Identification of Antibodies Against Neutrophil Surface Antigens in Two Iranian Patients with Autoimmune Neutropenia

Autoimmune neutropenia is a type of immune-mediated neutropenia, caused by antibody-induced neutrophil destruction. Here, we report two cases (a 3-year-old boy and a 9-year-old girl) with suspected autoimmune neutropenia. The presence of neutrophil antibodies in the sera of these two patients was investigated; using standard neutrophil antibody screening tests such as granulocyte immunofluorescence test (GIFT), granulocyte agglutination test (GAT), and lymphocyte immunofluorescence test (LIFT). A positive reactivity with two-panel cells was found in GIFT. No reactivities with panel cells were observed in GAT and LIFT. To the best of our knowledge, this is the first report for detecting the neutrophil reactive antibodies; using genotyped neutrophils in patients with autoimmune neutropenia in Iran. The final diagnosis of our patients was primary autoimmune neutropenia for the boy and autoimmune neutropenia associated with familial Mediterranean fever for the girl.

Spot the Difference: A Case of Hailey-Hailey Disease in a 64-year-old Filipino Female

Hailey-Hailey disease (HHD) is an uncommon acantholytic disorder of the skin. This is a case of a 64-year-old Filipino female with a chronic history of painful and malodorous intertriginous plaques. Histopathologic evaluation showed overlapping features of pemphigus vulgaris and Hailey-Hailey disease. A negative direct immunofluorescence test clinched the diagnosis of Hailey-Hailey disease. The patient was advised regarding preventive measures and treated with topical antibiotics and corticosteroids with improvement of her lesions.

Anticentromere antibody induced by immunization with centromere protein and Freund’s complete adjuvant may interfere with mouse early-stage embryo

Background Anticentromere antibody (ACA) is a member of the antinuclear antibody spectrum (ANAs) which has been speculated to be associated with subfertility. Thus, the present study aimed to investigate the induction of ACA production and its potential interference with early-stage embryos. Methods Recombinant centromere protein-A (CENP-A) or centromere protein-B (CENP-B) and complete Freund’s adjuvant (CFA) were used to immunize mice. Serum ACA level was then evaluated by using an indirect immunofluorescence test. Immunofluorescence assay was performed to detect IgG in follicles in ovarian tissues and early-stage embryos. Results Following treatment, serum positive ACA was observed in mice treated with CENP and CFA. Furthermore, IgG were detected in follicular fluid and early-stage embryos from mice treated with CENP and CFA. Conclusions This study preliminarily indicated that ACA induced by CENP and CFA may penetrate into the living embryos of early-stage in mice.

Effect of Zhuang Medicine Feilongzhangxue on the Expression of HMGB1-TLR4 / RANKL-NF-?B Signaling Pathway Related Factors in Osteoclasts

Objective: To study the regulatory effect of feilongzhangxue on the levels of HMGB1-TLR4 / RANKL-NF-?B signaling pathway related factors HMGB1, RANKL, rank, TRAF-6 and NF-?Bp65 in osteoclasts, so as to explore the mechanism of feilongzhangxue intervention in RA; Methods: The osteoclasts with good activity were randomly divided into blank group, methotrexate control group and Zhuang medicine feilongzhang blood containing serum treatment group, which were divided into OC + blank group, OC + methotrexate control group, OC + Zhuang medicine feilongzhang blood containing serum group; The expression of HMGB1, RANKL, rank, TRAF-6 and NF-?Bp65 mRNA was detected by RT-PCR; The protein expressions of HMGB1, RANKL, RANK, TRAF-6 and NF-?Bp65 were detected by immunofluorescence. Results: PCR results showed that: Compared with the blank group, feilongzhangxue could effectively inhibit the expression levels of HMGB1, RANKL, rank, TRAF-6 and NF-? B p65 mRNA in OC cells, and the inhibitory effect was stronger than methotrexate. Immunofluorescence test results showed that: Compared with the blank group and methotrexate group, feilongzhangxue could effectively inhibit the protein expression of HMGB1, RANKL, rank, TRAF-6 and NF - ? B p65 in OC cells. Conclusion: The effect of Zhuang medicine feilongzhangxue on hmgb1-tlr4 / rankl-nf - ? B signaling pathway of osteoclasts is through the regulation of related factors HMGB1, RANKL, rank, TRAF-6 and NF-?Bp65, which may be the key mechanism of Zhuang medicine feilongzhangxue on rheumatoid arthritis.

Neonatal Immune Neutropenia due to Isoantibodies against the Granulocyte Receptor FcγRIIIb

Introduction: Neonatal neutropenia is often secondary to sepsis, low birth weight, pregnancy-induced maternal hypertension, and other conditions. Case Report: We report a case of asymptomatic isoimmune neutropenia in a pair of preterm twins. Genotyping confirmed that the mother was negative for HNA-1a, 1b, and 1c, consistent with an FcγRIIIb deficiency. The father was 1(a+b+c–) and the neonates were 1(a–b+c–). A strongly positive result was observed in the granulocyte immunofluorescence test against paternal neutrophils (IgG antibodies). IgG anti-CD16b isoantibodies were detected in the mother’s breast milk. Neutropenia resolved after 28 days without requiring any specific treatments. Discussion: Even though neonatal alloimmune neutropenia (NAN) is usually benign and self-limiting, some patients pre­sent with delayed separation of the umbilical cord, mild skin infections, omphalitis, or severe infections like pneumonia, sepsis, and meningitis. Thus, it is important to rule out NAN in case of neonatal neutropenia.

Transcriptomic Analysis Reveals PTBP1 as a Key Molecular Signature of Sciatic Nerve Injury Recovery

Background: Peripheral nerves control motor, sensory, and autonomic functions, so injury can seriously affect a patient's quality of life. There have been studies that have shown that the repair factors are different at different ages, and we have identified a repair hub gene that plays a key role throughout the entire age group. Methods: From Gene Expression Omnibus database GSE4090, mice of 2 and 24 months of age after sciatic nerve injury were selected from mice transcriptome data of differentially expressed genes in common, and the hub genes were then determined using protein-protein network and MCODE analysis, DAVID biological process, molecular function, and cell component analysis, and the miRWalk analysis of hub genes was performed to verify the key molecule. In mice aged eight weeks of sciatic 2 nerve clamps damage building, on days 0, 1, 4, and 7, sciatic nerve motor and sensory function were evaluated, and sciatic nerve immunofluorescence test was performed to verify PTBP1 expression. The continuous data were expressed as the mean ±SD. An independent t-test was used to compare two groups. A p-value of less than 0.05 was considered statistically significant. Results: Bioinformatics analysis showed that PTBP1 is one of the key molecules in mouse sciatic nerve repair after injury. The immunofluorescence test verified that the number of positive cells reached a maximum value of 30.6 ±6.4/ROI on day 7 after injury and a minimum value of 17.4 ±7.0/ROI in the control group (p<0.001). However, the percentage of PTBP1 positive cells reached a peak of 90.8 ±16.9% at the early stage of injury, i.e., the first day, and then dropped to a minimum of 75.7 ±8.9% on the seventh day in the animal experiment as the repair time gradually increased (p<0.05). Conclusions: PTBP1 plays a key role in the repair of sciatic nerve injury, providing a new strategy for clinical treatment of patients of all ages.

Sensitivity of anti-SARS-CoV-2 serological assays in a high-prevalence setting

Evaluation and power of seroprevalence studies depend on the performed serological assays. The aim of this study was to assess four commercial serological tests from EUROIMMUN, DiaSorin, Abbott, and Roche as well as an in-house immunofluorescence and neutralization test for their capability to identify SARS-CoV-2 seropositive individuals in a high-prevalence setting. Therefore, 42 social and working contacts of a German super-spreader were tested. Consistent with a high-prevalence setting, 26 of 42 were SARS-CoV-2 seropositive by neutralization test (NT), and immunofluorescence test (IFT) confirmed 23 of these 26 positive test results (NT 61.9% and IFT 54.8% seroprevalence). Four commercial assays detected anti-SARS-CoV-2 antibodies in 33.3-40.5% individuals. Besides an overall discrepancy between the NT and the commercial assays regarding their sensitivity, this study revealed that commercial SARS-CoV-2 spike-based assays are better to predict the neutralization titer than nucleoprotein-based assays are.

Influence of polyelectrolytes on increasing sensitivity of an immunofluorescent analysis based on plasmon silver nanoparticles

It was studied the dependence of the interaction of the components of the immunofluorescence test system for the quantitative determination of prostate specific antigen based on plasmon silver nanoparticles on the physicochemical nature of various polyelectrolytes used to coat films of silver nanoparticles. It has been shown that the use of a weakly charged polycationic polyelectrolyte poly-L-lysine can increase the antigenic binding of the test system by 2.34 times, and the use of a highly charged polycationic polyelectrolyte polydiallyldimethylammonium chloride increases the binding affinity of prostatic specific antigen by 5 times. When developing various immunochemical test systems using films of silver nanoparticles, an important parameter is the choice of a polyelectrolyte for coating a silver nanolayer, since the physicochemical and electrostatic properties of the polyelectrolyte can significantly affect both the sorption capacity of the solid phase and the conformational state functional activity of immobilized protein molecules. Both specificity and sensitivity of the immunochemical test system, as well as the minimum possible detectable concentration of bioanalyte, largely depend on these parameters.