OBJECTIVE: To establish a flow cytometric immunobead array assay (FCIA) to quantify platelet antibodies and to explore its application in the diagnosis and treatment of ITP. METHODS: The guantitative standard curve was established by binding the human IgG of known concentration on antibody-coated microbeads; at the same time, the platelet-specific antigen and antibody complex was captured and levels of platelet antibodies were detected using the microbeads coated by 5 kinds of antibodies against platelets suca as GPIX (SZ1), GPⅠb (SZ2), GpⅢa (SZ21), GPⅡb (SZ22) and p-selection (SZ51). The fluorescence signal detected by flow cytometry were transformed into the conentration of platelet antibodies in samples through the quantitative standard curve, thereby establishing the method for quantititive detection of platelet-specific antibodies in plasm samples (FCIA), moreover the property, efficiency and clinical application of establishod FCIA method were evaluated. RESULTS: The FCIA could detect 5 kinds of antibodies against GPIX, GPⅠb, GpⅢa, GPⅡb and β-selection within a broad range of 33.29-1280 ng/ml, 45.17-1280 ng/ml, 42.07-1280 ng/ml, 46.40-1280 ng/ml, 42.48-1280 ng/ml and 42.48-1280 ng/ml respectively, and their recovery rates were 115.23%, 112.58%, 117.47%, 107.64% and 112.67% respectively. The intra-assay coefficient of variation (CV) for anti- GPIX, -GPⅠb, -GpⅢa, -GPⅡb and p-selection antibodies was 3.54%, 3.63%, 4.66%, 6.43% and 6.67% respectively, and the inter-assay CV for above mentioned antibodies were 10.89%, 7.57%, 10.34%, 6.95% and 10.72% respectively. The detection showed that the levels of 5 kinds of platelet-specific antibodies in ITP group all were higher than those in non-ITP and healthy control groups (P＜0.01). The sensitivity, specificity and accuracy of quantitatively detecting 5 kinds of antibodies for diagnosis of ITP by FCIA were 68.29%, 84.98% and 78.95% respectively, while the sensitivity, specificity and accuracy of detecting 5 kinds of antibodies by modified indirect MAIPA were 41.46%, 90.41% and 72.81% respectively. CONCLUSION The established quantitative FCIA for detection of antibodies provides a powerful tool for diaghosis and evaluation of therapeutic efficacy and prognosis of ITP patients.
Antibodies ; Autoantibodies ; Blood Platelets ; Flow Cytometry ; Humans ; Purpura, Thrombocytopenic, Idiopathic

OBJECTIVE: To explore the clinical application of screening cell combination method in the prediction of red blood cell alloantibody, so as to provide basis for clinical diagnosis. METHODS: From October 2018 to April 2020, 9 680 samples were screened with automatic blood group instrument, 79 patients with positive alloantibodies were identified by 4 sets of screening cells from different manufacturers (referred to as combined method). At the same time, cell panel Panocell-16 was used for comparative analysis. Meanwhile, the combined method was also used to identify the antibodies of 20 samples from National Center for Clinical Laboratories external quality assessment (EQA) in China and 12 samples from WHO EQA. RESULTS: The 79 alloantibodies included anti-Mia antibody (7 cases), anti-M antibody (13 cases), anti-Le CONCLUSION The combined method can identify the alloantibodies of red blood cells in Chinese population. The screening cells can be used for screening of irregular antibodies without wasting reagents at the same time.
Autoantibodies ; Blood Group Antigens ; China ; Erythrocytes ; Humans ; Isoantibodies

No abstract available.
Adult ; Autoantibodies/*blood ; Gangliosides/blood/*immunology ; Humans ; Male ; Ophthalmoplegia/blood/*diagnosis/immunology ; Recurrence ; Syndrome

Immunotherapy has become the fourth cancer therapy after surgery, chemotherapy, and radiotherapy. In particular, immune checkpoint inhibitors are proved to be unprecedentedly in increasing the overall survival rates of patients with refractory cancers, such as advanced melanoma, non-small cell lung cancer, and renal cell carcinoma. However, inhibitor therapies are only effective in a small proportion of patients with problems, such as side effects and high costs. Therefore, doctors urgently need reliable predictive biomarkers for checkpoint inhibitor therapies to choose the optimal therapies. Here, we review the biomarkers that can serve as potential predictors of the outcomes of immune checkpoint inhibitor treatment, including tumor-specific profiles and tumor microenvironment evaluation and other factors.
Autoantibodies ; blood ; immunology ; Biomarkers, Tumor ; blood ; immunology ; Humans ; Immunotherapy ; Neoplasms ; blood ; therapy ; Tumor Microenvironment

Allergic response to common environmental agents has been regarded as a main pathogenetic mechanism of bronchial asthma. However, allergic sensitization (atopy) can not be detected in a siginificant number of adult asthmatic patients. The etiology of nonatopic asthma has not yet been defined. To evaluate the possible involvement of autoimmune response against bronchial mucosa in the pathogenesis of nonatopic asthma, we performed indirect immunofluorescence staining of fresh frozen human bronchial mucosa tissue using serum samples from patients with atopic and nonatopic asthma, healthy controls, and patients with systemic lupus erythematosus. On immunostaining, circulating IgG autoantibodies against bronchial mucosa were detected in 2 (9.1%) of 22 patients with nonatopic asthma and in none of 22 patients with atopic asthma and of 22 healthy controls. IgG autoantibodies from the two patients with nonatopic asthma predominantly stained the cytoplasmic membrane of basal cells in bronchial epithelium. Serum samples from 10 patients with systemic lupus erythematosus immunostained the nucleus of epithelial cells in whole layer of bronchial epithelium. This study showed the presence of circulating IgG autoantibodies against the bronchial epithelial cell in a small portion of patients with nonatopic asthma. Further studies may be necessary to evaluate the possible involvement of autoimmune mechanism in the pathogenesis of nonatopic asthma.
Asthma/immunology* ; Autoantibodies/immunology* ; Autoantibodies/blood ; Bronchi/immunology* ; Epithelial Cells/immunology ; Human ; Immunity, Mucosal/immunology ; Respiratory Mucosa/immunology*

Vitiligo is a disease in which melanocytes are selectively destroyed. The disease is thought to be an autoimmune process being there are antibodies to pigment cells in the sera of patients and animals with vitiligo. In the present study, sera from vitiligo patients were examined for reactivity with the human melanoma cell line, SK-Mel-28, by Western blot analysis of solubilized membrane antigens of these cells to identify the pigment cell antigens defined by antibodies in the patients with vitiligo. Antibody reactivity to human melanoma cells (SK-Mel-28) was investigated in 14 patients with vitiligo, and 16 with normal control individuals. Antibodies to the 116-113, 60, 40 KD antigens were associated with vitiligo being present in 79%, 86%, and 43% respectively of the patients with vitiligo, but in only 6%, 38% and 6% of the normal controls. In contrast, antibodies to the 160-155, 78 and 64 KD antigens were equally common in vitiligo and in normal individuals. The results suggest that autoreactivity to pigment cells occurs more commonly in patients with vitiligo than in the normal control and high autoreactivity to pigment cells in the vitiligo sera might be an impertinent epiphemenon to destroyed pigment cell.
Antibodies, Neoplasm/*blood ; Antigens, Neoplasm/immunology ; Autoantibodies/blood ; Blotting, Western ; Human ; Melanoma/*immunology ; Vitiligo/*immunology

OBJECTIVETo establish the method of enzyme-linked immunosorbent assay (ELISA) for measuring human IgM autoantibody to folate receptor.

METHODSFolate receptor was extracted and purified from the healthy woman placenta. The protein was coated on 96-well plates with a concentration of 5 ng/Μl. Goat monoclonal antibody was used for detecting antibody. Pooled plasma from healthy donors was used to plot the standard curve and the IgM concentration of pooled plasma was defined as 1. We set up an ELISA procedure to measure human IgM autoantibody to folate receptor. The sensitivity, precision, and stability of the method were evaluated. Further, the folate receptor and bovine folate-binding protein were used as the antigen, respectively, to determine the autoantibody levels in 24 healthy individuals and 20 individuals once gave birth to baby with neural tube defects.

RESULTSThe measuring range of the method was from 6.25 × 10⁻⁴ to 8.00 × 10⁻². The lowest IgM level that can be detected was 3.12 × 10⁻⁴. The inter-assay coefficients of variations for samples with high, medium, and low IgM levels were 6.61%,3.50%, and 5.12%, respectively. The intra-assay coefficients of variations were 4.54%, 5.49%, and 5.44%, respectively. The stability test results were considered within acceptable limits. The data from folate receptor-ELISA was significantly higher than that from bovine folate binding protein-ELISA, both in the healthy group (t=-11.9, P<0.001) and in the neural tube defect group (t = 7.35, P<0.001).

CONCLUSIONSThe folate receptor-ELISA method for measuring human IgM autoantibody to folate receptor was successfully established. The method is sensitive, repeatable, and stable.

Autoantibodies ; blood ; Enzyme-Linked Immunosorbent Assay ; methods ; Folate Receptor 2 ; immunology ; Humans ; Immunoglobulin M ; blood

OBJECTIVETo assess the clinical features of primary Sjgren's syndrome (pSS) in childhood and adult patients to understand the differences between them.

METHODData of 21 childhood and 400 adult patients with definite primary Sjgren's syndrome were analyzed retrospectively.

RESULTSIn adult patients, initial clinical symptoms were various, dry mouth and dry eyes, arthritis and arthralgia, parotid swelling, renal tubular abnormalities (RTA) were more common. Compared with adult patients, RTA, parotid swelling and rash were more common in childhood, pulmonary abnormalities and neurologic system involvement were less common. The frequency of renal tubular abnormalities (52.4%) and rash (47.6%) in childhood pSS were higher than those of adults, but the frequencies of dry eyes (61.0%) and pulmonary interstitial fibrosis (25.2%) in adults pSS were higher than those in childhood (P < 0.01); serum rheumatoid factor (RF) and gamma-globulin were higher in all childhood patients (P < 0.01). Anti-nuclear antibody (ANA), anti-SSA and anti-SSB antibodies, and IgG were significantly higher in childhood cases than in adult patients (P < 0.05 or 0.01).

CONCLUSIONThe major clinical characteristics of childhood pSS cases included: (1) RTA, parotid swelling and rash which appeared earlier, but dry mouth and dry eyes appeared later and were mild. (2) The frequency of pulmonary abnormalities, nervous system involvement and Raynaud's phenomenon were less commonly seen. (3) Positivity of RF, anti-SSA and anti-SSB antibodies, and serum IgG were higher.

Adult ; Autoantibodies ; blood ; Child ; Humans ; Retrospective Studies ; Sjogren's Syndrome ; blood ; complications ; diagnosis

Autism spectrum disorders (ASD) are a group of neuro-developmental disorders in early childhood which are defined by social difficulties, communication deficits and repetitive or restrictive interests and behaviours. The etiology of ASD remains poorly understood. Much research has shown that children with ASD suffer from immunological dysfunction. This article reviews the current research progress on immunological dysfunction in children with ASD, including abnormalities in immune cells, antibodies, complements, cytokines, major histocompatibility complex and their potential association with ASD, and explores the impacts of maternal immunological activation on the immune dysfunction of children with ASD.
Autoantibodies ; blood ; Child ; Child Development Disorders, Pervasive ; etiology ; immunology ; Cytokines ; physiology ; Humans ; Immunoglobulins ; blood ; Lymphocytes ; immunology

We selected 12 antigens corresponding to commonly used autoantibodies in clinical practice to prepare antigen microarray. We chose NBT/BCIP color reaction as the end detection strategy to develop a new autoantibody protein chip detection system. Using this system, we optimized the best spotting solution, spotting concentration of the 12 antigens and the dilution of serum. We prepared a protein chip that could detect 12 autoantibodies simultaneously using the optimized antigen concentration. We established a new method to determine the cutoff of each autoantibodies by evaluation of 678 positive and 120 negative serum of clinical sample. We also evaluated the sensitivity and specificity of our new detection system. The optimal spotting solution was 0.1% TBST, the dilution of serum was 1:4 and the best spotting concentration of the 12 antigens were ANA 520 microg/mL, Ro-60/SSa 465 microg/mL, La/SSb 530 microg/mL, Jo-1 530 microg/mL, Scl-70 525 microg/mL, Sm 520 microg/mL, Ro-52/SSa 615 microg/mL, RF 340 microg/mL, CCP 465 microg/mL, ulRNP 410 microg/mL, CENP-B 490 microg/mL and dsDNA 580 microg/mL respectively. It had higher coincidence rate compared to current clinical used methods. We have developed a 12 antigens protein chip for the detection of autoantibodies based on the NBT/BCIP color reaction system. This system was fast, convenient, efficient, and cost-effective.
Autoantibodies ; blood ; immunology ; Autoimmune Diseases ; blood ; immunology ; Humans ; Protein Array Analysis ; instrumentation ; methods ; Sensitivity and Specificity