The tumor antigens RHAMM and G250/CAIX are expressed in head and neck squamous cell carcinomas and elicit specific CD8+ T cell responses.

Despite advances in surgery, radio- and chemotherapy, therapeutic approaches for patients with head and neck squamous carcinoma (HNSCC) need to be improved. Immunotherapies eliciting tumor specific immune responses might constitute novel treatment options. We therefore investigated the expression and immunogenicity of two tumor-associated antigens (TAA) the receptor for hyaluronic acid mediated motility (RHAMM) and carboanhydrase IX (G250/CAIX) in HNSCC patients. Twenty-two HNSCC samples were examined for the expression of RHAMM and G250 by Western blotting and immunohistochemistry, 14/22 samples were tested for HLA-A2 expression by flow cytometry. For 8/22 samples single tumor-cell suspensions were generated, and mixed lymphocyte peptide cultures (MLPC) were performed to evaluate the frequencies of cytotoxic T cells specifically recognizing RHAMM and G250 using Tetramer staining/multi-color flow cytometry and enzyme linked immunosorbent spot (ELISPOT) assays. RHAMM and G250 were expressed in 73 and 80% of the HNSCC samples at the protein level. A co-expression of both TAAs could be detected in 60% of the patients. In 4/8 HLA-A2+ patients, 0.06-0.13% of CD8+ effector T cells recognized Tetramers for RHAMM or G250 and secreted IFNgamma and granzyme B in ELISPOT assays. RHAMM and G250 are expressed at high frequency and high protein level in HNSCCs and are recognized by cytotoxic CD8+ effector T cells. Therefore both TAAs constitute interesting targets for T cell based immunotherapies for HNSCC.

Phase I trial with the CD44v6-targeting immunoconjugate bivatuzumab mertansine in head and neck squamous cell carcinoma.

CD44v6 is a tumor associated antigen abundantly expressed in head and neck squamous cell carcinomas (HNSCC) and in normal squamous epithelium. The immunoconjugate bivatuzumab mertansine (BIWI 1) consists of a highly potent antimicrotubule agent coupled to a monoclonal antibody against CD44v6. The maximum tolerated dose (MTD), safety and efficacy of BIWI 1 administered IV in patients with HNSCC has not been determined. In a clinical phase I trial, adult patients with recurrent or metastatic HNSCC were treated intravenously with BIWI 1. Starting with 25mg/m(2), the dose was escalated in steps of 25mg/m(2) until dose limiting toxicity was observed. Six women and 25 men were included. The MTD was 300 mg/m(2). Twelve patients were treated with at least the MTD. The principal toxic effects were maculopapular rashes, focal blister formation and skin exfoliation. Three patients had partial responses at doses of 200, 275 and 325 mg/m(2). The concept that bivatuzumab can direct mertansine activity to CD44v6 expressing tumors was confirmed. Although CD44v6 was abundantly expressed in all tumors, the response to BIWI 1 was variable. Binding to CD44v6 on skin keratinocytes mediated serious skin toxicity with a fatal outcome in a parallel trial, which led to the termination of the development program of bivatuzumab mertansine and the present study.

Pharmacokinetics, immunogenicity and safety of bivatuzumab mertansine, a novel CD44v6-targeting immunoconjugate, in patients with squamous cell carcinoma of the head and neck.

The prodrug bivatuzumab mertansine (BIWI 1) is a novel CD44v6-targeting humanized monoclonal antibody coupled to the toxin mertansine. In a phase I dose escalation trial 31 patients with squamous cell carcinomas of the head and neck were treated with doses of 25-325 mg/m2 as a 30-min infusion. Thirteen patients received a second infusion after 3 weeks. Serial serum samples were collected to determine the pharmacokinetic parameters of the prodrug BIWI 1 and of deconjugated BIWI 1 as well as the occurrence of anti-BIWI 1 antibodies. The maximum tolerated dose was reached at 300 mg/m2 attributable to skin toxicity. No immune response was observed in any patient. For BIWI 1 and deconjugated BIWI 1, clearance values were low and distribution was limited resulting in half-lives of approximately 3-3.5 days and approximately 6-7 days, respectively, for single and repeated dosing after three weeks. Overall, interindividual variability of the pharmacokinetic parameters was low. In general, the pharmacokinetics of both compounds after single and repeated dosing was comparable across the entire dose range and no significant accumulation took place. Over the dose range investigated, a dose proportional increase in the exposure of BIWI 1 and deconjugated BIWI 1 was observed. Dose individualization according to body size (weight or body surface area) was found to be appropriate and is recommended for the novel immunoconjugate.

Adoptive therapy of head and neck squamous cell carcinoma with antibody coated immune cells: a pilot clinical trial.

BACKGROUND: Catumaxomab is an antibody that binds with one arm epithelial cell adhesion molecule (EpCAM) positive tumors and with the other arm CD3+ T cells. Intravenous application of therapeutic antibodies may result in intravascular cytokine release. AIM: In this pilot trial we assessed whether cytokine release can be controlled by ex vivo cell opsonization and cytokine wash-out before administration of catumaxomab, preserving its anti-cancer activity. In addition, preliminary data on safety of and clinical response to catumaxomab coated autologous immune cells were acquired. METHODS: Peripheral blood mononuclear cells (PBMNC) of four patients with recurrent head and neck carcinoma were collected by leukapheresis, incubated ex vivo with catumaxomab for 24 h and cleared from released cytokines. Each patient received an escalated number of antibody-coated PBMNC equivalent to 1 x 10(4), 1 x 10(5), 1 x 10(6) and 1 x 10(7) CD3(+) cells/kgBW intravenously at bi-weekly intervals. RESULTS: After opsonization, PBMNC released substantial amounts of interferon gamma (IFNgamma) and tumor necrosis factor alpha (TNFalpha) in vitro, which were removed before administration. Catumaxomab up-regulated CD25, CD69, and CD83 on PBMNC, and catumaxomab loaded PBMNC released IFNgamma and granzyme B when coincubated with EpCAM(+) BHY cells, suggesting cell activation and target directed biological activity. During the study period, one patient died of aspiration pneumonia and one patient needed a tracheotomy. Treatment related adverse events (AE) occurred at the highest cell dose in two patients, whereas 1 x 10(6) loaded CD3(+) cells/kgBW were well tolerated by all patients. One patient showed stable disease for 6 months and one patient is in complete remission for 27 months. CONCLUSION: Ex vivo opsonization of PBMNC with catumaxomab provided biologically active, tumor targeting cells. Extracorporeal PBMNC coating may be an option to control intravascular cytokine release induced by therapeutic antibodies.

Radiofrequency-induced thermotherapy on nasal polyps: preliminary results.

BACKGROUND: The aim of this study was to determine the effects of radiofrequency-induced thermotherapy (RFITT) in patients with nasal polyps. METHODS: A retrospective analysis was performed of prospectively collected data from 17 consecutive patients (11 men and 6 women; mean age, 51.7 +/- 16.9 years) treated with RFITT from 2002 to 2003. The postoperative outcome was assessed using active anterior rhinomanometry, "sniffin' stick" test, and endoscopic nasal examination preoperatively and 4 weeks postoperatively. Subjective complaints were assessed with Likert scales. RESULTS: Transnasal airflow increased by 40.3% 4 weeks postoperatively (p = 0.029). Endoscopic appearance of nasal polyps indicated a nonsignificant reduction of 37.1%. Subjective complaints such as impaired nasal ventilation (p = 0.014), nasal discharge (p = 0.0007), postnasal drip (p = 0.0002), and hyposmia (p = 0.048) improved significantly 4 weeks after surgery. CONCLUSION: RFITT is well tolerated as a day case procedure under local anesthesia and might be a procedure for treating recurrence of NP after sinus surgery. It remains unclear at this point whether RFITT for nasal polyps results in a permanent reduction.

[Tonsillotomy]. / Die Tonsillotomie.

BACKGROUND: In contrast to tonsillectomy, only intracapsular parts of palatine tonsil are resected in tonsillotomy. This procedure has been revived for treatment of tonsillar hypertrophy causing obstructive sleep apnoea. Tonsillar hypertrophy is the most relevant cause for obstructive sleep apnoea in children often associated with malnutrition due to problems with swallowing. In these patients tonsillotomy is performed, because it is less painful, the children recover more quickly and the results on snoring, difficulty breathing and apnoea are equal in comparison to conventional tonsillectomy. In patients elder than 8 years or suffering from chronic or recurrent tonsillitis, tonsillectomy is the preferable surgical modality, because scarring and persistent inflammation in tonsillar remnants may be urge revision surgery. CONCLUSION: Tonsillotomy is a good alternative for tonsillectomy in children with symptomatic hypertrophy of the palatine tonsils.

Trifunctional bispecific antibody-induced tumor cell lysis of squamous cell carcinomas of the upper aerodigestive tract.

BACKGROUND: The trifunctional bispecific antibody Removab (tbAB) bridges and activates CD3 positive T cells to EpCAM on carcinoma cells and simultaneously binds to an accessory immune-cell inducing tumor cell lysis. tbAB-induced tumor cytotoxicity was assessed in an autologous human ex vivo system. METHODS: One hundred forty tumor samples and autologous peripheral blood mononuclear cells from a total of 36 patients with head and neck squamous cell carcinomas (HNSCCs) were incubated on a chicken embryo chorioallantois membrane with Removab. Tumor cells coincubated with cisplatin or cell culture medium served as positive and negative controls. Tumor cell lysis was assessed by acridine orange staining or by fluorescence-activated cell sorting of propidium iodide-marked cells after 24 and 48 hours (T24/T48) coincubation. RESULTS: Coincubation of HNSCC cells with tbAB and autologous peripheral blood mononuclear cells resulted in a 49% +/- 6% decrease of viable cells at T24 (p < .005) and in a decrease of 56% +/- 8% at T48 (p < .005) compared with the control. The tumor cytotoxicity was similar to that of cisplatin (49% +/- 7% decrease at T24 and 49% +/- 8% at T48). CONCLUSION: In an autologous human ex vivo system, the tbAB-induced tumor cell lysis was comparable to that by cisplatin.

A single-center 6-year experience with two types of pediatric tracheostomy.

OBJECTIVE: Among a wide variety of tracheostomy techniques, a vertical tracheal incision and a horizontal incision with creation of an inferior base tracheal flap have been favorized in children in the past. The aim of this study was to determine surgical and postoperative stoma complications after performing these two types of tracheal incision in tracheostomy in children. METHODS: A 6-year, prospective, observational cohort study was undertaken in 24 children (range, 0.03 month-15 years) at the Department of Otorhinolaryngology, University of Ulm. All children who underwent an elective tracheostomy were included in this study. Early and late local disorders in wound healing of the tracheal stoma and the clinical follow-up of both groups (Flap group and Vertical group) were analyzed. RESULTS: Formation of granulation tissue and tracheal stenosis were the most observed local disorders in both groups. Granulation tissue at the level of the stoma was the most frequently observed complication in the Flap group (4/12; 33%), whereas in the Vertical group only one child showed granulation tissue around the stoma (1/7; 14%). The difference between both groups was statistically not significant. The overall mortality rate 1 year after tracheostomy was 25% (6/24) and the tracheostomy-related death rate was 4% (1/24). An association of wound healing disorders with a feasible lethal outcome was not found after both tracheostomy types. There were no differences in the outcome between both tracheal incision types. CONCLUSIONS: Both types of tracheal incision proved to be a suitable surgical procedure for temporary or permanent tracheostomy in pediatric patients.

[Identification and evaluation of obstructive sleep apnea in children before adenotonsillectomy using evaluative surveys]. / Identifikation und Evaluation der kindlichen Schlafapnoe vor Adenotonsillektomie anhand evaluierter Fragebögen.

Sleep-disordered breathing (SDB) caused by enlarged tonsils and adnoids is common in early childhood. SDB without alterations in alveolar ventilation or sleep architecture is called primary snoring and affects up to 3-12 % of young children. In contrast, SDB with associated apneas or hypopneas affects between 0.7-3 % and is called obstructive sleep apnea syndrome (OSAS). OSAS and primary snoring should be distinguished with polysomnography (PSG). Despite the large number of patients undergoing tonsillectomy and adenoidectomy, the ability to assess patients-based preoperative diagnosis and surgical outcomes polysomnographically is limited. As a consequence it remains difficult which children will or will not benefit from surgical intervention. In the context of OSAS diagnosis, a discriminative survey is desired, one that cold classify differences before surgery as small, moderate or large. A promising office based surrogate measure are disease-specific quality-of-life surveys. Change within patients can be measured using an evaluative survey, administered before and after an intervention.

Nasal toxicity of benzalkonium chloride.

BACKGROUND: Benzalkonium chloride (BAC) is added to nasal preparations to prevent microbial contamination. Adverse effects of BAC on human nasal mucosa should be evaluated. METHODS: The ciliotoxicity of BAC was assessed in isolated human nasal epithelia from 15 donors. The effects of nasal BAC 0.05% (4 x 200 microL/day for 8 days) on nasal saccharin transport time, inflammatory cells and cytokine levels in nasal secretions, and nasal symptom scores were assessed in a randomized, double-blind crossover trial in 16 healthy volunteers. RESULTS: In vitro, BAC was ciliotoxic (p < 0.0001). In vivo, BAC did not alter saccharin transport time in healthy individuals (p > 0.8). No BAC-associated proinflammatory effects were observed. The staining index for myeloperoxidase was 4.8% in the placebo period and 6.3% (p = 0.42) in the BAC period. Also, nasal secretion levels of cytokines and the neuropeptide substance P revealed no BAC-associated differences. Concentrations for interleukin (IL)-6 in the placebo period were 41.5 pg/mL (0.9-91.7 pg/mL) and in the BAC period were 17.6 pg/mL (3.2-65.9 pg/mL; p = 0.46), and concentrations for substance P were 119 pg/mL (58-293 pg/mL) and 131 pg/mL (80-330 pg/mL; p = 0.31), respectively. Immediately after application, BAC caused nasal irritation (p = 0.001), a burning sensation (p = 0.0003), and hypersecretion (p = 0.006). Moreover, BAC caused a persistent sensation of nasal irritation (p < 0.01). CONCLUSION: BAC in concentrations used in nasal preparations is ciliotoxic. In healthy individuals, the ciliotoxic effect of BAC is neutralized, probably by components of nasal secretions. No BAC-related proinflammatory effects have been observed. At higher doses than normally used therapeutically, BAC caused significant nasal irritation.

Genotype and phenotype of glutathione-S-transferase in patients with head and neck carcinoma.

OBJECTIVE: Association of glutathione-S-transferase M1 (GSTM1) polymorphisms and cancer has been demonstrated. Possible underlying mechanisms and genotype-phenotype correlations are not adequately investigated. The aim of this study was to investigate the influence of the GSTM1-null-genotype on the level of GSTM enzyme concentration and on the enzyme activity of GST in patients with head and neck cancer (HNC). METHODS: We investigated in 83 patients and 91 healthy controls the GSTM1 polymorphisms, GSTM1 protein concentration, GSTM1 protein in tumor tissues, and total GST enzyme activity. RESULTS: Total GST enzyme activity was significantly lower in patients with HNC (208 +/- 9 micromol/min*l) than in controls (264 +/- 11 micromol/min*l, P< 0.0001) but did not depend on GSTM1-genotype (P = 0.1). GSTM protein concentration in null-genotype patients (3.6 +/- 2.5 microg/mL, mean +/- SE) was significantly lower than in GSTM1 allele carriers (26.7 +/- 9.6 microg/ml, P< 0.0001); GSTM protein expression did not depend on GSTM1-genotype (P> 0.5). CONCLUSION: GST enzyme activity in patients with HNC is suppressed, indicating impaired detoxification capacity of tobacco-smoke-related carcinogens. This suppression is not correlated with the GSTM1-genotype.

Early radiation-induced malignant fibrous histiocytoma of the oral cavity.

With an incidence of less than 0.3 per cent, post-radiation sarcomas are rare malignant neoplasms with a very poor prognosis. On average, they occur after a latency period of at least 15 years following radiation therapy with doses ranging from 24 to 80 Gy. We present the case of a post-irradiation malignant fibrous histiocytoma (MFH) on the floor of the mouth in a 79-year-old male patient arising only five and a half years after radiation therapy. The primary tumour was classified as a well differentiated squamous cell carcinoma of the right rim of the tongue. Primary therapy was surgical resection of the tumour and post-operative radiation with 50 Gy. Five and a half years later, the patient developed a rapidly progressing MFH within the field of radiation.

Immunohistomorphologic and molecular cytogenetic analysis of a carcinosarcoma of the urinary bladder.

Carcinosarcomas of the urinary bladder are malignant biphasic tumors with an epithelial and a spindle cell component. For the histogenesis of the two components, a biclonal and a monoclonal origin are discussed. We present the immunomorphology and molecular cytogenetics of such a case. The immunohistology of biopsies of the urinary bladder revealed a poorly differentiated urothelial carcinoma (GIII) and a co-existing pleomorphic, spindle cell leiomyosarcoma (GIII). The two components were microdissected and further analyzed for gains and losses of chromosomal material using comparative genomic hybridization. In addition, loss of heterozygosity analyses were included. The tumor components revealed as overlapping core aberrations losses on the short arm of chromosome 9 and on the long arm of chromosome 11. However, both components showed additional aberrations exclusively detected in one of the components. The occurrence of overlapping aberrations strongly argues for a monoclonal origin of this tumor with a common ancestor. The additional aberrations of the components point to an independent and divergent course of tumor progression in both components.