Immunoexpression of human leukocyte antigen-DR in actinic cheilitis and lower lip squamous cell carcinoma

Abstract The aim of this study was to evaluate the immunoexpression of human leukocyte antigen-DR (HLA-DR) in actinic cheilitis (AC) and lower lip squamous cell carcinoma (LLSCC), and to correlate the findings with clinical (tumor size/extent, regional lymph node metastasis, and clinical stage) and histopathological (grade of epithelial dysplasia and inflammatory infiltrate for AC and histopathological grade of malignancy for LLSCC) parameters. Twenty-four AC and 48 LLSCC cases (24 with regional nodal metastasis and 24 without regional nodal metastasis) were selected. The scores of immunopositive cells for HLA-DR in the epithelial component of the lesions were assessed and the results were analyzed statistically using the nonparametric Mann-Whitney test. Epithelial expression of HLA-DR was observed in only five (20.8%) cases of AC (two low-grade and three high-grade lesions), with a very low median score of immunopositivity. By contrast, expression of HLA-DR was found in most LLSCC (97.9%), with a relatively high median score of positive cells. The score of HLA-DR-positive cells tended to be higher in tumors with regional lymph node metastasis, tumors in advanced clinical stages, and low-grade tumors, but the difference was not statistically significant (p > 0.05). In addition, there was a tendency towards higher expression of HLA-DR in highly/moderately keratinized tumors, and tumors with little/moderate nuclear pleomorphism (p > 0.05). The results suggest a potential role of HLA-DR in lip carcinogenesis, particularly in the development and progression of LLSCC. The expression of this protein can be related to the degree of cell differentiation in these tumors.

Role of human leukocyte antigen-DR expression on predicting the prognosis of hepatocellular carcinoma / 国际肿瘤学杂志

Objective To explore the expression of human leukocyte antigen-DR (HLA-DR) in hepatocellular carcinoma and its clinical values in predicting prognosis.Methods The primary hepatocellular carcinoma tissues of 97 patients in our hospital from August 2012 to February 2014 were enrolled as subjects (case group),and the adjacent normal liver tissues were selected as control (control group,n =97).The expressions of HLA-DR in the two groups were detected by immunohistochemistry,and the relationships between HLA-DR expression and chnicopathological features,recurrence and prognosis were analyzed.Results The positive expression rate of HLA-DR in specimens of case group was 55.67％,which was higher than 21.65％ in comrol group,and the difference was statistically significant (x2 =23.671,P ＜ 0.001).The positive expression rates of HLA-DR in tumor tissues with diameter ≥4 cm,Ⅲ-Ⅳ stage,poorly differentiated,bile duct cell carcinoma,no lymph node metastasis were higher than those in tumor tissues with diameter ＜4 cm,Ⅰ-Ⅱ stage,medium and well differentiated,hepatocellular carcinoma,lymph node metastasis respectively,and the differences were statistically significant (x2=10.481,P =0.001;x2=18.854,P ＜ 0.001;x2 =9.876,P =0.002;x2 =6.834,P =0.009;x2=30.668,P ＜ 0.001).The recurrence rates of 2 years and 3 years were 27.91％ and 46.51％ in patients with negative expression of HLA-DR respectively,which were lower than 50.00％ and 70.37％ in patients with positive expression of HLA-DR respectively,and the differences were statistically significant (x2 =4.860,P =0.028;x2 =5.668,P =0.017).The median survival time of patients with HLA-DR positive expression was 33.64 months,which was shorter than 36.88 months in patients with negative expression,and the difference was statistically significant (x2 =10.356,P ＜ 0.001).TNM staging Ⅲ-Ⅳ (OR =0.899,95 ％ CI:0.721-0.995,P =0.012) and positive expression of HLA-DR (OR =1.125,95％ CI:1.025-2.568,P =0.018) were unfavorable to the prognosis of hepatocellular carcinoma patients.Conclusion HLA-DR expresses abnormally in patients with primary hepatocellular carcinoma,which may be involved in the occurrence and development of hepatocellular carcinoma,and the up-regulation of HLA-DR in patients with hepatocellular carcinoma indicates a high recurrence rate and short survival time.

Expressions of activation antigens CD69 and HLA-DR in peripheral blood mononuclear cells and skin lesions of patients with psoriasis vulgaris / 中华皮肤科杂志

Objective To investigate changes in expressions of activation antigens CD69 and HLA-DR in CD3+T lymphocytes in peripheral blood and skin lesions in patients with psoriasis vulgaris. Methods Peripheral blood samples were obtained from 20 patients with psoriasis vulgaris and 20 healthy controls, and skin specimens from the lesions of 15 out of the 20 patients and 10 healthy controls. Flow cytometry was performed to quantify the expressions of CD69 and HLA-DR in peripheral blood CD3+T cells, and an immunohistochemical study to measure the expression of HLA-DR in skin specimens. Statistical analysis was carried out by a two-sample t-test and Pearson correlation analysis with the SPSS 19.0 software. Results Compared with the healthy controls, the patients with psoriasis vulgaris showed increased expression rates of CD69 (4.70%± 1.90%vs. 1.56%± 0.95%, t=6.629, P<0.01)and HLA-DR (8.97%± 1.79% vs. 3.02% ± 1.15%, t= 6.204, P< 0.01)in peripheral blood. Pearson correlation analysis revealed that the percentage of CD3+HLA-DR+cells in peripheral blood was positively correlated with the psoriasis area and severity index (PASI)score (r=0.5626, P<0.05). The expression rate of HLA-DR was significantly higher in the dermis (64.87%± 17.31%vs. 19.80%± 5.69%, t=7.916, P<0.01), but lower in the epidermis(11.80%± 5.55%vs. 27.40%± 8.61%, t=5.479, P<0.01)in the psoriatic specimens compared with the control specimens. Immunohistochemically, HLA-DR was widely expressed in the dermis of psoriatic lesions, but mainly distributed around blood vessels in the control skin. Conclusions There is an aberrant activation of CD3+T cells in peripheral blood and inflammatory cells in skin lesions in patients with psoriasis vulgaris, and the percentage of CD3 +HLA-DR+ cells in peripheral blood is correlated with the severity of psoriasis vulagaris.

Frecuencia de alelos HLA de clase I y II en una cohorte de pacientes con espondiloartritis provenientes del noroccidente colombiano / Frequency of HLA alleles class I and II in a cohort of northwestern Colombian patients with spondyloarthritis

Introducción. Las espondiloartritis son enfermedades reumatológicas crónicas que afectan el esqueleto axial y las articulaciones periféricas, con varias manifestaciones extraarticulares. La asociación con el HLA-B27 sigue siendo uno de los vínculos más fuertes conocidos entre estas entidades y el complejo mayor de histocompatibilidad; sin embargo, la distribución mundial del HLA-B27 varía considerablemente y se han descrito asociaciones con genes no HLA-B27. Objetivo. Conocer la frecuencia de alelos HLA de clase I y II en pacientes con espondiloartritis provenientes del noroccidente colombiano y su frecuencia en las manifestaciones clínicas y radiológicas específicas. Materiales y métodos. Se condujo un estudio descriptivo, observacional, de corte transversal, retrospectivo y prospectivo entre 2005 y 2008 de 56 pacientes colombianos con espondiloartritis. Se identificaron los alelos correspondientes a los loci HLA de clase I y II (HLA-B, HLADQB1 y HLADRB). Se analizó su frecuencia con las manifestaciones clínicas axiales, periféricas, extraarticulares y radiológicas. Resultados. Se encontró una baja frecuencia de HLA-B27 en la población total (50 %), aunque fue el alelo más frecuente, junto con HLA-DRB4*01 (35,7 %) y HLA-DQB1*0501 (28,6 %), en todos los pacientes en general y en cada una de las manifestaciones clínicas y radiológicas. Se resalta la alta frecuencia de HLA-B27 y HLA-DRB4*01 (64,3 %) en pacientes con dactilitis, hallazgo novedoso sin previa descripción. Conclusión. Los alelos HLA-B27, HLA-DRB4*01 y HLA-DQB1*0501 fueron frecuentes en los diferentes subtipos de espondiloartritis y en las manifestaciones clínicas axiales, periféricas y extraarticulares específicas, además de la sacroiliítis radiológica.

Introduction. Spondyloarthritis is a chronic rheumatic disease that affect the axial skeleton and peripheral joints, along with several extra-articular manifestations. The association with HLA-B27 remains one of the strongest known links between these entities and the major histocompatibility complex. However, the global distribution of HLA-B27 varies considerably and furthermore, associations with non-HLA-B27 genes have been described. Objective. The frequency of HLA class I and II was determined in a population of patients with spondyloarthritis with respect to detection in the clinical setting and by radiology. Materials and methods. A descriptive, observational, cross-sectional, retrospective and prospective study was conducted in 56 patients from northwestern Colombia. Each was diagnosed with spondyloarthritis between 2005 and 2008. In each case, alleles were identified for the loci HLA class I and II (HLA-B; HLADQB1 and HLADRB). The frequency of these alleles in the axial, peripheral, extraarticular and radiological manifestations. Results.The frequency of HLA-B27 was 50% overall, and it was the most frequent allele. The two other alleles were HLA.DRB4*01 at 35.7% and HLA-DQB1*0501 at 28.6%, as detected in each of the clinical and radiological manifestations. A high frequency of HLA-B27 and HLA-DRB4*01 (64.3%) was noted in patients with dactylitis. Conclusion. The alleles HLA-B27, HLA-DRB4*01 and HLA-DQB1*0501 were common in the different subtypes of spondyloarthritis and were frequent in the specific clinical axial, peripheral and extraarticular clinical manifestations, as well as radiological sacroiliitis.

Clinical significance of monitoring of anti-HLA antibodies in kidney recipients after renal transplantation / 中华器官移植杂志

Objective To detect de novo development of anti-HLA antibodies after renal transplantation, and to investigate their influence on graft function. Methods 384 kidney recipients,who were negative for anti-HLA antibody before transplantation, were monitored for anti-HLA antibodies over a period of 3-96 months, and a sensitive enzyme-linked immunosorbent assay (ELISA) was used to detect anti-HLA antibodies. HLA antibody ＞10 % was defined as positive levels. Results Among 384 recipients tested, 318 recipients (82. 8 %) were negative for anti-HLA antibody after transplantation; 66 recipients (17. 2 %) developed de novo HLA antibodies, 3 recipients with HLA class Ⅰ, 61 with HLA class Ⅱ, 2 with both HLA class Ⅰ and Ⅱ. According to amino acid residue matching, 7 cases developed de novo antibodies among 92 recipients with 0 HLA-DR mismatches,compared with 59 cases among 292 recipients with 1-2 mismatches, which showed significant difference between two groups (P＜0. 01 ). 87. 4 % (278/318) recipients negative for HLA antibodies after transplantation achieved good graft function, in comparison with 65. 2 % (43/66) recipients positive for HLA antibodies (P＜0. 05). Conclusion De novo production of HLA antibodies posttransplantation may be closely associated with HLA-DR mismatch. De novo HLA antibodies posttransplantation might damage graft function and reduce graft survival rate. The detection of de novo development of anti-HLA antibodies after renal transplantation has clinical significance for assessing renal allograft function.

Study on the Langerhans cells with various surface marker infiltrating in the lesion of basal cell carcinoma / 肿瘤研究与临床

Objective Some features, as numbers, morphology and distribution, of the Langerhans cells (LC) with various surface marker infiltrating in the lesion of basal cell carcinoma (BCC) were explored in order to apply some experimental data for assessment of the roles of LCs in the cutaneous tumor immunity.Methods Expression of CD1a, S-100 and HLA-DR on the surface of infiltrating LC of 30 cases of BCC and 15 cases of normal skin tissues were investigated by immunohistochemistry. The expression qualities were studied with computer image analysis system. Results It was observed that CD1a+ LC distributed mainly in tumor nest compared with the control group, CD1a+ LC in BCC revealed morphological changes: the dendrites shortened or disappeared. The density of CD1a+ LC, detected by cell number counting as well as staining strength scanning, showed a statistic difference between the control group and BCC (P ＜0.05). S-100(+)LC spread evenly throughout BCC lesions. The number of cell dendrites as well as its length increased markedly.The cell number as well as the staining strength in BCC lesions increased markedly compared with the control group (P ＜0.05). HLA-DR(+)LC infiltrated mainly around the located tumor. This kind of LC in the lesion did not show significant differences compared with the controls (P ＞0.05). Conclusion LC showed differences between BCC lesion and normal skin tissues. The alterations in numbers, morphology and distribution of LC with various markers in the lesion of BCC supplied special local cellular immunity status, resulting in disable metastasis and good prognosis.

Effects of Leptin on the expression of CD80, CD86 and HLA-DR in human monocytes / 中国医师杂志

Objective To study the effects of Leptin on the expression of CD86 and HLA-DR in human monocytes.Methods The expression of CD86 and HLA-DR in THP-1 Cells and human primary monocytes were detected by flow cytometry.Results Expression of CD86 and HLA-DR in THP-1 cells was significantly increased after treatment with high-dose Leptin ( CD86Untreated group:8.78 ± 1.66,CD86leptin10:50.76 ± 4.29,CD86leptin100:95.20 ± 4.90; HLAUntreated group:20.75 ± 2.12,HLAleptin10:102.14 ± 5.75,HLAleptin100:104.32 ± 4.75;).The similar results were observed in human primary monocytes ( CD86Untreated group:17.91 ± 1.78,CD86leptin100:48.80 ± 3.60; HLAUntreated group:34.10 ± 2.76,HLAleptin100:88.86 ± 3.53).Conclusions By up-regulating CD86 and HLA-DR expression,Leptin might enhance the ability to present antigen in THP - 1 cells and human monocytes.

Correlation between immune response to hepatitis B vaccine and HLA DRB1 * 07 DRB1 * 04 DRB1 * 1001, DQB1 * 0401 genes in Han population in Ningbo / 第二军医大学学报

Objective: To study the correlation between responses to Hepatitis B and HLA DRB1 * 07 DRB1 * 04 DRB1 * 1001, DQB1 * 0401 genes in Han population in Ningbo. Methods: A total of 240 Han people living in Ningbo received the routine vaccination of recombinant hepatitis B vaccine. The serum levels of anti-HBs antibody were examined and the subjects were divided into 2 groups according to the results: negative responses group (n = 120, anti-HBs D< 0.1) and positive response group (n = 120, anti-HBs D≥0.1). The distribution and frequency of DRB1 * 07, DRB1 * 04, DRB1 * 1001, DQB1 * 0401 genes alleles were determined with PCR using sequence-specific primers (PCR-SSP). Results: The sex and age were compatible between the 2 groups. The frequency of DRB1 * 07 in negative response group was significantly higher than that in positive response group (20.3%, vs 10.3%, P<0.05); the failure of vaccination was correlated with DRB1 * 07 (OR=0.45, P<0.05). The frequencies of HLA- DRB1 * 04 DRB1 * 1001, DQB1 * 0401 showed no obvious difference between the 2 groups. Conclusion: Failure or deficiency response to HBV vaccine in the Han people in Ningbo is associated with the HLA-DR antigen; HLA-DRB1 * 07 might be a susceptible gene for failure of HBV vaccination.

Correlation of T-lymphocytes expressing HLA-DR antigen with serum HBV DNA and HBeAg levels in chronic hepatitis B / 中华临床感染病杂志

Objective To investigate the correlation of T-lymphocyte expressing HLA-DR with serum HBV DNA and HBeAg contents in chronic hepatitis B. Methods Totally 134 chronic hepatitis B patients and 36 healthy blood donors were enrolled in the study. The T-lymphocytes (CD3 + HLA-DR + ,CD4 + HLA-DR+ and CD8 + HLA-DR+ T) expressing HLA-DR were detected by flow cytometry, the serum HBV viral loads were detected by the real-time quantitative PCR and HBeAg was detected by chemiluminescence method. According to serum HBV DNA viral loads patients were defined as HBV DNA negative (≤ 103 copies/mL), low (＞ 103 - 105 copies/mL), medium (＞ 105 - 107 copies/mL) and high groups (＞ 107 - 109 copies/mL) ; according to serum HBeAg levels, patients were defined as HBeAg negative (≤1 PEIU/mL), low (＞ 1 - 100 PEIU/mL), medium (＞ 100-1 000 PEIU/mL) and high groups (＞ 1 000-10 000 PEIU/mL). T test and one-way ANOVA were performed. Results With HBV DNA loads, HBeAg levels increased, the percentage of CD3 + HLA-DR + , CD4 + HLA-DR + and CD8 + HLA-DR +decreased, especially CD8 + HLA-DR +. Compared with HBV DNA negative group, the percentages of CD3 +HLA-DR + , CD4 + HLA-DR + and CD8 + HLA-DR + were significantly reduced in high group (t = 3. 686,4. 592 and 3. 216, P ＜ 0. 0l); the percentages of CD4 + HLA-DR + and CD8 + HLA-DR + were also reduced in medium group (t = 3. 761 and 2.862, P ＜ 0.01); while in low group, only the percentage of CD8 + HLA-DR + was reduced (t = 2.215, P ＜ 0.05). Compared with HBeAg negative group, the percentages of CD3 +HLA-DR+, CD4 + HLA-DR+ and CD8 + HLA-DR+ were significantly reduced in medium and high groups (thigher =3. 144, 2.222 and 4.035; tmiddle =3.311, 2.362 and 3.374, P ＜0.05), while in the low group,only the percentage of CD8+HLA-DR+ was reduced (t=2.029, P＜0. 05). Conclusion The combined measurement of HBV DNA, HBeAg and T-lymphocytes expressing HLA-DR in chronic hepatitis B patients may not only help to evaluate the immune status of patients, but also can predict the disease progression and clinical outcomes.

Clinical Spectrum and Prognostic Factors of Acute Necrotizing Encephalopathy in Children

This study was conducted to investigate the etiology, the clinical characteristics and prognosis of acute necrotizing encephalopathy (ANE) in Korean children. Six children (1 yr to 7 yr) patients with ANE were enrolled. They were diagnosed by clinical and radiological characteristics and their clinical data were retrospectively analyzed. In a search of clinically plausible causes, brain MRI in all patients, mitochondrial DNA studies for mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes (MELAS) and myoclonus epilepsy and ragged red fibers (MERRF) in four patients, and genomic typing on HLA DRB/HLA DQB genes in three patients were performed. All had precedent illnesses and the main initial symptoms included mental change (83%), seizures (50%), and focal deficits (50%). MRI revealed increased T2 signal density in the bilateral thalami and/or the brainstem in all patients. Mitochodrial DNA studies for MELAS and MERRF were negative in those children and HLA-DRB1*1401, HLA-DRB3*0202, and HLA-DQB1*0502 seemed to be significant. A high dose steroid was given to all patients, which seemed to be partly effective except for 2 patients. In conclusion, ANE is relatively rare, but can result in serious neurological complication in children. Early detection and appropriate treatment may lead to a better neurological outcome.

New insights into Vogt-Koyanagi-Harada disease / Novos conhecimentos sobre a doença de Vogt-Koyanagi-Harada

Vogt-Koyanagi-Harada disease (VKH), a well-established multiorgan disorder affecting pigmented structures, is an autoimmune disorder of melanocyte proteins in genetically susceptible individuals. Several clinical and experimental data point to the importance of the effector role of CD4+ T cells and Th1 cytokines, the relevance of searching a target protein in the melanocyte, and the relevance of the HLA-DRB1*0405 in the pathogenesis of the disease. Vogt-Koyanagi-Harada disease has a benign course when early diagnosed and adequatey treated. Full-blown recurrences are rare after the acute stage of Vogt-Koyanagi-Harada disease is over. On the other hand, clinical findings, such as progressive tissue depigmentation (including sunset glow fundus) and uveitis recurrence, indicate that ocular inflammation may persist after the acute phase. Additionally, indocyanine green angiography findings suggest the presence of choroidal inflammation in eyes without clinically detectable inflammation. The aim of this paper is to review the latest research results on Vogt-Koyanagi-Harada disease pathogenesis and chronic/convalescent stages, which may help to better understand this potentially blinding disease and to improve its treatment.

A doença de Vogt-Koyanagi-Harada (VKH) afeta vários órgãos que têm em comum a presença de pigmento. É doença autoimune que agride os melanócitos de indivíduos geneticamente susceptíveis. Inúmeras evidências clínicas e experimentais demonstram a importância de células T CD4+ como células efetoras da resposta imune celular, das citocinas pró-inflamatórias Th1, da procura da proteína-alvo dentro do melanócito, e da relevância do HLA-classe II DRB1*0405 na patogênese desta doença. A doença de Vogt-Koyanagi-Harada apresenta bom prognóstico visual desde que o diagnóstico seja precoce e o tratamento instituído seja adequado. Recidivas com acometimento do segmento posterior são raras após a fase aguda da doença. No entanto, achados clínicos como a progressiva despigmentação do fundo, incluindo o aspecto em por do sol, e as recidivas da uveíte indicam que a inflamação ocular pode persistir mesmo após a fase aguda da doença. Os achados da angiografia com indocianina verde também sugerem a presença de inflamação da coróide mesmo em olhos sem inflamação clinicamente detectável. O objetivo do presente trabalho é rever os mais recentes estudos sobre a patogênese da doença Vogt-Koyanagi-Harada e sobre os aspectos clínicos da fase crônica e/ou convalescente da doença, permitindo melhores conhecimentos sobre esta doença potencialmente mórbida e oferecendo terapias mais adequadas.

Flow cytometry analysis of unusual peripheral monocyte population / 中华检验医学杂志

Objective To analyze a population of cells on the right lower lateral of monocyte population in forward scatter/side scatter(FSC/SSC)(X-axis/Y-axis)scatterplot of peripheral blood leucocyte by flow cytometry(FCM)and its influencing factors.Methods The type of cells were identified based on cluster of differentiation antigen(CD)by FCM.The impact of temperature,hemolysin concentration,and incubation time was evaluated.Blood lipid tests were performed to observe the relation between them by statistical methods.Results (1) Phenotypo of this population of cells on the right lower lateral of monocytes in FSc/SSC scatterplot is CD+45 CD+13 CD+14 CD3- CD-19 ,which was the same as monocyte cells:(2)The monocytes in FSC/SSC scatterplot shifted to left side after using haemolysin;(3)The monocytes showed less resistance to antihemolysin in 37℃ than that in 220C:There were more monocytes shifting to left side with the increase of haemolysis time:(4)The swarming ratio of monocytes in patients (31.5%,40/126) Was higher than it in normal controls (5.1%,5/98)(x2=22.74,P<0.01);(5)The levels of total serum cholesterol(TC),triglyeride(TG),low density lipoprotein cholesterol(LDL-C), apoprotein B100(Apo B100) in patients with swarming monocytes were lower than that in the patients without swarming monocytes,(P<0.05).There was no statistical significance between the two groups with respect to levels of total bilirubin(TBIL),albumin(Alb),hish density lipoprotein cholesterol(HDL-C),apoprotein A I(Apo A I),lipoprotein(Lpa).Conclusions Peripheral blood monocytes can be divided in two groups in FSC/SSC scatterplot when analyzed with FCM.The presence of this population of cell Was related to resistance to hemolysin.It can be influenced by haemolysis time and incubation temperature.Therefore,the effect of swarming monocytes and abnormal cell membrane should be taken into consideration when the markers and function of monocytes are detected by FCM.

Síndrome de Vogt-Koyanagi-Harada incompleta associada a HLA DRB1*01 em criança de quatro anos de idade: relato de caso / Incomplete Vogt-Koyanagi-Harada syndrome associated with HLA DRB1*01 in a 4-year-old child: case report

A síndrome de Vogt-Koyanagi-Harada é doença crônica, sistêmica e auto-imune, com manifestações oculares, nervosas, auditivas e tegumentares. Descrevemos aqui o caso de uma criança com início dos sintomas aos quatro anos e dois meses de idade, com positividade para o HLA DRB1*01.

Vogt-Koyanagi-Harada syndrome is chronic systemic autoimmune disease with ocular, nervous, auditory and tegumental manifestations. We report here the case of a child with onset of symptoms at four years and two months of age, with positive HLA DRB1*01.

Role of Staphylococcal Superantigen in Atopic Dermatitis: Influence on Keratinocytes

Staphylococcus aureus may perform an crucial function in atopic dermatitis (AD), via the secretion of superantigens, including staphylococcal enterotoxins (SE) A or B, and toxic shock syndrome toxin-1 (TSST-1). Dysregulated cytokine production by keratinocytes (KCs) upon exposure to staphylococcal superantigens (SsAgs) may be principally involved in the pathophysiology of AD. We hypothesized that lesional KCs from AD may react differently to SsAgs compared to nonlesional skin or normal skin from nonatopics. We conducted a comparison of HLA-DR or CD1a expression in lesional skin as opposed to that in nonlesional or normal skin by immunohistochemistry (IHC). We also compared, using ELISA, the levels of IL-1alpha, IL-1beta, and TNF-alpha secreted by cultured KCs from lesional, nonlesional, and normal skin, after the addition of SEA, SEB and TSST-1. IHC revealed that both HLA-DR and CD1a expression increased significantly in the epidermis of lesional skin versus nonlesional or normal skin in quite a similar manner. IL-1alpha, IL-1beta, and TNF-alpha secretion was also significantly elevated in the cultured KCs from lesional skin after the addition of SsAgs. Our results indicated that KCs from lesional skin appear to react differently to SsAgs and increased proinflammatory cytokine production in response to SsAgs may contribute to the pathogenesis of AD.

HLA genotyping by oligoneucleotide chip technique in kidney transplantation / 中华器官移植杂志

Objective To compare oligoneucleotide arrays with sequence specific primer polymerase chain reaction (PCR-SSP) for HLA-DR genotyping in order to develop a new technique of genotyping for donors and recipients in kidney transplantation. Methods Sixty DNA samples of donors and recipients were subjected to HLA-DR typing by oligoneucleotide arrays and PCR-SSP simultaneously. The results of the two typing techniques were analyzed.Results Of 60 samples using the two above-mentioned typing techniques, the results in 56 samples were identical with the accordance rate being 93 %, and in the remaining 4 unidentified samples verified by the other laboratory, oligoneucleotide arrays made 1 allele miss typing for 2 samples , 1 allele mistaking for 1 and PCR-SSP made 1 allele miss typing for 1 sample. Among the total, 20 samples retyping was made and its reproduction rate was 96 %. Conclusion The oligoneucleotide arrays technique for HLA genotyping has advantage of high sensitivity, high efficiency, high level standard and it is incomparable.

Molecular Analysis of HLA Class ll-Associated Susceptibility to Neuroinflammatory Diseases in Korean Children

The work was done to study immunogenetic peculiarities of neuroinflammatory diseases among Korean children. A total of 13 children with neuroinflammatory diseases (8 males and 5 females; mean age 4.6+/-2.6 yr) were consecutively recruited. Genomic typing was performed on their HLA DRB/HLA DQB genes using PCR-SSOP/ SSP techniques with gel immunoelectrophoresis. The frequencies of HLA-DR1* 15 in children with acute disseminated encephalomyelitis (ADEM) (31%) and DQB1* 06 in other neuroinflammatory diseases (38%) were significantly increased compared with control subjects. The frequencies of HLA-DRB3*0202 (100%), HLA-DRB1*1302 (67%), HLA-DRB3*0301 (67%), and HLA-DQB1*0301 (67%) were significantly increased in children with multiple sclerosis and the frequencies of HLA-DRB1*1501 (40%) and HLA-DRB5*0101 (40%) were significantly increased in children with ADEM. HLA-DRB1*1401, HLA- DRB3*0202, and HLA-DQB1*0502 were found in children with acute necrotizing encephalopathy. In conclusion, HLA-DR1*15 and DQB1*06 may be involved in susceptibility to inflammation in Korean children. The frequencies of HLA-DRB1*1501, HLA-DRB5*0101, HLA-DRB3*0301, and HLADQB1* 0602 were not as high in Korean children with multiple sclerosis as in western children. However, HLA-DRB3*0202 was seen in all children with multiple sclerosis. Our data may provide further evidence that the immunogenetic background of neuroinflammatory diseases in Korean is distinctly different from the ones in western countries. Further studies are necessary to confirm this finding.

Effects of Tazarotene Combined with Interferon-?on the Expression of HLA-DR in Cultured Hu-man Keratinocytes / 中华皮肤科杂志

Objective To investigate the effects of tazarotene on the proliferation of cultured human keratinocytes and IFN-?-induced expression of HLA-DR in those cells.Methods Keratinocytes were cul-tured from normal human skin in vitro,and were treated with various concentrations of tazarotene(10 -5 ,10 -6 ,10 -7 mol/L).At24h,48h after treatment,the effects on cell proliferation were assessed by MTT method.The expression of HLA-DR was determined using immunocytochemistry techniques in cultured human ker-atinocytes incubated with tazarotene,IFN-?or both for24h.Results①The proliferation of keratinocytes was decreased when exposed to10 -7 -10 -5 of tazarotene as compared to non-exposed keratinocytes after24h and48h.Moreover,the effects on cell proliferation by tazarotene were dose-dependent;②There was rare expression of HLA-DR in normal human keratinocytes.③HLA-DR expression was inducible significantly with500u/mL of IFN-?,but failed to be induced with10 -6 mol/L of tazarotene,in keratinocytes at24h af-ter treatment.④After24h combined treatment of10 -7 -10 -5 mol/L of tazarotene and IFN-?,the induction of HLA-DR expression was significantly stronger,in a dose-dependent manner,than IFN-?alone(P

Effects of host factors and virus factors on interferon treatment in patients with chronic hepatitis B / 中华传染病杂志

Objective To investigate the effects of host factors and hepatitis B virus (HBV) factors on interferon treatment in patients with chronic hepatitis B(CHB). Methods Polymerase chain reaction sequence specific primers (PCR SSP) technology was used to determine human leukocyte antigen (HLA) DR and DQ alleles in 32 patients with CHB. Serum HBV DNA level was quantified by a PCR assay with a low limit of detection of 1000 copies/ml. HBV genotypes, precore(A 1896 ) and core promoter(T 1762 A 1764 ) mutations was determined by PCR restriction fragment length polymorphism(PCR RFLP) analysis. Results The frequency of HLA DRB1*14 allele was higher in 10 responders to IFN treatment than in 22 non responders(20.0% v.s. 2.3%, relative risk(RR)=10.750, P =0.030); whereas that of HLA DQB1*07 is lower(10.0% v.s. 38.6%, RR=0.176, P =0.022). Pretreatment serum HBV DNA level in 10 responders was lower than that in 22 non responders (6.71?1.06 v.s. 7.59?0.56log 10 copies/ml, P =0.030). Patients with HBV genotype B infection had a higher response rate to IFN than those with HBV genotype C infection(53.8% v.s. 15.8%, P =0.049). Pretreatment serum ALT level, presences of A 1896 mutation and T 1762 A 1764 mutations had no difference between responders and non responders. Logistic multivariate analysis identified low pretreatment HBV DNA level(RR=0.212, P =0.034), presence of A 1896 mutation(RR=0.063, P =0.049) and HLA DQB1*07(RR=13.358, P =0.045) as independent factors associated with IFN antiviral response. Conclusions Low load of pretreatment serum HBV DNA and T 1762 A 1764 mutation in HBV genome or HBV of genotype B are associated with the response rate to IFN treatment in patients with CHB.