Elevated Antibody Levels Research Articles

ОСОБЕННОСТИ КЛИНИЧЕСКИХ И ЛАБОРАТОРНЫХ ПОКАЗАТЕЛЕЙ У ПАЦИЕНТОВ С ИНФЕКЦИЕЙ SARS-COV-2 В ЗАВИСИМОСТИ ОТ РЕЗУЛЬТАТОВ СЕРОЛОГИЧЕСКОЙ ДИАГНОСТИКИ

To date, over 270 million people have been ill with COVID-19 and more than 2 million have died. Tens of thousands of new cases of the disease are still registered in the world every day, so the assessment of the methods for diagnosing SARS-CoV-2 associated pathology remains highly relevant. The aim of the study was to evaluate the variability of clinical and laboratory parameters in patients with SARS-CoV-2 infection depending on the results of serological diagnostics. Material and methods. 170 patients (88 women, 82 men aged 23 to 90 years) diagnosed with community-acquired interstitial pneumonia following SARS-CoV-2 infection were examined in accordance with the clinical protocol, in 80 of them the blood levels of IL-1β, IL-6, IL-18, CXCL8, and interferon-α were determined. Results. It was found that only in 34.09% of cases a negative PCR result for the presence of the genetic material of SARS-CoV-2 viruses in patients diagnosed with interstitial pneumonia coincided with a negative result of testing for IgG and/or IgM to the viral antigen. A positive PCR result was confirmed by the presence of elevated levels of antibodies in 64.29% of cases. In PCR-positive patients, stage II III respiratory failure was determined significantly more often than in PCR-negative ones (in 40.28% and 16.32% of cases, respectively, p<0.01), and their condition was more often regarded as severe and extremely severe (in 31.94% and 10.0% of cases, respectively, p<0.05). The level of antibodies was associated with the severity of the immune response to a viral infection: the volume of lung damage, the blood level of the pro-inflammatory cytokine IL-18, acute phase inflammation proteins, and the activity of intracellular enzymes were significantly higher in patients with elevated levels of IgG and/or IgM than in patients with negative results of these tests. Conclusions. The determining of IgM and IgG specific to the Spike antigen of the SARS-CoV-2 virus does not duplicate, but complements the molecular genetic study. PCR helps to detect the presence of the genetic material of the virus. Detection of the level of antibodies can be used to make a conclusion about the intensity and stage of the immune response to a viral infection.

HLA Typing and Donor-Specific Antibody Screening in Kidney Transplantation: Bridging the Past to the Future

Human leukocyte antigens (HLA) are unique proteins expressed on the surface of human cells, playing a pivotal role in the immune system, particularly in the contexts of infection, cancer, and transplantation. The widespread adoption of HLA typing methods has become an essential component in assessing donor-recipient compatibility, a crucial limiting factor in solid organ transplantation. In general, the greater the disparity between a donor's and recipient's HLA types, the higher the likelihood of provoking an alloimmune response, which frequently results in alloimmune graft rejection. With significant advancements in organ transplantation techniques, immunosuppressive medications, and surgical procedures, attention has increasingly turned toward understanding and managing humoral rejection processes. Pre-transplant antibody screening plays a critical role in identifying individuals with elevated levels of antibodies against potential donor antigens. This screening aids in risk assessment and planning to mitigate the risk of antibody-mediated rejection (AbMR). Several methods are available for assessing circulating antigen-specific antibodies and HLA tissue typing, including cell-based assays like serological assays, complement-dependent cytotoxicity, and flow cytometry. However, non-cell-based approaches, such as molecular methods, HLA imputation techniques and high-throughput HLA-matchmaker assays have gained significant popularity due to their ability to provide higher resolution and robust donor-recipient matching. Despite the advancements in precision and sensitivity observed in HLA cutting-edge technologies, numerous challenges still persist. These challenges involve complexities linked to allelic ambiguities, the differentiation of closely related alleles, and the ongoing effort to establish a standardized HLA testing methodology across diverse laboratories. Additionally, correlating the HLA crossmatch results with the clinical outcomes for transplant donors poses another important aspect that warrants attention and requires expert analysis. In this review, we will enumerate the different methods of HLA typing and DSA screening and discuss the unmet needs and future directions for HLA typing methods.

Investigating the association between hyperprolactinemia and thyroid autoimmunity in women

Aims: Hyperprolactinemia (HPRL), a condition characterized by elevated levels of prolactin (PRL) hormone, is considered to be related to various autoimmune diseases, such as thyroid autoimmunity (TAI). The association between HPRL and TAI still evokes controversy. Our study intends to examine the possible relationship between HPL and TAI in young women. Methods: This retrospective study was done between January 2018 and December 2022, including 90 women with HPRL and 90 control subjects of similar age and gender. A retrospective examination was conducted on the medical records of 90 patients with HPRL to gather relevant data. Information regarding related parameters was analyzed and compared to the control group data. Results: Our results found that thyroid stimulating hormone (TSH) levels were notably elevated in patients with HPRL compared to healthy women (2.24 vs. 1.86). Also, a statistically significant association between the case and control regarding serum free triiodothyronine (fT3) (p<0.05). No statistically significant difference was found between the case and control regarding serum free thyroxin (fT4) (p>0.05). Patients with HPRL demonstrated considerably higher anti thyroperoxidase antibody (anti-TPO) levels (46.08 vs. 43.95). Significantly elevated antithyroglobulin antibody (anti-Tg) levels were observed in patients with HPRL (1.07 vs. 0.84). Conclusion: The study suggests a potential link between HPRL and TAI. Further research is needed to explore this relationship and its implications for patient care.

Dynamics of temporal immune responses in nonhuman primates and humans immunized with COVID-19 vaccines.

We assessed the humoral immune responses to a COVID-19 vaccine in a well-controlled rhesus macaque model compared to humans immunized with two mRNA vaccines over several months post-second dose. The plasma IgG levels against seven coronaviruses (including SARS-CoV-2) and antibody subtypes (IgG 1-4 and IgM) against SARS-CoV-2 were evaluated using multiplex assays. The neutralization capacity of plasma antibodies against the original SAR-CoV-2 isolate and nine variants was evaluated in vaccinated humans and non-human primates. Immunization of macaques and humans with SARS-CoV-2 vaccines induced a robust neutralizing antibody response. In non-SIV-infected adult macaques immunized with an adenoviral vector expressing S-RBD (n = 7) or N protein (n = 3), elevated levels of IgG and neutralizing antibodies were detected 2 weeks post-second dose. Immune responses to the S-RBD vaccine in SIV-infected adult macaques (n = 2) were similar to the non-SIV-infected animals. Adult humans immunized with Pfizer (n = 35) or Moderna (n = 18) vaccines developed IgG and neutralizing antibodies at 4 weeks post-second dose. In both vaccine groups, IgG 1 was the predominant subtype, followed by IgG 3. The IgG levels, including total and IgG 1,2,3 elicited by the Moderna vaccine, were significantly higher than the corresponding levels elicited by the Pfizer vaccine at 4 weeks post-second dose. A significant correlation was observed between the plasma total IgG antibody levels and neutralization titers in both macaques and humans. Furthermore, broad-spectrum neutralization antibodies against several variants of SARS-CoV-2 were detected in the plasma of both macaques and humans after two vaccinations.

Heterologous prime-boost immunization induces protection against dengue virus infection in cynomolgus macaques.

Currently licensed dengue vaccines do not induce long-term protection in children without previous exposure to dengue viruses in nature. These vaccines are based on selected attenuated strains of the four dengue serotypes and employed in combination for two or three consecutive doses. In our search for a better dengue vaccine candidate, live attenuated strains were followed by non-infectious virus-like particles or the plasmids that generate these particles upon injection into the body. This heterologous prime-boost immunization induced elevated levels of virus-specific antibodies and helped to prevent dengue virus infection in a high proportion of vaccinated macaques. In macaques that remained susceptible to dengue virus, distinct mechanisms were found to account for the immunization failures, providing a better understanding of vaccine actions. Additional studies in humans in the future may help to establish whether this combination approach represents a more effective means of preventing dengue by vaccination.

Ophthalmologic Manifestations of Ocular Myasthenia Gravis

Purpose: We investigated the ophthalmic manifestations observed in patients diagnosed with ocular myasthenia gravis.Methods: In total, 58 patients diagnosed with ocular myasthenia gravis visited the ophthalmology and neurology departments between January 2011 and August 2022. Patients were followed for > 6 months; their ophthalmic manifestations were analyzed retrospectively.Results: The study included 29 males and 29 females with a mean age of 55.52 ± 15.60 years. Among the patients, 31 tested positive for anti-acetylcholine receptor antibodies, resulting in a positive rate of 53.45%. Abnormalities in repeated nerve stimulation tests were observed in 33.33% of the patients. The antibody-positive group exhibited a higher frequency of progression to generalized myasthenia gravis (p = 0.011) and a higher incidence of chest abnormalities (p < 0.001) compared to the antibody-negative group. Horizontal and vertical diplopia were commonly observed in a complex group of patients with diplopia and blepharoptosis (p < 0.001); there was a difference in the pattern of strabismus between the diplopia single group with esotropia and the complex group with exotropia (p = 0.029). In addition, the combination group demonstrated a higher antibody titer (p = 0.034), a higher frequency of abnormalities in repeated nerve stimulation tests (p = 0.022), and a higher incidence of chest abnormalities (p = 0.022).Conclusions: The anti-acetylcholine receptor antibody-positive group had a higher incidence of progression to generalized myasthenia gravis; moreover, the complex group of patients with diplopia and ptosis exhibited elevated levels of anti-acetylcholine receptor antibodies and frequently accompanied both horizontal and vertical strabismus.

FRI559 An Uncommon Association Of Two Common Etiologies Of Hyperthyroidism

Abstract Disclosure: C. Chew: None. Introduction: Subacute thyroiditis and Graves’ disease are two common etiologies of hyperthyroidism. Association of these two diseases in patient presented with recurrent hyperthyroidism has rarely been reported. Case Report: A 35 years old women, with no known medical illness, was referred to Endocrine Clinic for painful anterior neck swelling associated with palpitation for 2 weeks. The symptoms were preceded by 3 days history of sore throat and mild fever She denied other symptoms of hyperthyroidism. Her last menses was 3 weeks prior. On physical examination, she was tachycardic with heart rate of 115 beats per minute. Her blood pressure was 135/90 mmHg and she was afebrile. There was a tender and diffusely enlarged goiter. However, there were no signs of thyroid eye disease and other signs of hyperthyroidism. Examinations of her throat, lung, abdomen, cardiovascular system and neurological system were unremarkable. There was also absence of lymphadenopathy. She denied family history of thyroid disease. Results of laboratory investigations were suggestive of hyperthyroidism secondary to subacute thyroiditis: Free T4 52 pmol/L (8-16), TSH <0.01 mIU/L (0.45-4.50), TRAb 0.8 IU/L (0-1.0) and ESR 108 mm/hr (1-10). The technetium scan showed mildly enlarged gland with decreased uptake of Tc-99m tracer suggestive of thyroiditis. She was treated with Ibuprofen and Propranolol, and her symptoms improved. At 6 weeks follow up, her thyroid function normalized with Free T4 of 13 pmol/L (8-16), TSH of 1.28 mIU/L (0.45-4.50). However, she complained of weight loss and palpitation at 6 months follow up. Examination revealed a small painless diffuse goiter and she was tachycardic with heart rate of 115 beats per minute. Thyroid function test confirmed recurrence of hyperthyroidism: free T4 45.8 pmol/L (8-16) and TSH <0.01 mIU/L (0.45-4.50). Elevated TSH receptor antibody (TRAb) level at 16.8 IU/L confirmed the diagnosis of Graves’ disease. Carbimazole was then started. Discussion: Subacute thyroiditis and Graves’ disease are two common etiologies of hyperthyroidism. The incidence of association of these two diseases had been reported to be rare at 0.15% in patients with Graves’ disease and 0.76% in patients with subacute thyroiditis (1). The postulated mechanism of the association is that the release of thyroid antigens and corresponding activation of the immune system due to viral invasion of the thyroid gland in subacute thyroiditis may cause the subsequent Graves’ disease (1,2). Conclusion: Graves’ disease should be suspected in patient with persistent or recurrent hyperthyroidism after an episode of subacute thyroiditis although it is rare. Reference: 1. Nakano Y et al. Graves’ disease following subacute thyroiditis. Tohoku J. Exp. Med. 2011; 225:301-309. 2. Bartalena L et al. Graves’ disease occurring after subacute thyroiditis. Report of a case and review of the literature. Thyroid. 1996; 4:345-348. Presentation: Friday, June 16, 2023

SAT460 Higher TSH Receptor Antibody At Diagnosis And Younger Age Predict Recurrence of Graves' Thyrotoxicosis After Thionamide Withdrawal At 10 Years

Abstract Disclosure: F.W. Gibb: None. N. Tun: None. Background: Anti-thyroid drug (ATD) therapy in Graves’ disease is generally accepted to be a safe and effective means of restoring euthyroidism. Treatment is typically administered for 12 to 18 months followed by cessation subject to certain conditions such as achieving euthyroidism on a low dose and, in some centers, exclusion of persistently elevated TSH receptor antibody (TRAb) levels. We and others have previously demonstrated the prognostic utility of TRAb at diagnosis and cessation of ATDs in predicting recurrence risk. However, very few studies have assessed recurrence risk in the long-term. Methods: Prospective observational study of thyroid status at 10 years in 231 people who completed a physician directed course of ATD for Graves’ disease (first episode). We also assessed clinical predictors of recurrence at 10 years post ATD-cessation. Results: Median age was 43 years (IQR 35 - 53) and 181/231 (78%) were female. 10 years after cessation of ATD 124/231 (53.7%) had experience a relapse of thyrotoxicosis. The following parameters were associated with recurrence at 10 years: 1. TRAb category at diagnosis: <5 IU/L (relapse 43.2%), 5 - 12 IU/L (relapse 50.0%) and >12 IU/L (relapse 71.9%) (p = 0.003). Median TRAb in those with relapse was 8.7IU/L (4.9 - 16.9) vs. 5.9 IU/L (4.1 - 9.9) in those in remission at 10 years (p = 0.002). 2. TRAb at cessation of ATD was 1.3 IU/L (<0.6 - 2.2) in relapse and 1.0IU/L (<0.6 - 1.4) in remission (p = 0.006). 3. Median age in those with relapse was 40 years (32 - 49) vs. 47 (39 - 56) in remission (p = 0.001). 4. Median fT4 at diagnosis was 33pM (26 - 42) in those with subsequent relapse vs. 30pM (24 - 36) in remission (p = 0.045) 5. Time to normalisation of fT4 was 2 months (1 - 2) in people with relapse vs. 1 month (1 - 2) in people in remission (p = 0.003). In logistic regression higher TRAb at cessation (p = 0.003) and younger age (p = 0.002) were independently associated with relapse at 10 years. Sex, smoking status and presence of ophthalmopathy did not influence recurrence risk. 73% of relapses occurred within 2 years and only 5% of relapses occurred between 5 and 10 years. At ten years, 16 patients (6.9%) had died (9 in remission and 7 after relapse), 45 were on long-term ATD (19.5%), 39 received radioiodine (16.9%), 13 had thyroidectomy (5.6%), 7 developed hypothyroidism (3.0%) and 111 were on no treatment (48.1%). Conclusions: Discussions with patients about risk of relapse should be individualised based on TRAb and age. In this cohort someone under 43 years with TRAb >12 IU/L at diagnosis had a 88% risk of relapse whilst someone older than 43 with TRAb <5 IU/L at diagnosis had a relapse rate of 38%. However, although the recurrence rate is lower in older individuals, the clinical risks of recurrent thyrotoxicosis are greater and may still support definitive therapy or long-term ATD use. Presentation: Saturday, June 17, 2023

Effect of physiotherapeutic treatment on the quality of life in a patient with antiphospholipid syndrome: A case report

Antiphospholipid syndrome (APS) is a systemic autoimmune disease with elevated levels of antiphospholipid antibodies (aPL). The chronic course, multisystem involvement, limitation of physical activity and frequent fatigue lead to reduced health-related quality of life. There is insufficient scientific evidence in the literature regarding the feasibility and safety of using physical rehabilitation factors and their impact on functional activity and quality of life in these patients.

Association of thyroid peroxidase antibody with the RNF213 p.R4810K variant in ischemic stroke/transient ischemic attack

Background and aimsRNF213 is a susceptibility gene for moyamoya disease and vasospastic angina, with a second hit considered necessary for their development. Elevated thyroid peroxidase antibody (TPO-Ab) levels have been observed in both diseases, suggesting a possible role of TPO-Ab as a second hit for developing RNF213-related vasculopathy. We investigated the association of TPO-Ab levels with RNF213-related ischemic stroke (IS)/transient ischemic attack (TIA), other than moyamoya disease. MethodsFrom the National Cerebral and Cardiovascular Center Genome Registry, a multicenter, prospective, observational study, we enrolled patients with IS/TIA who were admitted within 1 week of onset. Patients with IS/TIA due to definite moyamoya disease or hemorrhagic stroke were excluded. Participants underwent genotyping for RNF213 p. R4810K, and baseline characteristics and TPO-Ab levels were compared between RNF213 p. R4810K variant carriers and non-carriers. ResultsIn total, 2090 IS/TIA patients were analyzed [733 women (35.1%); median age 74 (interquartile range, 63–81) years, baseline NIHSS score 3 (2–6)], and 85 (4.1%) of them carried the variant. Median TPO-Ab levels were significantly higher in variant carriers (8.5 IU/mL vs. 2.1 IU/mL, p < 0.01), who also showed a higher frequency of elevated TPO-Ab levels (>16 IU/mL) (27.1% vs. 4.4%). In the multivariate analysis, presence of the RNF213 p. R4810K variant (adjusted odds ratio, 12.42; 95% confidential interval, 6.23–24.75) was significantly associated with elevated TPO-Ab levels. ConclusionsElevated TPO-Ab levels may be significantly associated with presence of the RNF213 p. R4810K variant in IS/TIA patients. Thus, TPO-Ab may inherently modify IS/TIA development in RNF213 p. R4810K variant carriers.

P06.19.A BEYOND T CELL TOXICITY - INTRATHECAL CXCL13 ELEVATION INDICATES B-CELL INVOLVEMENT IN NEUROLOGICAL IMMUNE-RELATED ADVERSE EVENTS FOLLOWING IMMUNE CHECKPOINT INHIBITION: 2 CASE OBSERVATIONS AND COMPREHENSIVE REVIEW

Abstract BACKGROUND Neurological immune-related adverse events (nirAE) can occur following cancer treatment with immune checkpoint inhibitors (ICI). Mainly attributed to autoreactive T-cells, nirAE usually respond to first line therapy with high-dose steroids (GC). Severe or fatal outcomes are rare &amp; most frequently reported in clinically overlapping syndromes with accumulating evidence of B cell autoimmunity. MATERIAL AND METHODS We present 2 patients who developed grade 3-4 nirAE with evidence suggestive for B cell involvement. RESULTS Case 1: A 75y female with metastatic melanoma developed double vision &amp; general weakness 2 weeks after the 1st cycle of ipilimumab &amp; nivolumab. Diagnostic workup showed pleocytosis (27 cells/µl) with elevated CXCL13 (316 pg/ml) in cerebrospinal fluid (CSF). Increases of creatine kinase, troponin T, elevated levels of anti-acetylcholine receptor antibodies &amp; positive anti-titin antibodies led to the diagnosis of myocarditis &amp; myositis/myasthenia gravis overlap syndrome. The patient received pulsed high-dose GC with tapering. CSF flow cytometry revealed intrathecal recruitment of CXCL13-responsive B cell helper T cells of a predominant inflammatory phenotype &amp; a highly increased pathological CD4/CD8 T cell ratio in peripheral blood. Clinically non-responding to GC, the patient received additional therapy with immunoglobulins (IVIG). 2 weeks later she was readmitted due to generalized myopathy with dropped head &amp; received a second cycle of high-dose GC since she refused plasmapheresis. Further escalation with rituximab could not be realized because the patient rapidly deteriorated &amp; deceased. Case 2: A 75y female with metastatic invasive enteric adenocarcinoma of the lung developed gait ataxia &amp; ataxia of the lower limbs 5 months after pembrolizumab therapy. CSF analysis showed a pleocytosis (248 cells/µl) with plasma cells, pronounced CXCL13 elevation (&amp;gt;488 pg/ml) &amp; positive screening for anti-neuronal cerebellar antibodies, suggesting ICI-mediated cerebellitis. The patient received high-dose GC &amp; IVIG for 5 days each, was discharged but readmitted 5 months later due to subacute onset of spastic paraparesis. Magnetic resonance imaging of the spine revealed thoracic myelopathy &amp; CSF showed mild pleocytosis (5 cells/µl), intrathecal immunoglobulin synthesis &amp; mild CXCL13 elevation (33 pg/ml). The patient completed a second cycle of steroids followed by rituximab targeting B-cell involvement &amp; currently 4 months later is clinically stable. CONCLUSION Intrathecal CXCL13 may serve as diagnostic tool of B cell involvement, here supported by intrathecal immunoglobulin synthesis, presence of plasma cells and/or severely altered immune cell compositions. Early treatment escalation with B cell depleting agents such as rituximab may help prevent fatal clinical courses.

The use of intravenous immunoglobulin in the treatment of Hashimoto's encephalopathy: case based review.

Hashimoto's encephalopathy (HE) is a controversial immunological neuropsychiatric disease, with a poorly understood pathogenesis. It is characterized by symptoms of acute or subacute encephalopathy which usually occur in the presence of elevated levels of antithyroid antibodies. Even though it is also known as steroid responsive encephalopathy associated with autoimmune thyroiditis (SREAT), some cases appear to be steroid-resistant. This review examined whether treatment of Hashimoto's encephalopathy with intravenous immunoglobulin (IVIG) is associated with better clinical outcomes than the standard therapy. Additionally, we presented a case of a 59-year-old man who presented with severe neurological manifestations and was successfully treated with intravenous immunoglobulin. The online databases PubMed and EMBASE were searched. A total of 1,365 articles were identified. After the deletion of 112 duplicates, 1,253 studies were screened by evaluating the title and abstract, focusing on Hashimoto's encephalopathy cases where IVIG were used. 846 studies were excluded because they were not relevant to the topic or included pediatric population. Therefore, 407 full-text articles were assessed for eligibility. The final analysis included 14 eligible articles after 393 were excluded (irrelevant texts, not written in English, full-text not available). In the majority of the selected case-reports, IVIG was associated with a good outcome, sometimes even with dramatic improvements in patient's status. In last years, intravenous immunoglobulin therapy proved its utility in Hashimoto's encephalopathy's treatment, being a well tolerated therapy associated with remarkable improvement in patient's status. Further research is still needed in order to define the optimal treatment protocol for Hashimoto's encephalopathy and to establish if intravenous immunoglobulin can also be used as a first-line therapy, alone or in combination with steroids.

Early Coronary Artery Disease in a Female Patient with Discoid Lupus Erythematosus and Hashimoto’s Thyroiditis

BACKGROUND: Atherosclerosis is a chronic inflammatory condition involving the endothelium of the blood vessels, predominant the coronary arteries. Main risk factors are dyslipidemia, hypertension, diabetes, smoking, obesity, and lack of physical activity. Patients with autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis etc., have a twofold increased risk of developing CAD at younger age, compared with the general population. CASE REPORT: A 41-year-old female patient, with history of smoking and dyslipidemia, was admitted to our hospital with acute inferoposterior myocardial infarction. Initially, coronary reperfusion therapy per protocol was administrated and primary percutaneous coronary intervention (PCI) was performed. Multivessel CAD was found, and two stents were implanted on obtuse marginal and circumflex artery. Transthoracic echocardiography (TTE) revealed left ventricle systolic and diastolic dysfunction with segmental hypokinesis. Additionally, the patient was first diagnosed with DLE at the age of 15, but the disease was uncontrolled in the last 7 years. She also has hypothyroidism, regularly treated with hormone replacement therapy. The patient was discharged with medicamentous therapy including dual antiplatelet agents, statin, beta-blocker, angiotensin-receptor blocker, potassium sparing diuretic and proton pump inhibitor. One month later, recoronarography was performed with stenting of left anterior descending artery. TTE showed improvement of the left ventricle systolic function with preserved ejection fraction. The blood test showed elevated levels of antithyroid antibodies. A rheumatologist was consulted, who recommended therapy with hydroxychloroquine and regular follow-ups. CONCLUSION: In younger patients with chronic inflammatory diseases, inflammatory mediators play a significant role in the development of the atherosclerotic plaques, regardless of co-existing risk factors. Therefore, an early cardiovascular assessment is required in these patients for preventing severe or life-threatening cardiovascular events.

Autoimmune gastritis as an unexpected cause of diarrhea in a young adult with type I diabetes: a case report

BackgroundType 1 diabetes mellitus (T1DM) is a lifelong diagnosis that involves immune-mediated damage of pancreatic beta cells and subsequent hyperglycemia, manifesting as: polyuria, polydipsia, polyphagia, and weight loss. Treatment of type 1 diabetes centers on insulin administration to replace or supplement the body's own insulin with the goal of achieving euglycemia and preventing or minimizing complications. Patients with T1DM are at risk for developing other autoimmune conditions, most commonly thyroid or celiac disease.Case presentationA 20-year-old African American female with T1DM was referred by her endocrinologist to pediatric gastroenterology for 2 months of nocturnal, non-bloody diarrhea, left lower quadrant pain, and nausea; she was also being followed by neurology for complaints of lower extremity paresthesias and pain. The patient’s initial lab-workup was remarkable for a low total Immunoglobulin A (IgA) level of < 6.7 mg/dL. As IgA deficiency is associated with an increased risk of celiac disease, the patient underwent upper and lower endoscopy, which was grossly unremarkable; however, histology revealed a pattern consistent with autoimmune gastritis. Subsequent serum evaluation was remarkable for an elevated fasting gastrin level and an elevated parietal cell antibody level without macrocytic anemia, iron deficiency, or vitamin B12 depletion. The patient was diagnosed with autoimmune gastritis (AIG) and subsequently initiated on parenteral B12 supplementation therapy with improvement in her neurologic and gastrointestinal symptoms.ConclusionThis case illustrates the importance of recognition of red flag findings in a patient with known autoimmune disease. Following well-established health maintenance recommendations for individuals with T1DM ensures that common comorbidities will be detected. Autoimmune gastritis, while a rarer pathology in the pediatric population, deserves consideration in patients with pre-existing autoimmune conditions and new gastrointestinal or neurologic symptoms, as AIG can be associated with poor outcomes and risk of malignancy. Initial lab findings associated with an eventual diagnosis of AIG typically include anemia, iron deficiency, or Vitamin B12 deficiency. However, as demonstrated in this case, symptoms of AIG can rarely present before anemia or Vitamin B12 deficiency develops. To prevent permanent neurological damage, parenteral Vitamin B12 therapy must be considered even in the absence of Vitamin B12 deficiency, especially in those patients already experiencing neurological symptoms.

Upregulation of IQGAP2 by EBV transactivator Rta and its influence on EBV life cycle.

Epstein-Barr virus (EBV) is a human oncogenic γ-herpesvirus that establishes persistent infection in more than 90% of the world's population. EBV has two life cycles, latency and lytic replication. Reactivation of EBV from latency to the lytic cycle is initiated and controlled by two viral immediate-early transcription factors, Zta and Rta, encoded by BZLF1 and BRLF1, respectively. In this study, we found that IQGAP2 expression was elevated in EBV-infected B cells and identified Rta as a viral gene responsible for the IQGAP2 upregulation in both B cells and nasopharyngeal carcinoma cell lines. Mechanistically, we showed that Rta increases IQGAP2 expression through direct binding to the Rta-responsive element in the IQGAP2 promoter. We also demonstrated the direct interaction between Rta and IQGAP2 as well as their colocalization in the nucleus. Functionally, we showed that the induced IQGAP2 is required for the Rta-mediated Rta promoter activation in the EBV lytic cycle progression and may influence lymphoblastoid cell line clumping morphology through regulating E-cadherin expression. IMPORTANCE Elevated levels of antibodies against EBV lytic proteins and increased EBV DNA copy numbers in the sera have been reported in patients suffering from Burkitt's lymphoma, Hodgkin's lymphoma, and nasopharyngeal carcinoma, indicating that EBV lytic cycle progression may play an important role in the pathogenesis of EBV-associated diseases and highlighting the need for a more complete mechanistic understanding of the EBV lytic cycle. Rta acts as an essential transcriptional activator to induce lytic gene expression and thus trigger EBV reactivation. In this study, scaffolding protein IQGAP2 was found to be upregulated prominently following EBV infection via the direct binding of Rta to the RRE in the IQGAP2 promoter but not in response to other biological stimuli. Importantly, IQGAP2 was demonstrated to interact with Rta and promote the EBV lytic cycle progression. Suppression of IQGAP2 was also found to decrease E-cadherin expression and affect the clumping morphology of lymphoblastoid cell lines.

Booster vaccinations and Omicron: the effects on SARS-CoV-2 antibodies in Dutch blood donors

IntroductionThe severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) booster vaccination campaign and the emergence of SARS-CoV-2 Omicron variants impact the prevalence and levels of SARS-CoV-2 antibodies in the Netherlands. In this study we determined antibody levels across age groups, the impact of Omicron variant infections, and the effect of booster vaccinations on antibody levels.MethodsIn September and December 2021 and in February 2022, over 2000 Dutch blood donors were tested for presence of SARS-CoV-2 antibodies. Donations were selected based on age, sex, and region of residence, to provide an optimal coverage and representation of the Dutch population.ResultsLevels of vaccination-induced spike antibodies decreased over time in all age groups. Donors vaccinated with Janssen or AstraZeneca had significantly lower antibody levels than donors vaccinated with Pfizer or Moderna vaccine. Boostering with an mRNA vaccine elevated antibody levels in all age-groups irrespective of the initial vaccine. In donors aged < 56 years, the proportion of infected donors almost doubled between December 2021 and February 2022.ConclusionThe booster vaccination campaign increased antibody levels in all age-groups. After a booster vaccination, donors initially vaccinated with AstraZeneca or Janssen vaccine showed antibody levels similar to donors initially vaccinated with an mRNA vaccine. The emergence of the SARS-CoV-2 Omicron variant in the Netherlands caused a substantial increase in donors with infection-induced antibodies, especially among younger donors.

Investigation of a subunit protein vaccine for HFRS based on a consensus sequence between envelope glycoproteins of HTNV and SEOV

Due to the global resurgence of hemorrhagic fever with renal syndrome (HFRS), more attention is being focused on this dangerous illness. In China and Korea, the only vaccines available are the virus-inactivated vaccine against Hantaan virus (HTNV) or Seoul virus (SEOV), but their efficacy and safety are inadequate. Therefore, it is important to develop new vaccines that are safer and more efficient to neutralize and regulate areas with a high prevalence of HFRS. We employed bioinformatics methods to design a recombinant protein vaccine based on conserved regions of protein consensus sequences in HTNV and SEOV membranes. The S2 Drosophila expression system was utilized to enhance protein expression, solubility and immunogenicity. After the Gn and Gc proteins of HTNV and SEOV were successfully expressed, mice were immunized, and the humoral immunity, cellular immunity, and in vivo protection of the HFRS universal subunit vaccine were systematically evaluated in mouse models. These results indicated that the HFRS subunit vaccine generated elevated levels of binding and neutralizing antibodies, particularly IgG1, compared to that of the traditional inactivated HFRS vaccine. Additionally, the spleen cells of immunized mice secreted IFN-r and IL-4 cytokines effectively. Moreover, the HTNV-Gc protein vaccine successfully protected suckling mice from HTNV infection and stimulated GC responses. In this research, a new scientific approach is investigated to develop a universal HFRS subunit protein vaccine that is capable of producing effective humoral and cellular immunity in mice. The results suggest that this vaccine could be a promising candidate for preventing HFRS in humans.

#5555 TAKAYASU ARTERITIS IN THE TIME OF SARS-COV-2

Abstract Background and Aims SARS CoV 2 infection is characterized by pulmonary, cardiovascular, neurological and other complications. New onset hypertension after SARS Cov 2 infection was observed in various studies with an unclear mechanism. Vessel inflammation is a common finding in these patients. Case A young woman (24 years old) referred to our Unit for high blood pressure levels appeared few days after SARS COVID 2 infection. She had a positive familiar history of hypertension. A 24 hours blood pressure monitoring confirmed hypertension and the patient was treated with a beta-blocker and referred to the nephrologist. Physical examination was negative and peripheral pulses were palpable. Blood pressure was 150/90 mmHg without significative discrepancy (&amp;lt;10 mmHg) between right and left arm and between upper and lower limbs. Carotid-femoral and carotid-radial pulse wave velocity was in the upper limit of the normal range (9.8 and 8.0 m/sec respectively), renal function was preserved (serum creatinine 0.6 mg/dl, eGFR 154 ml/min) and proteinuria was 450 mg/die. C-reactive protein was 13 mg/L. Complement components C3 and C4 as well as IgG and anti-neutrophil cytoplasmic antibody (ANCA) levels were in the normal range. IgA and IgM were slightly elevated and anti-nuclear antibody (ANA) levels were 1:160. TSH was in the normal range. Patient was switched to calcium channel blockers and a screening to exclude secondary hypertension was performed. Hormonal profile showed hypersecretion of cortex and medullary adrenal gland (high renin and aldosterone, plasma and urinary cortisol, and epinephrine levels). Patient was treated with ACE inhibitor and showed an optimal blood pressure profile. Abdominal-Chest CT angiography detected no increase in adrenal gland dimension. Conversely, a left renal artery stenosis and a mild enlargement of the para-aortic tissue, suggestive of a retroperitoneal fibrosis, was described (Figure 1). A Doppler ultrasound examination confirmed a high systolic peak velocity in left renal artery and a low resistive index in the left kidney. According to the diagnosis of renal fibrosis, patient was treated with oral prednisone at a dosage of 1 mg/Kg/BW. Three months later, Doppler ultrasound and CT were materially unchanged. After a case revision and a negative evidence of inflammation at PET-FDG examination, the Takayasu Arteritis diagnosis was formulated and the patient underwent left renal artery angioplasty. Oral prednisone was tapered and methotrexate was started. One month later blood pressure and Doppler ultrasound velocimetric parameters were normalized. Conclusion This case report suggests that Takayasu arteritis may occur after SARS COVID 2 infection.

Autoantigen-selected B cells are bystanders in spontaneous T cell-driven experimental autoimmune hepatitis.

Autoreactive B cells are considered pathogenic drivers in many autoimmune diseases; however, it is not clear whether autoimmune B cells are invariably pathogenic or whether they can also arise as bystanders of T cell-driven autoimmune pathology. Here, we studied the B cell response in an autoantigen- and CD4+ T cell-driven model of autoimmune hepatitis (AIH), the Alb-iGP_Smarta mouse in which expression of a viral model antigen (GP) in hepatocytes and its recognition by GP-specific CD4+ T cells causes spontaneous AIH-like disease. T cell-driven AIH in Alb-iGP_Smarta mice was marked by autoantibodies and hepatic infiltration of plasma cells and B cells, particularly of isotype-switched memory B cells, indicating antigen-driven selection and activation. Immunosequencing of B cell receptor repertoires confirmed B cell expansion selectively in the liver, which was most likely driven by the hepatic GP model antigen, as indicated by branched networks of connected sequences and elevated levels of IgG antibodies to GP. However, intrahepatic B cells did not produce increased levels of cytokines and their depletion with anti-CD20 antibody did not alter the CD4+ T cell response in Alb-iGP_Smarta mice. Moreover, B cell depletion did not prevent spontaneous liver inflammation and AIH-like disease in Alb-iGP_Smarta mice. In conclusion, selection and isotype-switch of liver-infiltrating B cells was dependent on the presence of CD4+ T cells recognizing liver antigen. However, recognition of hepatic antigen by CD4+ T cells and CD4+ T cell-mediated hepatitis was not dependent on B cells. Thus, autoreactive B cells can be bystanders and need not be drivers of liver inflammation in AIH.

GADD34 activates p53 and may have utility as a marker of atherosclerosis.

We previously identified growth arrest and DNA-damage-inducible gene 34 (GADD34) as a marker of ischemic stroke. In the present study, serum levels of anti-GADD34 antibodies were found to be significantly higher in patients with acute ischemic stroke or chronic kidney disease compared to healthy donors. We then examined the biological function of GADD34 by transfection into U2OS human osteosarcoma and U87 human glioblastoma cells. Knockdown of GADD34 by siRNA resulted in enhanced cell proliferation, which was reversed by co-knockdown of MDM2. Luciferase reporter assays revealed that the transactivation ability of p53 enhanced by genotoxic anticancer drugs such as camptothecin and etoposide was further potentiated by enforced expression of GADD34 but attenuated by co-transfection with p53 shRNA expression plasmids. Western blotting demonstrated increased p53 protein levels after treatment with camptothecin, which was also potentiated by GADD34 but suppressed by GADD34 siRNA, ATM siRNA, and ATM inhibitor wortmannin. GADD34 levels also increased in response to treatment with camptothecin or adriamycin, and this increase was attenuated by MDM2 siRNA. Immunoprecipitation with anti-GADD34 antibody followed by Western blotting with anti-MDM2 antibodies indicated ubiquitination of GADD34 is mediated by MDM2. Accordingly, GADD34 may function as a ubiquitination decoy to reduce p53 ubiquitination and increase p53 protein levels. Increased neuronal cell death due to activation of p53 by GADD34 may account for the elevated serum levels of anti-GADD34 antibodies observed in patients with acute ischemic stroke.