Clinical analysis of sIgG antibody in children with food intolerance in Qingdao / 中华健康管理学杂志

Objective:To analyze food intolerance status in children in Qingdao by detecting the serum levels of food-specific IgG (sIgG).Methods:In this cross-sectional study, a total of 4 249 children aged 0 to 14 years (all were permanent residents of Qingdao City) admitted to Women and Children′s Hospital Affiliated to Qingdao University from May 2017 to December 2020 for suspected food intolerance were selected as the study objects with the whole sampling method. According to the age, the objects were divided into 4 groups: 0-<1 year group (440 cases), 1-<3 years group (1 761 cases), 3-<6 years group (1 193 cases), and ≥6 years group (855 cases). Positive condition of serum sIgG antibodies of 14 kinds of food in the children were detected by enzyme-linked immunosorbent assay. Chi-square test was used to compare the positive rate of the antibodies among different foods, gender and age groups.Results:The total positive rate of food sIgG antibody in 4 249 children was 95.32% (4 050/4 249), the highest positive rate was found in eggs (81.50%) and the lowest positive rate was found in pork (1.15%). The positive rates of sIgG antibody in milk (54.98% vs 49.69%, χ2=11.627), crab (5.59% vs 3.71%, χ2=8.049) and shrimp (4.62% vs 2.75%, χ2=9.784) in boys were significantly higher than those in girls, and the positive rates of sIgG antibody in tomato (49.19% vs 45.54%, χ2=5.510), cod (8.53% vs 5.96%, χ2=10.512) and beef (2.58% vs 1.70%, χ2=3.959) in girls were significantly higher than those in boys (all P<0.05). The total positive rate of sIgG antibody in 14 foods was the lowest in 0-<1 year group (89.09%), and it was the highest in 3-<6 years group (96.98%) ( χ2=63.950, P<0.001). The highest positive rate in 0-<1 year group was found in tomato (56.36%), and it was eggs (85.29%, 88.94%, 85.50%) in all the other 3 groups. The positive rates of corn and beef decreased with age ( χ2=44.098, 20.106, P<0.001), while those of cod and mushroom increased with age ( χ2=32.315, 40.338, P<0.001). The positive rate of wheat (57.13%, χ2=42.273), tomato (57.01%, χ2=209.862), soybean (24.99%, χ2=92.580), crab (6.81%, χ2=33.201), shrimp (6.25%, χ2=47.863) were all the highest in 1-<3 years group among the 4 groups (all P<0.001), and the positive rate of chicken was the highest in 3-<6 years group (7.88%, χ2=29.875; P<0.001). Conclusions:Children in Qingdao have a high level of food intolerance, and the highest positive rate is for eggs. Milk, crab and shrimp should be focused on for boys, while tomatoes, cod and beef shoud be paid more attention to for girls. Children of different ages have different kinds of food intolerance, and their diets should be adjusted reasonably according to the characteristics.

Factors affecting the efficacy of platelet transfusion and specificity of HLA-Ⅰ antibodies in tumor patients / 中国输血杂志

【Objective】 To analyze the factors affecting the efficacy of platelet transfusion and the specificity of HLA-Ⅰ antibodies in tumor patients. 【Methods】 Tumer patients applied for platelet transfusion from November 2019 to July 2021 in Sichuan Cancer Hospital province were screened for platelet antibodies. The transfusion efficacy was evaluated and its influencing factors were analyzed, Antibody positive patients were detected for HLA-Ⅰ antibodies. 【Results】 The positive rates of platelet antibody and HLA-Ⅰ antibody, as well as the platelet transfusion refractoriness rate were 35.2%(50/142), 32.4%(46/142) and 16.9%(24/142), respectively. The platelet refractoriness rate of platelet antibody positive patients was higher than that of platelet antibody negative patients [30.0%(15/50) vs 9.8%(9/92)] (χ2=9.428, P<0.05). Multivariate logistic results showed that gender (χ2=12.608, P<0.001, OR=3.800, 95%CI: 1.819-7.940) was an independent risk factor for platelet antibody production, and platelet antibody(χ2=8.648, P<0.05, OR=3.952, 95%CI: 1.581~9.878) was the independent risk factor for transfusion efficacy. The detection rates of strong positive, positive and weak positive HLA-I antibodies were 69.6% (32/46), 80.4% (37/46) and 97.8% (45/46), respectively. The antibodies with high detection rate were anti-B*15∶12, anti-B*57∶03, anti-B*57∶01, anti-B*13∶02, anti-B*49∶01, anti-B*50∶01, anti-A*25∶01, anti-B*15∶01, anti-B*15∶02 and anti-B*44∶02. There were 19 kinds of HLA-Ⅰ antibodies with MFI≥10 000, including anti-A*02∶01, anti-A*02∶03 and anti-A*02∶06. The CCI values were lower in HLA-Ⅰ antibody positive patients(n=46) than in HLA-I antibody negative patients(n=96) by Wilcoxon test (P<0.05). 【Conclusion】 Among tumor patients who received platelet transfusion, gender(female) was the risk factor for platelet antibody production. Platelet antibody positive patients were prone to platelet transfusion refractoriness. HLA-Ⅰ antibodies were the main platelet antibodies, suggesting that HLA-Ⅰ gene matching platelets could improve the efficacy of platelet transfusion.

Comparative study of alloimmunization against red cell antigens in sickle cell disease & thalassaemia major patients on regular red cell transfusion

Background & objectives: Sickle cell disease (SCD) patients require red cell transfusion during different clinical complications of the disease. Such patients are at a high risk for developing alloantibody against red cell antigens. From India, there are limited data available on alloantibody formation in multiply transfused SCD patients. The present study was thus undertaken to fill up this lacunae by looking at the development of red cell alloantibodies in SCD and ?-thalassaemia patients on regular transfusion. Methods: All sickle cell disease patients undergoing red cell transfusion between 2008 and 2016, were included. During this period, a large number of ?-thalassaemia major patients also underwent regular red cell transfusion. These thalassaemia patients were also included to compare the tendency of antibody formation between SCD and ?-thalassaemia major patients. All patients before regular transfusion were regularly assessed for the development of red cell antibody. Red cell antigen, antibody screen crossmatch and antibody identification were done using the standard technique. Results: A total of 138 patients with SCD aged between 4 and 53 yr (mean 17.6 yr) consisting of 83 males and 55 females (male:female, 1.5:1) along with 333 transfusion-dependent ?-thalassaemia patients were studied. Over the last eight years, 15 patients with SCD and four patients with thalassaemia developed alloantibody (P <0.001). Antibody specificity of their alloantibodies was against Rhc, RhE, Kell, Fya and Fyb only. Sickle cell disease patients with and without alloantibody required on the average 11.8 and 8.6 units of red cell concentrate, respectively (P <0.05). Interpretation & conclusions: About 11 per cent of the transfused sickle cells patients developed alloantibodies. The antibody specificity was restricted to Rh, Kell and Duffy blood group systems. Extended antigen matching involving Rh, Kell and Duffy antigens may prevent alloantibody in such patients.

Progress in the study of organ specific autoantibodies / 中华检验医学杂志

Organ specific autoantibodies have important clinical value in the diagnosis , differential diagnosis, disease assessment, treatment monitoring and prognosis of organ specific autoimmune disease .In recent years , many organ specific autoantibodies have been discovered , and although there are many challenges in clinical application , its important clinical value has been gradually recognized .

The profile and clinical significance of myositis-specific autoantibodies in Chinese patients with der-matomyositis / 中华风湿病学杂志

Objective The aim of this study is to analyze the prevalence of myositis-specific autoantibodies (MSAs) and to elucidate their associations with clinical features in Chinese dermatomyositis (DM) patients. Methods Four hundreds and twenty-seven DM patients were enrolled in this retrospective study. Clinical features and sera were collected. Twelve subtypes of MSAs were detected by commercial test kits. The correlations between MSAs and clinical phenotypes in DM patients were calculated by t test, Mann-Whitney U test or χ2 test. In order to clarify whether MSAs subsets would be independent factors of certain clinical feature or not, separate models were established to test the correlation via the Logistic regression analysis. Results The positivity of MSAs was 69.8% in 427 patients with DM. Anti-ARS, anti-MDA5 and anti-TIF1-γ antibodies were the three most common MSAs in the DM patients with positivity of 19.9%, 17.6%and 17.1% respectively. Different kinds of rash associated with MSAs subtypes by χ2 test. Certain MSAs subtype might be an independent factor for clinical features via logistic regression analysis. Interstitial lung disease (ILD) was observed more frequently in patients carrying anti-MDA5 [OR=5.266, 95%CI (2.522, 10.996), P<0.01] and anti-Jo-1 [OR=6.232, 95%CI (1.674, 23.199), P=0.006]. On the contrary, anti-Mi2 [OR=0.208, 95%CI (0.074, 0.580, P=0.003] and anti-TIF1-γ [OR=0.189, 95%CI (0.096, 0.370), P<0.01] were protective factors against developing ILD. Anti-TIF1-γ was an independent risk factor for cancer-associated myositis [OR=5.907, 95%CI (2.868, 12.168), P<0.01]. Anti-TIF1-γ[OR=2.789, 95%CI (1.594, 4.880) P<0.01], anti-NXP2 [OR=2.983, 95%CI (1.274, 6.982), P=0.012] and anti-SAE1 [OR=4.815, 95%CI (1.082, 21.424), P=0.039] could worsen dysphagic tendencies. In contrast, anti-MDA5 [OR=0.349, 95%CI (0.169, 0.720), P=0.004] might decrease the prevalence of this manifestation. Conclusion Patients with DM have a high frequency of MSAs. Some subtypes of MSAs are correlated with and may be independent factors of different clinical phenotypes. These indicated that MSAs can be useful biomarkers in monitoring the extramuscular features in DM patients.

The profile and clinical significance of myositis-specific autoantibodies in Chinese patients with der-matomyositis / 中华风湿病学杂志

Objective The aim of this study is to analyze the prevalence of myositis-specific autoantibodies (MSAs) and to elucidate their associations with clinical features in Chinese dermatomyositis (DM) patients. Methods Four hundreds and twenty-seven DM patients were enrolled in this retrospective study. Clinical features and sera were collected. Twelve subtypes of MSAs were detected by commercial test kits. The correlations between MSAs and clinical phenotypes in DM patients were calculated by t test, Mann-Whitney U test or χ2 test. In order to clarify whether MSAs subsets would be independent factors of certain clinical feature or not, separate models were established to test the correlation via the Logistic regression analysis. Results The positivity of MSAs was 69.8% in 427 patients with DM. Anti-ARS, anti-MDA5 and anti-TIF1-γ antibodies were the three most common MSAs in the DM patients with positivity of 19.9%, 17.6%and 17.1% respectively. Different kinds of rash associated with MSAs subtypes by χ2 test. Certain MSAs subtype might be an independent factor for clinical features via logistic regression analysis. Interstitial lung disease (ILD) was observed more frequently in patients carrying anti-MDA5 [OR=5.266, 95%CI (2.522, 10.996), P<0.01] and anti-Jo-1 [OR=6.232, 95%CI (1.674, 23.199), P=0.006]. On the contrary, anti-Mi2 [OR=0.208, 95%CI (0.074, 0.580, P=0.003] and anti-TIF1-γ [OR=0.189, 95%CI (0.096, 0.370), P<0.01] were protective factors against developing ILD. Anti-TIF1-γ was an independent risk factor for cancer-associated myositis [OR=5.907, 95%CI (2.868, 12.168), P<0.01]. Anti-TIF1-γ[OR=2.789, 95%CI (1.594, 4.880) P<0.01], anti-NXP2 [OR=2.983, 95%CI (1.274, 6.982), P=0.012] and anti-SAE1 [OR=4.815, 95%CI (1.082, 21.424), P=0.039] could worsen dysphagic tendencies. In contrast, anti-MDA5 [OR=0.349, 95%CI (0.169, 0.720), P=0.004] might decrease the prevalence of this manifestation. Conclusion Patients with DM have a high frequency of MSAs. Some subtypes of MSAs are correlated with and may be independent factors of different clinical phenotypes. These indicated that MSAs can be useful biomarkers in monitoring the extramuscular features in DM patients.

The serological characteristic and transfusion strategy in patients who got positive both in autoantibody test and direct Coombs test / 国际检验医学杂志

Objective To reduce the risk of blood transfusion ,and discuss the serological characteristic and cross‐match test of those patients who got positive both in autoantibody test and direct coombs test .Methods With absorption‐elution testing ,antibody screening cells and panel cells belong to different manufactures and batch numbers were applied to distinguish autoantibody from al‐loantibody within serum and reagent red cells for absorption test .Appropriate donors were selected to do cross matching test ;the specificity of autoantibody and alloantibody and the relationship between ABO blood type ,diseases ,anemic and the efficacy of blood transfusion were analysed .Results Among the 139 study subjects ,all of them were identified positive both in autoantibody and di‐rect antibody test ,including 20 cases just have autoantibody ,59 cases accompanied with Rh system or MNs system or Kidd system antibody ,47 with autoantibody except for alloantibody ,13 with drug resistant or other system antibody .Within the 221 cases of blood transfusion ,none of them has hemolytic transfusion reaction .We found a positive correlation(P0 .05) .Conclusion To those cases which were identified positive both in autoantibody and direct antibody test ,the degree of anaemia ,the numbers of transfusion and the efficacy of transfusion were associated with the intensity of agglutination of autoantibody and direct antibody .In order to de‐crease the risk of blood transfusion and make it highly efficient ,we should affirmed it if there is any autoantibody .In this process , an appropriate procedure and assay must be adopted .Besides ,relative antigen‐negative donors or high frequency local autoantibody need to be matched with corresponding the donors who have the same type of antigen .

Transfusion efficacy and safety evaluation in patients with non-specificity irregular antibody / 国际检验医学杂志

Objective To evaluate the transfusion efficacy and safety of patients with non‐specificity irregular antibodies .Meth‐ods A total of 19 cases of patients with non‐specificity irregular antibodies were analyzed ,then the transfusion efficacy and the in‐cidences of adverse transfusion reactions were investigated .Blood donor samples with the same ABO blood type of patients were randomly collected ,then blood cross‐matching was carried out .And the transfusion efficacy and safety of patients with non‐specific‐ity irregular antibody were evaluated when compatible blood were found by using both micro‐column gel technique and saline meth‐od .Results The non‐specificity irregular antibodies of the 19 cases of patients all were IgG irregular antibodies .The erythrocyte transfusion was carried out in the 19 cases of patients ,the total efficiency rate ,partial efficiency rate and inefficiency rate was 73 .7% ,26 .3% and 0 .0% ,respectively .No delayed hemolytic transfusion reactions were observed in any patient .Conclusion For patients with non‐specificity irregular antibodies ,the blood transfusion could be efficacy and safety when compatible blood samples are confirmed by using both micro‐column gel technique and saline method .

Interlaboratory Comparison of the Results of Lifecodes LSA Class I and Class II Single Antigen Kits for Human Leukocyte Antigen Antibody Detection

BACKGROUND: Although single antigen bead assays (SAB) are approved qualitative tests, the median fluorescence intensity (MFI) values obtained from SAB are frequently used in combination with quantitative significances for diagnostic purposes. To gauge the reproducibility of SAB results, we assessed the interlaboratory variability of MFI values using identical kits with reagents from the same lot and the manufacturer's protocol. METHODS: Six serum samples containing HLA-specific antibodies were analyzed at five laboratories by using Lifecodes LSA Class I and Class II SAB kits (Immucor, USA) from the same lot, according to the manufacturer's protocol. We analyzed the concordance of qualitative results according to distinct MFI cutoffs (1,000, 3,000, 5,000, and 10,000), and the correlation of quantitative MFI values obtained by the participating laboratories. The CV for MFI values were analyzed and grouped by mean MFI values from the five laboratories ( or =10,000). RESULTS: The categorical results obtained from the five laboratories exhibited concordance rates of 96.0% and 97.2% for detection of HLA class I and class II antibodies, respectively. The Pearson correlation coefficients for MFI values of class I and class II antibodies were between 0.947-0.991 and 0.992-0.997, respectively. The median CVs for the MFI values among five laboratories in the lower MFI range (<1,000) were significantly higher than those for the other MFI ranges (all P<0.01). CONCLUSIONS: Analysis of SAB performed in five laboratories using identical protocols and reagents from the same lot resulted in high levels of concordance and strong correlation of results.

Cutaneous melanoma: new advances in treatment

Cutaneous melanoma is a challenge to treat. Over the last 30 years, no drug or combination of drugs demonstrated significant impact to improve patient survival. From 1995 to 2000, the use of cytokines such as interferon and interleukin become treatment options. In 2011, new drugs were approved by the U.S. Food and Drug Administration, including peginterferon alfa-2b for patients with stage III disease, vemurafenib for patients with metastatic melanoma with the BRAF V600E mutation, and ipilimumab, a monoclonal antibody directed to the CTLA-4 T lymphocyte receptor, to combat metastatic melanoma in patients who do not have the BRAF V600E mutation. Both ipilimumab and vemurafenib showed results in terms of overall survival. Other trials with inhibitors of other genes, such as the KIT gene and MEK, are underway in the search for new discoveries. The discovery of new treatments for advanced or metastatic disease aims to relieve symptoms and improve patient quality of life.

Dynamic monitoring of donor specific antibodies in living-relative renal transplantation and early intervention / 中华器官移植杂志

Objective To analyze the clinical application of HLA donor specific antibodies (DSAs) detected by Luminex single antigen beads,and to discuss the impact of early intervention on renal function.Method In 64 cases of living-relative renal transplantation,DSA was detected using a Luminex single antigen assay before and after transplantation.The positive recipients were given large doses of intravenous irnmunoglobulin (IVIG) and increased doses of mycophenolate mofetil (MMF).The relationship between DSA and renal function was analyzed.Result DSA was negative in all recipients before transplantation.Ten cases of DSA positive recipients were found in HLA mismatch after transplantation.After the intervention,two cases of DSA positive recipients became negative,immunofluorescence intensity was decreased by more than 50％ in 6 cases,and no significant reduction was found in the other two cases.Antibody-mediated rejection (AMR) occurred in two cases of intervention ineffective recipients after 3 to 6 months and the renal function was impaired.Conclusion Dynamic monitoring of DSA using Luminex single antigen beads may timely predict changes of renal function.Early application of large doses of IVIG and increasing doses of MMF can reduce the incidence of AMR.

Protective role of triptolide in a rat model of antibody-mediated rejection in kidney transplantation / 中华器官移植杂志

Objective To investigate the protective effect and the mechanisms of triptolide on antibody-mediated rejection (AMR) in kidney transplantation.Method A rat model of AMR in kidney transplantation was constructed by sensitizing major histocompability complex (MHC) completely incompatible species of rats via blood transfusion before kidney transplantation.The protective effect of Triptolide on the renal function and pathologic injury was studied in this rat model.The levels of donor specific antibodies were measured by flow cytometry.C4d expression and macrophage infiltration were examined by immunohistochemistry and immunofluorescence.Result Triptolide could improve the function of transplanted kidney significantly (P＜0.05) and alleviate the pathologic injury.Triptolide can also reduce the levels of IgM,IgG2b and IgG2c in the serum of the recipient rats and inhibit the macrophage infiltration as well (P＜0.05).Conclusion Triptolide plays protective and therapeutic roles in AMR of kidney transplantation,which may be contributed to the inhibition of the production of donor specific antibody and the macrophage infiltration.

Smooth muscle antibodies and cryoglobulinemia are associated with advanced liver fibrosis in Brazilian hepatitis C virus carriers

Cryoglobulinemia and non-organ-specific-autoantibody are biomarkers of autoimmunity of the chronic infection caused by hepatitis C virus (HCV). In this work, we report the association between the presence of smooth muscle antibodies (SMA) and cryoglobulinemia and chronic liver disease in HCV carriers. Sixty-five untreated HCV patients, 38 women and 27 men were included in this study. Cryoglobulinemia was tested by cryoprecipitation, SMA by indirect fluorescent antibody test, and liver fibrosis and hepatocellular inflammation activity was investigated by histology of liver biopsy using the METAVIR score. The prevalence of SMA in the patients was 33.8 percent and cryoglobulinemia was demonstrated in 36.9 percent patients. Cryoglobulinemia and SMA seropositivity was associated with advanced fibrosis (p < 0.05). The presence of SMA and cryoglobulinemia was not associated with hepatocellular inflammation activity, age, carrier gender or HCV genotype. We concluded that liver biopsy should be recommended for HCV carriers that are seropositive for SMA or cryoglobulinemia.

Establishment of platelet antigen panel and its application in the identification of platelet specific antibodies / 中华检验医学杂志

Objective To establish the platelet antigen panel for identifying the specificity of platelet antibodies which cause platelet transfusion refractoriness and neonatal alloimmune thrombocytopenia and provide evidence for clinical therapy and platelet genotyping research.Methods Based on the frequency distribution of human platelet alloantigen (HPA)-1 to HPA-16 gene in China, the frequencies of HPA-1 to HPA-6,HPA-15 alleles in blood group O donors were genotyped by the polymerase chain reaction with sequence-specific primers (PCR-SSP) method, and suitable donors were chosen to establish platelet-specific antigen panel.Using the established platelet-specific antigen panel, the specificity of platelet antibodies caused by alloimmune reaction was identified by using simplified sensitized erythrocyte platelet serology assay (SEPSA).Results Eleven ptatelet donors with blood group O were chosen to establish platelet-specific antigen panel which can identify specificity of HPA-1 to HPA-6, HPA-15 antibodies.One case of HPA-4b (Penb) and two cases of HPA-15a (Govb) platelet specific antibodies were detected in 1 120 samples.Conclusion Identifying the specific platelet antibodies using platelet specific antigen panel has profound significance on increasing the safety and effectiveness of clinical platelet transfusion and prevention of neonatal alloimmune thrombocytopenia.

Characterization of a novel anti-human TNF-alpha murine monoclonal antibody with high binding affinity and neutralizing activity

In order to develop an anti-human TNF-alpha mAb, mice were immunized with recombinant human TNF-alpha. A murine mAb, TSK114, which showed the highest binding activity for human TNF-alpha was selected and characterized. TSK114 specifically bound to human TNF-alpha without cross-reactivity with the homologous murine TNF-alpha and human TNF-beta TSK114 was found to be of IgG1 isotype with kappa light chain. The nucleotide sequences of the variable regions of TSK114 heavy and light chains were determined and analyzed for the usage of gene families for the variable (V), diversity (D), and joining (J) segments. Kinetic analysis of TSK114 binding to human TNF-alpha by surface plasmon resonance technique revealed a binding affinity (KD) of ~5.3 pM, which is about 1,000- and 100-fold higher than those of clinically relevant infliximab (Remicade) and adalimumab (Humira) mAbs, respectively. TSK114 neutralized human TNF-alpha-mediated cytotoxicity in proportion to the concentration, exhibiting about 4-fold greater efficiency than those of infliximab and adalimumab in WEHI 164 cells used as an in vitro model system. These results suggest that TSK114 has the potential to be developed into a therapeutic TNF-alpha-neutralizing antibody with picomolar affinity.

Experience on Identification of Cross-Reactive Group Specificity Performed by Anti-human Globulin Panel Reactive Antibody Tests / 대한진단검사의학회지

BACKGROUND: Panel reactive antibody (PRA) is to screen and identify HLA antibody. Majority of antibody specificities in high-PRA are directed against cross reactive group (CREG). Thus, this study was to know the advantage of identifying CREG specificity and whether antibody specificities are changed according to CREG classification. METHODS: HLA class I antibodies were identified from 159 sera from 108 patients in Asan Medical Center, who had shown more than 5% PRA by anti-human globulin (AHG)-complement-dependent cytotoxicity (CDC). Tail analysis-based computer program was developed to identify specificities, applying both Rodey (R-ABC) and Takemoto (T-ABC) classification. The results were also compared with those obtained when without CREG application (ABC). RESULTS: Among 151 cases in which HLA specificities was identified, the frequency of CREG specificity was 22.5% in R-ABC and 27.2% in T-ABC. Eleven cases showed CREG specificities only in one classification. However, the individual antigen specificities in one hand were all included in the CREG identified in the other hand. CREG specificities in samples with PRA >50% (60%) were more frequently identified than those in samples with PRA < or =50% (9%) (in R-ABC, P<0.0001). Without applying CREG to interpretation, specificity was not identified in 9 cases. CONCLUSIONS: Application of CREG enhanced the rate of antibody identification. Antibody specificities of those cases where CREG specificities were different between Rodey and Takemoto classifications were almost the same when compared at the individual antigen level. Therefore, it was thought that it makes no difference to use any one of these two classifications in interpreting PRA.

Development of a Web-based Program for the Identification of Human Leukocyte Antigen Antibody Specificities / 대한진단검사의학회지

BACKGROUND: Panel reactive antibody (PRA) test is used to determine whether a patient awaiting transplantation is previously sensitized. Tail analysis algorithm is widely used to identify antibody specificities, but it is very difficult to perform manually. METHODS: To develop a web-based program, PHP (5.1.2), Apache (2.0.55), and MySQL (5.0.22) were used. Tail analysis algorithm was applied to identify specificities, which analyzed statistically 2 x 2 tables representing reactivities to broad antigens, splits and cross reactive groups (CREG). Exploiting two CREG classifications of Rodey (R) and Takemoto (T), antibody specificities were identified by 3 methods (ABC, R-ABC, T-ABC) simultaneously. Performance of the system was evaluated using 159 samples that showed > or =6 PRA% by a lymphocytotoxicity assay. RESULTS: A web-based system that can identify HLA antibody specificities was implemented on www.koreanhla.com. Among 159 samples tested, antibody specificities were identified in 151 (95.0 %), but not in 8 samples with PRA >97%. Among the 151 samples, 110 showed broad or split specificities and 41 CREG specificities. CONCLUSIONS: We developed a web-based computer program for the identification of HLA antibody specificities. Accessible to everyone on the internet, this program should be of help in sharing PRA results among laboratories.

The epitope analysis of HLA class I specific antibody in highly sensitized renal transplant recipients / 中华器官移植杂志

Objective To investigate the epitope of HLA class I specific antibody in the serum of highly sensitized patients (HSP). Methods The specificities and epitopes of HLA class I specific antibody from 54 HSP were monitored continually by enzyme linked immunosorbent assay (ELISA). Results 90.7 % of HSP had public epitopes of HLA class I antibody. The common public epitopes were 163E, 142 145TTKH, 83R, 71 74AQTD, 127K, 163R, 163L, 41T, 62 66RNTRN and 41 45ASPRT. The public epitopes were stable in the course of continual monitoring, and it could reflect antibody spectrum more accurately than antibody specificity analysis. Conclusion Most HSP produce HLA class I antibodies in a fairly predictable fashion. These antibodies direct against a small number of high frequency public amino acid residues with high immunogenicity. Analysis of public epitopes of HLA antibodies is very useful to predict acceptable mismatches.

Preparation and identification of Nogo-66 polyclonal antibodies / 中华检验医学杂志

Objective To prepare polyclonal antibodies of anti Nogo-66, the extracellular region of one central nervous system neurite regeneration inhibitor Nogo, which could be used to further identification and functional study of Nogo molecule.Methods Preparing rabbit anti rat Nogo-66 polyclonal antibodies with a purified Nogo-66 fusion protein expressed in E.coli system. Studying its specificity by Western-blot and immuno-histochemical techniques and identifying its biological activity in PC12 cells.Results The high titer (1∶[KG-*2]10 000) anti rat Nogo-66 polyclonal antibodies were obtained.This antibody could specifically recognize the Nogo protein expressed in E.coli system.Immuno-histochemical staining indicated that the Nogo was widely expressed in rat spinal cord neurons and oligodendrocytes.It could effectively block the neurite extensioninhibition of Nogo protein in PC12.Conclusion Successful preparation of anti rat Nogo polyclonal antibodies provides a useful tool in identification or further functional study of Nogo molecule.

The Frequency of Unexpected Antibodies in Blood Donors and Transfusion Candidates in Korea / 대한수혈학회지

BACKGROUND: A retrospective study was performed to estimate the frequency of red cell antibodies in blood donors (n=1,620,023) and transfusion candidates (SNUH n=12,111, YUMH n=26,665) for last 2 years (2000~2001). The results of the antibody screening and identification tests, the frequency and specificities of antibodies identified compared with blood centers and two hospitals had been used the different test methods each others. METHOD: Blood centers had been used tube and micro-plate method simultaneously with an in house and commercial panels. SNUH had been used micro-plate method using V plate with an in house and commercial panels. YUMH had been used gel agglutination technique (DiaMed ID System : DiaMed, Murten; Switzerland) since 1998. RESULTS: The frequencies of red blood cell antibodies were 0.26% (4,204 / 1,620,023 donor sera ), 0.11% (135 / 12,111 patient sera in SNUH) and 0.48% (128 / 26,665 patient sera in YUMH). Female donors and old ages showed higher frequency of red cell antibodies than male and young ages. Most of antibodies detected in donors were clinically less relevant antibodies such as Anti-Lea and Leb (38.9%), anti-P1 (18.1%), anti-H(IH) (8.4%), anti-M (6.2%) and so on. Clinically significant antibodies including Rh system antibodies (2.0%) were few, and composed only 12% of all the antibodies detected. In patients, clinically relevant antibodies including Rh antibodies (40.4% in SNUH, 71.9% in YUMH) were more frequently observed comparing with in donors. CONCLUSION: Antibodies found in donors were mostly clinically less relevant. Antibody screening method used in blood centers would be standardized. Blood banks using gel technique showed high detection rate of clinically significant antibodies comparing with facilities using other methods.