AIM: To perform a meta-analysis of the prevalence of anti-ribosomal P (aRP) antibodies in lupus psychosis, and the odds of psychosis in aRP-positive subjects.METHODS: We identifed articles by searching PubMed, Psy...AIM: To perform a meta-analysis of the prevalence of anti-ribosomal P (aRP) antibodies in lupus psychosis, and the odds of psychosis in aRP-positive subjects.METHODS: We identifed articles by searching PubMed, PsychInfo, and ISI, and the reference lists of identifed studies.RESULTS: Twenty-four studies met the inclusion criteria. Positive aRP antibodies were found in 51% (91 of 179 total cases) of cases of lupus psychosis. There was an almost 3.5-fold increased odds of psychosis in aRP-positive patients (OR = 3.46, 95%CI: 1.97-6.09, P 〈 0.001). The population attributable risk percentage was 36% for aRP antibodies.CONCLUSION: aRP antibodies are common in lupus psychosis, although the potential mechanism(s) underlying this association remain unclear. Given the overlap between the clinical presentation and risk factors for lupus psychosis and schizophrenia, further investigation of aRP antibodies in schizophrenia is warranted.展开更多
With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driv...With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driven by the deeper understanding of structure/function relationships.In monoclonal antibodies,many functions are regulated by N-glycans present in the constant region of the heavy chains and their mechanisms of action are not completely known.The importance of their function focuses analytical research efforts on the development of robust,accurate and fast methods to support drug development and quality control.Released N-glycan analysis is considered as the gold standard for glycosylation characterisation;however,it is not the only method for quantitative analysis of glycoform heterogeneity.In this study,ten different analytical workflows for N-glycan analysis were compared using four monoclonal antibodies.While observing good comparability between the quantitative results generated,it was possible to appreciate the advantages and disadvantages of each technique and to summarise all the observations to guide the choice of the most appropriate analytical workflow according to application and the desired depth of data generated.展开更多
A double antibodies additivity ELISA test was employed to identify the epltopes which can be recognized by monoclonal antibodies (McAbs) against IgM from B chronic lymphocyte leukemia (B-CLL). The computer grouping pr...A double antibodies additivity ELISA test was employed to identify the epltopes which can be recognized by monoclonal antibodies (McAbs) against IgM from B chronic lymphocyte leukemia (B-CLL). The computer grouping programme analysis showed that 4 and- isotypic MaAbs could be divided into two groups and 10 anti- idiotype McAbs could be divided into four groups. The result was consistent with that of the indirect sandwich ELISA and inhibition ELISA test. It suggested that there were at least 6 distinct IgM epitopes which can react specifically with 14 McAbs. Our study indicated that the combination of the additivity ELISA test and the computer grouping programme analysis is of help in studying the relationship of the structure and function of antigen.展开更多
In this review,we focus on providing basics and examples for each component of the protein therapeutic specifications to interested pharmacists and biopharmaceutical scientists with a goal to strengthen understanding ...In this review,we focus on providing basics and examples for each component of the protein therapeutic specifications to interested pharmacists and biopharmaceutical scientists with a goal to strengthen understanding in regulatory science and compliance.Pharmaceutical specifications comprise a list of important quality attributes for testing,references to use for test procedures,and appropriate acceptance criteria for the tests,and they are set up to ensure that when a drug product is administered to a patient,its intended therapeutic benefits and safety can be rendered appropriately.Conformance of drug substance or drug product to the specifications is achieved by testing an article according to the listed tests and analytical methods and obtaining test results that meet the acceptance criteria.Quality attributes are chosen to be tested based on their quality risk,and consideration should be given to the merit of the analytical methods which are associated with the acceptance criteria of the specifications.Acceptance criteria are set forth primarily based on efficacy and safety profiles,with an increasing attention noted for patient-centric specifications.Discussed in this work are related guidelines that support the biopharmaceutical specification setting,how to set the acceptance criteria,and examples of the quality attributes and the analytical methods from 60 articles and 23 pharmacopeial monographs.Outlooks are also explored on process analytical technologies and other orthogonal tools which are on-trend in biopharmaceutical characterization and quality control.展开更多
The ongoing coronavirus disease 2019(COVID-19)pandemic is a global public health crisis,causing social and economic disasters in many countries.In China,two-consecutive negative results of nucleic acid tests for SARS-...The ongoing coronavirus disease 2019(COVID-19)pandemic is a global public health crisis,causing social and economic disasters in many countries.In China,two-consecutive negative results of nucleic acid tests for SARS-CoV-2 from the respiratory samples are required to end the quarantine of COVID-19 patients.However,clinicians face a dilemma in case of patients with long-term viral shedding.This report described an unusual COVID-19 case who had persistent viral RNA positivity for more than 4 months after initial illness in the presence of low neutralizing antibodies,but without prolonged clinical symptoms.Multiple anti-viral drug treatments had no impact and there was no evidence of re-infection.When the patient was self-quarantined at home,no infection occurred to the three family members living with her for 15 to 19 days.Sputum viral culture in BSL-3 laboratory on the 102nd day after symptom onset was negative.From the 129th day on,8 continuous nucleic acid tests of sputum samples showed negative results.The patient was discharged on 137th days since symptom onset.In conclusion,viral RNA shedding in the sputum of the COVID-19 patient may last over 4 months.As no evidence shows the existence of infectious virus,two-consecutive negative nucleic acid tests may not be the prerequisite for ending quarantine of COVID-19 patients with prolonged viral shedding.展开更多
Positive nucleic acid(NA)results have been found in recovered and discharged COVID-19 patients,but the proportion is unclear.This study was designed to analyze the recurrent positive rate of NA results after consecuti...Positive nucleic acid(NA)results have been found in recovered and discharged COVID-19 patients,but the proportion is unclear.This study was designed to analyze the recurrent positive rate of NA results after consecutively negative results,and the relationship between the specific antibody production and positive NA rate.According to Strengthening the Reporting of Observational Studies in Epidemiology guidelines,data of inpatients in Sino-French New City Branch of Tongji Hospital between Jan.28 and Mar.6,2020 were collected.A total of 564 COVID-19 patients over 14 years old who received the examinations of NA and antibodies against SARS-CoV-2 were included.Days of viral shedding and specific antibodies were recorded and assessed.Among NA tests in respiratory samples(throat swabs,nasopharyngeal swabs,sputum and flushing fluid in alveoli),the patients with all-negative NA results accounted for 17.20%,those with single-positive results for 46.63%,and those with multiple-positive results for 36.17%respectively.Besides,the recurrent positive NA results after consecutively negative results appeared in 66 patients(11.70%).For multiple-positive patients,median viral shedding duration was 20 days(range:1 to 57 days).Of the 205 patients who received 2 or more antibody tests,141(68.78%)had decreased IgG and IgM concentrations.IgM decreased to normal range in 24 patients,with a median of 44 days from symptom onset.Viral shedding duration was not significantly correlated with gender,age,disease severity,changes in pulmonary imaging,and antibody concentration.It is concluded that antibody level and antibody change had no significant correlation with the positive rate of NA tests and the conversion rate after continuous negative NA tests.In order to reduce the recurrent positive proportion after discharge,3 or more consecutive negative NA test results with test interval more than 24 h every time are suggested for the discharge or release from quarantine.展开更多
Background: Measuring fasting C-peptide (FCP) and antibodies against Glutamic acid decarboxylase (GADA) and Islet cell antibodies (ICA) are not so commonly explored in children and young adults. Objectives: To assess ...Background: Measuring fasting C-peptide (FCP) and antibodies against Glutamic acid decarboxylase (GADA) and Islet cell antibodies (ICA) are not so commonly explored in children and young adults. Objectives: To assess the levels of FCP, GADA and ICA in subjects below the age of 25 years with DM and compare their levels to differentiate between Autoimmune and Non-Autoimmune Type 1 DM. Methodology: Blood samples of 93 subjects diagnosed with DM, reporting to the tertiary care hospital, were analysed for ICA, GADA and FCP. Receiver operating characteristics (ROC) curves were analysed to check the ability of autoimmune markers, BMI and C-peptide to differentiate between Autoimmune (Ai) and Non-Autoimmune (NonAi) diabetes. Results: 30/93 (32.2%) were positive for anti-GAD ab and/or ICA and categorised as Autoimmune (Ai), the most common antibody being, anti-GAD ab (80%) in them. The level of FCP among Ai compared to NonAi, was significantly low (p 20.75 nmol/l) as a very dependable test for diagnosing Ai, Type 1 DM, in children and young adults. Its sensitivity and specificity are in the range of 86.2% and 96.8% respectively. Low level of C-peptide (Conclusion: This study revealed predominant positivity for anti-GAD ab (80%) among Ai+ patients. ROC analysis shows GADA above 20.75 nmol/l and Fasting C-peptide < 0.36 nmol/l as a good indicator for diagnosing Ai in children and young adults.展开更多
AIM To prepare hybridoma cell lines which secrete anti HCV recombinant NS3 and NS5 proteins′ monoclonal antibodies, and to evaluate their usage in the study of the distribution of HCV NS3 and NS5 antigen in liver t...AIM To prepare hybridoma cell lines which secrete anti HCV recombinant NS3 and NS5 proteins′ monoclonal antibodies, and to evaluate their usage in the study of the distribution of HCV NS3 and NS5 antigen in liver tissues. METHODS The hybridoma cell lines were raised using the spleen cells of BALB/C mouse immunized with recombinant NS3 and NS5 proteins according to the conventional protocols. The antibody secreting cells were screened using solid phase ELISA and cloned by limited dilution method. In order to determine the specificity of these hybridoma cell lines, the culture supernatant of these cells was western blot assayed with expression and nonexpresion E. coli and ELISA with other antigens, including HCV core and NS3 and HBsAg. Immunohistochemistry of 51 cases paraffin embedded liver tissues was performed to determine the distribution of HCV NS5 antigen as well as NS3 antigen in liver tissues. RESULTS Eight hybridoma cell lines secreting monoclonal antibodies against HCV NS3 and NS5 proteins were raised. They are named 2B6, 2F3, 3D8, 3D9, 8B2, 6F11, 4C6 and 7D9. Among them only 2B6 against NS3 protein can react with the polypipetides of C7 that is another recombinant polypipetides of NS3 gene. Others have no reaction with HCV core and HBsAg of HBV, and there is no cross reaction between NS3Ag and anti NS5Ag McAb and between NS5Ag and anti NS3 McAb. The immunohistochemistry results indicate that no HCV antigen was detected in the specimens of HBV infection in 20 cases. In 31 HCV infected specimens the positive rate of NS3Ag and NS5Ag are 51 6% (16/31) and 54 9% (17/31), respectively. There were six pure HCV infected specimens in these 31 specimens and half of them were HCV NS3Ag and NS5Ag positive. In the co infection of HBV and HCV group the positive rate of NS3Ag and NS5Ag were 52% (13/25) and 56% (14/25), respectively, almost the same with that of pure HCV infected group. The positive rates of HCV antigens were 70 6% (12/17) and 76 5% (13/17) in CAC patients. CONCLUSION The monoclonal antibodies we prepared are specific to the recombinant HCV NS3 and NS5 proteins and can be used in the clinical immunohistochemistry diagnosis.展开更多
Hepatitis C virus (HCV) is a major cause of hepatitis world-wide. The majority of infected individuals develop chronic hepatitis which can then progress to liver cirrhosis and hepatocellular carcinoma. Spontaneous vir...Hepatitis C virus (HCV) is a major cause of hepatitis world-wide. The majority of infected individuals develop chronic hepatitis which can then progress to liver cirrhosis and hepatocellular carcinoma. Spontaneous viral clearance occurs in about 20%-30% of acutely infected individuals and results in resolution of infection without sequaelae. Both viral and host factors appear to play an important role for resolution of acute infection. A large body of evidence suggests that a strong, multispecific and long-lasting cellular immune response appears to be important for control of viral infection in acute hepatitis C. Due too the lack of convenient neutralization assays, the impact of neutralizing responses for control of viral infection had been less defined. In recent years, the development of robust tissue culture model systems for HCV entry and infection has finally allowed study of antibody-mediated neutralization and to gain further insights into viral targets of host neutralizing responses. In addition, detailed analysis of antibody-mediated neutralization in individual patients as well as cohorts with well defined viral isolates has enabled the study of neutralizing responses in the course of HCV infection and characterization of the impact of neutralizing antibodiesfor control of viral infection. This review will summarize recent progress in the understanding of the molecular mechanisms of antibody-mediated neutralization and its impact for HCV pathogenesis.展开更多
AIM: To establish a convenient immunoassay method based on recombinant antigen preS1(21-119aa) to detect anti-preS1 antibodies and evaluate the clinical significance of antibodies in hepatitis B. METHODS: The expressi...AIM: To establish a convenient immunoassay method based on recombinant antigen preS1(21-119aa) to detect anti-preS1 antibodies and evaluate the clinical significance of antibodies in hepatitis B. METHODS: The expression plasmid pET-28a-preS1 was constructed, and a large quantity of preS1(21-119aa) fragment of the large HBsAg protein was obtained. The preS1 fragment purified by Ni(2+)-IDA affinity chromatography was used as coated antigen to establish the indirect ELISA based on streptavidin-biotin system for detection of the anti-preS1 antibodies in sera from HBV-infected patients. For follow-up study, serial sera were collected during the clinical course of 21 HBV-infected patients and anti-preS1 antibodies, preS1 antigen, HBV-DNA and other serological HBV markers were analyzed. RESULTS: preS1(21-119aa) fragment was highly expressed from the plasmid pET-28a-preS1 in a soluble form in E.Coli (30mg.L(-1)), and easily purified to high purity over 90% by one step of Ni(2+)-IDA-sepharose 6B affinity chromatography. The purity and antigenicity of the purified preS1(21-119aa) protein was determined by 150g.L(-1) SDS-PAGE, Western blot and a direct ELISA. Recombinant preS1(21-119aa) protein was successfully applied in the immunoassay which could sensitively detect the anti-preS1 antibodies in serum specimens of acute or chronic hepatitis B patients. Results showed that more than half of 19 acute hepatitis B patients produced anti-preS1 antibodies during recovery of the disease, however, the response was only found in a few of chronic patients. In the clinical follow-up study of 11 patients with anti-preS1 positive serological profile, HBsAg and HBV-DNA clearance occurred in 6 of 10 acute hepatitis B patients in 5-6 months, and seroconversion of HBeAg and disappearance of HBV-DNA occurred in 1 chronic patients treated with lavumidine, a antiviral agent. CONCLUSION: The high-purity preS1(21-119aa) coated antigen was successfully prepared by gene expression and affinity chromatography. Using this antigen, a conveniently detective system of anti-preS1 antibodies in sera was established. Preliminarily clinical trial the occurrence of anti-preS1 antibodies in acute hepatitis B patients suggests the clearance of HBV from serum in a short-term time, and anti-preS1 positive in chronic patients means health improvement or recovery from the disease.展开更多
The results of research work completed in recent years in the authors’ laboratory withmonoclonal antibodies (McAbs) against herpes simplex virus (HSV) are reported in this paper as fol-lows.(1) Eighteen hybridoma cel...The results of research work completed in recent years in the authors’ laboratory withmonoclonal antibodies (McAbs) against herpes simplex virus (HSV) are reported in this paper as fol-lows.(1) Eighteen hybridoma cell lines steadily producing McAbs against HSV were established byhybridoma technic.(2) The mechanisms of neutralization in vitro and animal protection in vivo medi-ated by different McAbs were investigated.(3) A method of detecting virus antigua was developedwith the McAbs.(4) Tne isolates of virus were typed and antigenically analysed by type-specificand type-common McAbs.(5) HSV antigen in clinical spectimens was detected and directly typed byan ELISA method coating with type-specific and type-common McAb.(6) The target antigens ofMcAbs were identified,purified,and their genes were localized.(7) Experimental rabbitHSV keratitis was treated with McAbs.展开更多
Japanese encephalitis (JE) is a central nervous system disease caused by Japanese encephalitis virus (JEV), which can infect human and a variety of animals and cause irreversible nerve damages. NS3 protein plays a...Japanese encephalitis (JE) is a central nervous system disease caused by Japanese encephalitis virus (JEV), which can infect human and a variety of animals and cause irreversible nerve damages. NS3 protein plays an important role in the process of JEV polyprotein hydrolysis, which is essential for JEV replication. Therefore, NS3 protein may be used as a potential drug target to treat Japanese encephalitis. In this study, the pET-28a-NS3 plasmid was successfully constructed and expressed in E. coli BL21 ( DE3 ) under IPTG induction. The molecular weight of the expressed recombinant protein was 55 ku, which was consistent with the expected result. The positive serum was prepared by immunizing BALB/c mice with NS3 protein and identified by indirect immunofluorescence (IFA). The results showed that there was a fluorescence reaction between the prepared positive serum of NS3 protein and cells infected with JEV.展开更多
Monoclonal antibodies (McAb) to Hantavirus (HTV)were derived from the hybridoma fused with Lou/c plasmocytoma rat cells (IR983F) and spleen cells from lou/c inbred rat immunized with HTV Chen strain. Eleven cell lines...Monoclonal antibodies (McAb) to Hantavirus (HTV)were derived from the hybridoma fused with Lou/c plasmocytoma rat cells (IR983F) and spleen cells from lou/c inbred rat immunized with HTV Chen strain. Eleven cell lines producing monoclonal antibodies direc展开更多
To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from...To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from the total RNA of H9 cells. The resulting DNA construct was cloned into a prokaryotic expression vector (pET-32a). Recombinant pET-vif and pET-APOBEC3G were expressed respectively in Eserichia coli BL21 (DE3) as an insoluble protein. The vector also contained a six-histidine tag at the C-terminus for convenient purification and detection. To express and purify the HIV-1 Vif and hAPOBEC3G in E.coli cells, the accuracy of inserted gene and specificity of proteins were detected by the two enzyme digestion method, SDS-PAGE, and Western blotting. Rabbits were then immunized by Vif or APOBEC3G protein and serum samples were tested by indirect ELISA to determine the level of antibodies. Immunoenzyme and immunofluorescence assays were performed to identify the specificity of polyclonal antibodies. The titer of the anti-Vif antibodies was 1:204800, and that of the anti-APOBEC3G antibodies was 1:102400. Thus the antibodies could detect the antigen expression in the cells, demonstrating that fusion proteins with high purity and their corresponding polyclonal antibodies with high titer and specificity were achieved.展开更多
Objective To investigate the characteristics and outcome of glomerulonephritis in patients with both antineutrophil cytoplasmic antibody and anti-glomerular basement membrane antibody.Methods The sera of 23 antiGBM gl...Objective To investigate the characteristics and outcome of glomerulonephritis in patients with both antineutrophil cytoplasmic antibody and anti-glomerular basement membrane antibody.Methods The sera of 23 antiGBM glomerulonephritis patients were collected and were tested for ANCA respectively.Characteristics and outcome of patients with coexisting anti-GBM antibody展开更多
Objective To define risk stratification and guide optimal surgical timing of perioperative viral respiratory infection(VRI)in children with cardiac surgery.Methods Retrospective study with propensity score-matched ana...Objective To define risk stratification and guide optimal surgical timing of perioperative viral respiratory infection(VRI)in children with cardiac surgery.Methods Retrospective study with propensity score-matched analysis.A total of 2,831 patients had performed RespPCR testing,and finally there were 2,740 negative RespPCR patients and 91 positive RespPCR patients.展开更多
AIM To provide a clear understanding of viral hepatitis epidemiology and their clinical burdens in Somalia.METHODS A systematic review and meta-analysis was conducted as Preferred Reporting Items for Systematic Review...AIM To provide a clear understanding of viral hepatitis epidemiology and their clinical burdens in Somalia.METHODS A systematic review and meta-analysis was conducted as Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. A comprehensive literature search of published studies on viral hepatitis was performed from 1977-2016 in Pub Med, Google Scholar, Science Direct, World Health Organization African Index Medicus and the Africa Journals Online databases, as well as on the Ministry of Health website. We also captured unpublished articles that were not available on online systems.RESULTS Twenty-nine studies from Somalia and Somali immigrants(United Kingdom,United States,Italy,Libya)with a combined sample size for each type of viral hepatitis[hepatitis A virus(HAV):1564,hepatitis B virus(HBV):8756,hepatitis C virus(HCV):6257,hepatitis D virus(HDV):375 and hepatitis E virus(HEV):278]were analyzed.The overall pooled prevalence rate of HAV was 90.2%(95%CI:77.8%to 96%).The HAV prevalence among different age groups was as follows:<1 year old,61.54%(95%CI:40.14%to79.24%);1-10 years old,91.91%(95%CI:87.76%to94.73%);11-19 years old,96.31%(95%CI:92.84%to 98.14%);20-39 years old,91.3%(95%CI:83.07%to 95.73%);and>40 years old,86.96%(95%CI:75.68%to 93.47%).The overall pooled prevalence of HBV was 18.9%(95%CI:14%to 29%).The overall pooled prevalence among subgroups of HBV was20.5%(95%CI:5.1%to 55.4%)in pregnant women;5.7%(95%CI:2.7%to 11.5%)in children;39.2%(95%CI:33.4%to 45.4%)in patients with chronic liver disease,including hepatocellular carcinoma(HCC);7.7%(95%CI:4.2%to 13.6%),12.4%(95%CI:6.3%to 23.0%)and 11.8%(95%CI:5.3%to 24.5%)in age groups<20 years old,20-39 years old and>40years old,respectively.The HBV prevalence among risk groups was 20%(95%CI:7.19%to 44.64%)in female prostitutes,21.28%(95%CI:7.15%to48.69%)in hospitalized adults,5.56%(95%CI:0.99%to 25.62%)in hospitalized children,60%(95%CI:31.66%to 82.92%)in patients with acute hepatitis,33.55%(95%CI:14.44%to 60.16%)in patients with ancylostomiasis,12.34%(95%CI:7.24%to 20.26%)in patients with leprosy and 20.19%(95%CI:11.28%to33.49%)in schistosomiasis patients.The overall pooled prevalence of HCV was estimated as 4.84%(95%CI:3.02%to 7.67%).The prevalence rates among blood donors,risk groups,children and patients chronic liver disease(including HCC)was 0.87%(95%CI:0.33%to 2.30%),2.43%(95%CI:1.21%to 4.8%),1.37%(95%CI:0.76%to 2.46%)and 29.82%(95%CI:15.84%to 48.98%),respectively.The prevalence among genotypes of HCV was 21.9%(95%CI:15.36%to 30.23%)in genotype 1,0.87%(95%CI:0.12%to 5.9%)in genotype 2,25.21%(95%CI:18.23%to 33.77%)in genotype 3,46.24%(95%CI:37.48%to 55.25%)in genotype 4,2.52%(95%CI:0.82%to7.53%)in genotype 5,and 1.19%(95%CI:0.07%to16.38%)in genotype 6.The overall pooled prevalence of HDV was 28.99%(95%CI:16.38%to 45.96%).The HDV prevalence rate among patients with chronic liver disease,including HCC,was 43.77%(95%CI:35.09%to 52.84%).The overall pooled prevalence of HEV was46.86%(95%CI:5.31%to 93.28%).CONCLUSION Our study demonstrates a high prevalence of all forms of viral hepatitis in Somalia and it also indicates that chronic HBV was the commonest cause of chronic liver disease.This highlights needs for urgent public health interventions and strategic policy directions to controlling the burden of the disease.展开更多
Donor human leukocyte antigen(HLA)-specific antibodies(DSA) play an important role in solid organ transplantation. Preexisting IgG isotype DSA are considered a risk factor for antibody mediated rejection, graft failur...Donor human leukocyte antigen(HLA)-specific antibodies(DSA) play an important role in solid organ transplantation. Preexisting IgG isotype DSA are considered a risk factor for antibody mediated rejection, graft failure or graft loss. The post-transplant development of DSA depends on multiple factors including immunogenicity of mismatched antigens, HLA class Ⅱ typing of the recipient, cytokine gene polymorphisms, and cellular immunoregulatory mechanisms. De novo developed antibodies require special attention because not all DSA have equal clinical significance. Therefore, it is important for transplant clinicians and transplant immunologists to accurately characterize DSA. In this review, the contemporary immunological techniques for detection and characterization of anti-HLA antibodies and their pitfalls are described.展开更多
<strong>Background:</strong> Worldwide, prostatic adenocarcinoma is the most common tumour type among men. <strong>Aim:</strong> The aim of the present investigation was to develop a computer p...<strong>Background:</strong> Worldwide, prostatic adenocarcinoma is the most common tumour type among men. <strong>Aim:</strong> The aim of the present investigation was to develop a computer program to identify normal prostate biopsies and distinguish them from biopsies showing premalignant alterations (LGPIN, HGPIN) and adenocarcinoma. <strong>Method:</strong> Prostate biopsies (n = 2094) taken from 191 consecutive men during 2016 were stained with triple immunehistochemisty (antibodies to AMACRA, p63 and CK 5). Digital images of the biopsies were obtained with a scanning microscope and used to develop an automatic computer program (CelldaTM), intended to identify the morphological alterations. Visual microscopic finding was used as a reference. <strong>Result:</strong> Of the 191 men, 121 (63.4%) were diagnosed as having prostate adenocarcinoma and 70 (36.6%) as having no malignancy on the basis of the visual microscopy. In comparison, computer analysis identified 134 (70.2%) men with malignant disease and 57 (29.8%) with non-malignant disease after exclusion of artifacts, which constituted 10.4% of areas (indicated as malignant disease). Discrepant results were recorded in 15 (7.9%) men, and in 14 of these cases, HGPIN and areas suggestive of early invasion were common. Thus, it was uncertain whether these cases should be regarded as malignant or not. The agreement between the visual examination and the computer analysis was 92.1% (kappa value 0.823, sensitivity 99.2 and specificity was 0.80). <strong>Conclusion:</strong> It seems that computer analysis could serve as an adjunct to simplify and shorten the diagnostic procedure, first of all by ensuring that normal prostate biopsies are sorted out from those sent for visual microscopic evaluation.展开更多
ELISA method was used to detect antisperm antibodies in cervix mucus. Out of 400 women whose husbands had sperms, 92 were antisperm antibody positive (23% ). 42 of 200 women with azoospermic partners were antisperm an...ELISA method was used to detect antisperm antibodies in cervix mucus. Out of 400 women whose husbands had sperms, 92 were antisperm antibody positive (23% ). 42 of 200 women with azoospermic partners were antisperm antibody positive (21% ). No difference was found between the two groups. Hence, antisperm antibodies in azoospermic men's wives may be caused by sperm'sadhering antigens in semina.展开更多
基金Supported by The National Institute of Mental Health(1K23MH098014)Georgia Regents Universityhonoraria from Medscape,Insight Consulting,and Decision Resources Group
文摘AIM: To perform a meta-analysis of the prevalence of anti-ribosomal P (aRP) antibodies in lupus psychosis, and the odds of psychosis in aRP-positive subjects.METHODS: We identifed articles by searching PubMed, PsychInfo, and ISI, and the reference lists of identifed studies.RESULTS: Twenty-four studies met the inclusion criteria. Positive aRP antibodies were found in 51% (91 of 179 total cases) of cases of lupus psychosis. There was an almost 3.5-fold increased odds of psychosis in aRP-positive patients (OR = 3.46, 95%CI: 1.97-6.09, P 〈 0.001). The population attributable risk percentage was 36% for aRP antibodies.CONCLUSION: aRP antibodies are common in lupus psychosis, although the potential mechanism(s) underlying this association remain unclear. Given the overlap between the clinical presentation and risk factors for lupus psychosis and schizophrenia, further investigation of aRP antibodies in schizophrenia is warranted.
文摘With the size of the biopharmaceutical market exponentially increasing,there is an aligned growth in the importance of data-rich analyses,not only to assess drug product safety but also to assist drug development driven by the deeper understanding of structure/function relationships.In monoclonal antibodies,many functions are regulated by N-glycans present in the constant region of the heavy chains and their mechanisms of action are not completely known.The importance of their function focuses analytical research efforts on the development of robust,accurate and fast methods to support drug development and quality control.Released N-glycan analysis is considered as the gold standard for glycosylation characterisation;however,it is not the only method for quantitative analysis of glycoform heterogeneity.In this study,ten different analytical workflows for N-glycan analysis were compared using four monoclonal antibodies.While observing good comparability between the quantitative results generated,it was possible to appreciate the advantages and disadvantages of each technique and to summarise all the observations to guide the choice of the most appropriate analytical workflow according to application and the desired depth of data generated.
文摘A double antibodies additivity ELISA test was employed to identify the epltopes which can be recognized by monoclonal antibodies (McAbs) against IgM from B chronic lymphocyte leukemia (B-CLL). The computer grouping programme analysis showed that 4 and- isotypic MaAbs could be divided into two groups and 10 anti- idiotype McAbs could be divided into four groups. The result was consistent with that of the indirect sandwich ELISA and inhibition ELISA test. It suggested that there were at least 6 distinct IgM epitopes which can react specifically with 14 McAbs. Our study indicated that the combination of the additivity ELISA test and the computer grouping programme analysis is of help in studying the relationship of the structure and function of antigen.
基金supported by the Grant for Development of New Faculty Staff,Ratchadaphiseksomphot Endowment Fund,Chula-longkorn University,Thailand(Grant No.:DNS64_047_33_003_1 to Patanachai K.Limpikirati)Grant for Development of New Scholar,Office of the Permanent Secretary,Ministry of Higher Ed-ucation,Science,Research and Innovation,Thailand(Grant No.:RGNS64_012 to Patanachai K.Limpikirati).
文摘In this review,we focus on providing basics and examples for each component of the protein therapeutic specifications to interested pharmacists and biopharmaceutical scientists with a goal to strengthen understanding in regulatory science and compliance.Pharmaceutical specifications comprise a list of important quality attributes for testing,references to use for test procedures,and appropriate acceptance criteria for the tests,and they are set up to ensure that when a drug product is administered to a patient,its intended therapeutic benefits and safety can be rendered appropriately.Conformance of drug substance or drug product to the specifications is achieved by testing an article according to the listed tests and analytical methods and obtaining test results that meet the acceptance criteria.Quality attributes are chosen to be tested based on their quality risk,and consideration should be given to the merit of the analytical methods which are associated with the acceptance criteria of the specifications.Acceptance criteria are set forth primarily based on efficacy and safety profiles,with an increasing attention noted for patient-centric specifications.Discussed in this work are related guidelines that support the biopharmaceutical specification setting,how to set the acceptance criteria,and examples of the quality attributes and the analytical methods from 60 articles and 23 pharmacopeial monographs.Outlooks are also explored on process analytical technologies and other orthogonal tools which are on-trend in biopharmaceutical characterization and quality control.
文摘The ongoing coronavirus disease 2019(COVID-19)pandemic is a global public health crisis,causing social and economic disasters in many countries.In China,two-consecutive negative results of nucleic acid tests for SARS-CoV-2 from the respiratory samples are required to end the quarantine of COVID-19 patients.However,clinicians face a dilemma in case of patients with long-term viral shedding.This report described an unusual COVID-19 case who had persistent viral RNA positivity for more than 4 months after initial illness in the presence of low neutralizing antibodies,but without prolonged clinical symptoms.Multiple anti-viral drug treatments had no impact and there was no evidence of re-infection.When the patient was self-quarantined at home,no infection occurred to the three family members living with her for 15 to 19 days.Sputum viral culture in BSL-3 laboratory on the 102nd day after symptom onset was negative.From the 129th day on,8 continuous nucleic acid tests of sputum samples showed negative results.The patient was discharged on 137th days since symptom onset.In conclusion,viral RNA shedding in the sputum of the COVID-19 patient may last over 4 months.As no evidence shows the existence of infectious virus,two-consecutive negative nucleic acid tests may not be the prerequisite for ending quarantine of COVID-19 patients with prolonged viral shedding.
文摘Positive nucleic acid(NA)results have been found in recovered and discharged COVID-19 patients,but the proportion is unclear.This study was designed to analyze the recurrent positive rate of NA results after consecutively negative results,and the relationship between the specific antibody production and positive NA rate.According to Strengthening the Reporting of Observational Studies in Epidemiology guidelines,data of inpatients in Sino-French New City Branch of Tongji Hospital between Jan.28 and Mar.6,2020 were collected.A total of 564 COVID-19 patients over 14 years old who received the examinations of NA and antibodies against SARS-CoV-2 were included.Days of viral shedding and specific antibodies were recorded and assessed.Among NA tests in respiratory samples(throat swabs,nasopharyngeal swabs,sputum and flushing fluid in alveoli),the patients with all-negative NA results accounted for 17.20%,those with single-positive results for 46.63%,and those with multiple-positive results for 36.17%respectively.Besides,the recurrent positive NA results after consecutively negative results appeared in 66 patients(11.70%).For multiple-positive patients,median viral shedding duration was 20 days(range:1 to 57 days).Of the 205 patients who received 2 or more antibody tests,141(68.78%)had decreased IgG and IgM concentrations.IgM decreased to normal range in 24 patients,with a median of 44 days from symptom onset.Viral shedding duration was not significantly correlated with gender,age,disease severity,changes in pulmonary imaging,and antibody concentration.It is concluded that antibody level and antibody change had no significant correlation with the positive rate of NA tests and the conversion rate after continuous negative NA tests.In order to reduce the recurrent positive proportion after discharge,3 or more consecutive negative NA test results with test interval more than 24 h every time are suggested for the discharge or release from quarantine.
文摘Background: Measuring fasting C-peptide (FCP) and antibodies against Glutamic acid decarboxylase (GADA) and Islet cell antibodies (ICA) are not so commonly explored in children and young adults. Objectives: To assess the levels of FCP, GADA and ICA in subjects below the age of 25 years with DM and compare their levels to differentiate between Autoimmune and Non-Autoimmune Type 1 DM. Methodology: Blood samples of 93 subjects diagnosed with DM, reporting to the tertiary care hospital, were analysed for ICA, GADA and FCP. Receiver operating characteristics (ROC) curves were analysed to check the ability of autoimmune markers, BMI and C-peptide to differentiate between Autoimmune (Ai) and Non-Autoimmune (NonAi) diabetes. Results: 30/93 (32.2%) were positive for anti-GAD ab and/or ICA and categorised as Autoimmune (Ai), the most common antibody being, anti-GAD ab (80%) in them. The level of FCP among Ai compared to NonAi, was significantly low (p 20.75 nmol/l) as a very dependable test for diagnosing Ai, Type 1 DM, in children and young adults. Its sensitivity and specificity are in the range of 86.2% and 96.8% respectively. Low level of C-peptide (Conclusion: This study revealed predominant positivity for anti-GAD ab (80%) among Ai+ patients. ROC analysis shows GADA above 20.75 nmol/l and Fasting C-peptide < 0.36 nmol/l as a good indicator for diagnosing Ai in children and young adults.
文摘AIM To prepare hybridoma cell lines which secrete anti HCV recombinant NS3 and NS5 proteins′ monoclonal antibodies, and to evaluate their usage in the study of the distribution of HCV NS3 and NS5 antigen in liver tissues. METHODS The hybridoma cell lines were raised using the spleen cells of BALB/C mouse immunized with recombinant NS3 and NS5 proteins according to the conventional protocols. The antibody secreting cells were screened using solid phase ELISA and cloned by limited dilution method. In order to determine the specificity of these hybridoma cell lines, the culture supernatant of these cells was western blot assayed with expression and nonexpresion E. coli and ELISA with other antigens, including HCV core and NS3 and HBsAg. Immunohistochemistry of 51 cases paraffin embedded liver tissues was performed to determine the distribution of HCV NS5 antigen as well as NS3 antigen in liver tissues. RESULTS Eight hybridoma cell lines secreting monoclonal antibodies against HCV NS3 and NS5 proteins were raised. They are named 2B6, 2F3, 3D8, 3D9, 8B2, 6F11, 4C6 and 7D9. Among them only 2B6 against NS3 protein can react with the polypipetides of C7 that is another recombinant polypipetides of NS3 gene. Others have no reaction with HCV core and HBsAg of HBV, and there is no cross reaction between NS3Ag and anti NS5Ag McAb and between NS5Ag and anti NS3 McAb. The immunohistochemistry results indicate that no HCV antigen was detected in the specimens of HBV infection in 20 cases. In 31 HCV infected specimens the positive rate of NS3Ag and NS5Ag are 51 6% (16/31) and 54 9% (17/31), respectively. There were six pure HCV infected specimens in these 31 specimens and half of them were HCV NS3Ag and NS5Ag positive. In the co infection of HBV and HCV group the positive rate of NS3Ag and NS5Ag were 52% (13/25) and 56% (14/25), respectively, almost the same with that of pure HCV infected group. The positive rates of HCV antigens were 70 6% (12/17) and 76 5% (13/17) in CAC patients. CONCLUSION The monoclonal antibodies we prepared are specific to the recombinant HCV NS3 and NS5 proteins and can be used in the clinical immunohistochemistry diagnosis.
基金Inserm, France Université Louis Pasteur, France+3 种基金the European Union (Virgil Network of Excellence)the DeutscheForschungsgemeinschaft (Ba1417/11-1), Germanythe ANRchair of excellence program and ANRS, FranceInserm "PosteVert" research fellowship in the framework of Inserm EuropeanAssociated Laboratory Inserm U748-Department of Medicine Ⅱ,University of Freiburg, Germany
文摘Hepatitis C virus (HCV) is a major cause of hepatitis world-wide. The majority of infected individuals develop chronic hepatitis which can then progress to liver cirrhosis and hepatocellular carcinoma. Spontaneous viral clearance occurs in about 20%-30% of acutely infected individuals and results in resolution of infection without sequaelae. Both viral and host factors appear to play an important role for resolution of acute infection. A large body of evidence suggests that a strong, multispecific and long-lasting cellular immune response appears to be important for control of viral infection in acute hepatitis C. Due too the lack of convenient neutralization assays, the impact of neutralizing responses for control of viral infection had been less defined. In recent years, the development of robust tissue culture model systems for HCV entry and infection has finally allowed study of antibody-mediated neutralization and to gain further insights into viral targets of host neutralizing responses. In addition, detailed analysis of antibody-mediated neutralization in individual patients as well as cohorts with well defined viral isolates has enabled the study of neutralizing responses in the course of HCV infection and characterization of the impact of neutralizing antibodiesfor control of viral infection. This review will summarize recent progress in the understanding of the molecular mechanisms of antibody-mediated neutralization and its impact for HCV pathogenesis.
基金the grants No.KY951-Al-301 and No.KY95T-06-03 from the 9th Five Years Plan Key Research Programs of the Chinese Academy of Sciences.
文摘AIM: To establish a convenient immunoassay method based on recombinant antigen preS1(21-119aa) to detect anti-preS1 antibodies and evaluate the clinical significance of antibodies in hepatitis B. METHODS: The expression plasmid pET-28a-preS1 was constructed, and a large quantity of preS1(21-119aa) fragment of the large HBsAg protein was obtained. The preS1 fragment purified by Ni(2+)-IDA affinity chromatography was used as coated antigen to establish the indirect ELISA based on streptavidin-biotin system for detection of the anti-preS1 antibodies in sera from HBV-infected patients. For follow-up study, serial sera were collected during the clinical course of 21 HBV-infected patients and anti-preS1 antibodies, preS1 antigen, HBV-DNA and other serological HBV markers were analyzed. RESULTS: preS1(21-119aa) fragment was highly expressed from the plasmid pET-28a-preS1 in a soluble form in E.Coli (30mg.L(-1)), and easily purified to high purity over 90% by one step of Ni(2+)-IDA-sepharose 6B affinity chromatography. The purity and antigenicity of the purified preS1(21-119aa) protein was determined by 150g.L(-1) SDS-PAGE, Western blot and a direct ELISA. Recombinant preS1(21-119aa) protein was successfully applied in the immunoassay which could sensitively detect the anti-preS1 antibodies in serum specimens of acute or chronic hepatitis B patients. Results showed that more than half of 19 acute hepatitis B patients produced anti-preS1 antibodies during recovery of the disease, however, the response was only found in a few of chronic patients. In the clinical follow-up study of 11 patients with anti-preS1 positive serological profile, HBsAg and HBV-DNA clearance occurred in 6 of 10 acute hepatitis B patients in 5-6 months, and seroconversion of HBeAg and disappearance of HBV-DNA occurred in 1 chronic patients treated with lavumidine, a antiviral agent. CONCLUSION: The high-purity preS1(21-119aa) coated antigen was successfully prepared by gene expression and affinity chromatography. Using this antigen, a conveniently detective system of anti-preS1 antibodies in sera was established. Preliminarily clinical trial the occurrence of anti-preS1 antibodies in acute hepatitis B patients suggests the clearance of HBV from serum in a short-term time, and anti-preS1 positive in chronic patients means health improvement or recovery from the disease.
文摘The results of research work completed in recent years in the authors’ laboratory withmonoclonal antibodies (McAbs) against herpes simplex virus (HSV) are reported in this paper as fol-lows.(1) Eighteen hybridoma cell lines steadily producing McAbs against HSV were established byhybridoma technic.(2) The mechanisms of neutralization in vitro and animal protection in vivo medi-ated by different McAbs were investigated.(3) A method of detecting virus antigua was developedwith the McAbs.(4) Tne isolates of virus were typed and antigenically analysed by type-specificand type-common McAbs.(5) HSV antigen in clinical spectimens was detected and directly typed byan ELISA method coating with type-specific and type-common McAb.(6) The target antigens ofMcAbs were identified,purified,and their genes were localized.(7) Experimental rabbitHSV keratitis was treated with McAbs.
基金Supported by Youth Fund of Hubei Academy of Agricultural Sciences(2013NKYJJ12)
文摘Japanese encephalitis (JE) is a central nervous system disease caused by Japanese encephalitis virus (JEV), which can infect human and a variety of animals and cause irreversible nerve damages. NS3 protein plays an important role in the process of JEV polyprotein hydrolysis, which is essential for JEV replication. Therefore, NS3 protein may be used as a potential drug target to treat Japanese encephalitis. In this study, the pET-28a-NS3 plasmid was successfully constructed and expressed in E. coli BL21 ( DE3 ) under IPTG induction. The molecular weight of the expressed recombinant protein was 55 ku, which was consistent with the expected result. The positive serum was prepared by immunizing BALB/c mice with NS3 protein and identified by indirect immunofluorescence (IFA). The results showed that there was a fluorescence reaction between the prepared positive serum of NS3 protein and cells infected with JEV.
文摘Monoclonal antibodies (McAb) to Hantavirus (HTV)were derived from the hybridoma fused with Lou/c plasmocytoma rat cells (IR983F) and spleen cells from lou/c inbred rat immunized with HTV Chen strain. Eleven cell lines producing monoclonal antibodies direc
基金National Natural Science Foundation ofChina(30400368)The Natural Science foundation ofBeijing(5072003)Beijing Natural Science foundationProgram and Scientific Research Key Program of BeijingMunicipal commission of Education(KZ20051005001).
文摘To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from the total RNA of H9 cells. The resulting DNA construct was cloned into a prokaryotic expression vector (pET-32a). Recombinant pET-vif and pET-APOBEC3G were expressed respectively in Eserichia coli BL21 (DE3) as an insoluble protein. The vector also contained a six-histidine tag at the C-terminus for convenient purification and detection. To express and purify the HIV-1 Vif and hAPOBEC3G in E.coli cells, the accuracy of inserted gene and specificity of proteins were detected by the two enzyme digestion method, SDS-PAGE, and Western blotting. Rabbits were then immunized by Vif or APOBEC3G protein and serum samples were tested by indirect ELISA to determine the level of antibodies. Immunoenzyme and immunofluorescence assays were performed to identify the specificity of polyclonal antibodies. The titer of the anti-Vif antibodies was 1:204800, and that of the anti-APOBEC3G antibodies was 1:102400. Thus the antibodies could detect the antigen expression in the cells, demonstrating that fusion proteins with high purity and their corresponding polyclonal antibodies with high titer and specificity were achieved.
文摘Objective To investigate the characteristics and outcome of glomerulonephritis in patients with both antineutrophil cytoplasmic antibody and anti-glomerular basement membrane antibody.Methods The sera of 23 antiGBM glomerulonephritis patients were collected and were tested for ANCA respectively.Characteristics and outcome of patients with coexisting anti-GBM antibody
文摘Objective To define risk stratification and guide optimal surgical timing of perioperative viral respiratory infection(VRI)in children with cardiac surgery.Methods Retrospective study with propensity score-matched analysis.A total of 2,831 patients had performed RespPCR testing,and finally there were 2,740 negative RespPCR patients and 91 positive RespPCR patients.
文摘AIM To provide a clear understanding of viral hepatitis epidemiology and their clinical burdens in Somalia.METHODS A systematic review and meta-analysis was conducted as Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. A comprehensive literature search of published studies on viral hepatitis was performed from 1977-2016 in Pub Med, Google Scholar, Science Direct, World Health Organization African Index Medicus and the Africa Journals Online databases, as well as on the Ministry of Health website. We also captured unpublished articles that were not available on online systems.RESULTS Twenty-nine studies from Somalia and Somali immigrants(United Kingdom,United States,Italy,Libya)with a combined sample size for each type of viral hepatitis[hepatitis A virus(HAV):1564,hepatitis B virus(HBV):8756,hepatitis C virus(HCV):6257,hepatitis D virus(HDV):375 and hepatitis E virus(HEV):278]were analyzed.The overall pooled prevalence rate of HAV was 90.2%(95%CI:77.8%to 96%).The HAV prevalence among different age groups was as follows:<1 year old,61.54%(95%CI:40.14%to79.24%);1-10 years old,91.91%(95%CI:87.76%to94.73%);11-19 years old,96.31%(95%CI:92.84%to 98.14%);20-39 years old,91.3%(95%CI:83.07%to 95.73%);and>40 years old,86.96%(95%CI:75.68%to 93.47%).The overall pooled prevalence of HBV was 18.9%(95%CI:14%to 29%).The overall pooled prevalence among subgroups of HBV was20.5%(95%CI:5.1%to 55.4%)in pregnant women;5.7%(95%CI:2.7%to 11.5%)in children;39.2%(95%CI:33.4%to 45.4%)in patients with chronic liver disease,including hepatocellular carcinoma(HCC);7.7%(95%CI:4.2%to 13.6%),12.4%(95%CI:6.3%to 23.0%)and 11.8%(95%CI:5.3%to 24.5%)in age groups<20 years old,20-39 years old and>40years old,respectively.The HBV prevalence among risk groups was 20%(95%CI:7.19%to 44.64%)in female prostitutes,21.28%(95%CI:7.15%to48.69%)in hospitalized adults,5.56%(95%CI:0.99%to 25.62%)in hospitalized children,60%(95%CI:31.66%to 82.92%)in patients with acute hepatitis,33.55%(95%CI:14.44%to 60.16%)in patients with ancylostomiasis,12.34%(95%CI:7.24%to 20.26%)in patients with leprosy and 20.19%(95%CI:11.28%to33.49%)in schistosomiasis patients.The overall pooled prevalence of HCV was estimated as 4.84%(95%CI:3.02%to 7.67%).The prevalence rates among blood donors,risk groups,children and patients chronic liver disease(including HCC)was 0.87%(95%CI:0.33%to 2.30%),2.43%(95%CI:1.21%to 4.8%),1.37%(95%CI:0.76%to 2.46%)and 29.82%(95%CI:15.84%to 48.98%),respectively.The prevalence among genotypes of HCV was 21.9%(95%CI:15.36%to 30.23%)in genotype 1,0.87%(95%CI:0.12%to 5.9%)in genotype 2,25.21%(95%CI:18.23%to 33.77%)in genotype 3,46.24%(95%CI:37.48%to 55.25%)in genotype 4,2.52%(95%CI:0.82%to7.53%)in genotype 5,and 1.19%(95%CI:0.07%to16.38%)in genotype 6.The overall pooled prevalence of HDV was 28.99%(95%CI:16.38%to 45.96%).The HDV prevalence rate among patients with chronic liver disease,including HCC,was 43.77%(95%CI:35.09%to 52.84%).The overall pooled prevalence of HEV was46.86%(95%CI:5.31%to 93.28%).CONCLUSION Our study demonstrates a high prevalence of all forms of viral hepatitis in Somalia and it also indicates that chronic HBV was the commonest cause of chronic liver disease.This highlights needs for urgent public health interventions and strategic policy directions to controlling the burden of the disease.
文摘Donor human leukocyte antigen(HLA)-specific antibodies(DSA) play an important role in solid organ transplantation. Preexisting IgG isotype DSA are considered a risk factor for antibody mediated rejection, graft failure or graft loss. The post-transplant development of DSA depends on multiple factors including immunogenicity of mismatched antigens, HLA class Ⅱ typing of the recipient, cytokine gene polymorphisms, and cellular immunoregulatory mechanisms. De novo developed antibodies require special attention because not all DSA have equal clinical significance. Therefore, it is important for transplant clinicians and transplant immunologists to accurately characterize DSA. In this review, the contemporary immunological techniques for detection and characterization of anti-HLA antibodies and their pitfalls are described.
文摘<strong>Background:</strong> Worldwide, prostatic adenocarcinoma is the most common tumour type among men. <strong>Aim:</strong> The aim of the present investigation was to develop a computer program to identify normal prostate biopsies and distinguish them from biopsies showing premalignant alterations (LGPIN, HGPIN) and adenocarcinoma. <strong>Method:</strong> Prostate biopsies (n = 2094) taken from 191 consecutive men during 2016 were stained with triple immunehistochemisty (antibodies to AMACRA, p63 and CK 5). Digital images of the biopsies were obtained with a scanning microscope and used to develop an automatic computer program (CelldaTM), intended to identify the morphological alterations. Visual microscopic finding was used as a reference. <strong>Result:</strong> Of the 191 men, 121 (63.4%) were diagnosed as having prostate adenocarcinoma and 70 (36.6%) as having no malignancy on the basis of the visual microscopy. In comparison, computer analysis identified 134 (70.2%) men with malignant disease and 57 (29.8%) with non-malignant disease after exclusion of artifacts, which constituted 10.4% of areas (indicated as malignant disease). Discrepant results were recorded in 15 (7.9%) men, and in 14 of these cases, HGPIN and areas suggestive of early invasion were common. Thus, it was uncertain whether these cases should be regarded as malignant or not. The agreement between the visual examination and the computer analysis was 92.1% (kappa value 0.823, sensitivity 99.2 and specificity was 0.80). <strong>Conclusion:</strong> It seems that computer analysis could serve as an adjunct to simplify and shorten the diagnostic procedure, first of all by ensuring that normal prostate biopsies are sorted out from those sent for visual microscopic evaluation.
文摘ELISA method was used to detect antisperm antibodies in cervix mucus. Out of 400 women whose husbands had sperms, 92 were antisperm antibody positive (23% ). 42 of 200 women with azoospermic partners were antisperm antibody positive (21% ). No difference was found between the two groups. Hence, antisperm antibodies in azoospermic men's wives may be caused by sperm'sadhering antigens in semina.