Blockade of Rho-associated kinase prevents inhibition of axon regeneration of peripheral nerves induced by anti-ganglioside antibodies

Anti-ganglioside antibodies are associated with delayed/poor clinical recovery in Guillain-Barrèsyndrome,mostly related to halted axon regeneration.Cross-linking of cell surface gangliosides by anti-ganglioside antibodies triggers inhibition of nerve repair in in vitro and in vivo paradigms of axon regeneration.These effects involve the activation of the small GTPase Rho A/ROCK signaling pathways,which negatively modulate growth cone cytoskeleton,similarly to well stablished inhibitors of axon regeneration described so far.The aim of this work was to perform a proof of concept study to demonstrate the effectiveness of Y-27632,a selective pharmacological inhibitor of ROCK,in a mouse model of axon regeneration of peripheral nerves,where the passive immunization with a monoclonal antibody targeting gangliosides GD1a and GT1b was previously reported to exert a potent inhibitory effect on regeneration of both myelinated and unmyelinated fibers.Our results demonstrate a differential sensitivity of myelinated and unmyelinated axons to the pro-regenerative effect of Y-27632.Treatment with a total dosage of 9 mg/kg of Y-27632 resulted in a complete prevention of anti-GD1a/GT1b monoclonal antibody-mediated inhibition of axon regeneration of unmyelinated fibers to skin and the functional recovery of mechanical cutaneous sensitivity.In contrast,the same dose showed toxic effects on the regeneration of myelinated fibers.Interestingly,scale down of the dosage of Y-27632 to 5 mg/kg resulted in a significant although not complete recovery of regenerated myelinated axons exposed to anti-GD1a/GT1b monoclonal antibody in the absence of toxicity in animals exposed to only Y-27632.Overall,these findings confirm the in vivo participation of Rho A/ROCK signaling pathways in the molecular mechanisms associated with the inhibition of axon regeneration induced by anti-GD1a/GT1b monoclonal antibody.Our findings open the possibility of therapeutic pharmacological intervention targeting Rho A/Rock pathway in immune neuropathies associated with the presence of anti-ganglioside antibodies and delayed or incomplete clinical recovery after injury in the peripheral nervous system.

Autoantibodies related to ataxia and other central nervous system manifestations of gluten enteropathy

Gluten ataxia and other central nervous system disorders could be linked to gluten enteropathy and related autoantibodies.In this narrative review,we focus on the various neuro-logical manifestations in patients with gluten sensitivity/celiac disease,immunological and autoimmune mechanisms of ataxia in connection to gluten sensitivity and the autoantibodies that could be used as a biomarker for diagnosing and following.We focused on the anti-gliadin antibodies,antibodies to different isoforms of tissue transglutaminase(TG)(anti-TG2,3,and 6 antibodies),anti-glycine receptor antibodies,anti-glutamine acid decarboxylase antibodies,anti-deamidated gliadin peptides antibodies,etc.Most studies found a higher prevalence of these antibodies in patients with gluten sensitivity and neurological dysfunction,presented as different neurological disorders.We also discuss the role of a gluten-free diet on the clinical improvement of patients and also on imaging of these disorders.

A nanobody-based blocking enzyme-linked immunosorbent assay for detecting antibodies against pseudorabies virus glycoprotein E

Pseudorabies(PR)is an acute infectious disease of pigs caused by the PR virus(PRV)and results in great economic losses to the pig industry worldwide.PRV glycoprotein E(gE)-based enzyme-linked immunosorbent assay(ELISA)has been used to distinguish gE-deleted vaccine-immunized pigs from wild-type virus-infected pigs to eradicate PR in some countries.Nanobody has the advantages of small size and easy genetic engineering and has been a promising diagnostic reagent.However,there were few reports about developing nanobody-based ELISA for detecting anti-PRV-gE antibodies.In the present study,the recombinant PRV-gE was expressed with a bacterial system and used to immunize the Bactrian camel.Then,two nanobodies against PRV-gE were screened from the immunized camel by phage display technique.Subsequently,two nanobody-HRP fusion proteins were expressed with HEK293T cells.The PRV-gE-Nb36-HRP fusion protein was selected as the probe for developing the blocking ELISA(bELISA)to detect anti-PRV-gE antibodies.Through optimizing the conditions of bELISA,the amount of coated antigen was 200 ng per well,and dilutions of the fusion protein and tested pig sera were separately 1:320 and 1:5.The cut-off value of bELISA was 24.20%,and the sensitivity and specificity were 96.43 and 92.63%,respectively.By detecting 233 clinical pig sera with the developed bELISA and a commercial kit,the results showed that the coincidence rate of two assays was 93.99%.Additionallly,epitope mapping showed that PRV-gE-Nb36 recognized a conserved conformational epitope in different reference PRV strains.Simple,great stability and low-cost nanobody-based bELISA for detecting anti-PRV-gE antibodies were developed.The bELISA could be used for monitoring and eradicating PR.

Establishment of an indirect immunofluorescence assay for the detection of African swine fever virus antibodies

African swine fever(ASF)continues to cause enormous economic loss to the global pig industry.Since there is no safe and effective vaccine,accurate and timely diagnosis of ASF is essential to implement control measures.Indirect immunofluorescence assay(IFA)is a gold standard serological method recommended by the World Organization for Animal Health(WOAH).In this study,we used primary fetal kidney cells to establish a wild boar cell line(BK2258)that supported the efficient replication of ASF virus(ASFV)SD/DY-I/21 and showed visible cytopathic effect(CPE).Moreover,using BK2258,we established a sensitive and specific IFA for ASFV antibody detection.To standardize and evaluate the performance of this assay,we used serum samples from pigs infected with the low virulent genotype I SD/DY-I/21 and genotype II HLJ/HRB1/20,and immunized with the vaccine candidate HLJ/18-7GD,field samples,and negative serum samples.The IFA reacted with the ASFV-positive sera and displayed bright fluorescence foci.There was no non-specific green fluorescence due to cellular senescence or other cell damage-causing factors.Compared to a commercial indirect enzyme-linked immunosorbent assay(iELISA),ASFV antibodies were detected 1–4 days earlier using our IFA.The detection limits of the IFA and iELISA for the same ASFV-antibody positive serum samples were 1:25,600 and 1:6,400,respectively,indicating that the IFA is more sensitive than iELISA.The newly established IFA was highly specific and did not cross-react with sera positive for six other important porcine pathogens(i.e.,Classical swine fever virus(CSFV),Porcine reproductive and respiratory syndrome virus(PRRSV),Porcme circovirus type 2(PCV2),Pseudorabies virus(PRV),Foot-and-Mouth disease virus type O(FMDV/O),and Porcine epidemic diarrhea virus(PEDV)).This study thus provides a sensitive,specific,and reliable detection method that is suitable for the serological diagnosis of ASF.

New glucocerebrosidase antibodies can advance research in the field of neurodegenerative disorders

In medical research,there are times when the introduction of a new tool can launch scientific discovery in new directions.While antibody development may be considered mundane,in the field of glucocerebrosidase(GCase)research,the dearth of validated antibodies for different applications has impeded progress in studies of disease pathogenesis and therapeutic development.The recent introduction of new,rigorously evaluated antibodies can now propel research into the link between glucocerebrosidase and Parkinson’s disease(PD)as well as aspects of the pathobiology of Gaucher disease(Jong et al.,2024).

Allogenic Stem Cell Transplantation and COVID-19 Antibodies: Mechanistic Insights and Recipient Concerns

Collecting umbilical cord stem cells is widely practiced due to its numerous benefits. Over the past decade, umbilical cord stem cells (UCSCs) have shown effectiveness in treating various conditions, such as bone pathologies, neuropsychiatry disorders, hereditary diseases, and metabolic disorders. However, factors like immunization affect the quantity and quality of cord harvesting. Studies suggest that antibodies from the mother pass through the umbilical cord to protect the infant against infections. Cleaning the umbilical cord before stem cell extraction is crucial to maintain sterility and cell integrity. Vaccinating a female donor, including for COVID-19, typically does not directly affect the stem cells. Although vaccines aim to trigger an immunological response, they generally do not affect the donor’s stem cells.

Prevalence and Factors Associated with Positivity of Antinuclear Antibodies (ANA) Patterns, Native Anti-DNA and Extractable Nuclear Antigens (ENA) Antibodies: Experience from a Laboratory in Dakar

Background: Diagnosis of autoimmune diseases (AID) is challenging, due to overlapping features with other non-immune disorders. Anti-nuclear antibodies (ANA) are sensitive screening tests but anti-deoxyribonucleic acid-antibody (anti-DNA), and anti-extractable nuclear antigens (anti-ENA) are specific for AIDs. We aimed to look at ANA patterns in our patients and correlated them with anti-ENA for proper interpretation and better patient management cost-effectively. Methods: A retrospective study was conducted over 1 year from January to December 2022 who were tested for ANA at biology medical laboratory of Pasteur Institute of Dakar. Anti-ENA and anti-DNA results were also analyzed for ANA-positive patients. Statistical analysis was performed using STATA 14.0, p Results: 216 patients were analyzed. Women predominated at 79.2% and mean age was 48 years [CI 95%, 46 - 50], with extremes of 10 and 89. Most represented age group was [41 - 60] with 38%. ANA was positive in 27 (12.5%) of patients, 59.2% of whom were strongly positive (titer of 1/1000, 1/3200 or 1/6400). The most common pattern was nuclear speckled, which was found in 77.8% of samples. Anti-ENA and anti-DNA positivity in ANA-positive patients was found respectively in 63% (17/27) and 1.4% (3/27) of the samples analyzed. Most commonly identified anti-ENA was anti-Sm 29.6%, anti-SSA 29.6%, anti-Ro-52 25.9%, anti-RNP 18.5% and anti-SSB 14.8% which was associated with speckled pattern. Association results indicated a significant relationship between both tests and between ANA titer in the anti-ENA- and ANA-positive patients (p 0.001). Conclusions: ANA, Anti-ENA and anti-DNA antibodies are essential for AIDS diagnosis. However, the testing repertoire should follow an algorithm comprising of clinical features, followed by ANA results with nuclear, mitotic, and cytoplasmic patterns, anti-ENA, and anti-DNA for a more meaningful, and cost-effective diagnostic approach.

Antibodies elicited by Newcastle disease virus-vectored H7N9 avian influenza vaccine are functional in activating the complement system

H7N9 subtype avian influenza virus poses a great challenge for poultry industry.Newcastle disease virus(NDV)-vectored H7N9 avian influenza vaccines(NDV_(vec)H7N9)are effective in disease control because they are protective and allow mass administration.Of note,these vaccines elicit undetectable H7N9-specific hemagglutination-inhibition(HI)but high IgG antibodies in chickens.However,the molecular basis and protective mechanism underlying this particular antibody immunity remain unclear.Herein,immunization with an NDV_(vec)H7N9 induced low anti-H7N9 HI and virus neutralization titers but high levels of hemagglutinin(HA)-binding IgG antibodies in chickens.Three residues(S150,G151 and S152)in HA of H7N9 virus were identified as the dominant epitopes recognized by the NDV_(vec)H7N9 immune serum.Passively transferred NDV_(vec)H7N9 immune serum conferred complete protection against H7N9 virus infection in chickens.The NDV_(vec)H7N9 immune serum can mediate a potent lysis of HA-expressing and H7N9 virus-infected cells and significantly suppress H7N9 virus infectivity.These activities of the serum were significantly impaired after heat-inactivation or treatment with complement inhibitor,suggesting the engagement of the complement system.Moreover,mutations in the 150-SGS-152 sites in HA resulted in significant reductions in cell lysis and virus neutralization mediated by the NDV_(vec)H7N9 immune serum,indicating the requirement of antibody-antigen binding for complement activity.Therefore,antibodies induced by the NDV_(vec)H7N9 can activate antibody-dependent complement-mediated lysis of H7N9 virus-infected cells and complement-mediated neutralization of H7N9 virus.Our findings unveiled a novel role of the complement in protection conferred by the NDV_(vec)H7N9,highlighting a potential benefit of engaging the complement system in H7N9 vaccine design.

Dynamically changing antineutrophil cytoplasmic antibodies in granulomatosis with polyangiitis:A case report

BACKGROUND Granulomatosis with polyangiitis(GPA)is one of the most prevalent forms of the antineutrophil cytoplasmic antibody(ANCA)-associated vasculitis.GPA is characterized histologically by necrotizing granulomatous inflammation in addition to vasculitis.The diagnosis of GPA depends on clinical presentation,serological evidence of a positive ANCA,and/or histological evidence of necrotizing vasculitis or granulomatous destructive parenchymal inflammation.Cytoplasmic ANCA(c-ANCA)is positive in 65%-75% of GPA patients,accompanied by proteinase 3(PR3),the main target antigen of c-ANCA,another 5% of GPA patients had negative ANCA.CASE SUMMARY The patient,a 52-year-old male,presented with unexplained nasal congestion,tinnitus,and hearing loss.After a duration of 4 months experiencing these symptoms,the patient subsequently developed fever and headache.The imaging examination revealed the presence of bilateral auricular mastoiditis and partial paranasal sinusitis,and the ANCA results were negative.The anti-infective therapy proved to be ineffective,but the patient's symptoms and fever were quickly relieved after 1 wk of treatment with methylprednisolone 40 mg once a day.However,after continuous use of methylprednisolone tablets for 3 months,the patient experienced a recurrence of fever accompanied by right-sided migraine,positive c-ANCA and PR3,and increased total protein in cerebrospinal fluid.The and cyclophosphamide 0.8 g monthly,the patient experienced alleviation of fever and headache.Additionally,the ANCA levels became negative and there has been no recurrence.CONCLUSION For GPA patients with negative ANCA,there is a potential for early missed diagnosis.The integration of histopathological results and multidisciplinary communication plays a crucial role in facilitating ANCA-negative GPA.

Contribution of Anti-p63 Antibodies in the Interpretation of Benign Label Prostatic Biopsies

Introduction: Prostate cancer is the second most common cancer in men. The diagnosis is most often based on the prostate biopsies’ analysis and on histological criteria recognizable on standard coloring. In some cases, the use of immunohistochemistry is important. Objectives: This paper aims to specify the p63 phenotypic profile of lesions diagnosed benign, with minimal suspect foci, difficult to interpret, HGPIN (high grade intraepithelial neoplasia) and LGPIN (low-grade prostatic intraepithelial neoplasia) and evaluate the manual technique of p63 immunohistochemistry. Patients and Method: This was a retrospective, descriptive study of prostate biopsies recorded in the PAC service of the HALD from January 1st, 2018 to December 31st, 2018. It was completed by a manual immunohistochemical study of the blocks enrolled from November 19th to December 4th, 2020 in the PAC department of the HPD. The studied parameters were: registry number, age, clinical stage, prostate volume, PSA level, microscopic appearance and p63 immunohistochemical profile. Results: Our study included 60 prostate biopsies. The ages of our patients varied from 45 to 77 years, with an average of 64.2 years and a standard deviation of 6.2. The majority of patients were at clinical stage cT2b (33%) with a prostate volume varying between 33.15 and 169.4 cc. The minimum value of PSA in our series is 5 ng/ml, the maximum being 100 ng/ml with an average level of 24.1 ng/ml and a standard deviation of 21.2. Our series included 50 adenomyomatous hyperplasias, 7 adenomyomatous hyperplasias associated with chronic prostatitis, 2 HGPIN and 1 LGPIN. After re-reading we found 5 discordant cases, which corresponded to minimal suspect foci (kappa = 0.5098). The p63 marking was informative in 53 cases, i.e. 88%, and non-informative in 7 cases, i.e. 12%. Among the uninformative markings, 2 were due to lack of tissue adhesion to the slides. Among the informative markings, 11 were negative. p63 immunohistochemistry was useful in all suspected foci and detected 6 other minimal foci of adenocarcinoma. Conclusion: The immunostaining with the anti-p63 antibody in the prostate cancer diagnosis is of considerable benefit. It made it possible to correct 11.3% of benign diagnosis in minimal malignant focus in our context. Despite the difficulties associated with the manual technique, it is possible to have an informative rate, similar to the automatic technique.

Hepatitis B virus reactivation in patients treated with monoclonal antibodies

Hepatitis B virus(HBV)reactivation poses a significant clinical challenge,espe-cially in patients undergoing immunosuppressive therapies,including mono-clonal antibody treatments.This manuscript briefly explores the complex rela-tionship between monoclonal antibody therapy and HBV reactivation,drawing upon current literature and clinical case studies.It delves into the mechanisms underlying this phenomenon,highlighting the importance of risk assessment,monitoring,and prophylactic measures for patients at risk.The manuscript aims to enhance the understanding of HBV reactivation in the context of monoclonal antibody therapy,ultimately facilitating informed clinical decision-making and improved patient care.This paper will also briefly review the definition of HBV activation,assess the risks of reactivation,especially in patients treated with monoclonal antibodies,and consider management for patients with regard to screening,prophylaxis,and treatment.A better understanding of patients at risk can help clinicians provide optimum management to ensure successful patient outcomes and prevent morbidity.

Development of donor specific antibodies after SARS-CoV-2 vaccination:What do we know so far?

Vaccination against Coronavirus disease-19(COVID-19)was pivotal to limit spread,morbidity and mortality.Our aim is to find out whether vaccines against COVID-19 lead to an immunological response stimulating the production of de novo donor specific antibodies(DSAs)or increase in mean fluorescence intensity(MFI)of pre-existing DSAs in kidney transplant recipients(KTRs).This study involved a detailed literature search through December 2nd,2023 using PubMed as the primary database.The search strategy incorporated a combination of relevant Medical Subject Headings terms and keywords:"COVID-19","SARS-CoV-2 Vaccination","Kidney,Renal Transplant",and"Donor specific antibodies".The results from related studies were collated and analyzed.A total of 6 studies were identified,encompassing 460 KTRs vaccinated against COVID-19.Immunological responses were detected in 8 KTRs of which 5 had increased MFIs,1 had de novo DSA,and 2 were categorized as either having de novo DSA or increased MFI.There were 48 KTRs with pre-existing DSAs prior to vaccination,but one study(Massa et al)did not report whether pre-existing DSAs were associated with post vaccination outcomes.Of the remaining 5 studies,35 KTRs with pre-existing DSAs were identified of which 7 KTRs(20%)developed de novo DSAs or increased MFIs.Overall,no immunological response was detected in 452(98.3%)KTRs.Our study affirms prior reports that COVID-19 vaccination is safe for KTRs,especially if there are no pre-existing DSAs.However,if KTRs have pre-existing DSAs,then an increased immunological risk may be present.These findings need to be taken cautiously as they are based on a limited number of patients so further studies are still needed for confirmation.

Development of Non-ABO Red Blood Cell Alloantibodies in Patient Undergoing Allogeneic Haematopoietic Stem Cell Transplant

Alloantibodies that are non ABO Alloimmunization to protein antigens happens only after exposure, in contrast to ABO isohaemagglutinins, which are present naturally, even in the absence of prior exposure. It is recognized that while non-ABO RBC antibodies are less common than ABO antibodies, they generate essentially the same issues that lead to unfavorable clinical results. If non-ABO alloantibodies are identified early on, these issues related complications may be avoided This call for an in-depth understanding of the recipient and donor’s ABO-Rh grouping, antibody screening, and the phenotype of certain antigens. Equally important, the temporal association time between transplantation and hemolysis can help identify the underlying mechanism of hemolysis and direct appropriate management. Therefore, for that, it is crucial to identify the etiology of post-HSCT anemia for prevention and therapy, in addition to a thorough grasp of the mechanism of anemia in non-ABO-incompatible HSCT and the temporal link between HSCT and anemia. Finding the cause of post-HSCT anemia is essential for prevention and therapy, in addition to a thorough grasp of the mechanism of anemia in non-ABO-incompatible HSCT and the temporal link between HSCT and anemia. Therefore, for that, it is crucial to identify the etiology of post-HSCT anemia. In this case report review, we would like to highlight the vital role of transfusion medicine services and stem cell clinical teams in paying particular attention to the clinical significance of non-ABO alloantibodies involved to avoid causing overt hemolysis of incompatible donor RBCs or delayed erythropoiesis. Considering the fact that some of the Haematopoietic stem cell transplant centers do not give an attention to the other non-ABO RBC antigens.

Developability assessment of the therapeutic antibodies studies: are they druggable?

In recent years,with the continuous development of biotechnology and medical science and technology,therapeutic antibodies,as new biologics,have shown great potential and have become a research hotspot in the fields of oncology,autoimmune diseases,cardiovascular and neuroscience.Therapeutic antibodies refer to drugs or therapeutic methods for the treatment of diseases with specificity,high affinity and high efficiency antibody molecules prepared by biotechnological means.In this paper,we will systematically review the types of therapeutic antibodies,research progress,problems in research and development,mechanisms of action,and challenges faced in clinical applications.

原发与继发免疫性血小板减少症患者血小板膜糖蛋白特异性抗体及其与出血评分关系的研究

目的探讨原发与继发免疫性血小板减少症(ITP)患者抗GPⅡb/Ⅲa及GPΙb/Ⅸ抗体的表达、抗体阳性表达与出血评分的关系,以及不同抗体类型出血评分的差异,为原发与继发ITP患者的诊治和出血严重程度提供临床依据。方法选取2013年10月—2020年8月在新疆医科大学第一附属医院诊断为原发ITP患者(42例)及继发ITP患者(42例)为研究对象,所有患者入院时采用ITP出血评分量表行出血评分。应用改良MAIPA法检测患者抗GPⅡb/Ⅲa和抗GPΙb/Ⅸ抗体。结果原发与继发组患者抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体阳性率比较,差异无统计学意义(P>0.05),原发组患者抗体表达以抗GPⅡb/Ⅲa抗体为主,继发组抗体表达以抗GPΙb/Ⅸ抗体为主,两者抗体阳性构成比较,差异有统计学意义(P<0.05)。继发组患者整体出血程度较原发组重(P<0.05)。抗体阳性表达与出血程度的关系:(1)抗GPⅡb/Ⅲa抗体阳性患者出血程度较抗体阴性患者重(P<0.05)。(2)抗GPΙb/Ⅸ抗体阳性患者出血程度较抗体阴性患者重(P<0.05),双抗体阳性患者出血程度较抗体阴性患者重(P<0.05)。结论抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体阳性表达对原发与继发ITP无鉴别意义,但原发ITP患者抗体表达以抗GPⅡb/Ⅲa抗体为主,继发ITP患者抗体表达以抗GPΙb/Ⅸ抗体为主。继发ITP患者临床出血程度较原发ITP患者重。抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体及抗体双阳性患者出血程度较抗体阴性患者重。

孕妇抗核抗体谱与胎儿发育异常的关系

目的:探讨孕妇抗核抗体谱与胎儿发育异常的关系。方法:选择2020年1月至2023年12月在郑州大学第三附属医院行抗核抗体谱检测的孕妇2490例,通过胎儿四维彩超和超声心动图检测胎儿发育情况。使用免疫印迹法检测孕妇血清抗核抗体谱(抗双链DNA、核小体、组蛋白、核糖体蛋白P、SM、nRNP、SSA/Ro60、SSA/Ro52、SSB/La、着丝点、Scl-70和Jo-1抗体)。比较胎儿发育正常和发育异常孕妇血清抗核抗体谱阳性检出情况,胎儿心脏发育正常和发育异常孕妇血清抗着丝点抗体、抗SSA/Ro52抗体、抗着丝点或SSA/Ro52抗体和抗SSA/Ro60+SSA/Ro52+SSB/La抗体阳性检出情况。分析抗核抗体谱在胎儿心脏异常中的诊断价值。结果:2490例孕妇抗核抗体谱阳性检出率为5.38%。胎儿发育异常孕妇抗核抗体谱阳性检出率为7.2%(37/514),高于胎儿发育正常孕妇的4.9%(97/1976)(P=0.040)。胎儿发育正常和发育异常孕妇抗SSA/Ro60、SSA/Ro52、SSB/La和着丝点抗体阳性检出率比较,差异无统计学意义(P>0.05);78例胎儿心脏异常,其中心血管畸形51例、心律失常27例。胎儿心脏发育正常和发育异常孕妇抗着丝点抗体、抗SSA/Ro52抗体、抗着丝点或SSA/Ro52抗体、抗SSA/Ro60+SSA/Ro52+SSB/La抗体阳性检出率比较,差异有统计学意义(P<0.05)。孕妇抗着丝点或SSA/Ro52抗体预测胎儿心脏发育异常的敏感度和阴性预测值分别为0.090和0.971;抗着丝点抗体预测的特异度和阳性预测值分别为0.999和0.400。结论:孕妇血清抗着丝点抗体、抗SSA/Ro52抗体以及抗SSA/Ro60+SSA/Ro52+SSB/La抗体阳性,可能提示胎儿心脏发育异常。

假病毒中和试验对狂犬病毒中和抗体效价的检测效能分析

目的:组织不同的实验室分析假病毒中和法(Pseudo Virus-based Neutralization Assay,PBNA)对不同样品的狂犬病病毒中和抗体水平的检测能力,并比较其与经典的快速免疫荧光灶抑制试验(Rapid Fluorescent Focus Inhibition Test,RFFIT)以及小鼠中和试验法(Mouse Neutralization Test,MNT)的相关性。方法:共9家实验室,分别采用PBNA、RFFIT以及MNT法对6份样品进行狂犬病病毒中和抗体检测,每个实验室进行3~5次重复实验。采用配对t检验及Pearson相关系数对3种检测方法的结果进行统计学分析。结果:PBNA与RFFIT、PBNA与MNT、RFFIT与MNT Pearson相关系数分别为0.985、0.988和0.974,抗体几何平均滴度进行t检验,差异均无统计学意义(P>0.05)。结论:3种方法重复检测6份样品的定量结果具有高度的相关性,PBNA法可作为《中华人民共和国药典》方法RFFIT、MNT的一种替代或补充方法,用于人用狂犬病疫苗临床血清、狂犬病人免疫球蛋白及抗狂犬病血清的狂犬病病毒中和抗体效价检测。

益气养阴化痰祛瘀法对甲状腺机能亢进症激素抗体及氧化应激因子表达的影响

目的研究益气养阴化痰祛瘀法对甲状腺机能亢进症(Hyperthyroidism,简称甲亢)患者激素抗体和氧化应激因子表达的影响。方法选取2022年1月—2023年12月收治的76例甲亢患者,按照随机数字表法分组,对照组(38例)采用常规西药治疗,观察组(38例)在对照组基础上加用益气养阴化痰祛瘀方治疗;治疗2个月后对比两组临床疗效、中医证候积分、甲状腺功能(各项激素水平)、氧化应激因子、促甲状腺激素受体抗体(TRAb)、甲状腺过氧化物酶抗体(TPOAb)的表达。结果观察组治疗总有效率为94.74%(36/38),高于对照组(76.32%,29/38),差异有统计学意义(P<0.05)。治疗后,两组主症积分、次症积分和舌脉积分均降低,观察组低于对照组;两组促甲状腺激素(TSH)上升,游离三碘甲状腺原氨酸(FT3)、游离甲状腺素(FT4)下降(P<0.05),观察组改善高于对照组;两组超氧化物歧化酶(SOD)升高,观察组SOD水平高于对照组,差异有统计学意义(P<0.05)。结论益气养阴化痰祛瘀法用于甲亢患者的治疗取得确切治疗成果,可以有效改善甲状腺功能,通过调控甲状腺激素水平、TRAb、TPOAb以及氧化应激因子的表达以改善病情,缓解症状。

孕产妇不规则抗体血清学特征和临床意义分析

目的:明确不规则抗体在孕产妇人群中的血清学特征,探讨其临床意义。方法:选取2017年1月-2022年3月于重庆医科大学附属妇女儿童医院就诊的孕产妇151471例,采用微柱凝胶抗人球蛋白法进行不规则抗体筛查,对部分抗体筛查阳性标本进一步做抗体特异性鉴定。结果:孕产妇不规则抗体筛查阳性率为0.91%(1375/151471),其中0.23%(355/151471)在孕早期检出,0.05%(71/151471)在孕中期检出,0.63%(949/151471)在孕晚期检出,孕晚期抗体阳性率明显高于孕早期和孕中期,且孕晚期新增阳性例数明显高于孕中期新增例数;分析1375例不规则抗体阳性结果的凝集强度,其中凝集强度为弱阳性占比最高,为50.11%(689/1375),可疑阳性占18.69%(257/1375),阳性占31.20%(429/1375),孕中期凝集强度分布与孕早期相比差异无统计学意义,但孕晚期可疑阳性和弱阳性的比例低于孕早期,阳性比例高于孕早期,比较差异有统计学意义(P<0.001)。不规则抗体阳性孕产妇中,妊娠≥2次孕产妇的占比明显高于≤1次者;进行抗体鉴定的60例孕产妇中,Rh系统占25.00%(15/60),Lewis系统占43.33%(26/60),Kidd系统占3.33%(2/60),MNS系统抗体占16.67%(10/60),P1PK系统抗体占1.67%(1/60),自身抗体占1.67%(1/60),4例无法确认特异性占6.67%(4/60);特异性抗体中以抗-Lea(30.00%)最为常见,其次是抗-E(16.67%)和抗-M(16.67%)。结论:不同地区、具有不同遗传背景的孕产妇人群的不规则抗体血清学特征存在差异,掌握本地区孕产妇不规则抗体的特征,对保证孕产妇输血安全、防治新生儿溶血病具有十分重要的意义。

基于黏膜sIgA抗体的非洲猪瘟病毒感染早期血清学检测方法的建立

目前的非洲猪瘟病毒(ASFV)血清学检测方法存在因采血引起交叉感染风险、抗体转阳滞后影响检测敏感性等问题,因此建立一种无创采样且可实现早期诊断的血清学方法对于在临床上ASFV感染的诊断与监测有重要意义。本研究以人工合成的方式构建了pFastbac1-P30-His重组表达载体,利用杆状病毒-昆虫细胞真核表达系统表达ASFV重组P30蛋白(SUMO-P30),用Ni柱亲和纯化后作为包被抗原,经过一系列条件优化,建立一种以猪口腔液中黏膜sIgA抗体为靶标的ASFV抗体间接ELISA方法。结果显示,真核重组表达的P30蛋白(SUMO-P30)约为47 ku,Western blot鉴定显示其具有良好的反应原性;经优化确定了ELISA最佳反应条件:包被量为1.0μg·mL^(-1),5%脱脂乳为最佳样品稀释液,与待检口腔液的最佳混合比例为2∶8,样品反应时间为120 min,鼠抗猪IgA-HRP最佳稀释度为1∶5000,反应时间为60 min,最佳显色时间为15 min。该方法检测ASFV感染口腔液抗体效价可达1∶32,与猪瘟病毒、猪伪狂犬病病毒、猪繁殖与呼吸综合征病毒黏膜抗体阳性口腔液均无交叉反应,具有良好的敏感性和特异性。利用该方法和ASFV商品化ELISA血清抗体检测试剂盒,分别检测感染ASFV强毒株或人工致弱株后同一头猪不同时间点的口腔液和血清样品,口腔液中黏膜sIgA抗体在感染后3~5 d S/P值就显著提升,而血清抗体在监测期内未检测到转阳。检测人工感染或同圈感染自然变异株后不同时间点的口腔液和血清样品,本研究建立的方法与商品化非洲猪瘟病毒血清抗体检测试剂盒检测的阳性符合率为100%,阴性符合率为37.5%,总符合率为61.5%。综上,本研究建立了一种无需采血,以口腔液为样品的ASFV黏膜sIgA抗体ELSIA检测方法,可实现ASFV感染的无创、早期、敏感诊断,为ASFV的监测和防控提供新的技术支持和补充。