The BamHI Z EBV replication activator (ZEBRA) protein is involved in the switch from latency to productive cycle of Epstein-Barr virus. A recombinant ZEBRA protein was synthesized and assessed in enzyme-linked immunosorbent assay (ELISA) for serum IgG response in nasopharyngeal carcinoma (NPC) patients. In 100 NPC serum samples that were positive for IgA to the EBV viral capsid antigen (VCA), 75% had IgG anti-ZEBRA antibodies. In contrast, only 3/83 (3.6%) serum samples from healthy donors and 2/50 (4%) from other cancers were positive for IgG to ZEBRA. Interestingly, in a selected group of 100 NPC sera negative for IgA to VCA, 25% contained IgG anti-ZEBRA antibodies. This suggests that the ELISA for IgG anti-ZEBRA may also identify earlier cases of NPC not detected by the conventional immunofluorescence test for IgA to VCA.
Clinical synopsis and investigations on fourteen patients with blast injuries of the ear successfully treated at the ENT Unit of the University Hospital, Kuala Lumpur, revealed few interesting observations in addition to usual findings. The most outstanding one was the absence of sensorineural deafness, although it has been claimed as one of the most constant features. Conservative management alone was found to be of significant value.
Nasopharyngeal carcinoma (NPC) is one of the commonest presentation of head and neck cancers in Malaysia, especially in the Chinese. The standard treatment is radical radiotherapy to the post-nasal space and the neck. Chemotherapy is given to patients with primary advanced disease and to patients with recurrence. The study reviews results of chemotherapy given to 33 patients at the University Hospital, Kuala Lumpur, over the last four years.
BACKGROUND: The Epstein-Barr virus (EBV) is consistently detected in patients with nasopharyngeal carcinoma. To determine whether EBV infection is an early, initiating event in the development of this malignant tumor, we screened nasopharyngeal-biopsy samples, most of which were archival, for preinvasive lesions, including dysplasia and carcinoma in situ. Preinvasive lesions were found in 11 samples, which were tested for the presence of EBV.
METHODS: EBV infection was detected with in situ hybridization for EBV-encoded RNAs (EBERs) and by immunohistochemical staining for latent membrane protein 1 (LMP-1). The larger samples were also tested for the EBV genome with the use of Southern blotting. The expression of specific EBV RNAs was determined by the amplification of complementary DNA with the polymerase chain reaction.
RESULTS: Evidence of EBV infection was detected in all 11 tissue samples with dysplasia or carcinoma in situ. EBERs were identified in all eight samples tested, and LMP-1 was detected in all six of the tested samples. Six of the seven samples tested for the EBV termini contained clonal EBV DNA: Transcription of the latent EBV gene products, EBV nuclear antigen 1, LMP-1, LMP-2A, and the BamHI-A fragment, was detected in most of the samples. Viral proteins characteristic of lytic lesions were not detected.
CONCLUSIONS: Preinvasive lesions of the nasopharynx are infected with EBV. The EBV DNA is clonal, indicating that the lesions represent a focal cellular growth that arose from a single EBV-infected cell and that EBV infection is an early, possibly initiating event in the development of nasopharyngeal carcinoma. Preinvasive lesions contain EBV RNAs that are characteristic of latent infection but not the viral proteins that are characteristic of lytic infection. The detection of the EBV-transforming gene, LMP-1, in all the neoplastic cells suggests that its expression is essential for preinvasive epithelial proliferations associated with nasopharyngeal carcinoma.
Nasopharyngeal carcinoma (NPC) is one of the most common cancers in Malaysia, a country in Southeast Asia with a multiracial population. While hospital-based data on NPC and data from a few states have been reported, a comprehensive study involving every known NPC patient in the whole of Peninsular Malaysia in one particular year had never been done. In the present study, the computed incidence rate was not only adjusted for age, sex, ethnicity, and place of residence, but also direct standardization methods of Rothman and Dever were used to reduce any distortion. The mean age of the 365 new cases of NPC registered in 1988 was 46.8 years (SD +/- 12.2 years). The ages of patients ranged from 10 to 80 years. The incidence in both sexes rose after the age of 20-29 years and reached a plateau between 40 and 49 years. No further rise was exhibited after age 60 years. The Chinese had the highest age-adjusted incidence rates, particularly for the age group 40-49 years, where the incidence rate was 40.1 per 100,000 for males and 14.9 for females. The average age-adjusted male/female ratio was 2.8:1. Age-adjusted incidence varied by place of residence. The pattern that emerged from the data indicated the possibility of interaction between genetic susceptibility and environmental cofactors in the etiology of NPC.
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.
Epstein-Barr virus (EBV) receptors (EBV/C3d receptors) were detected, using the monoclonal antibody HB5, on 23 ectocervical and 5 endocervical biopsies of the uterine cervix. Elevated IgA titers against the viral capsid antigen and early antigen of EBV were also found in the cervical secretions from cervical carcinoma patients (83%), compared with samples from patients with cervical intraepithelial neoplasia (75%), herpes simplex virus-infected patients (0%), and gynecologic patients with nonmalignant conditions (0%). EBV DNA was present in 63% of cervical carcinoma biopsies detected by in situ hybridization. These observations suggest a positive association between EBV and carcinoma of the cervix.
HLA associations were observed in unrelated Malay patients with nasopharyngeal carcinoma (NPC). HLA-B18 was observed in 18/45 (40%) Malay NPC patients compared to 22/167 (13%) Malay normals (P = 0.0001; Pc = 0.0027, RR = 4.4). The frequency of HLA-B17, one of the antigens associated with Chinese NPC, was also increased among Malay NPC (13/45 29%) compared to controls (18/167 11%; P = 0.003, Pc = 0.07 RR = 3.4). Similar to the findings among Chinese NPC, the frequency of B17 was higher in early onset (less than or equal to 30 years) Malay NPC resulting in a higher relative risk (RR = 5.0).
Titers of IgA/VCA from 92 nasopharyngeal carcinoma (NPC) patients were monitored for 3 to 11 years from the time of diagnosis. The fluctuations in the IgA/VCA titers during follow-up did not correlate with the clinical status of the patients, suggesting that IgA/VCA is of marginal significance in the monitoring of NPC patients during follow-up. In addition, the frequency of recurrence of NPC was independent of presence or absence of elevated IgA/VCA at diagnosis.
Serum antibodies against Epstein-Barr virus (EBV)-determined antigens have traditionally been titrated by the indirect immunofluorescence (IIF) technique. The avidin-biotin complex (ABC) immunocytochemical technique was used to determine the serum levels of IgA against EBV viral capsid antigen (IgA/VCA) and IgA against EBV early antigen (IgA/EA) in sera of 106 nasopharyngeal carcinoma (NPC) patients prior to treatment and 100 normal individuals. The sensitivity of the ABC technique is enhanced by an amplification of the antigen-antibody reaction, which involves the binding of the enzyme-linked ABC to the second biotinylated antibody. There was a good correlation (r = 0.9988) between ABC and IIF-determined IgA/VCA-positive titres, with the ABC technique being more sensitive than IIF in the detection of IgA/VCA in NPC sera: 94% (99/106) and 76% (80/106), respectively. The frequency of IgA/EA reactivity in NPC sera was also markedly increased by immunodetection with the ABC technique as compared with IIF technique: 63% (69/106) and 28% (30/106) respectively. Both the immunocytochemical techniques were equally specific in discriminating between elevated serum titres of IgA/VCA and IgA/EA in NPC sera from normal human sera.
The linear antigenic epitopes of the Epstein-Barr virus replication activator protein (ZEBRA), recognised by specific serum IgG in nasopharyngeal carcinoma (NPC), were determined. This was achieved by synthesizing the entire amino acid sequence of ZEBRA as a set of 29, 22-residue peptides with an overlap of 14 amino acids. The ZEBRA peptides were tested in enzyme-linked immunosorbent assay (ELISA) for IgG binding in sera from 37 selected NPC patients who had IgG antibodies to the native ZEBRA protein. The most immunogenic epitope was peptide 1 at the amino-terminal end with 36 of the sera reactive against it. Further analysis of peptide 1, using the multipin peptide-scanning technique, defined a 10-amino-acid sequence FTPDPYQVPF, which was strongly bound by IgG. Two other regions of ZEBRA were also identified as immunodominant IgG epitopes, namely peptide 11 (amino acids 82-103) and peptide 19/20 (amino acids 146-175) with 8-13 of the NPC sera reactive against the peptides. The number of peptides reactive with individual NPC serum varies from 1 to 6 or more and there is some correlation between a greater number of peptide (at least 4) bound and a higher (at least 1:40) titre of serum IgA to viral capsid antigen. The immunodominant ZEBRA peptide 1 could be utilised in IgG ELISA for the detection of NPC.
To assess the long-term survival of patients with nasopharyngeal carcinoma (NPC) who were treated with conventional radical radiotherapy (RT) followed by adjuvant chemotherapy.
The antibody titres to P. falciparum and Epstein-Barr Virus-associated antigens were assayed in 22 patients with NPC and 43 controls. All, but one patient had antimalarial titres; 14 had titres greater than 80 and 4 patients greater than 640. Compared to controls the mean anti-malarial titre for most age groups were higher in the patients. Those patients with high anti-malarial titres also had high IgA anti-VCA titre, an antibody which has been demonstrated to be diagnostic for NPC. The peak anti-VCA (IgG) and anti-EA (IgG) antibody titres were associated with anti-falciparum titres of 320-640 and 80-160, respectively. The results are discussed in relation to the possible association between malarial infection and etiology of NPC.
A retrospective study of 1000 cases of microscopically diagnosed head and neck cancers in the University Hospital, Kuala Lumpur was done. Head and neck neoplasms comprise 7.1% of all tumours diagnosed in this Hospital and the commonest sites of involvement are the nasopharynx (29.1%), cervical lymph nodes (22.6%), oral cavity (10.2%), thyroid (8.2%) and skin (6.5%). Histologically, 87% of tumours are epithelial in nature. The results of this study show that nasopharyngeal carcinoma is commonest in Chinese males, while oral and laryngeal malignancies occur more frequently in Indians.
New data are presented concerning the relationship between NPC and HLA antigens among Chinese. When attention is confined to newly diagnosed cases, it can be shown that, apart from the increased risk associated with the joint occurrence of A2 and B-Sin 2, there is also an increased risk associated with BW17 and a decrease in risk associated with A11. Among long-term survivors, however, BW17 is appreciably decreased, whereas A2 in the absence of B-Sin 2 or BW17 is increased. Among Malays, a non-Chinese group, there is an excess among NPC patients of a locus A blank, a blank which is probably associated with the AW19 complex.
One hundred consecutive newly diagnosed patients with nasopharyngeal carcinoma (NPC) since January 1994 were the subjects for studying various factors that contribute to the delay in the confirmation of the diagnosis. Seventy-nine of them were males and the peak age of incidence was the fifth decade. Ninety two percent were Chinese, 7% Malay and 1% Indian. Seventy six percent were agriculture workers or labourers with 66% having either no formal education (16%) or only primary level education (50%). In 50% of patients neck swelling was the first presenting symptom, 26% had nasal symptoms, 12% ear symptoms and 11% has symptoms due to intracranial extension of tumour. As many as 80% were at UICC Stage IV at the time of diagnosis. While the median delay, on the part of patients, in consulting a doctor was 2.5 days, the median delay on the part of the doctors to confirm the diagnosis of NPC was 127 days, the delays was particularly worse when the patients presented with ear symptoms (266 days) followed by those with neck swelling (94 days). For those patients who were required to undergo more than one nasopharyngoscopy and biopsy the median doctor's delay was 144 days. Since 82% of patient's had first consulted general practitioners, it is suggested that their level of awareness with regards to the diagnosis of NPC be significantly raised so that the delay on their part be greatly minimized.
Five rare cases of nasopharyngeal carcinoma with optic nerve involvement are reported. Computerised Tomographic Scan (CT Scan) studies were performed in four of them. Evidence of intracranial spread of the tumour, from the roof of the fossa of Rosenmuller to the apex of the orbit through the cavernous sinus, was noted in three patients. In one of them there was extracranial extension of the tumour, to the orbit through the posterior ethmoid.
Papillomas of nose and paranasal sinuses are uncommon tumours. Based on the detailed clinical and
histopathological examination of seven cases of papillomas, the authors would accept and recommend
t~e. su~ested unifying name of transitional cell papilloma for these lesions which may further be sub·
dIvIded onto Type I and Type II. The rationale behind this classification is discussed at length. There is
a greater tendency for Type II papillomas to recur and undergo malignant change. It is suggested that
Type I papillomas be managed by relatively simple surgical procedures whereas Type II cases be dealt with more radically.