Purpose To investigate the pathological characteristics of loco-regional recurrent nasopharyngeal carcinoma ( rNPC ) . Methods Nasopharyngeal biopsy specimens of 46 rNPCs and 63 primary NPCs were collected. HE staining, immunohistochemistry and EBV small RNAs ( EBERs) in-situ hybridization were performed. Results The over-expression rates of both p63 and CK5/6 in rNPC were significantly higher than those of primary NPCs (P=0. 005, P=0. 026), while no statistical significance of Ki-67 over-ex-pression existed between the two groups ( P=0. 387 ) . More necrotic tissues, inflammatory exudates, giant bizarre carcinoma cells, desmoplastic stroma, giant bizarre tumor cells and higher degree of squamous differentiation were found in rNPCs. The carcinoma cells of 5 rNPCs were negative for both EBERs in-situ hybridization and LMP-1 immunohistochemical staining. Conclusion The loco-re-gional rNPC has two peaks of latency interval:2~5 and 9~11 years. The loco-regional rNPC cells have higher degree of squamous differentiation with higher expression of p63 and CK5/6, as well as more invasive ability. In addition, both EBERs in-situ hybridization and LMP-1 immunostaining are negative in 10. 87% (5/46) of loco-regional rNPC.

Purpose To investigate the relationship between Epstein-Barr virus (EBV) and lymphoepithelioma-like carcinoma (LELC) of the parotid gland and detect the gene expression products of EBV harbouring in LELC cells. Methods Thirty-two parotid LELCs were collected from the Departments of Pathology, Sun Yat-sen University of Medical Sciences, Guangzhou, China during the period of January 1986 and December 1995. All the 32 formalin-fixed paraffin-embedded blocks had been consecutively re-sectioned again. Immunohistochemical and in situ nucleic acid hybridization methods for detection of EBV gene encoded products were performed. Results (1) 32 LELCs were found out of 125 parotid gland carcinomas, the frequency was 25.6% (32/125). (2) All of the 32 specimens contained a variable number of EBNA-1 and EBERs positive neoplastic cells. (3) Twenty-seven out of 32 specimens (27/32, 84.4%) had a portion of carcinoma cells expressing LMP-1. (4) No ZEBRA positive cell could be found. (5) EA-D, VCA and MA positivity rates for these 32 parotid LELCs reached to 71.9%(23/32), 68.8%(22/32), and 12.5%(4/32), respectively. Conclusions (1) The parotid gland LELC is frequently to be seen in Guangzhou locale of China, where is a high-incidence area of nasopharyngeal carcinoma (NPC). The parotid gland LELC and NPC are co-prevalent in Guangzhou locale. (2) This disease is also consistently associated with EBV infection. (3) The EBV infection of the parotid gland LELCs is essentially the type of latency Ⅱ, expressing EBNA-1, EBERs and LMP-1. (4) The latent infected EBV harbouring in LELC cells could in part be switched over to lytic cycle, producing EA-D, VCA or/and MA.

[Objective] To investigate the variation of Q promoter (Qp) in nasopharyngeal carcinoma (NPC) cells, and to compare the existing two mutant sites [62 225 site(g→a)and 62 422 site (g→c) ] Qp in NPC cells with the Qp in B95.8 cell line in the functional and biological difference. [Methods] The Qp sequence was amplified in the samples from 29 cases of paraffin-embedded tissues of NPC suffers and 14 cases of peripheral blood of healthy adults by polymerase chain reaction (PCR) method (totally 43 cases). The point mutations on specified sites were analyzed and statistically compared from sequencing results. The sequences of variant and prototype Qp were amplified by PCR and cloned into luciferase reporter vector (pGL3-basic), then transfected into HaCat cells respectively. The transcriptional activity was compared between variant and prototype Qp using luciferase reporter system. The DNA binding affinity of mutant and prototype Qp to Sp1 was compared through chromatin immunoprecipitation (CHIP) method since mutation of nt 62 225 located in a Spl binding site. [Results] The mutation rate of Qp was significantly higher in NPC compared with healthy controls (P=0.039 5, <0.05), which suggested the variant Qp was closely associated with NPC. The transcription of the luciferase gene promoted by variant Qp was significant more than that of prototype Qp in transient transfection assay (2.5:1, P<0.05). The binding affinity of variant Qp to Sp1 was about 1.52 times higher than that of prototype Qp as determined by quantitative ChIP assay. [Conclusions] The transcriptional activity was enhanced in variant Qp in NPC cells compared with prototype, which possibly through the higher binding affinity to Sp1. We suggest that the mutated Qp may play an important role during the EBV infection and transformation of nasopharyngeal epithelium.

Objective: To investigate whether the nasopharyngeal squamous cell carcinoma or keratinizing squamous cell carcinoma (KSCC) is also closely associated with Epstein-Barr virus infection as nasopharyngeal non-keratinizing carcinoma does. Methods: The Epstein-Barr virus (EBV) DNA and its gene expression products, including EBNA-1, EBNA-2, EBERs, LMP-1, ZEBRA, EA-D, VCA and MA were detected in 38 nasopharyngeal KSCCs using PCR Southern blotting hybridization, in situ hybridization and immunohistochemistry. Results:(1) The EBERs positive cancer cells could be found in 37 KSCCs (97.4% , 37/38). (2) Most undifferentiated and poorly differentiated cancer cells of these 37 KSCCs expressed EBNA-1 and EBERs; and LMP-1 positive cells could be found in 21 specimens (56.8% , 21/37). (3) A few of ZEBRA positive or/and EA- D positive cancer cells were found in 23 out of 37 KSCCs (62.2% , 23/37). (4) Several of more differentiated or keratinizing cancer cells expressed VCA or/and MA in 22 of 37 KSCCs (59.5% , 22/37). Conclusions:(1) KSCCs of the nasopharynx in the south of China are also consistently, not occasionally, associated with EBV infection as non-keratinizing carcinomas (NKCs) do; (2) The majority of undifferentiated and poorly differentiated cancer cells in KSCCs are always infected with EBV in type II latency, mainly expressing EBNA-1, EBERs, and LMP-1, and a small number of them may express the abortive immediate early and early viral proteins (ZEBRA ＆ EA-D); (3) The more differentiated or keratinizing cancer cells infected with EBV in KSCCs might complete its production cycle and express late viral proteins (VCA or/and MA).

OBJECTIVETo investigate the efficiency of concurrent application of VCA-IgA, EA-IgA and EA-IgG serological tests in diagnosing nasopharyngeal carcinoma (NPC).

METHODSThe sera of 266 untreated NPC patients and 347 healthy adults were collected. In addition to the conventional immunoenzymatic method of VCA-IgA test, enzyme-linked immunosorbent assay (ELISA) was adopted as an alternative to test the antibody level of EA-IgG and EA-IgA. A new statistical formula was used to evaluate the odds ratio of different combinations of these three tests.

RESULTSThe sensitivity and specificity of VCA-IgA, EA-IgG and EA-IgA concurrently were as high as 95.11% and 97.41%, respectively, which were higher than those of single test (90.60% and 94.52% for VCA-IgA, 93.98% and 93.66% for EA-IgG, 89.84% and 88.18% for EA-IgA). Furthermore, the odds ratio of 3-test positivity (1 912.5) was higher than those of 2-test positivity (27.903 2 for VCA-IgA and EA-IgG, 11.169 0 for EA-IgG and EA-IgA, 8.032 8 for VCA-IgA and EA-IgA), which were even higher than those of 1-test positivity (0.121 4 for VCA-IgA, 0.170 5 for EA-IgG and 0.048 8 for EA-IgA).

CONCLUSIONELISA is more accurate in reflecting the antibody level of EA-IgG and EA-IgA than the conventional immunoenzymatic method. The concurrent application of VCA-IgA, EA-IgG and EA-IgA test can markedly improve the sensitivity, specificity and odds ratio as well, thus resulting in enhancing the efficiency of diagnosing nasopharyngeal carcinoma serologically.

Adult ; Antibodies, Viral ; blood ; Antigens, Viral ; immunology ; Capsid Proteins ; immunology ; Diagnostic Errors ; Epstein-Barr Virus Infections ; blood ; diagnosis ; immunology ; virology ; Herpesvirus 4, Human ; immunology ; isolation & purification ; Humans ; Immunoglobulin A ; blood ; Nasopharyngeal Neoplasms ; blood ; immunology ; virology ; Sensitivity and Specificity ; Serologic Tests

OBJECTIVESTo investigate the immunophenotypes of primary nasopharyngeal non-Hodgkin lymphoma (NPL) and their relationship to Epstein-Barr virus (EBV) infection.

METHODSThe clinical data and biopsies of 73 patients with NPL were collected in Guangzhou. In situ hybridization was performed to detect the EBV-encoded small non-polyadenylated nuclear RNAs (EBERs) on biopsy slides. Immunohistochemistry was used to classify the immunophenotypes of NPL and detect EBV antigen expression.

RESULTSForty-four (60.27%) of the 73 NPLs were of B cell lineage (CD79alpha(+)/CD3(-)/CD56(-)) while the 29 others (39.73%) were of non-B cell lineage. Seventy-three NPLs could be classified into 3 major immunophenotypes: B cell (CD79alpha(+)/CD3(-)/CD56(-), 44 cases), peripheral T cell (CD79alpha(-)/CD3(+)/CD56(-), 22) and NK/T cell (CD79alpha(-)/CD3(+)/CD56(+), 7). The percentages of EBV infection differed among the 3 major immunophenotypes (B cell: 11.36%, 5/44; peripheral T cell: 81.82%, 18/22; NK/T cell: 100%, 7/7). Both CD56(-) positive and CD56(-) negative immunophenotypes could further be divided into 4 subtypes: CD8(-)/CD4(-), CD8(+)/CD4(-), CD8(-)/CD4(+) and CD8(+)/CD4(+). All the CD8(-)/CD4(-) NPLs with CD56(-) positivity (7) or CD56(-) negativity (2) were infected with EBV. The neoplastic cells of a nasopharyngeal Burkitt's lymphoma expressed EBV nuclear antigen 1 (EBNA1) and EBV RNA (EBERs) only. In the other 29 EBV-infected NPLs, most of the lymphoma cells harboring EBV also expressed EBNA1 and EBERs; 21 of the 29 NPLs had a considerable number of neoplastic cells expressing latent membrane protein 1 (LMP1) (21/29, 72.41%) and 23 of 29 NPLs expressed latent membrane protein 2A (LMP2A) (23/29, 79.31%). A few lymphoma cells in 17 (17/29, 58.62%), 23 (23/29, 79.31%) and 22 NPLs (22/29, 75.86%) expressed Zta (Bam HI Z transactivator), viral capsid antigen (VCA) and membrane antigen (MA), respectively.

CONCLUSIONSThe prevalence ratio of the 3 immunophenotypes, namely, B cell, peripheral T cell and NK/T cell lymphoma, is about 6:3:1. However, the EBV infection ratio is reversed, 1:8:10. All the NK/T cell (CD56(+)) and peripheral immature T cell (CD3(+)/CD8(-)/CD4(-)) NPLs were EBV-infected. Except for one Burkitt's lymphoma, the EBV harbored in both B cell and non-B cell NPLs was mainly latent infection, type II, expressing EBNA1, LMP1 and LMP2A. However, the EBV found in a few lymphoma cells could become replicative, expressing lytic proteins.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; CD56 Antigen ; analysis ; Child ; Epstein-Barr Virus Infections ; complications ; Epstein-Barr Virus Nuclear Antigens ; analysis ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunophenotyping ; Lymphoma, Non-Hodgkin ; etiology ; immunology ; virology ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; etiology ; immunology ; virology ; RNA, Viral ; analysis

OBJECTIVETo compare the detection rates of Epstein-Barr virus (EBV) DNA in the serum/plasma between apparently healthy adults (AHAs) and nasopharyngeal carcinoma (NPC) patients in attempt to evaluate the efficiency of EBV DNA assay for serodiagnosis of NPC.

METHODSThe plasma and serum were obtained from 58 AHAs and 66 untreated NPC patients. EBV DNA W-fragment was detected using nested ploymerase chain reaction (PCR). Immunoenzymatic assay for titration of IgA-VCA was also adopted.

RESULTSEBV DNA detection rate (84.85%) in the plasma/serum of 66 NPC patients was significantly higher than that (10.34%) in 58 AHAs. The sensitivity of plasma/serum EBV DNA assay (0.8485) was higher than that (0.8030) of titrating IgA-VCA (positive criterion >/= 1:40) though the specificities of these two tests were the same (0.8966). The correct rate, predictive value of a positive test, and Odds ratio of dual positivity (0.8387, 0.9792 and 141.0, respectively) were higher than those of single positivity either to plasma/serum EBV assay (0.5242, 0.7333 and 1.1423, respectively) or to IgA-VCA >/= 1:40 test (0.4839, 0.5385 and 1.0480, respectively).

CONCLUSIONThe EBV DNA detection in the plasma/serum using nested PCR may be a useful indicator for serodiagnosis of NPC.

Antigens, Viral ; blood ; DNA, Viral ; blood ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunoglobulin A ; blood ; Nasopharyngeal Neoplasms ; diagnosis ; virology ; Serologic Tests

Female ; Humans ; Infant, Newborn ; Thoracic Surgical Procedures ; methods ; Twins, Conjoined ; surgery

OBJECTIVETo evaluate the risk of nasopharyngeal carcinoma (NPC) through EB virus antibody profile by enzyme linked immunosorbent assay (ELISA).

METHODSEBNA 1/IgA, EBNA 1/IgG and zta/IgG by ELISA and VCA/IgA by immmunoenzymatic method were detected in 121 NPC patients and 332 healthy subjects (HS) in the Pearl river estuary.

RESULTSThe sensitivity rates were 85%, 83% and 79% for EBNA 1/IgA, EBNA 1/IgG and zta/IgG, all three of which if combined was the highest 92%. The specificity rates were 86%, 86% and 80% for EBNA 1/IgA, EBNA 1/IgG and zta/IgG, all three of which if combined was also the highest 93%. According to the level of odds ratio, nasopharyngeal carcinoma risk could be divided into 3 groups: low, moderate and high-risk groups. 93% of HS had low risk of NPC with the odds ratio 0.0 to 0.3. 0.4% of HS had high risk of NPC with the odds ratio of 137.9%.

CONCLUSIONELISA is more objective than the traditional immunoenzymatic method in the detection and diagnosis of NPC. The combination of EBNA 1/IgA, EBNA 1/IgG and zta/IgG is able to evaluate the risk of NPC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Viral ; analysis ; Enzyme-Linked Immunosorbent Assay ; Epstein-Barr Virus Nuclear Antigens ; analysis ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnosis ; virology ; Risk Factors