喉咽癌树突状细胞表型抗原表达

目的对喉咽癌中树突状细胞(Dendriticcells,DCs)的表型抗原的表达进行研究,探讨DC表型抗原的表达与喉咽癌各临床因素尤其是转移和预后的关系。方法对45例喉咽癌标本,用EnvisionHIS的方法检测DC的表型抗原S-100、CD1a、CD83的表达,并观察CD45RO T细胞与DC的关系。结果S-100 DCs在高分化组以及生存组中的数目明显多于中低分化组及死亡组,具有显著性差异。CD83 DCs的表达与喉咽癌的分化程度、远处转移、生存以及是否复发有关,差异具有显著性;CD83 DCs在癌巢中的数目较癌旁中的少,差异具有显著性。CD1a DCs的表达与喉咽癌的各临床因素间未发现明显相关性。CD45RO T细胞主要分布在CD83 DCs的周围。结论表达不同表型抗原的DC具有不同的功能,CD83 DCs被认为是成熟DC分布在癌旁与其激活T细胞的抗原提呈功能有关,对于肿瘤免疫防御机制的建立起着重要作用。DC在喉咽癌中的浸润程度及表型抗原的表达情况是反映宿主肿瘤免疫状况的重要指标,也是预测喉咽癌转移和预后的一项重要指标。     

Head and neck squamous carcinomas with exophytic and endophytic type of growth have the same prognosis after surgery and adjuvant radiotherapy

Head and neck squamous cell carcinomas (HNSCC) are characterized by exophytic or endophytic growth. We hypothesized that the growth pattern predicts outcome and associates with distinct clinical and immunological profiles. Tumors obtained from 60 HNSCC patients treated with surgery and adjuvant radiotherapy were identified as exophytic or endophytic. Recurrence-free survival (RFS) at 42 months was determined. In a subsets of 30 patients (22 exophytic and 8 endophytic) tumor stroma and parenchyma were evaluated for infiltrating CD4⁺ and CD8⁺ T, dendritic, myeloid and FOXP3⁺ regulatory T cells (Treg) and expression of immunosuppressive cytokines by immunohistochemistry. The localization and frequency of positive cells were determined microscopically and analyzed by hierarchical clustering to distinguish exophytic versus endophytic tumors. 34/60 patients had exophytic and 26/60 endophytic tumors. No differences in clinicopathologic data, disease progression or RFS were seen between the two cohorts. Infiltrates of CD3⁺CD8⁺ T cells were larger in endophytic than exophytic tumors, while FOXP3⁺ Treg, TGF-β⁺, IL-10⁺, Arg-1⁺, CD11b⁺ cells were equally prominent in both. FOXP3⁺ Treg accumulated in endophytic tumor nests, while the exophytic tumor stroma was enriched in IL-10⁺ cells (both at p < 0.05). Hierarchical clustering based on immunophenotyping failed to identify different clusters in these two tumor types. However, CD68⁺ macrophages and FOXP3⁺ Treg showed a distinct distribution. The HNSCC growth pattern did not predict RFS. Although higher numbers and differences in localization of immunosuppressive cells in endophytic versus exophytic tumors were observed, no significant relationship was established between the growth pattern and the immune profile of infiltrating lymphocytes.     

Expression of intercellular adhesion molecule (ICAM)-1 in adenoid cystic carcinoma of the head and neck

OBJECTIVES/HYPOTHESIS: Adenoid cystic carcinoma of the head and neck (ACCHN) is characterized by late recurrence and frequent distant metastasis. Tumor attack by cytotoxic T lymphocytes and macrophages is mediated by the interaction of leukocyte function-associated antigen (LFA)-1 on lymphocytes with intercellular adhesion molecule (ICAM)-1 on the tumor surface. Thus, the reduced expression of ICAM-1 on tumor cells could contribute to their escape from host immune surveillance. To investigate the relationship between the clinical features of ACCHN and host immune surveillance, the expression of ICAM-1 and infiltration of T/natural killer (NK) cells and macrophages were immunohistochemically examined. STUDY DESIGN: Retrospective analysis of immunohistochemical tumor characteristics and clinical outcome. METHODS: Immunohistochemical study of ICAM-1, T/NK cells, and macrophages was performed on paraffin sections of 42 patients with ACCHN. The expression of T/NK cells and macrophages was represented by T-cell-restricted antigen (TIA)-1 and CD68 expression, respectively. The expression of these molecules and clinical features were analyzed. RESULTS: Of 42 ACCHN cases, 15, 9, and 15 patients were classified as ICAM-1 high, TIA-1 high, and CD68 high, respectively. The TIA-1 expression scores in ICAM-1-low patients were significantly lower than those in ICAM-1-high patients (1.3 +/- 3.7 vs. 8.3 +/- 12.7, P =.0031). The CD68 expression scores in ICAM-1-low patients were also significantly lower than those in ICAM-1-high patients (9.6 +/- 9.6 vs. 21.1 +/- 17.6, P =.0047). Moreover, ICAM-1-high patients had a significantly better disease-free survival rate (P =.043). CONCLUSIONS: Reduced expression of ICAM-1 may promote immune evasion and metastasis, resulting in poor prognosis in ACCHN.     

Histologic and immunohistochemical characterization of tumor and inflammatory infiltrates in oral squamous cell carcinomas treated with local multikine immunotherapy: the macrophage at the front line

Squamous cell carcinomas of the head and neck (SCCHN) are excellent candidates for local immunotherapy owing to their accessibility and their infiltration by mononuclear cells that are susceptible to immunomodulation. A response rate of 25–60% has been reported for treatment with natural IL-2 or a mixture of natural lymphokines. In the present study, biopsies and posttreatment excision specimens from nine patients with operable SCCHN treated systemically with a variety of immunomodulators and locally with natural lymphokines (multikine, CelSci) were analyzed in an attempt to correlate clinical response to histopathological and immunohistochemical changes. Formalin-fixed, paraffin-embedded tissues were stained with antibodies against lymphocytes (CD45, CD3, CD4, CD8, CD20), macrophages (CD68) including dendritic cells (S-100), markers for lymphocyte activation (CD30, HLA-DR), natural killer cells (CD56 and CD57), beta-2-microglobulin and keratin. One patient showed a complete response to treatment and two a partial response. Tumor size was significantly smaller after therapy. Clinical and pathological regression were more prominent in the smaller tumors. Numerous macrophages, both mononucleated and multinucleated, were present along the tumor-stroma interface in the posttreatment specimens of seven patients, most prominently in the three patients with tumor regression. The increase in the number of CD68+ and S-100+ macrophages after treatment was statistically significant. Lymphocytic infiltrates, which showed some increase following treatment, were composed of a mixture of T and B lymphocytes, the former mostly in contact with the tumor and the latter placed more peripherally. CD8+ lymphocytes extended into the tumors, whereas CD4+ lymphocytes showed minimal extension. Intensity of beta-2-microglobulin staining in tumors was significantly higher following therapy and associated with a better outcome. The marked increase in macrophages following treatment may indicate that the macrophage plays a major role in tumor recognition, destruction and clearance. An increase in the number of macrophages in a posttreatment specimen may indicate immunoresponsiveness.This study is a CISEPO project and was supported in part by the Saul A. Silverman Family Foundation.     

树突状细胞表型抗原在喉咽癌颈部淋巴结中的表达

目的：探讨喉咽癌颈部淋巴结中树突状细胞（Dc）表型抗原的表达与喉咽癌转移及预后的关系。方法：对37例喉咽癌144枚淋巴结DC的表型抗原S-100、CD1a、CD83的表达用Envision HIS的方法检测,并观察CD45RO^＋T细胞与DC的关系。结果：S-100^＋DC、CD1a^＋DC、CD83^＋DC在转移组淋巴结中的数目较非转移组淋巴结中减少,差异有统计学意义。CD83^＋DC在转移淋巴结的癌巢中的数目较癌旁中的少,差异有统计学意义。在生存组淋巴结中的浸润数目较死亡组多,差异有统计学意义。CD45RO^＋T细胞主要分布在CD83^＋DC的周围。结论：表达不同表型抗原的DC具有不同的功能,CD83^＋DCs被认为是成熟DC分布在癌旁与其激活T细胞的抗原提呈功能有关,对于肿瘤免疫防御机制的建立起着重要作用。DC在淋巴结中的浸润程度及表型抗原的表达情况是反映宿主肿瘤免疫状况的重要指标,也是预测喉咽癌转移和预后的一项重要指标。     

Functional and phenotypic analysis of lymphocytes in head and neck cancer.

We compared the phenotype and antitumor effector function of lymphocytes obtained from tumor tissues, lymph nodes, and the peripheral blood of patients with head and neck cancer. Freshly isolated tumor-infiltrating lymphocytes were deficient in CD4+ T cells in comparison with lymph node lymphocytes (LNL) and peripheral blood lymphocytes. A significantly higher CD4/CD8 ratio observed in LNL vs tumor-infiltrating lymphocytes and peripheral blood lymphocytes was attributable to both a significant enrichment in CD4+ T cells as well as a decrease in CD8+ T cells. The percentage of natural killer cells (CD3-CD56+) was uniformly low in both tumor-infiltrating lymphocytes and LNL. In patients with cervical metastases, LNL contained an increased proportion of CD16+ cells. Tumor-involved lymph nodes were not enriched in the CD8+C11b+ subset of T "suppressor" lymphocytes compared with uninvolved lymph nodes. Also, tumor-involved lymph nodes had significantly fewer CD4+ T cells than did uninvolved lymph nodes. In comparison with peripheral blood lymphocytes, freshly isolated tumor-infiltrating lymphocytes and LNL were depleted of cytotoxic effector cells, as indicated by low or absent cytotoxic activity against tumor cell targets. The ability to generate lymphokine-activated killer cells was significantly reduced in LNL in comparison with peripheral blood lymphocytes. In patients with head and neck cancer, depressed local and regional antitumor responses are associated with a deficiency of functional cytotoxic effector cells rather than an increase in suppressor T lymphocytes.     

Anti-tumor immunity against nasopharyngeal carcinoma by means of LMP2A-specific cytotoxic T lymphocytes induced by dendritic cells

OBJECTIVE: Adoptive immunotherapy with specific cytotoxicity T lymphocytes (CTLs) induced by dendritic cells (DCs) pulsed with tumor antigens plays a crucial role in the immunity against tumor. The purpose of this study was to assess the feasibility and efficacy of latent membrane protein 2A (LMP2A)-specific CTLs immunity against nasopharyngeal carcinoma (NPC) in vitro and in vivo. METHODS: DCs were generated by culturing the monocytes purified from human peripheral blood mononuclear cells (PBMCs) in cytokine cocktail containing granulocyte/macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4), and tumor necrosis factor-alpha (TNF-alpha). The phenotype of mature DCs was analyzed by fluorescence activated cell sorter (FACS). Mature DCs transfected with EBV-LMP2A recombinant adenovirus were co-cultured with autologous PBMCs to induce LMP2A-specific CTLs. The expression of the surface antigens such as CD3, CD4, CD8 and CD56 on CTLs were detected by FACS. The specific cytotoxicity of CTLs on target CNE cells expressing EBV-LMP2A was confirmed using cytotoxicity assay. The anti-tumor effect of LMP2A-specific CTLs in vivo was assessed in the mice tumor models implanted with CNE cells expressing EBV-LMP2A. RESULTS: Mature DCs expressed typically morphologic characteristics and high level of surface markers (CD1a, CD83, CD86, CD80 and HLA-DR). LMP2A-transfected DCs could induce LMP2A-specific CTLs consisting of a majority of CD4+ and CD8+ T cells. Cytotoxicity assay confirmed that the LMP2A-specific CTLs displayed significant cytotoxicity on target CNE cells compared with the controls. The study in vivo demonstrated that the treatment using these specific CTLs retarded the growth of established tumor in the treated mice. CONCLUSION: These findings suggest that DCs transfected with EBV-LMP2A recombinant adenovirus can elicit LMP2A-specific CTLs that have a specific killing effect on NPC in vitro and in vivo. The results provide experimental basis for the further immunotherapy of NPC in clinical trails.     

Administration of a prostaglandin synthetase inhibitor associated with an increased immune cell infiltrate in squamous cell carcinoma of the head and neck.

Squamous cell carcinoma of the head and neck produces a prostaglandin, PGE2, a potent inhibitor of cellular immune responses. We tested the effects of prostaglandin synthetase inhibition on the infiltration of squamous cell carcinoma of the head and neck with host lymphocytes. Tumor tissue samples were obtained from six patients (age range, 51 to 72 years) who presented with squamous cell carcinoma of the head and neck before and after 14 days of treatment with indomethacin (50 mg administered orally three times a day). Tumor-infiltrating immune cells were assayed in frozen tissue samples by indirect immunofluorescence. An eightfold increase in CD2+ lymphocytes compared with pretreatment tissue was observed. The number of CD4 and CD8 lymphocytes increased similarly. CD57 lymphocytes increased 15-fold and CD11b cells increased 11-fold. No infiltrating B-cell populations were evident. Double-labeling studies revealed that the mononuclear cells were located outside blood vessel walls, indicating that they had infiltrated the tumor parenchyma. Our findings demonstrate that the administration of indomethacin is associated with the increased immune cell infiltration of squamous cell carcinoma of the head and neck. This suggests that inhibition of PGE2 synthesis as it occurs in the tumor and or systemically may contribute to the homing of mononuclear cells to the tumor. These data suggest a mechanism to account for the clinical response to indomethacin previously reported in squamous cell carcinoma.     

Autologous mixed lymphocyte reaction in head and neck cancer

The proliferative response of T cells cultured with autologous non-T cells is known as the autologous mixed lymphocyte reaction (AMLR). The AMLR in patients with head and neck tumors was found to be significantly decreased in comparison with that in normal donors and patients without evidence of recurrent disease more than 3 years after treatment. Responder T cells in AMLR were CD4+ T cells but not CD8+ T cells. In one patient, AMLR were performed between lymphocytes in condition with tumor burden and those in condition with tumor free by using cryopreserved lymphocytes. T cells in tumor burden state poorly responded to both of these non-T cells. On the other hand, T cell in tumor free state vigorously responded to non-T cells in the same state, but not to non-T cells in tumor burden state. These data indicated that impairment of AMLR in patients with cancer was due to deficit of both responder T cells and autologous stimulator non-T cells.     

Immunologically activated cells in aural cholesteatoma.

In this immunohistochemical study, we characterized the cells infiltrating the stroma of acquired aural cholesteatomas in detail, using a panel of monoclonal antibodies directed against immune cell type-specific antigens, HLA class II antigens, and interleukin-2 receptor. For all antibodies used, normal ear skin was stained for comparison. The vast majority of the infiltrating cells was CD45-positive, ie, derived from bone marrow. Reactivity with anti-CD3 and anti-CD6 antibodies revealed an abundant infiltration of T lymphocytes beneath the squamous epithelium of cholesteatoma. The B lymphocyte-specific anti-CD19 and anti-CD22 antibodies detected only occasional positive cells. Hence, the cellular infiltrate in the stroma of aural cholesteatoma is made up primarily of T cells with macrophages scattered between them. Expression of HLA-DR was almost as high as that of CD45, whereas CD25-positive cells were detected in lower amounts. We infer that the majority of T cells and macrophages in the stroma of cholesteatoma are in an immunologically activated state. The characteristics of the infiltrating cell population suggest an antigen-driven process in cholesteatoma.     

An immunohistologic study of the distribution and status of activation of head and neck tumor infiltrating leukocytes

We examined tumor infiltrating leukocytes (TIL) in frozen sections of 28 biopsies from squamous cell carcinomas of the head and neck (SCCHN). In so doing, we used monoclonal antibodies (MoAb) directed against various leukocyte antigens. As defined by HLe-1+ cells, leukocyte infiltration was present in all biopsies. The amount of HLe-1+ cells was more often greater in stage III than in stage IV lesions. Most of the TIL were identified as CD5+ T-lymphocytes. In contrast, CD19+ B-cells were sparse in most biopsies. CD14+ monocytes/macrophages were found in only a few specimens. The relative proportion of CD4+ T-helper cells was higher than or at least equal to CD8+ suppressor/cytotoxic cells in all samples tested. Interleukin-2 (IL-2) receptor+ lymphocytes were evident in 13 of 22 biopsies stained for CD25 reactivity, and were more often observed in stage III than in stage IV tumors. All biopsies from recurrent tumors had no detectable IL-2 receptor+ cells. Our findings provide evidence for a positive correlation between a greater amount of TIL in earlier stages of SCCHN. The presence of IL-2+ lymphocytes suggests that SCCHN may be capable of activating resting lymphocytes for further IL-2-induced proliferation.     

An immunohistologic study of the distribution and status of activation of head and neck tumor infiltrating leukocytes

Summary We examined tumor infiltrating leukocytes (TIL) in frozen sections of 28 biopsies from squamous cell carcinomas of the head and neck (SCCHN). In so doing, we used monoclonal antibodies (MoAb) directed against various leukocyte antigens. As defined by HLe-1+ cells, leukocyte infiltration was present in all biopsies. The amount of HLe-1+ cells was more often greater in stage III than in stage IV lesions. Most of the TIL were identified as CD5+ T-lymphocytes. In contrast, CD19+ B-cells were sparse in most biopsies. CD14+ monocytes/ macrophages were found in only a few specimens. The relative proportion of CD4+ T-helper cells was higher than or at least equal to CD8+ suppressor/ cytotoxic cells in all samples tested. Interleukin-2 (IL-2) receptor+ lymphocytes were evident in 13 of 22 biopsies stained for CD25 reactivity, and were more often observed in stage III than in stage IV tumors. All biopsies from recurrent tumors had no detectable IL-2 receptor+ cells.Our findings provide evidence for a positive correlation between a greater amount of TIL in earlier stages of SCCHN. The presence of IL-2+ lymphocytes suggests that SCCHN may be capable of activating resting lymphocytes for further IL-2¡nduced proliferation.     

Combination nonviral cytokine gene therapy for head and neck cancer

OBJECTIVE: To establish the feasibility and efficacy of combination nonviral murine interferon-alpha (mIFN-alpha)and murine interleukin-2 (mIL-2) or murine interleukin-12 (mIL-12) gene therapy for head and neck squamous cell carcinoma in a murine model. STUDY DESIGN: Randomized controlled studies in a murine head and neck cancer model were performed to assess antitumor responses, secondary cytokine expression, and both natural killer (NK) cell and cytolytic T-cell (CTL) activity. METHODS: Tumors were established in the floor of mouth in C3H/HeJ immunocompetent mice. Established tumors were directly injected with polymer-formulated murine interferon-alpha (mIFN-alpha), lipid-formulated mIL-2, and polymer-formulated mIL-12 alone or in combination. Primary and secondary cytokine expression,NK cell activity, and CTL activity were assayed. RESULTS: The use of mIFN-alpha gene therapy in combination with either mIL-2 or mIL-12 resulted in significant antitumor effects as compared with each of the single cytokine and control treatment groups (P = .002).Increased levels of NK cell activity and tumor specific CD8+ cytotoxic T-lymphocyte activity were found in the combination mIFN-alpha and mIL-2 or mIL-12 groups. Augmented immune responses correlated with clinical antitumor effects. CONCLUSIONS: The present study demonstrates that mIL-2 or mIL-12 augments tumor inhibition from mIFN-alpha and increases activation of NK and CD8+ T cells. These data support further investigation of polymer and lipid mediated delivery of cytokine genes for head and neck cancer.     

c-myc oncogene copy number in squamous carcinoma of the head and neck.

PURPOSE: Altered resident cellular genetic sequences (oncogenes) may result in malignant transformation, maintenance of tumor growth, and metastatic propensity. In this pilot study, we have elected to probe c-myc oncogene in evaluating specimens from human squamous cell carcinoma. MATERIALS AND METHODS: Samples were obtained from 24 patients with squamous cell carcinoma of the head and neck. The ratio of tumor DNA values to that of control DNA was used to estimate the c-myc copy number. RESULTS: Data from material obtained from eight patients was analyzed to the point of c-myc copy number. Tumors varied from stage II through IV. Five originated in the oral cavity and three in the larynx. Analysis of primary tumors demonstrated that two of eight had increased c-myc copy numbers. Histologically positive neck specimens were encountered in five of the study patients. Three demonstrated elevated c-myc copy numbers, two of which had had increased copy number at the primary site. CONCLUSION: This study confirms that c-myc amplification can be present in squamous cell carcinoma of the head and neck. c-myc Amplification may also be present in neck metastasis. Oncogene amplification in neck metastasis may indicate an increased metastatic propensity for individual tumor cells demonstrating c-myc amplification.     

Immunotype findings in macrophages in aural cholesteatomas

Summary Since a heavy cellular infiltrate is seen in the stroma of most aural cholesteatomas, we attempted to characterize this cell population in more detail using monocyte/macrophage-specific monoclonal antibodies. KiM1+ (specific for CD 11c antigen, the 150kDa -chain of a leukocyte integrin), and KiM6+ phagocytes were present in two- or fourfold higher numbers in the stroma of the six excised cholesteatomas than in the control tissues. Since the stroma of the cholesteatoma is devoid of microvessles, the typical perivascular localization of dermal macrophages was not seen in the cholesteatomas studied. The density of the macrophages in the normal ear skin was much higher in the upper dermis than in the lower dermis. In the cholesteatomatous specimens, the phagocytes were evenly scattered within the connective tissue and the cellular infiltrate. In contrast to diseased skin, no Mac 387+ macrophages were detected in the cholesteatomas. A great number of phagocytic cells closely resembling dermal macrophages was found in the stroma of the cholesteatomas and probably contributes to an active autoimmune process. Offprint requests to: B. Negri     

Immunohistochemical identification of squamous carcinoma of the head and neck with monoclonal antibody SQM1

Frozen tissue sections of biopsies from head and neck squamous cancer lesions were examined for immunohistochemical staining with a recently developed monoclonal antibody, designated as SQM1 antibody and directed against the surface membrane of squamous carcinoma cells. SQM1 antibody stained selectively squamous carcinoma, while normal mucosa and cells of the stroma were non-reactive. Positive staining of tumor was found in 33/35 specimens obtained from several major sites of the head and neck area and was observed in primary manifestations and lymph node metastases as well as in recurrences. The most consistent reactivity was seen with carcinomas of the tongue. Well differentiated squamous carcinomas contained a higher proportion of SQM1 positive tumor cells than poorly differentiated carcinomas. We suggest that the SQM1 antibody may aid in the immunohistochemical identification of squamous carcinoma of the head and neck area.     

Antitumor and antiangiogenic effects of interleukin 12 gene therapy in murine head and neck carcinoma model

Interleukin-12 (IL-12) plays a critical role in producing an immune response, as indicated in many ways, e.g., induction of interferon-gamma (IFN-gamma), and augmentation of the cytotoxic activity of resting activated T cells and natural killer (NK) cells. In this study, we examined whether intratumoral injection of a recombinant retrovirus vector expressing IL-12s induce antitumor and antiangiogenic effects in a murine model using a murine head and neck squamous cell carcinoma (NR-S1). In vitro the levels of vascular endothelial growth factor (VEGF) mRNA and protein expression were decreased in IL-12 gene transfected NR-S1 cell. in vivo direct IL-12 gene therapy resulted in significantly remarkable inhibition of tumor growth compared to the control group. The tumor regression by direct IL-12 gene therapy was also associated with decreased vessel density, and apoptosis and increased infiltration of CD8(+) T cells and CD56(+) NK cells in the tumor increased. Also, the number of IFN-gamma expressed cells of spleen cells was increased in the treatment group compared with the control group. These results suggested that direct IL-12 gene therapy appears to be effective in reducing tumor growth by triggering both antiangiogenic effects and an immunological enhancing mechanism through induction of IFN-gamma.     

Immunolocalization of dendritic cells and pattern recognition receptors in chronic rhinosinusitis

BACKGROUND: Dendritic cell (DC) activation and antigen presentation to T cells are critical to innate and adaptive immunity. Toll-like receptors (TLRs) are known to bind pathogen-associated molecular patterns in addition to sinonasally secreted surfactant proteins (SP) such as SP-A and SP-D. TLR binding is known to activate DCs. Based on these observations, we sought to establish the presence, in sinonasal mucosa, of DC and the pattern recognition receptors (PRRs), CD14, TLR2, and TLR4. METHODS: Sinonasal biopsy specimens were taken from patients with eosinophilic nonatopic nasal polyposis (n = 4), allergic fungal sinusitis (n = 1), and nondiseased patients undergoing cerebrospinal fluid leak repair or pituitary tumor resection (n = 2). Tissue samples were stained immunohistochemically for PRR (CD14, TLR2, and TLR4), mature DC marker (CD208), iDC marker (CD209), or isotype controls. RESULTS: Immature and mature DC were immunolocalized to the subepithelial stroma and ciliated epithelial surface, respectively. Diffuse staining of CD14 was observed throughout the stroma with additional staining in the ciliated epithelium. The TLR markers showed no staining in the ciliated epithelium. TLR2 primarily localized in stroma immediately deep to the ciliated epithelial surface. TLR4 immunolocalized to submucosal seromucinous gland ductal epithelium. Data from nondiseased patients were mixed, with one patient showing minimal staining of any of the tested cellular markers. CONCLUSION: This study indicates progressive DC activation and emigration of mature antigen-presenting cells from the epithelial surfaces of sinonasal mucosa. The presence of TLR known to bind SP-A and SP-D suggests a link between SP expression and immune response in sinonasal mucosa.     

T-cells activated with a trifunctional bi-specific antibody in head and neck cancer

BACKGROUND: The trifunctional bi-specific antibody Removab bridges and activates CD3 positive T cells to EpCAM on carcinoma cells and simultaneously binds to an accessory immune cell, thus inducing tumor cell lysis. Following intravenous application, Removab may induce cytokine-related side effects resulting in a sepsis like syndrom. It was questioned, if peripheral mononuclear blood cells (PBMN) already opsonized with Removab could retain antitumor activity and induce less cytokine release than the mere antibody. METHODS: PBMN of patients with head and neck cancer were incubated with Removab and the released cytokines were washed out. Then the Removab-opsonized PBMN were coincubated with genuine tumor cells of the same patient on a chorioallantois membrane for 24 h (T 24) and 48 h (T 48). Tumor cells coincubated with Cisplatin or solely cell culture medium served as control. RESULTS: Coincubation of tumor cells with opsonized PBMN resulted in a 32 % decrease of viable cells at T 24 and a 37 % decrease at T 48, whereas viable cells increased by 10 % at T 24 or 3 % at T 48 when incubated with medium alone (p < 0.05). This tumor cytotoxicity was similar to that of Cisplatin (35 % at T 24/37 % at T 48). CONCLUSION: In an autologous human ex vivo tumor system, Removab-opsonized PBMN induce tumor cell lysis with significantly reduced cytokine release after i. v. application.     

Tumor-associated macrophages (TAM) in the stroma of squamous cell carcinomas of the ENT area studied using monoclonal antibodies

Monoclonal antibodies were used to examine the presence and nature of tumor associated macrophages and T-lymphocytes in 26 cases of squamous cell carcinoma of head and neck. The content of macrophages varied according to the differentiation of the carcinoma: high differentiation was evidenced by a labelling index by an index between 13.5% and 23.7%, poor differentiation from 27.4% to 35.8%. T-lymphocytes were found where tumors were inflamed, but there was no significant correlation between tumor differentiation and infiltrating T-lymphocytes. The present results that prognosis decreases with the increasing number of tumor associated macrophages in squamous cell carcinoma.