Local infiltration of T-lymphocyte subsets as a prognostic indicator in patients with nasopharyngeal carcinoma.

We studied tumor-host interactions in 47 patients with NPC. The local infiltration of T-lymphocyte subsets was investigated by an immunoperoxidase technique using monoclonal antibodies. Biopsy specimens of patients without cervical metastasis showed more T-lymphocyte (T11) infiltration. The amount of Leu-3a (helper/inducer) and T8 (cytotoxic/suppressor) cell infiltration did not correlate with the age, sex, clinical stage, and peripheral blood T4 and T8 cells of the patients. A higher incidence of Leu-3a cell infiltration was found in patients with high serum IgA antibody titers to EBV VCA. A trend of better prognosis was revealed in those cases with no or slight stromal T8 cell infiltration. A local immune response was found to exist which may prevent the spread of NPC to the cervical nodes, but this needs further study to evaluate the local infiltration of T-lymphocyte subsets as a prognostic indicator.

[DNA content and its distribution pattern of poorly differentiated nasopharyngeal carcinoma measured by Feulgen cytophotometry].

Nuclear area and DNA content of the normal columnar cells and cancer cells which were small, fusiform, irregular, vesicular-nuclear, giant malformed-nuclear and giant vesicular-nuclear were measured by Leitz MPV-III microscope photometer with HP-85 microcomputer in 32 poorly differentiated nasopharyngeal carcinoma (NPC) and 5 chronic nasopharyngitis sections. Based on different DNA contents and distribution patterns, it was divided into 4 cancer cell populations: small, fusiform and/or irregular, vesicular-nuclear and giant tumor cell populations. It should be noted that the nuclear DNA pattern of vesicular-nuclear cancer cell population had pathognomonic characteristics. The nuclear unit DNA content ratio of vesicular-nuclear cancer cells to lymphocytes was under 0.4. This cancer cell population was sensitive to irradiation. If the biological characteristics of these four cancer cell populations could be clarified not only in nuclear DNA pattern but also in biochemistry at molecular level, it will be possible to design the different treatment trials on the different cancer cell populations, then resulting in better prognosis of NPC patients.

Detection of Epstein-Barr virus DNA in nasopharyngeal carcinoma using a non-radioactive digoxigenin-labelled probe.

The presence of Epstein Barr virus (EBV) DNA in biopsies from the post-nasal space (PNS) of patients suspected of nasopharyngeal carcinoma (NPC) was detected by in situ cytohybridization with an EBV DNA probe labelled with the novel labelling compound digoxigenin. The digoxigenin probe was hybridised to cryostat sections of NPC biopsies and subsequently detected by an enzyme immunoassay procedure. It was found that in situ cytohybridization using the digoxigenin probe was much more rapid and sensitive (96 h compared to five weeks) than the current method of using 3H-labelled probe. Using the digoxigenin EBV probe, it was found that in all the eighteen NPC biopsies tested, EBV DNA was detected in malignant epithelial cells and infiltrating lymphocytes. EBV DNA was also detected in some normal epithelial cells in these NPC biopsies. EBV DNA was not detected in epithelial cells of non-malignant biopsies.

EBV gene expression in an NPC-related tumour.

A nasopharyngeal carcinoma tumour (designated C15) propagated in nude mice has been used to generate a large cDNA library that we have analysed for Epstein-Barr virus (EBV) gene expression. No gross alterations exist in viral DNA from C15 relative to other human isolates and the large deletion present in the B95-8 'prototype' viral strain established in marmoset cells is not found; C15 contains no linear virion DNA. In the cDNA library, of the six EBV nuclear antigens (EBNAs) expressed in latently infected B-lymphocytes, only clones for EBNA-1 are found. These data are confirmed by immunoblotting. Sequence analysis shows the EBNA-1 mRNA splicing pattern in the carcinoma to differ from that observed in B-lymphocytes. Further, contrary to observations with B-cell lines, most viral transcription in the tumour is localized onto the 'rightmost' region of the conventional EBV physical map. Transcripts identified corresponding to known genes include those for the latent membrane protein (LMP), the alkaline DNA exonuclease and probably the terminal protein; major transcripts are also derived from the BamHI D fragment and the region deleted in B95-8 EBV DNA. Novel transcripts have also been identified that proceed in an anti-sense direction to genes encoding functions associated with replication, such as the viral DNA polymerase. They contain a large, hitherto unidentified, open reading frame in the viral genome that is complementary to the putative function known as BALF3 and a smaller open reading frame complementary to BALF5 (the DNA polymerase gene). From the present studies we can conclude that: (i) EBV transcription patterns in the epithelial cells vary markedly from those identified previously in B-cells, reflecting differential use of promoters or splicing patterns. (ii) Transcription is tightly regulated and restricted in the C15 tumour with many latent genes, notably EBNAs 2-6, being 'switched off.' (iii) A family of cytoplasmic RNAs are transcribed in an antisense direction to a number of existing open reading frames in the EBV genome. (iv) There are a number of mutations in C15 transcripts relative to the B95-8 genome, some of which could result in amino acid alterations in proteins.

Polymorphous low-grade adenocarcinoma of seromucous glands of the nasopharynx. A report of a case and a discussion of the morphologic and immunohistochemical features.

Polymorphous low-grade adenocarcinoma (PLGA) is a minor salivary gland neoplasm that is characterized by morphologic variability, cytologic uniformity, and an infiltrating growth pattern. To date, this entity has been identified either within the confines of the oral cavity or, rarely, in the nasal cavity. The authors report a case of PLGA that arose in the nasopharynx of a 44-year-old female, representing the first documented occurrence of this tumor outside the oral cavity or the sinonasal tract. The histopathologic and immunohistochemical differentiation from pleomorphic adenoma and adenoid cystic carcinoma is discussed.

DNA flow cytometric analysis of human nasopharyngeal carcinoma in nude mice.

The tissue from a patient with nasopharyngeal carcinoma has been transferred to nude mice (BALB/c), and has successfully been growing through twenty passages. The tumors in the nude mice and primary human tumor have been examined for the cellular DNA content by FCM and also conventional pathological examination, chromosome analysis, and EBV test. The tumor take rate varied markedly in different passages with a mean value of approximately 70%, and showing a tendency to increase. The tumor doubling time within 6-12 weeks after transplantation of six tumors in 18 and 20 passages were 14.8 and 9.3 days respectively. However, the tumor volume at 12 weeks varied significantly, ranging from 438 to 1998 mm3. By FCM, it has been found that the values of DNA index were about the same in both primary tumor and the tumors in nude mice. The distribution of various phase cells in cell cycle was also about the same in both. In conclusion, the application of FCM to examine the cellular DNA content of the tumor in nude mice is a rapid and sensitive method, useful in the investigation on the stability of biological characteristics of human NPC in nude mice and in further studies on the effects by various therapeutic methods.

Malignant rhabdoid tumor.

Immunocytochemistry and electron microscopy have allowed more precise identification and classification of neoplasms formerly considered as representatives of other groups. The malignant rhabdoid tumor is one example. Principally neoplasms of the kidney, rhabdoid tumors present uncommonly in the head and neck, where because of the age of the patients and the neoplasms' resemblance to rhabdomyosarcoma, misdiagnosis is possible and likely. Wherever it arises, the lesion is usually fatal.

[Particle induced X-ray emission for trace element analysis of hairs from patients with cancer].

In this paper, trace element analysis of the hairs from the patients with nasopharyngeal carcinoma (NPC) and osteoma was made using particle induced X-ray emission (PIXE) technique in order to obtain some information on the correlation between trace element and these two tumors. The hair samples of 34 NPC patients were each collected before and three months after radiotherapy (60Co) (group 1). The hair specimens from 34 osteoma patients were collected preoperatively and postoperatively. For comparison, 65 hair samples were collected from 40 normal subjects (group 2) and 25 NPC patients, who had received radiotherapy (60Co) five years before without any recrudescence (group 3). The trace element analysis of all specimens was made by PIXE. The results show that there is a significant difference between groups 1 and 2 (P less than 0.01) but no difference is found between groups 2 and 3 for Mn, Cu, Zn, As etc. content in the hairs. Cu and Zn content in the hairs of the osteoma patients is much lower than that of the normal subjects (P less than 0.01) while for K, Ti and Mn content, the former is higher than the latter. The above experimental results are discussed in detail.

Diagnosis, staging, and treatment of juvenile nasopharyngeal angiofibroma (JNA).

Nineteen patients with juvenile nasopharyngeal angiofibroma (JNA) were surgically treated with different techniques from January 1968 through December 1985. Two patients had undergone a previous operation at another hospital; all patients were males (mean age 15.4), and the most common symptom was nasal obstruction (84.2%). Lateral extension into the pterygomaxillary fossa occurred in 14 patients (73.6%), and 2 also had intracranial invasion (10.5%). In five cases, the tumor's cytosol was analyzed for hormonal receptors. Negative values for estrogen and progesterone receptors were obtained, although the content of dehydrotestosterone receptors was highly positive. These results tend to support the hypothesis of JNA's androgen-dependence. The authors emphasize the need of a preoperative staging classification based on clinicoradiological data in selecting the most adequate surgical approach. Tumors with lateral extension into the pterygomaxillary fossa can be easily removed through a midface degloving; large involvement of the infratemporal fossa requires, also, a transzygomatic dissection. In JNAs with intracranial extension a combined intracranial-extracranial approach is advisable.

Hormonal receptors in juvenile nasopharyngeal angiofibroma.

Specific thermostable androgen receptors were detected in the tissues of nasopharyngeal angiofibroma. The receptors seemed to be specific with high affinity toward DHT more than testosterone. No abnormalities in serum levels of DHT, testosterone, and estradiol-17 beta could be detected in the patients studied. A concept of pathogenesis of the tumor in relation to that reported in literature recently is interpreted in the text.

[Nuclear DNA content in nasopharyngeal carcinoma (NPC) cells].

Cancer cell nuclear DNA content of 40 biopsy imprints of NPC was measured by Leitz MPV3 microspectrophotometer using scanning method. The pathological diagnosis and histological classification were done on the same specimens. By morphology, the poorly differentiated squamous carcinoma (34 cases) could be divided into four subtypes: ordinary nucleus type (14), spindle nucleus type (5), large and pleomorphic nucleus types (12 and 3). The first two types revealed a bimodal peak DNA histogram (2C and 4C) and slightly scattering toward the right to the 8C region in some nuclear DNA. The large nucleus type showed the bimodal peaks with a broad 4C and many nuclear DNA located in the 4C-8C range. The pleomorphic nucleus type appeared as multiple low peaks diffusing between 2C-12C. The vesicular nucleus cell carcinoma (3 cases) had a double peak DNA histogram similar to the ordinary nucleus type. The undifferentiated cell carcinoma (1 case) showed a broad unimodal peak DNA, ranging from 2C to 6C. The well differentiated squamous carcinoma (2 cases) had a double peak DNA, which was close to the normal diploid pattern but each peak was relative wide and a few DNA value located beyond the 4C region. The results indicate that DNA values showing a predominance of double peak histogram and diploid pattern imply the low grade malignancies in its biological behavior. In contrast, those showing the aneuploid and multiploid patterns in the majority of cancer cells point to the high grade malignancies. Hence, the supplementary DNA measurement of NPC cells can judge exactly the malignant degree of the tumor and provide evidence for prognosis.

The isolation, characterization, and comparison of the membrane-associated and soluble folate-binding proteins from human KB cells.

Human nasopharyngeal epidermoid carcinoma (KB) cells contain a membrane-associated particulate folate-binding protein which is important in the cellular accumulation of physiologic folates (Antony, A. C., Kane, M. A., Portillo, R. M., Elwood, P. C., and Kolhouse, J. F. (1985) J. Biol. Chem. 260, 14911-14917) and in the binding of methotrexate (Kane, M. A., Portillo, R. M., Elwood, P. C., Antony, A. C., and Kolhouse, J. F. (1986) J. Biol. Chem. 261, 44-49). A soluble folate-binding protein appears in media exposed to proliferating KB cells. We have purified to homogeneity both the membrane-associated and the soluble folate-binding proteins from the KB cell tissue culture system. The purified membrane-associated and soluble folate-binding proteins give single bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with apparent Mr values of 50,000 and 40,000, respectively. The membrane-associated folate-binding protein contains 45,000 g of amino acids and the soluble folate-binding protein contains 24,000 g of amino acids per mole of folate bound. Each of the purified proteins has a single folate-binding site, and the carbohydrate content is approximately 25% for each species of protein. The affinity constants for 5-methyltetrahydrofolate of the membrane-associated and soluble folate-binding proteins are 0.3 and 2.5 X 10(9) liters/mol, respectively. The affinities of various polyglutamated forms of methotrexate are similar for each protein, increase as the chain length of the polyglutamate increases (from approximately 0.004 X 10(9) liters/mol for methotrexate to 0.3 X 10(9) liters/mol for methotrexate heptaglutamate), are equal to the affinity for 5-methyltetrahydrofolate, and exceed the reported increase in affinity of methotrexate polyglutamates for dihydrofolate reductase.

Immunohistochemical distribution of monoclonal antibodies against keratin in papillomas and carcinomas from oral and nasopharyngeal regions.

Papillomas (40) and squamous cell carcinomas (75) were examined for the presence of three keratin proteins with the use of an immunohistochemical technique. Polyclonal keratin antibody (TK, detecting 41 to 65 kDa keratin) and monoclonal antibodies KL1 and PKK1 (55 to 57 kDa and 41 to 56 kDa, respectively) were used. Squamous epithelium in normal oral mucosa showed marked TK staining in cells of upper strata and relatively slight staining in basal layer cells, moderate KL1 staining in spinous and granular layers and was negative in basal cells. Positive PKK1 staining was noted in cells of the basal layer. Columnar epithelium in the nasal mucosa showed TK staining in all layers, KL1 staining on the apical side of epithelial cells and trace or negative staining in basal layer cells. There was moderate PKK1 staining along the apical side of cells and variable staining in basal cells. Keratin distribution in oral papillomas was similar to that in normal oral epithelium, whereas in nasal and nasopharyngeal papillomas, keratin distribution was restricted to the upper layers. Tonsillar papillomas showed a strong TK reaction, negative KL1 in upper layer cells, and marked PKK1 staining in basal cells. Well-keratinized squamous carcinomas indicated an irregular TK distribution and decreased KL1 and negative PKK1 stainings. Intermediate and poorly differentiated keratinizing squamous carcinoma showed irregular staining patterns for the three classes of keratins studied. Immunohistochemically detectable keratins utilizing monoclonal antibodies were described as useful markers of epithelial tumors of squamous origin. Keratin expression within benign tumors was related to normal regional distribution, whereas in malignant tumors, keratin distribution was irregular in its distribution profile.

Adenoid squamous cell carcinoma of the nasopharynx and neck region.

Adenoid squamous cell carcinoma (ASCC) is a rare variant of squamous cell carcinoma. This tumor most commonly occurs in the skin of the head and neck and rarely in the vermillion of the lip, vulva, and oral cavity. This report of two patients includes the first (to our knowledge) case of ASCC in the nasopharynx. It also describes the immunocytochemical and ultrastructural features of this tumor. Our findings and a review of the literature indicate that ASCC of the upper aerodigestive tract is associated with an aggressive behavior and poor prognosis, in contrast with ASCC of the skin.

Nasopharyngeal carcinoma: immunohistochemical study for keratin, secretory component and leukocyte common antigen.

Immunohistochemical studies using antisera to keratin and secretory component (SC) and monoclonal antibody against leukocyte common antigen (LCA) were performed on 92 biopsy and autopsy specimens taken from 65 patients with nasopharyngeal carcinoma. Five biopsy specimens from malignant lymphoma of the nasopharynx and tonsil, and 20 biopsy specimens of nasopharyngeal epithelium were also included in the study. Keratin was positively stained in all squamous cell carcinomas and nonkeratinizing carcinomas, and 38 of 46 undifferentiated carcinomas (82.6%), but in no malignant lymphomas. LCA was intensely stained in all malignant lymphoma, but in no nasopharyngeal carcinomas. SC was positively stained in two of the 46 undifferentiated carcinomas (4.3%). Nasopharyngeal carcinoma is a definite malignant epithelial neoplasm and can be distinguished from malignant lymphoma by immunostaining for keratin and LCA. Some undifferentiated carcinoma may show cellular differentiation toward ciliated epithelium.

Estrogen and progesterone receptors in head and neck cancer.

The existence of hormone receptors on or within neoplastic tissue has potential diagnostic, therapeutic, and prognostic importance. It was the purpose of this study to measure estrogen and progesterone receptors in a large group of patients with head and neck cancer to determine their frequency. Sixty-five patients with head and neck tumors underwent a total of 75 estrogen and 50 progesterone receptor assays. In this group, 87.7% were squamous cell carcinoma. In the estrogen receptor assays, 89.3% (67/75) were negative, 8% (6/75) were borderline, and only 2.7% (2/75) were positive. In the progesterone receptor assays, 78% (39/50) were negative, 22% (11/50) were borderline, and there were no positive results. There were no changes in assays of tissue removed at biopsy v tissue removed during surgery. There was no impact with chemotherapy. In conclusion, head and neck cancers do not appear to possess estrogen or progesterone receptors and can be considered to be hormonally independent.