Immunohistology of tumor tissue in local administration of recombinant interleukin-2 in head and neck cancer

Biopsied specimens from patients treated by local administration of rIL-2 were examined immunohistologically. Lymphocytes infiltrating into the cancer tissue before and after administration of rIL-2 were observed on paraffin embedded sections by Hematoxilin Eosin (HE) stain in relation to the clinical effect. And their subsets were identified on frozen sections by Avidin Biotin peroxidase Complex (ABC) method using monoclonal antibodies to examine what kinds of effector cells were induced and increased in the tumor site by rIL-2 in vivo. The intensity of increasing of tumor infiltrating lymphocytes (TIL) was correlated with the clinical effect of local administration of rIL-2. Tumor Infiltrating Lymphocytes were mainly T lymphocytes, of which Leu-2a+ cells (suppressor/cytotoxic T cells) and Leu-3a + 3b+ cells (helper/inducer T cells) were almost equally observed. Leu-2a+ cells were considered to be mainly cytotoxic T lymphocytes because few Leu-2a+ cells were stained with anti-Leu-15 antibody. After administration of rIL-2, marked infiltration of T lymphocytes was observed and there was no obvious different response in degree of infiltration between Leu-2a+ cells and Leu 3a + 3b+ cells. Furthermore, IL-2 receptor+ T lymphocytes (activated T lymphocytes) were increased after administration of rIL-2. And natural killer (NK) cells were slightly observed and also increased after administration of rIL-2. These findings suggest local administration of rIL-2 has possibility to induce cytotoxic T lymphocytes, to increase NK cells and to activate these lymphocytes in vivo.     

Identification of lymphocyte subsets and natural killer cells in head and neck cancers. An immunohistological study using monoclonal antibodies

We investigated host-immune defenses against head and neck cancer cells by using various monoclonal antibodies with an immunoperoxidase technique to define lymphocyte subsets and natural killer (NK) cells. By so doing, we were able to identify lymphocyte subsets and NK cells in various head and neck cancers. We found that the majority of these cells infiltrate in or around nests of cancer cells and are stained with Leu-1 antibody. They include both Leu-2a and Leu-3a positive cells, which show equally intense levels of infiltration. Leu-7 positive cells were only scattered in the peripheral portion of the cancer nests in some cases. We also found a tendency for T-cells to infiltrate more intensely in poorly differentiated squamous cell carcinomas (SCC) than in moderately or well-differentiated SCC. Similarly, T-cells were more prevalent in maxillary carcinomas than in laryngeal carcinomas. These findings suggest the presence of a host-immune defense mechanism against cancer cells in patients with head and neck cancers.     

Identification of lymphocyte subsets and natural killer cells in head and neck cancers

Summary We investigated host-immune defenses against head and neck cancer cells by using various monoclonal antibodies with an immunoperoxidase technique to define lymphocyte subsets and natural killer (NK) cells. By so doing, we were able to identify lymphocyte subsets and NK cells in various head and neck cancers. We found that the majority of these cells infiltrate in or around nests of cancer cells and are stained with Leu-1 antibody. They include both Leu-2a and Leu-3a positive cells, which show equally intense levels of infiltration. Leu-7 positive cells were only scattered in the peripheral portion of the cancer nests in some cases. We also found a tendency for T-cells to infiltrate more intensely in poorly differentiated squamous cell carcinomas (SCC) than in moderately or well-differentiated SCC. Similarly, T-cells were more prevalent in maxillary carcinomas than in laryngeal carcinomas. These findings suggest the presence of a host-immune defense mechanism against cancer cells in patients with head and neck cancers.     

Lymphocytic infiltration in undifferentiated nasopharyngeal cancer

BACKGROUND: Undifferentiated nasopharyngeal carcinoma (NPC) is characterized by prominent lymphocytic infiltration. Although the lymphoid infiltrate in NPC has been examined extensively in morphologic and immunocytochemical studies, the significance of this lymphoid infiltrate and its correlation with prognosis has been a subject of controversy for years. OBJECTIVE: To elucidate the significance of lymphoid infiltration in undifferentiated NPC. DESIGN: Evaluation of the relationship between lymphocytic infiltration in NPC and cervical lymph node status, ultrastructural examination of the lymphoid infiltrate, and assessment of lymphocytic infiltration as an independent prognosticator of regional node metastasis. MATERIALS AND METHODS: Lymphocytic infiltration was evaluated quantitatively in 20 cases of undifferentiated NPC using light microscopy. Four cases of undifferentiated NPC were processed for conventional electron microscopy. The effects of degree of lymphocytic infiltration, age, and tumor stage on cervical nodal metastasis were analysed using the logistic regression model. RESULTS: The degree of lymphoid infiltration correlated with cervical nodal metastasis (P<.001). Ultrastructural evidence of lymphocytes destroying cancer cells was seen. Lymphocytic infiltration was found to be an independent factor affecting cervical nodal metastasis (P =.02, univariate analysis; P =.03, multivariate analysis). CONCLUSIONS: The lymphoid infiltrate is beneficial in undifferentiated NPC, and its presence may deter regional metastasis of cancer cells to the cervical nodes.     

Significance of the involvement of parapharyngeal space cervical nodes and cranial nerves in nasopharyngeal carcinoma

Two hundred cases of nasopharyngeal carcinoma (NPC) admitted to this department from Feb. 1985 to May. 1988 were analysed according to the CT scanning and clinical findings of the primary lesions prior to radiotherapy. The results showed that involvement of parapharyngeal space was very common in NPC, about 80% (160/200 cases); particularly unilateral or bilateral retro-styloid spaces, about 69.5% (139/200 cases). It was proposed that patients with NPC had a high incidence of ipsilateral cervical node metastasis. Contralateral cervical node metastasis was rare. The development of cervical node metastasis in NPC has to modes: one is direct infiltration of the retro-styloid space by the lesion; the other is along the nasopharyngeal lymphatic rete. The data also showed that patients with NPC who presented symptoms of IX-XII cranial nerve paralyses always had ipsilateral or bilateral retro-styloid space infiltrations.     

T-cell subsets and natural killer cell function with squamous cell carcinoma of the head and neck

Twenty-two patients with squamous cell carcinoma of the head and neck were studied to assess potential abnormalities in total T lymphocytes, T4/T8 ratios, and natural killer cell function. Total T-cell counts were reduced for the entire group with the greatest reduction occurring in individuals with hypopharyngeal tumors and/or stage IV disease. Helper to suppressor ratios were within the norm for the total group, but specific site variances were detected in patients with oral cavity and hypopharyngeal primary carcinomas. Similarly altered T4/T8 ratios were noted with stage III disease. Substantive differences in natural killer cell activity were not observed. These data suggest that perturbations in T-lymphocyte subsets can be associated with decreased survival.     

咽喉鳞状细胞癌患者外周血T淋巴细胞亚群和NK细胞活性研究

目的:研究咽喉癌患者外周血T淋巴细胞亚群分布和NK细胞活性及其意义。方法:应用流式细胞仪检测123例咽喉癌患者及36例正常志愿者空腹外周静脉血淋巴细胞亚群和NK细胞活性。结果:咽喉癌患者外周血总T淋巴细胞百分比低于正常人(P<0.05),辅助T淋巴细胞百分比略低于正常人(P>0.05),抑制T淋巴细胞百分比高于正常人(P<0.01),辅助/抑制T淋巴细胞比值低于正常人(P<0.05)。外周血总B淋巴细胞咽喉癌患者略低于正常人(P>0.05),活化T细胞正常人略低于咽喉癌患者(P>0.05),NK细胞活性比较咽喉癌患者低于正常人(P<0.01)。Ⅰ～Ⅱ期外周血总T淋巴细胞略低于Ⅲ～Ⅳ期患者(P>0.05),Ⅰ～Ⅱ期辅助T淋巴细胞含量略高于Ⅲ～Ⅳ期患者(P>0.05),Ⅰ～Ⅱ期抑制T淋巴细胞比值低于Ⅲ～Ⅳ期患者(P<0.01),Ⅰ～Ⅱ期辅助/抑制T淋巴细胞比值高于Ⅲ～Ⅳ期患者(P<0.01)。Ⅰ～Ⅱ期外周血总B淋巴细胞相对值高于Ⅲ～Ⅳ期患者(P<0.05),活化T细胞在Ⅰ～Ⅱ期和Ⅲ～Ⅳ期患者变化不大(P>0.05);Ⅰ～Ⅱ期外周血NK细胞比值略高于Ⅲ～Ⅳ期患者(P>0.05)。结论:咽喉癌患者存在免疫功能紊乱或低...     

鼻咽癌放化疗局部失败的相关因素分析

目的：探讨鼻咽癌放化疗后局部失败（残留或复发）的相关影响因素。方法：对308例鼻咽部原发癌患者的临床病理资料进行回顾分析。选择性别、年龄、T分级、N分级、原发癌病理类型、有无颈淋巴结转移、颈转移淋巴结大小、颈淋巴结转移侧数、颈淋巴结转移累及区域、放疗方法、有无同步化疗等临床病理因素,用x^2检验和Logistic回归进行单因素和多因素分析,并用Kaplan-Meier法对残留和复发患者进行生存分析。结果：在308例头颈部原发鳞状细胞癌患者中,93例（30．2％）发生原发灶和颈部的残留或复发。单因素分析显示,T分级（P〈0.01）、N分级（P〈0.01）、有无颈淋巴结转移（P〈0.05）、颈转移淋巴结大小（P〈0.05）、颈淋巴结转移侧数（P〈0.01）与残留或复发有关。多因素分析结果表明,仅T分级与残留或复发明显相关。用Kaplan-Meier法进行生存分析显示71例残留或复发患者再次治疗的1年、3年、5年生存率分别为77．2％、40．4％、22．4％。结论：原发癌T分级是鼻咽癌治疗局部失败的决定性因素。而有无颈淋巴结转移、原发癌N分级、颈淋巴结转移侧数、颈淋巴结大小是影响因素和T分级的协同因素,但不是导致残留和复发的的初始和根本因素。鼻咽癌侵犯骨时易导致治疗失败。治疗失败者经再次治疗可以提高生存率。     

Definite vs adjuvant radiotherapy. Comparative effects on lymphocyte subpopulations in patients with head and neck squamous carcinoma

The recent association of alterations in T-lymphocyte subpopulations and impaired cellular immunity prompted an investigation of the effects of radiotherapy (RT) on serial levels of lymphocyte subsets in 30 patients with head and neck squamous carcinoma. Percentage and absolute levels of T3, T4, T6, T8, T10, T11, and Leu 7 cells were measured before, during, and after RT at monthly intervals for six months and compared with levels in 40 normal subjects. Sixteen patients received curative and 14 postoperative adjuvant RT. Before treatment, mean subset levels were similar among the patient and normal groups except for elevated Leu 7 (natural killer) cells in patients with stage I and II disease. There were profound decreases in absolute levels of each subpopulation during and after RT. The percentage of T4 (helper/inducer) cells decreased, whereas that of T8 (cytotoxic/suppressor) and Leu 7 cells tended to increase. Compared with normal values, the mean T4/T8 ratio decreased significantly by six months after RT, when absolute levels of the subsets had rebounded to pretreatment levels in the definitive RT group but remained profoundly decreased in the adjuvant group. The differing recovery patterns suggest that factors other than RT may contribute to persistent immunosuppression following RT.     

鼻咽癌患者外周血γδ T细胞的体外扩增及对鼻咽癌细胞的体外杀伤活性

目的：通过体外扩增鼻咽癌（NPC）患者外周血γδT细胞，研究对NPC细胞体外细胞毒作用，以探讨γδT细胞在NPC免疫机制中的作用。方法：用抗体固相法体外扩增7例NPC患者（NPC组）及6例健康者（对照组）的外周血γδT细胞，以流式细胞仪检测γδT细胞亚型，对MTT法观察不同来源的γδT细胞在不同效靶比条件下，对Daudi、CNE1和CNE2细胞的细胞毒作用。结果：对照组与NPC组外周血γδT细胞经体外扩增，纯度可达到82％以上，其中90％为Vγ/Vδ2亚型；γδT细胞对Daudi、CNE1和CNE2细胞显示出作用强度相同的细胞毒作用（P＞0．05）。随效靶比升高，γδT细胞对CNE1和CNE2的细胞毒作用明显增强，在相同效靶比时对Daudi细胞的细胞毒作用最强。结论：NPC患者外周血γδT细胞对NPC细胞有明显体外杀伤作用。     

Soluble Adhesion Molecules and Cytokines in Tumor-associated Tissue Eosinophilia of Nasopharyngeal Carcinoma

The phenomenon of tumor-associated tissue eosinophilia (TATE) is seen in some cases of nasopharyngeal carcinoma (NPC) and is characterized by the eosinophils breaking through the vascular wall and pervading the tumor stroma. The margination and trans-endothelial migration of eosinophils in a typical inflammatory reaction depend on the activating effects of certain cytokines and the expression of adhesion molecules on the eosinophils and endothelial cells. In order to investigate whether the adhesion molecules and activating cytokines play a role in eosinophil tumor infiltration, we measured the serum levels of 3 adhesion molecules, intercellular adhesion molecule-1, E-selectin and vascular cell adhesion molecule-1, and 2 cytokines, IL-3 and IL-5, in 60 NPC patients and 40 normal healthy subjects. We found that the NPC patients had higher serum levels of all three soluble adhesion molecules than the normal subjects but the levels of adhesion molecules failed to correlate with the TATE phenomenon. The levels of IL-3 and IL-5 appeared not to differ between the NPC and control groups. We postulate that the three soluble adhesion molecules do not play a major role in TATE and that their elevation in serum may be due to local and/or systemic immune responses.     

Soluble adhesion molecules and cytokines in tumor-associated tissue eosinophilia of nasopharyngeal carcinoma

The phenomenon of tumor-associated tissue eosinophilia (TATE) is seen in some cases of nasopharyngeal carcinoma (NPC) and is characterized by the eosinophils breaking through the vascular wall and pervading the tumor stroma. The margination and trans-endothelial migration of eosinophils in a typical inflammatory reaction depend on the activating effects of certain cytokines and the expression of adhesion molecules on the eosinophils and endothelial cells. In order to investigate whether the adhesion molecules and activating cytokines play a role in eosinophil tumor infiltration, we measured the serum levels of 3 adhesion molecules, intercellular adhesion molecule-1, E-selectin and vascular cell adhesion molecule-1, and 2 cytokines, IL-3 and IL-5, in 60 NPC patients and 40 normal healthy subjects. We found that the NPC patients had higher serum levels of all three soluble adhesion molecules than the normal subjects but the levels of adhesion molecules failed to correlate with the TATE phenomenon. The levels of IL-3 and IL-5 appeared not to differ between the NPC and control groups. We postulate that the three soluble adhesion molecules do not play a major role in TATE and that their elevation in serum may be due to local and/or systemic immune responses.     

Nasopharyngectomy for recurrent nasopharyngeal carcinoma: a review of 53 patients and prognostic factors

CONCLUSIONS: Salvage surgery is a justified treatment for primary recurrence of nasopharyngeal carcinoma (NPC). Skull base surgery can play a role in rescuing patients with more advanced local recurrence of NPC. OBJECTIVES: The purpose of this study was to report the local control and overall survival outcome of patients with (NPC) with local failure who received salvage nasopharyngectomy and to identify prognostic factors. PATIENTS AND METHODS: Fifty-three consecutive patients who had primary recurrence of NPC and underwent salvage surgery with curative intention from July 1993 to December 2006 were retrospectively reviewed. The follow-up time ranged from 5.1 to 142.2 months. The numbers of cases of recurrent NPC stage were as follows: stage I, 26; stage II, 9; stage III, 10 and stage IV, 8. Fifty patients had one course of radiation therapy while 3 had two courses of radiation therapy before the salvage surgery. For the nasopharyngectomy, 2 patients underwent endoscopic approach and 33 underwent facial translocation, while 18 had craniofacial resection. Postoperative adjuvant treatment included radiation therapy, 4 cases; radiosurgery, 8 cases; concurrent chemoradiation therapy, 7 cases; and chemotherapy, 2 cases. RESULTS: The 5-year local control rates were T1, 58.3%; T2, 27.8%; T3, 53.3%; T4, 75.0%; and all stages, 53.6%. The 5-year overall survival rates were stage I, 64.8%; stage II, 38.1%; stage III, 25.9%; stage IV, 46.9%; and all stages, 48.7%. Multivariate analysis revealed that gender, margin status, adjuvant treatment type and parapharyngeal space involvement were significant impact factors of local control, whereas dura or brain involvement, local recurrence and adjuvant treatment type were significant impact factors of survival.     

胸大肌肌皮瓣在鼻咽癌放疗后颈部挽救性手术中的应用

目的总结胸大肌肌皮瓣在修复鼻咽癌放疗后颈部转移灶残留或复发根治术后缺损中的应用。方法回顾性分析27例鼻咽癌颈部转移灶残留或复发的病人,行根治性颈清扫术,对颈部皮肤缺损用胸大肌肌皮瓣修复。结果1例病人术后3 h死亡,26例病人均一期修复成功。1例术后发生乳糜漏,3例远端吻合口处有感染,5例肌皮瓣远端部分坏死,8例延迟愈合。3年生存率为52.4%。结论鼻咽癌放疗后颈部转移灶残留或复发的挽救性手术应选择根治性颈清扫术;胸大肌肌皮瓣血供丰富,易成活,可取瓣面积大,能满足修复术后缺损的需要。     

Histological examination in Sj?gren's syndrome. Relationship between sialography and histological examination

Although the etiology of Sj?gren's syndrome (SJS) remains unknown, Sj?gren's syndrome can be diagnosed by means of clinical features and sialographic and histopathological examinations. In this study of 35 patients with primary SJS and 21 with secondary SJS, two parameters, (the findings of sialography of parotid gland and the histopathology of labial minor salivary gland), were evaluated. Then it was concluded that grade of lesion in sialography was parallel to infiltrative grade of round cells around labial salivary gland in primary SJS but not in secondary SJS. Using monoclonal antibodies to cell surface antigens, lymphocyte subsets in 16 patients with primary SJS and 4 patients with secondary SJS were studied. A predominance of T cells was found and there was no definitive characterization of lymphocyte subset infiltration, between CD4 positive cell and CD8 positive cell. This result was different from the previous reports, and needs further studies.     

EB病毒潜伏膜蛋白2A诱导细胞毒性T细胞抗鼻咽癌的体内外实验

目的观察EB病毒（Epstein—Barr virus，EBV）-潜伏膜蛋白2A（1atent membrane protein2A，LMP2A）转染人树突状细胞（dendritic cell，DC）诱导特异性细胞毒性T细胞（cytotoxicity T lymphocyte，CTL）的体外生物学特性；建立表达EBV—LMP2A的裸鼠鼻咽癌动物模型，探讨LMP2A特异性CTL在荷瘤鼠体内的抗肿瘤效应。方法分离人外周血单核细胞（pripheral blood mononuclear cell，PBMC），以粒细胞-巨噬细胞集落刺激因子（granulocyte—monocyte colony stimulating factor，GM—CSF）、白细胞介素4（interleukin-4，IL4）及肿瘤坏死因子（tumor necrosis factor-α，TNF—α）诱导培养获得DC，荧光激活细胞分选仪（fluorescence activated cell sorter，FACS）检测成熟DC的表面分子表达。用LMP2A重组腺病毒转染成熟DC，将转染DC与自体PBMC混合培养，在白细胞介素2（interleukin-2，IL-2）作用下诱导针对LMP2A的特异性CTL，FACS检测CTL群体中阳性细胞的组成。将鼻咽癌CNE细胞接种BALB／c裸鼠，建立鼻咽癌动物模型；在裸鼠肿瘤局部注射LMP2A特异性CTL，观察治疗后移植瘤生长情况及病理变化。结果人外周血PBMC体外在GM—CSF、IL4、TNF—α诱导下获得典型形态及表型特征的成熟DC。FACS检测表明，以LMP2A重组腺病毒转染DC诱导的CTL细胞群体以CIM、CD8阳性细胞组成为主。在接种CNE细胞3周后建立表达EBV—LMP2A的裸鼠鼻咽癌动物模型；动物体内实验表明注射CTL的裸鼠肿瘤生长缓慢，体积明显小于对照组（P〈0．01）；病理检查显示肿瘤局部发生液化性坏死并有淋巴细胞浸润。结论人外周血单核细胞体外经细胞因子诱导，可生成具典型特征的成熟DC。利用LMP2A重组腺病毒将LMP2A基因转入DC，可在同一个体体外诱导出针对LMP2A的特异性CTL。CNE细胞接种裸鼠，可成功构建鼻咽癌动物模型；LMP2A特异性CTL在动物体内可明显抑制鼻咽癌肿瘤的生长，发挥有效的抗肿瘤作用。     

Alterations in T-lymphocyte subpopulations in patients with head and neck cancer. Correlations with prognosis

Impaired cell-mediated immunity has been consistently demonstrated in patients with advanced head and neck squamous cancer (HNSC); however, the results of prior studies of correlations of cellular immune parameters with treatment outcome have been inconsistent, and routine assessment of immune parameters has been of limited clinical use. To determine the prognostic importance of alterations in the proportions of various T-lymphocyte subpopulations in the peripheral blood of patients with HNSC, levels of T3, T4, T6, T8, T9, T10, T11, and Leu 7 cells were quantitated by flow cytometry in 80 previously untreated patients and prospectively correlated with tumor characteristics and clinical course (median length of follow-up, 27 months). The mean helper/suppressor cell ratio (T4/T8) increased progressively with increasing tumor stage and was significantly elevated among patients with cancer as a group and in patients with advanced (stage III or IV) disease compared with 40 normal subjects. Decreased disease-free survival was significantly associated with elevated T4/T8 ratios and low percent T8 and T11 cell levels. The prognostic significance of percent T8 (cytotoxic/suppressor) cell levels persisted even after adjusting for known prognostic factors of tumor stage, T class, N class, and tumor site. These correlations provide new insight into immune alterations in HNSC that may prove useful in identifying patients with early clinical disease who have a poor prognosis.     

Immunofluorescent assessment of tumour infiltrating cells in laryngeal carcinoma. Application of monoclonal antibodies

In order to gain some insight into host cell accumulations within primary tumour, frozen sections from surgical specimens of laryngeal carcinoma were subjected to indirect immunofluorescence using a panel of monoclonal antibodies against various human lymphocyte subsets as well as macrophages. In addition, polyclonal antibodies against Ig were used in order to trace B cells. Numerous host cell infiltrates seen at the tumour periphery were composed of T4 (helper) lymphocytes and macrophages. Lymphocytes of OKT8 (suppressor/cytotoxic) and Leu-7 (NK cells) series were intermingled with tumour cells in the case of scanty infiltrates. Infiltrating cells were also linked to the presence of metastases in regional lymph nodes. OKT4-positive abundant infiltrates were usually accompanied by uninvolved nodes, while scanty ones with OKT8 specificity were relatively frequently seen in the patients with evidence of nodal metastases. These differences were not statistically significant, however, B cells as well as plasma cells were infrequently observed and were encountered both in tumour samples with intensive cellular infiltrates as well as in those with scanty ones.