.02; RR = 0.07), V alpha 13 (Pc = 0.002; RR = 0), V alpha 14 (Pc = 0.04; RR = 0.05), V beta 14 (Pc = 0.001; RR = 0.03) and V beta 20 (Pc = 0.001; RR = 0.03) remained significantly reduced after correction for the number of families typed. The frequency of V alpha 17 was higher in NPC biopsies than in NPC PBLs (P = 0.05), and the frequency of V beta 15 was lower in NPC biopsies than in NPC PBLs (P = 0.02). The frequencies of V alpha 17 and V alpha 18 in HLA-B46+ patients were significantly lower (P = 0.009; P = 0.044) than in B46+ controls. The results suggest that the restriction of TCR gene use in NPC patients may be important in NPC pathogenesis.     

乳腺癌患者肿瘤组织及外周血多药耐药基因表达的临床意义

目的:检测多药耐药基因在乳腺癌患者肿瘤组织及外周血中的表达,探讨其临床意义。方法:荧光定量RT-PCR法检测40例原发性乳腺癌患者肿瘤组织及外周血、13例辅助化疗后乳腺癌患者外周血及10例乳腺良性疾病组织与外周血中mdr1的表达情况。结果:40例原发性乳腺癌患者肿瘤组织mdr1阳性表达率为67.5%(27/40),与对照组组织(无阳性表达)相比差异有统计学意义,χ2=14.380,P=0.001;乳腺癌患者外周血mdr1阳性表达率为22.5%(9/40),高于对照组(无阳性表达),χ2=4.486,P=0.034;化疗后乳腺癌患者外周血中mdr1的表达率为53.8%(7/13),高于化疗前外周血mdr1的表达,二者差异有统计学意义,χ2=12.345,P=0.001。结论:乳腺癌患者存在先天性和获得性耐药性,检测肿瘤组织和外周血mdr1表达对制定个体化化疗方案有指导意义。     

肿瘤患者自然杀伤细胞活性水平及其临床意义

目的　研究恶性肿瘤患者外周血自然杀伤 (NK)细胞活性及其临床意义。方法　采用LDH释放法对 43名健康成年人及 2 6 7例肿瘤患者进行了外周血NK细胞活性测定。结果　恶性肿瘤患者NK细胞活性显著低于正常对照组 (P <0 .0 0 1)。伴转移患者NK细胞活性降低更明显 ,与不伴转移者相比 ,差异显著 (P <0 .0 0 0 5 )。化疗及放疗后NK细胞活性显著降低 (P <0 .0 1) ,而IL 2治疗后NK细胞活性显著升高 (P <0 .0 1) ,但低于正常对照组 (P <0 .0 1)。结论　NK细胞活性显著降低与恶性肿瘤发生、发展及转移相关 ,监测NK细胞活性有助于评价肿瘤患者治疗过程中的免疫功能。