Enzyme-linked immunosorbent assay of 3 Screen Islet Cell Autoantibody in patients with autoimmune thyroid disease

BACKGROUND In recent years,the emergence of multiplex technology that can simultaneously measure multiple anti-islet autoantibodies has become particularly valuable for the staging and early diagnosis of immune-mediated type 1 diabetes(T1D).While it has been established that 20%-30%of T1D patients suffer from autoimmune thyroid disease(AITD),there is limited available data regarding the presence of anti-islet autoantibodies in AITD patients.Among commercially available anti-islet autoantibodies,glutamic acid decarboxylase 65 autoantibodies(GADAs)are often the first marker measured in general clinical practice.AIM To investigate the frequency of anti-islet autoantibodies in AITD patients.METHODS Our study involved four hundred ninety-five AITD patients,categorized into three distinct groups:AITD with T1D(n=18),AITD with phenotypic type 2 diabetes(T2D)(n=81),and AITD without diabetes(n=396),and the enzyme-linked immunosorbent assay(ELISA)was employed to determine the frequencies of 3 Screen Islet Cell Autoantibody(3 Screen ICA),GADA,insulinoma-associated antigen-2 autoantibodies(IA-2As),and zinc transporter 8 autoantibodies(ZnT8As)within these groups.RESULTS The frequency of 3 Screen ICA in AITD patients with T1D,T2D,and those without diabetes were 88.9%,6.2%,and 5.1%,respectively,with no significant difference seen between the latter two groups.Notably,the frequency of 3 Screen ICA was 11.1%higher in AITD patients with T1D,1.3%higher in AITD patients with T2D,and 1.1%higher in AITD patients without diabetes compared to GADA,respectively.Furthermore,12.5%,20.0%,and 20.0%of the 3 Screen ICA-positive patients were negative for GADA.Additionally,1.3%of the AITD patients who tested negative for 3 Screen ICA in both the AITD with T2D and non-diabetic AITD groups were found to be positive for individual autoantibodies.Among the 3 Screen ICA-positive patients,there was a significantly higher proportion of individuals with multiple autoantibodies in AITD patients with T1D compared to those without diabetes(37.5%vs 5.0%,P<0.05).However,this proportion was similar to that in AITD patients with T2D(20.0%).Nevertheless,there was no significant difference in 3 Screen ICA titers between AITD patients with T1D and those without diabetes(436.8±66.4 vs 308.1±66.4 index).Additionally,no significant difference in 3 Screen ICA titers was observed between Graves’disease and Hashimoto’s thyroiditis in any of the groups.CONCLUSION Our findings reveal that some AITD patients without diabetes exhibit 3 Screen ICA titers comparable to those in AITD patients with T1D.Thus,3 Screen ICA outperforms GADA in identifying latent anti-islet autoantibody-positive individuals among AITD patients.

Autoantibody profiles in autoimmune hepatitis and chronic hepatitis C identifies similarities in patients with severe disease

To determine how the auto-antibodies (Abs) profiles overlap in chronic hepatitis C infection (CHC) and autoimmune hepatitis (AIH) and correlate to liver disease.METHODSLevels of antinuclear Ab, smooth muscle antibody (SMA) and liver/kidney microsomal-1 (LKM-1) Ab and markers of liver damage were determined in the sera of 50 patients with CHC infection, 20 AIH patients and 20 healthy controls using enzyme linked immunosorbent assay and other immune assays.RESULTSWe found that AIH patients had more severe liver disease as determined by elevation of total IgG, alkaline phosphatase, total serum bilirubin and serum transaminases and significantly higher prevalence of the three non-organ-specific autoantibodies (auto-Abs) than CHC patients. Antinuclear Ab, SMA and LKM-1 Ab were also present in 36% of CHC patients and related to disease severity. CHC cases positive for auto-Abs were directly comparable to AIH in respect of most markers of liver damage and total IgG. These cases had longer disease duration compared with auto-Ab negative cases, but there was no difference in gender, age or viral load. KLM-1+ Ab CHC cases showed best overlap with AIH.CONCLUSIONAuto-Ab levels in CHC may be important markers of disease severity and positive cases have a disease similar to AIH. Auto-Abs might have a pathogenic role as indicated by elevated markers of liver damage. Future studies will unravel any novel associations between these two diseases, whether genetic or other.

Experimental Study of Serum Substantia Nigra Neuron Autoantibody and Its Effect in Parkinson Disease Patients

To investigate the serum substantia nigra neuron autoantibody and its effect in the patients with Parkinson disease (PD), substantia nigra slices and a rat model of injection of serum from PD patients in unilateral side substantia nigra were applied. The results showed that the positive rate of substantia nigra neuron autoantibody in PD patients was significantly higher than in the healthy control group (36.67 % vs 6.67 %, P <0.01), but no significant difference was found between PD group and myasthenia gravis (MG) group (26.67 %, P >0.05). The sera from PD patients positive for substantia nigra neuron autoantibody could decrease the number of the dopaminergic neurons more seriously than those from MG and the healthy once respectively (both P <0.01). The results suggested that the immunological mechanism might partly play a role in the development of PD.

Anti-ENA autoantibody profiles and humoral immune activation

Objective: To study the clinical significance of anti-ENA autoantibody profiles and its relationship withother immune markers. Methods: AntiuENA autoantibodies were tested with Western--blot while immunoglobulinsand complements were detected quantitatively by turbidimetry. Results: 1. High positive rate of anti--ENAautoantibody was detected but not specific in several autoimmune diseases; 2. lgG and IgA was increased in antiENA antibody positive sera while IgM, C3 and C4 was decreased; 3. Elevation of IgG and decrease of IgM, C3and CHS, were correlated with the number of bands of anti-- ENA autoantibodies by immunoblot; 4. Associationbetween clinical activity of diseases and C3, C4 and CH50 were tested. Conclusion: Humoral immune activation wasdemonstrated in patients with positive ENA autoantibody and positively correlated with the number of bands ofanti-- ENA autoantibodies.

Purification and identification of natural antikeratin autoantibody from normal human sera and its effects on the cultured keratinocytes

Objective: To purify the natural antikeratin autoantibody (AK auto-Ab) and observe its effects on the prolif eration of the cultured keratinocytes. Methods: Natural AK auto-Ab was purified by using keratin affinity column, and then the titre and specificity of the Abs were studied by ELISA, immunoperoxidase staining and immuno-electronicmicroscope. The effect of the purified Abs on the cultured keratino-cytes was assayed by 3H-TdR incorporation. Results: Natural AK auto-Ab was obtained. The binding activity of IgG AK auto-Ab in purified Ah remained similar to that in pooled human sera. and the specificity of the obtained antibody is strong. The purified antibody could decrease the Il-TdR incorporation of the cultured keratinocytes in a dose-dependent manner. Conclusion: The method of punning AK auto-Ab is simple, practicable and reliable. Natural AK auto-Ab, existing in normal human individuals, has inhibitory etiect on the proliferation of the cultured keratinocytes.

Immunohistochemical Studies on Anti-keratin Autoantibody

On the basis of the detection of IgG anti-keratin autoantibody (AK auto Ab)in human body fluids,it was shown that IgG AK auto Ab could hardly pass the blood-brain barrier,but it could easily penetrate the capillary walls into epithelial tissues.Un-der electron microscope,the interaction between AK auto Ab and keratinocyte is largelylocated in tonofilaments and desmosomes and is irrelevant to other organellae.Dynamicobservations of AK auto Ab suggest that the accumulation of AK auto Ab in keratinocy-te cytoplasm and transitory decrease of IgG AK auto Ab in serum of rabbits with sodiumsulfide dermatitis are related predominantly to keratinocytic injury.

A Multiplex Autoantibody Panel for Early Detection of Autoimmune Disease Activity

Background: Detection of autoantibodies has played a consolidate role in diagnosis of systemic autoimmune disorders. A cascade autoantibody testing is usually performed by employing antinuclear antibodies (ANA) test as a first screening test and the other tests as second level determinations. Here, we present that supplementing extractable nuclear antigens (ENA) tests to the ANA test may capture more autoimmunity and provide critical medical information at an early stage. In this study, we?evaluated the clinical significance of a multiplex ANA + ENA panel. Methods: A cohort of 110 subjects, identified as ANA negative but ENA positive, were followed up for two years. The detection of their ANA and anti-ENA autoantibodies was assessed with a multiplex ANA + ENA panel at Vibrant America Clinical Laboratory. Results: During two years of multi-visit follow-up, 23 out of 110 subjects (20.9%) were found to become ANA positive within an average of 385 (±144) days. Histone (50/110, 45.5%) and Chromatin (25/110, 22.7%) antibodies were the most frequently found antibodies at their first visits. The subjects who were positive for RNP (5/8, 62.5%) and SSA (Ro) (10/22, 45.5%) have the highest ratio of conversion to positive ANA. No significant correlation was observed between the conversion frequency and the number of anti-ENA antibodies being carried. Conclusion: This study, which followed up on the subjects who had disparate ANA and ENA test results, showed that anti-ENA antibodies may exist years earlier than ANA. Combining ENA tests with ANA screening may reduce false negatives and improve sensitivity.

An Evans Syndrome Case Expressing Anti-Jk^aAutoantibody under Condition of Primary IgA Immunodeficiency

Autoimmune haemolytic anaemia is a haemolytic disease resulting from an autoimmune reaction to the surface of red blood cells. A part of autoantibody is known to react with the blood type antigen. This is the case of a 14 years old female with Evans syndrome in which autoimmune haemolysis may cause from anti-Jka autoantibody reaction. As this case is complicated with primary IgA immunodeficiency syndrome, anti-Jka autoantibody may occur under the condition of primary immunodeficiency status, in which autoantibody production is accelerated. Considering the co-occurrence of autoimmune haemolytic anaemia and primary IgA immunodeficiency syndrome, analysis focusing on specificity for red blood cells antigens will be required in IgA immunodefi-ciency syndrome patients.

Keratin 18 is an autoantibody of rheumatic heart disease

Objectives Autoantibodies play a key role in mechanism of rheumatic heart disease(RHD) and in this research,we focus on identifying the autoantibody in Chinese RHD patients which can be a potential molecular biomarker. Methods To construct RHD expression library,the total RNA of heart tissue of RHD patients was extracted and mRNA was isolated,purified and reverse-transcripted to long cDNA.Phage was used to recombining cDNA to be expression library.The library was immunoscreened by serum of active rheumatic fever patients,An autoantigen positive clone was immunoscreened.To identify the autoantibody,autoantibody gene was analysed by PCR,sequencing and bioinfor-matics. It also was subcloned and expressed in vitro.Western blotting was used to identify the expression protein and test cross-reaction effect with serum.Results An expression library with heart tissue of RHD patients was successfully established. The titer of the primary library was 3.3×10~6 pfu/ml, recombinant rate of it was 99%and 81%inserted segments were larger than 1 kb.A positive antoantibody gene was sreened and it is homologous to keratin 18.The recombined vector could expression objective protein in vitro which could be reacted with sera of active rheumatic fever patients and rheumatic heart disease patients,but could not be detected by sera of health persons.Conclusions It is an effective strategy to investigate autoantigens of Chinese RHD that constructing and immunoscreening heart tissues expression library. This research implied that keratin 18 was a candidate molecular biomarker of RHD.

Progress in research on the relationship between autism and folate receptor autoantibody expression

Autism spectrum disorder(ASD)is a serious neurodevelopmental disorder,the etiology and mechanism of which are not yet clear.Studies have shown that folate deficiency can lead to abnormalities in the de-velopment of the central nervous system.Patients with autism spectrum disorders develop folate-alpha recep-tor autoantibodies.Folate-alpha receptor autoantibodies block folate transport,leading to a deficiency of folate in nerve tissues.Folate is effective in treating patients with folate-alpha receptor autoantibodies.

Anti-rods/rings autoantibody generation in hepatitis Cpatients during interferon-α/ribavirin therapy

Chronic inflammation associated with hepatitis C virus(HCV) infection can lead to disabling liver diseases with progression to liver cirrhosis and hepatocellular carcinoma. Despite the recent availability of more effective and less toxic therapeutic options, in most parts of the world the standard treatment consists of a weekly injection of pegylated interferon α(IFN-α) together with a daily dose of ribavirin. HCV patients frequently present circulating non-organ-specific autoantibodies demonstrating a variety of staining patterns in the indirect immunofluorescence assay for antinuclear antibodies(ANA). Between 20% to 40% of HCV patients treated with IFN-α and ribavirin develop autoantibodies showing a peculiar ANA pattern characterized as rods and rings(RR) structures. The aim of this article is to review the recent reports regarding RR structures and anti-rods/rings(antiRR) autoantibody production by HCV patients after IFN-α/ribavirin treatment. Anti-RR autoantibodies first appear around the sixth month of treatment and reach a plateau around the twelfth month. After treatment completion, anti-RR titers decrease/disappear in half the patients and remain steady in the other half. Some studies have observed a higher frequency of anti-RR antibodies in relapsers, i.e., patients in which circulating virus reappears after initially successful therapy. The main target of anti-RR autoantibodies in HCV patients is inosine-5'-monophosphate dehydrogenase 2(IMPDH2), the rate-limiting enzyme involved in the guanosine triphosphate biosynthesis pathway. Ribavirin is a direct IMPDH2 inhibitor and is able to induce the formation of RR structures in vitro and in vivo. In conclusion, these observations led to the hypothesis that anti-RR autoantibody production is a human model of immunologic tolerance breakdown that allows us to explore the humoral autoimmune response from the beginning of the putative triggering event: exposure to ribavirin and interferon.

Cathepsin D: Autoantibody profiling as a diagnostic marker for cancers

Current diagnostic assays for many cancers are antigen-based and rely on the detection of circulating proteins that are associated with a particular cancer.These assays depend on the expression,synthesis,and release of specific proteins by cells(e.g.,tumor cells)through either active secretion or shedding,or as a consequence of cell death(either necrosis or apoptosis).As such,these antigenic proteins must“escape”the primary site of disease,saturate the antigen-processing capacity of the individual’s immune components,gain access to the circulation,and reach a sufficient steady-state concentration to be detected by enzyme-or radiolabel-based immunoassays.These events usually occur after the initial establishment of disease.Thus,and despite the fact that certain specific antigenic epitopes exhibit common recognition among patients with the same tumor types,the use of these antigen-based cancer assays has not been widely accepted in clinical practice,and many individual countries differ in the use of these potential diagnostic factors.Lately,an increasing number of studies demonstrated that procathepsin D secreted from cancer cells,acts as a mitogen on cancer cells and stimulates their pro-invasive and pro-metastatic properties.In this report,we focused on the possibility to use antiprocathepsin D autoantibodies as a diagnostic and/or predictive marker for cancers.

Negative conversion of autoantibody profile in chronic hepatitis B:A case report

BACKGROUND Autoimmune antibodies are detected in many diseases.Viral infections are accompanied by several immunopathological manifestations.Some autoimmune antibodies have been associated with the immune response induced by virus or drugs.Thus,a comprehensive diagnosis of chronic hepatitis B combined with autoimmune hepatitis is required,and immunosuppressant or antiviral therapy should be carefully considered.CASE SUMMARY We present a case of a patient who had negative transformation of autoimmune antibodies during chronic active hepatitis B.A 50-year-old female who had a history of asymptomatic hepatitis B virus carriers for more than 10 years presented to the hospital with the complaint of weakness for 1 wk.Blood tests revealed elevated liver enzymes;the detection of autoantibodies was positive.Hepatitis B viral load was 72100000 IU/mL.The patient started tenofovir alafenamide fumigate 25 mg daily.Liver biopsy was performed,which was consistent with chronic active hepatitis B.The final diagnosis of the case was chronic active hepatitis B.The autoimmune antibodies turned negative after 4 wk of antiviral therapy.The patient recovered and was discharged with normal liver function.There was no appearance of autoantibodies,and liver function was normal at regular follow-ups.CONCLUSION Autoimmune antibodies may appear in patients with chronic active hepatitis.It is necessary to differentiate the diagnosis with autoimmune hepatitis.

An Autoantibody Based Protein Microarray Blood Test to Enhance the Specificity of a Negative Screening Mammogram

Background: Current screening mammography for breast cancer is associated with misdiagnosis in as many as 30% of cases. Objectives: To develop and clinically evaluate a unique autoantibody based protein microarray blood test to improve the accuracy of breast cancer screening. Materials and Methods: A microarray was constructed from commercial antigens and antigens selected from screened cDNA libraries of breast cancer tissue samples. A training set containing 439 healthy controls and 276 biopsy proven breast cancer cases was used to establish a set of separating models between the two groups. These models were used to assign a diagnosis to 285 blind samples from 120 breast cancer patients and 165 healthy controls. Results: The test identified 82 of the 120 breast cancer patients and 160 of the 165 healthy controls. These results can be translated into a sensitivity of 68.3% [CI: 59% -77%] and a specificity of 97% [CI: 93% -99%], with a PPV for this validation set of 94.3% (CI: 87.10% -98.11%), NPV of 80.81% [CI: 74.62% -86.05%] and an AUC of 89.2% [CI: 78% -87%]. Conclusions: The protein microarray can be utilized to reduce the false negative rate of routine screening mammography. Women with a negative mammography and a negative blood test can be reassured and encouraged to continue routine breast cancer screening. A positive test should alert the physician about the possible presence of a breast cancer not detected by routine screening mammography and drive to perform additional investigation, such as breast ultrasound and MRI.

Application of autoantibody detection in chronic liver disease

Autoantibody(AAb)detection has become one of the standards of diagnosis for autoimmune liver disease(AILD),and some AAbs have become specific biomarkers of AILD.In addition,AAbs can be detected in patients with non-AILDs,such as viral hepatitis and alcoholic liver disease.However,the distribution characteristics and pathogenic mechanisms of AAbs in patients with non-AILD are unclear.This article summarizes the characteristics of AAbs in several common clinical chronic liver diseases(CLDs)and discusses the value of AAb analysis in CLD.

Biomarkers for neuromyelitis optica:a visual analysis of emerging research trends

Neuromyelitis optica is an inflammatory demyelinating disease of the central nervous system that differs from multiple sclerosis.Over the past 20 years,the search for biomarke rs for neuromyelitis optica has been ongoing.Here,we used a bibliometric approach to analyze the main research focus in the field of biomarkers for neuromyelitis optica.Research in this area is consistently increasing,with China and the United States leading the way on the number of studies conducted.The Mayo Clinic is a highly reputable institution in the United States,and was identified as the most authoritative institution in this field.Furthermore,Professor Wingerchuk from the Mayo Clinic was the most authoritative expe rt in this field.Keyword analysis revealed that the terms "neuro myelitis optica"(261 times), "multiple sclerosis"(220 times), "neuromyelitis optica spectrum disorder"(132 times), "aquaporin4"(99 times),and "optical neuritis"(87 times) were the most frequently used keywords in literature related to this field.Comprehensive analysis of the classical literature showed that the majority of publications provide conclusive research evidence supporting the use of aquaporin-4-IgG and neuromyelitis optica-IgG to effectively diagnose and differentiate neuromyelitis optica from multiple sclerosis.Furthermore,aquaporin-4-IgG has emerged as a highly specific diagnostic biomarker for neuromyelitis optica spectrum disorder.Myelin oligodendrocyte glycoprotein-IgG is a diagnostic biomarke r for myelin oligodendrocyte glycoprotein antibody-associated disease.Recent biomarkers for neuromyelitis optica in clude cerebrospinal fluid immunological biomarkers such as glial fibrillary acidic protein,serum astrocyte damage biomarkers like FAM19A5,serum albumin,and gammaaminobutyric acid.The latest prospective clinical trials are exploring the potential of these biomarkers.Preliminary results indicate that glial fibrillary acidic protein is emerging as a promising candidate biomarker for neuromyelitis optica spectrum disorder.The ultimate goal of future research is to identify non-invasive biomarkers with high sensitivity,specificity,and safety for the accurate diagnosis of neuro myelitis optica.

Unmet needs in biomarkers for autoimmune pancreatitis diagnosis

Autoimmune pancreatitis(AIP)is a rare chronic autoimmune disorder.The diagnosis of AIP mainly depends on histopathology,imaging and response to treatment.Serum immunoglobulin 4(IgG4)is used only as collateral evidence in diagnostic criteria for AIP because of its moderate sensitivity.Serum IgG4 levels are normal in 15%-37%of type 1 AIP and most of type 2 AIP patients.In these patients,the indeterminate imaging and histopathology may lead to the difficulty in definitive diagnosis of AIP.Therefore,discovery of new biomarkers is impor-tant for AIP diagnosis.Here,we provide some views on the progression and challenges in identifying novel serological biomarkers in AIP diagnosis.

Autoantigen Microarray for High-throughput Autoantibody Profiling in Systemic Lupus Erythematosus

Systemic lupus erythematosus （SLE） is a complex autoimmune disease characterized by the production of autoantibodies to a broad range of self-antigens. Profiling the autoantibody repertoire using array-based technology has emerged as a powerful tool for the identification of biomarkers in SLE and other autoimmune diseases. Proteomic microarray has the capacity to hold large number of self-antigens on a solid surface and serve as a high-throughput screening method for the determination of autoantibody specificities. The autoantigen arrays carrying a wide variety of self-antigens, such as cell nuclear components （nucleic acids and associated proteins）, cytoplas- mic proteins, phospholipid proteins, cell matrix proteins, mucosal/secreted proteins, glomeruli, and other tissue-specific proteins, have been used for screening of autoantibody specificities associated with different manifestations of SLE. Arrays containing synthetic peptides and molecular modified proteins are also being utilized for identification of autoantibodies targeting to special antigenic epi- topes. Different isotypes of autoantibodies, including IgG, IgM, IgA, and IgE, as well as other Ig subtypes, can be detected simultaneously with multi-color labeled secondary antibodies. Serum and plasma are the most common biologic materials for autoantibody detection, but other body fluids such as cerebrospinal fluid, synovial fluid, and saliva can also be a source of autoantibody detection.

Adult-onset immunodeficiency due to anti-interferon-gammaautoantibody in mainland Chinese

Immunodeficiency due to anti-interferon-gamma(anti-IFN-γ)autoantibody is an emerging adult-onsetimmunodeficiency syndrome associated with severeor disseminated infections caused by non-tuberculous

Exogenous insulin autoimmune syndrome:A case report and review of literature

BACKGROUND Insulin autoimmune syndrome(IAS)is a severe manifestation of spontaneous hypoglycemia.It is characterized by elevated levels of immune-reactive insulin and highly potent insulin autoantibodies(IAAs),which are induced by endogenous insulin circulating in the bloodstream.It is distinguished by recurring instances of spontaneous hypoglycemia,the presence of IAA within the body,a substantial elevation in serum insulin levels,and an absence of prior exogenous insulin administration.Nevertheless,recent studies show that both conventional insulin and its analogs can induce IAS episodes,giving rise to the notion of nonclassical IAS.Therefore,more attention should be paid to these diseases.CASE SUMMARY In this case report,we present a rare case of non-classical IAS in an 83-year-old male patient who present with symptoms of a psychiatric disorder.Upon symptom onset,the patient exhibited Whipple's triad(including hypoglycemia,blood glucose level less than 2.8 mmol/L during onset,and rapid relief of hypoglycemic symptoms after glucose administration).Concurrently,his serum insulin level was significantly elevated,which contradicted his C-peptide levels.After a comprehensive examination,the patient was diagnosed with exogenous insulin autoimmune syndrome.Considering that the patient had type 2 diabetes mellitus and a history of exogenous insulin use before disease onset,it was presumed that non classical IAS was induced by this condition.The PubMed database was used to search for previous cases of IAS and non-classical IAS to analyze their characteristics and treatment approaches.CONCLUSION The occurrence of non-classical IAS is associated with exogenous insulin or its analogs,as well as with sulfhydryl drugs.Symptoms can be effectively alleviated through the discontinuation of relevant medications,administration of hormones or immunosuppressants,plasma exchange,and lifestyle adjustments.