OBJECTIVES: The purpose of this study is to compare newly developed assay for identification of ENA antibody, Phadia EliA ENA with Euroimmun line immunoassay by analyzing the degree of agreement and the individual antibodies between two methods. METHODS: A total of 82 patient samples were used. Indirect immunofluorescence assay using Hep-2 cell was performed to screen the antinuclear antibody (ANA). Euroimmun line immunoassay (LIA) and Phadia EliA ENA assay were tested to identify the antibodies against extractable nuclear antigens (ENAs). Kappa statistics was used to evaluate the degree of agreement. RESULTS: Mean age of patients was 41.0 (8-79), and the M:F ratio was 21:61. ANA was positive in 74 samples, and negative were 8 samples. Kappa analysis of the 82 tested samples showed a moderate strength of agreement (kappa = 0.521, P = 0.000). There were differences in the order of identified individual antibodies between two methods (Ro > La = RNP > Centromere > Sm > Scl-70 in Phadia EliA ENA, Ro > RNP > Sm>La > Scl-70 > Centromere=Jo-1 in Euroimmun LIA). Ro antibody was most frequently identified in Phadia EliA ENA negative-Euroimmun LIA positive specimens (Ro > RNP = Jo-1 > La = Sm = Centromere > Scl-70). CONCLUSIONS: A moderate strength of agreement was observed between the Phadia EliA ENA and the Euroimmun LIA. There seemed to be a significant difference in the ratio of individual antibodies, especially in the anti-Ro and Sm antibodies.
Antibodies ; Antibodies, Antinuclear ; Antigens, Nuclear ; Centromere ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoassay

OBJECTIVE: Systemic Lupus Erythematosus(SLE) is characterized by various autoantibodies to a variety of nuclear antigens. Certain extractable nuclear antigens(ENA) have been served as extremly useful aids in differentiation of clinical subset, diagnostic marker and in the detection of early forms of systemic rheumatic diseases. This study was to employ immunoblot to determine the prevalences of antibodies to ENA in SLE compared with immunodiffusion. METHODS: Sera were obtained from 127 SLE patients. Antibodies to ENA were assessed by double immunodiffusion (DID) and immunoblot method. RESULTS: Using the immunblot method, the prevalences of antibodies to ENA were as follows: The antibody to Sm was 27%, UIRNP 33.6%, Ro 41.8%, La 14%, ribosomal P 14% and Ku 4%. The prevalences of antibodies to ENA by DID were as follows: The antibody to Sm was 15%, RNP 24.5%, Ro 54.3% and La 9.4%. CONCLUSIONS: Compared with immunodiffusion, results using immunoblot showed greater sensitivity in the detection of autoantibodies to ENA in SLE.
Antibodies* ; Antigens, Nuclear ; Autoantibodies ; Humans ; Immunoblotting* ; Immunodiffusion* ; Lupus Erythematosus, Systemic* ; Prevalence ; Rheumatic Diseases

Pigs are considered as ideal donors for xenotransplantation because they have many physiological and anatomical characteristics similar to human beings. However, antibody-mediated immunity, which includes both natural and induced antibody responses, is a major challenge for the success of pig-to-primate xenotransplantation. Various genetic modification methods help to tailor pigs to be appropriate donors for xenotransplantation. In this study, we applied transcription activator-like effector nuclease (TALEN) to knock out the porcine α-1, 3-galactosyltransferase gene GGTA1, which encodes Gal epitopes that induce hyperacute immune rejection in pig-to-human xenotransplantation. Meanwhile, human leukocyte antigen-G5 gene HLA-G5, which acts as an immunosuppressive factor, was co-transfected with TALEN into porcine fetal fibroblasts. The cell colonies of GGTA1 biallelic knockout with positive transgene for HLA-G5 were chosen as nuclear donors to generate genetic modified piglets through a single round of somatic cell nuclear transfer. As a result, we successfully obtained 20 modified piglets that were positive for GGTA1 knockout (GTKO) and half of them expressed the HLA-G5 protein. Gal epitopes on the cell membrane of GTKO/HLA-G5 piglets were completely absent. Western blotting and immunofluorescence showed that HLA-G5 was expressed in the modified piglets. Functionally, the fibroblasts from the GTKO/HLA-G5 piglets showed enhanced resistance to complement-mediated lysis ability compared with those from GTKO-only or wild-type pigs. These results indicate that the GTKO/HLA-G5 pigs could be a valuable donor model to facilitate laboratory studies and clinics for xenotransplantation.
Animals ; Animals, Genetically Modified ; Gene Knockout Techniques ; HLA Antigens ; Humans ; Nuclear Transfer Techniques ; Swine ; Transplantation, Heterologous

The proliferative activity of gastric adenomas from 18 patients (42 endoscopic procedures) was compared with follow-up results. These cases were gastric adenomas proven by follow-up with repeated endoscopic procedures for more than 2 years, or were confirmed as gastric adenocarcinoma thereafter by histopathologic examination. Among the eighteen cases, nine showed carcinoma in the subsequent biopsies (group 1) and the remaining nine did not result in carcinoma (group 2). The proliferating cell nuclear antigen (PCNA) positivity rates of the two groups were significantly different (P < 0.01). The average PCNA positivity in group 1 was 33.1%, while it was 10.0% in group 2. The risk of developing carcinoma increased as the PCNA positivity increased: 0% in the low PCNA positivity group, 41% in the mid-positivity group and 89% in the high positivity group. We concluded that growth fraction could be taken into account as one of the most important prognostic factors for gastric adenoma, and accordingly repeated endoscopic biopsies with close follow-up should be carried out especially in the high PCNA positivity group.
Adenoma/*immunology ; Antigens, Neoplasm/immunology/*metabolism ; Carcinoembryonic Antigen/metabolism ; Cell Cycle ; Follow-Up Studies ; Gastroscopy ; Humans ; Nuclear Proteins/*metabolism ; Prognosis ; Proliferating Cell Nuclear Antigen ; Stomach Neoplasms/*immunology

To characterize cellular responses during hepatic regeneration, we examined 13 explant livers and 5 liver allografts by immunohistochemistry for cytokeratin 7, HepPar1, CD68, alpha-smooth muscle actin (alpha-SMA) and proliferating cell nuclear antigen as well as reticulin and Masson-trichrome staining. Within a week after liver damage, elongated CD68-positive cells were detected along the border of necrotic area. The number of alpha-SMA-positive cells was slightly increased along the sinusoids. Ductular proliferation or fibrosis was negligible. After one or two weeks, the size and number of CD68-positive cells were markedly increased. alpha-SMA-positive cells increased in number within lobules and portal tracts. Ductular proliferation occurred predominantly at the limiting plate or along the border of necrotic areas. After one month, necrotic parenchyma was replaced by many ductules, CD68-positive cells, alpha-SMA-positive cells. Nodules of regenerating hepatocytes and irregular fibrosis were diffusely present. Other nonparenchymal cells were not significantly changed. These observations indicate that chronological interaction between nonparenchymal and parenchymal cells occur during the course of human hepatic regeneration and suggest extensive porto-periportal fibrosis more than a few months after the onset of fulminant hepatitis is a major indicator of chronic functional impairment necessitating liver transplantation.
Actins/analysis ; Antigens, CD/analysis ; Antigens, Differentiation, Myelomonocytic/analysis ; Human ; Immunohistochemistry ; Keratin/analysis ; Liver/*cytology ; *Liver Regeneration ; Proliferating Cell Nuclear Antigen/analysis

OBJECTIVETo evaluate the value of EBNA1-IgA and EA-IgG in serological diagnosis of nasopharyngeal carcinoma (NPC).

METHODSThe serum EBNA1-IgA and EA-IgG of 56 patients with NPC and 58 healthy adults were detected by ELISA. The sensitivity, specificity, positive predictive value, accuracy rate and odds ratio of the two tests used singly or in combination were compared with each other.

RESULTSThe sensitivity of EBNA1-IgA (91.07%) was higher than that of EA-IgG (87.50%), while the specificity of EA-IgG (87.93%) was higher than that of EBNA1-IgA (84.48%). The combination of EBNA1-IgA and EA-IgG could enhance the specificity (94.83%), positive predictive value (0.9375), likelihood ratio (15.5435) and odds ratio (75.0000) for serological diagnosis of NPC. Forty-five patients showed both positive EBNA1-IgA and positive EA-IgG. A positive EA-IgG was detected in 4 out of 5 patients with negative EBNA1-IgA and a positive EBNA1-IgA was founded in 6 out of 7 patients with negative EA-IgG.

CONCLUSIONAlthough relatively high sensitivity and specificity could be obtained by either EBNA1-IgA or EA-IgG test alone, the combination of these two tests with a complementary effect is able to enhance the reliability of serological diagnosis of NPC as most patients have positive ENBA1-IgA and EA-IgG concurrently.

Adult ; Antigens, Viral ; immunology ; Enzyme-Linked Immunosorbent Assay ; Epstein-Barr Virus Nuclear Antigens ; immunology ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Nasopharyngeal Neoplasms ; diagnosis ; immunology ; Sensitivity and Specificity

OBJECTIVETo investigate the relationship between changes in high-risk populations and screening detected nasopharyngeal carcinoma (NPC) during the three-year follow-up of high-risk and moderate-risk groups at initial EB virus serology screening.

METHODSWe tested EB virus VCA-IgA and EBNA1-IgA antibody to identify the probability of suffering from NPC of the crowd. The high-risk and moderate-risk groups at initial screening in one county during 2009 to 2010 were followed-up once a year with EB virus serology testing. All the high-risk people during initial screening and follow-up were conducted with nasopharyngeal fiber endoscopy. Through the follow-up of three years, we analyzed changes in the number of high-risk group, detection rate of NPC in high-risk group, and tumor staging. Firstly detected NPC by screening was defined as screening group, and detected by following-up was defined as following-up group.

RESULTSA total of 404 participants were at high-risk and 1 041 participants were at moderate-risk group, 1 445 persons were in the group. All 404 persons were at high-risk at initial screening, the number of high-risk people during follow-up decreased from 371 to 187, 853 people of the all high-risk group were conducted with nasopharyngeal fiber endoscopy, and 38 cases of NPC were detected. NPC detection rate of high-risk group was 6.2% (25/404), 3.2% (12/371), 0.5% (1/188) and 0 (0/187) during the initial screening and three years follow-up respectively. The cumulative incidence of NPC in the high-risk and moderate-risk group were 7.7% (31/404) ,0.8% (8/1 041) . The early diagnosis rate of NPC in screening group and following-up group was 80% (20/25)and 11/13, respectively. With the primary tumor, the rate of T1 in screening group was higher than following-up group (80% to 38%, 20/25 to 5/13; P = 0.028). However, compared with following-up group, the rate of regional lymph node metastasis in screening group was higher (19/25 to 5/13; P = 0.035 ).

CONCLUSIONAlong with the high detection rate of early staging NPC in screening group and following-up group, the detection of NPC in high risk people is mainly at initial screening and the first year following-up and NPC detection rate thereafter is dropping significantly.

Antibodies, Viral ; Antigens, Viral ; Capsid Proteins ; Carcinoma ; Early Detection of Cancer ; Epstein-Barr Virus Nuclear Antigens ; Follow-Up Studies ; Herpesvirus 4, Human ; Humans ; Nasopharyngeal Neoplasms ; Neoplasm Staging ; Risk Factors

BACKGROUND: Identification of antibodies recognizing extractable nuclear antigens (ENAs) is use-ful in the diagnosis and characterization of a variety of connective tissue diseases. Recently, LG ENA Immunoblot (LGCI, Seoul, Korea) was introduced for detecting various autoantibodies to ENAs simultaneously. Performance of this kit was evaluated in this study. METHODS: Sera from 108 SLE patients and 103 RA patients were tested for the presence of spe-cific autoantibodies to ENAs by LG ENA Immunoblot and DID. Concordance rates in each autoan-tibody were obtained. After discordant results were resolved by EIA (ENA ELISA TEST SYSTEM, Zeus Scientific Inc., NJ, USA) and western blot (ANA Western Blot Immunoassay, IMMCO Diag-nostics Inc., NY, USA), sensitivity and specificity of LG ENA Immunoblot were evaluated. Between-day precision was also tested. RESULTS: Concordance rates in each autoantibody in two methods were as follows: anti-RNP (88.0%, 95/108; 100%, 103/103), anti-Sm (87.0%, 94/108; 97.1%, 100/103), anti-SSA (94.4%, 102/108; 99.0%, 102/103), anti-SSB (97.2%, 105/108; 98.1%, 101/103), anti-Scl70 (99.1%, 107/108; 100%, 103/103) in SLE and RA patients, respectively. Sensitivity and specificity of Immunoblot were 92.0% and 99.6% for anti-RNP, 100% and 99.6% for anti-Sm, 100% and 98.6% for anti-SSA, 90.0% and 98.5% for anti-SSB, and 100% and 100% for anti-Scl70, respectively. Between-day precisions were 100% in all anti-ENA antibodies. CONCLUSIONS: LG ENA Immunoblot showed good concordance rates with the conventional DID method and high sensitivity (>90%) and specificity (>98.5%) in detecting all kinds of anti-ENA autoantibodies. LG Immunoblot has another merit in that it can detect several autoantibodies simul-taneously. It is suggested that LG ENA Immunoblot can replace DID for anti-ENA detection without any problem.
Antibodies ; Antigens, Nuclear* ; Autoantibodies* ; Blotting, Western ; Connective Tissue Diseases ; Diagnosis ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoassay ; Sensitivity and Specificity ; Seoul

BACKGROUND: Antibodies to Bo antigen are present in most patients with subacute cutaneous lupus erythematosus and in about 50% of Korean patients with systemic lupus erythematosus (LE). However, the pattern of in vivo epidermal deposits of anti Ro antibodies has not been widely recognized. OBJECTIVE: The purpose of thjs study was to define characteristic findings of direct and indirect immunofluorescence(IF) in patients with high-titer anti-Ro or anti-Ro/La positive LE. METHODS: Lupus band test (riirect IF with normal appearing forearm skin specimens) and indi- rect IF with normal skin substr ates were performed with 3 patients of systemic LE who have high titers( >1: 640) of anti-Ro or anti-Ro/La antibodies but have no antibodies against other nuclear antigens such as nDNA/Sm/nRNP/Scl-70. RESULTS: An identical pattern of immune deposits was observed in the epidermis in all 3 pa- tients through direct and indirect, IF examinations. The characteristic pattern recognized was "fine speckling" of IgG (or IgG/IgM) mainly at the nuclei on the basal keratinocytes or keratinocytes throught the epidermis. In the immunoblot assay performed with one patient, IgG anti-Ro/La anti- bodies were identified to recognize the 52/42kD antigens (probably, the Ro/La antigens) in the cultured keratinocyte extracts. CONCLUSION: Most direct IF studies in patients with systemic LE(lupus band test) have shown granular depositions of immunoglabulins and complement components along the dermoepidermal junction, however, the staining patterns as observed in this study may have been overlooked. The recognizable fine speckled patteen of immune deposits at the epidermal keratinocytes could he taken into account as a positivi. finding in the broad category of "lupus band", seen with the normal appearing skin in patient s with systemic LE, especially, who have high titers of anti-Ro/ La antibodies.
Antibodies ; Antigens, Nuclear ; Complement System Proteins ; Epidermis ; Forearm ; Humans ; Immunoglobulin G ; Keratinocytes ; Lupus Erythematosus, Cutaneous ; Lupus Erythematosus, Systemic ; Skin

OBJECTIVEThe objective of this study is to investigate the effect of epidermal growth factor receptor (EGFR) monoclonal antibody MAb225 on repair of DNA double strand break (DNA-DSB) after radiation in tongue squamous cell carcinoma cell.

METHODSThe single cell gel electrophoresis (SCGE) was performed to estimate the repair of DNA-DSB induced by radiation in human tongue carcinoma cells Tca8113 treated with or without MAb225. Expression of Ku70 and Ku80 were detected by semiquantitative reverse transcription polymerase chain reaction and Western bolt.

RESULTSComet tail moment of MAb225 treated cell was significantly higher than untreated cell (P < 0.05). The expression of Ku70 and Ku80 were inhibited by MAb225.

CONCLUSIONMAb225 can inhibit repair of DNA-DSB induced by down-regulated expression of Ku70 and Ku80.

Antibodies, Monoclonal ; Antigens, Nuclear ; DNA Repair ; DNA-Binding Proteins ; Humans ; Ku Autoantigen ; Receptor, Epidermal Growth Factor ; Tongue Neoplasms