Autoantibodies elicited with SARS-CoV-2 infection are linked to alterations in double negative B cells.

Double negative (DN) B cells (CD27-IgD-) comprise a heterogenous population of DN1, DN2, and the recently described DN3 and DN4 subsets. In autoimmune disease, DN2 cells are reported to be precursors to autoreactive antibody secreting cells and expansion of DN2 cells is linked to elevated interferon levels. Severe SARS-CoV-2 infection is characterized by elevated systemic levels of pro-inflammatory cytokines and serum autoantibodies and expansion of the DN2 subset in severe SARS-CoV-2 infection has been reported. However, the activation status, functional capacity and contribution to virally-induced autoantibody production by DN subsets is not established. Here, we validate the finding that severe SARS-CoV-2 infection is associated with a reduction in the frequency of DN1 cells coinciding with an increase in the frequency of DN2 and DN3 cells. We further demonstrate that with severe viral infection DN subsets are at a heightened level of activation, display changes in immunoglobulin class isotype frequency and have functional BCR signaling. Increases in overall systemic inflammation (CRP), as well as specific pro-inflammatory cytokines (TNFα, IL-6, IFNγ, IL-1ß), significantly correlate with the skewing of DN1, DN2 and DN3 subsets during severe SARS-CoV-2 infection. Importantly, the reduction in DN1 cell frequency and expansion of the DN3 population during severe infection significantly correlates with increased levels of serum autoantibodies. Thus, systemic inflammation during SARS-CoV-2 infection drives changes in Double Negative subset frequency, likely impacting their contribution to generation of autoreactive antibodies.

Rational Design and Development of SARS-CoV-2 Serological Diagnostics by Immunoprecipitation-Targeted Proteomics.

Current design of serological tests utilizes conservative immunoassay approaches and is focused on fast and convenient assay development, throughput, straightforward measurements, and affordability. Limitations of common serological assays include semiquantitative measurements, cross-reactivity, lack of reference standards, and no differentiation between human immunoglobulin subclasses. In this study, we suggested that a combination of immunoaffinity enrichments with targeted proteomics would enable rational design and development of serological assays of infectious diseases, such as COVID-19. Immunoprecipitation-targeted proteomic assays allowed for sensitive and specific measurements of NCAP_SARS2 protein with a limit of detection of 313 pg/mL in serum and enabled differential quantification of anti-SARS-CoV-2 antibody isotypes (IgG, IgA, IgM, IgD, and IgE) and individual subclasses (IgG1-4 and IgA1-2) in plasma and saliva. Simultaneous evaluation of the numerous antigen-antibody subclass combinations revealed a receptor-binding domain (RBD)-IgG1 as a combination with the highest diagnostic performance. Further validation revealed that anti-RBD IgG1, IgG3, IgM, and IgA1 levels were significantly elevated in convalescent plasma, while IgG2, IgG4, and IgA2 were not informative. Anti-RBD IgG1 levels in convalescent (2138 ng/mL) vs negative (95 ng/mL) plasma revealed 385 ng/mL as a cutoff to detect COVID-19 convalescent plasma. Immunoprecipitation-targeted proteomic assays will facilitate improvement and standardization of the existing serological tests, enable rational design of novel tests, and offer tools for the comprehensive investigation of immunoglobulin subclass cooperation in immune response.

Plasmablast-like Phenotype Among Antigen-Experienced CXCR5-CD19low B Cells in Systemic Lupus Erythematosus.

OBJECTIVE: Altered composition of the B cell compartment in the pathogenesis of systemic lupus erythematosus (SLE) is characterized by expanded plasmablast and IgD-CD27- double-negative B cell populations. Previous studies showed that double-negative B cells represent a heterogeneous subset, and further characterization is needed. METHODS: We analyzed 2 independent cohorts of healthy donors and SLE patients, using a combined approach of flow cytometry (for 16 healthy donors and 28 SLE patients) and mass cytometry (for 18 healthy donors and 24 SLE patients) and targeted RNA-Seq analysis. To compare B cell subset formation during the acute immune response versus that during autoimmune disease, we investigated healthy donors at various time points after receipt of the BNT162b2 messenger RNA COVID-19 vaccine and patients with acute SARS-CoV-2 infection, using flow cytometry. RESULTS: We found that IgD-CD27+ switched and atypical IgD-CD27- memory B cells, the levels of which were increased in SLE patients, represented heterogeneous populations composed of 3 different subsets each. CXCR5+CD19intermediate , CXCR5-CD19high , and CXCR5-CD19low populations were found in the switched memory and double-negative compartments, suggesting the relatedness of IgD-CD27+ and IgD-CD27- B cells. We characterized a hitherto unknown and antigen-experienced CXCR5-CD19low subset that was enhanced in SLE patients, had a plasmablast phenotype with diminished B cell receptor responsiveness, and expressed CD38, CD95, CD71, PRDM1, XBP1, and IRF4. Levels of CXCR5-CD19low subsets were increased and correlated with plasmablast frequencies in SLE patients and in healthy donors who received BNT162b2, suggesting their interrelationship and contribution to plasmacytosis. The detection of CXCR5-CD19low B cells among both CD27+ and CD27- populations calls into question the role of CD27 as a reliable marker of B cell differentiation. CONCLUSION: Our data suggest that CXCR5-CD19low B cells are precursors of plasmablasts. Thus, cotargeting this subset may have therapeutic value in SLE.

The Impact of Pandemic-Related Life Stress on Internet Gaming: Social Cynicism and Gaming Motivation as Serial Mediators.

A heightened interest in online gaming has emerged during COVID-19, and people have become increasingly vulnerable to internet gaming disorder (IGD). However, playing video games can also have a positive effect; gaming has been recognized as an efficient coping strategy. Currently, relatively little is understood about how online gaming can turn from an efficient coping strategy into an addiction disorder. This study investigated the mediating roles of social cynicism, escape and coping motives on the association between daily disruption during COVID-19 and IGD, seeking to reveal the underlying mechanism that influences the effects of gaming. A total of 203 participants in Hong Kong who reported having played electronic games during COVID-19 were surveyed. We conducted three hierarchical multiple regressions, then tested a serial mediation model using path analysis with structural equation modeling. The results revealed that escape motives significantly mediated the relationship between daily disruption related to COVID-19 and IGD, but no such effect was found for coping motives. Social cynicism alone was not a significant mediator, but social cynicism and escape motives in series mediated the relationship between daily disruption and IGD. These difference outcomes suggested different underlying mechanisms of escape and coping motives.

Internet gaming disorder in an adolescent during the COVID-19 pandemic: a case report.

The internet has become an indispensable tool in people´s daily lives during the COVID-19 pandemic. Internet and video game use are experiencing rapid growth in the youth and adult populations as a major source of entertainment. However, excessive gaming may cause addiction and negatively impact mental health, entailing low psychosocial well-being, poor social skills, and decreased academic achievement. We report the case of a 16-year-old student with a "typical" pattern of internet gaming disorder (IGD) developed during the pandemic, which improved after weeks of treatment with pharmacotherapy and psychosocial interventions. This case highlights that it is essential for the mental health professionals to know the psychopathology of IGD and multimodal approaches to treat it.

Structural Equation Modeling (SEM): Gaming Disorder Leading Untreated Attention-Deficit/Hyperactivity Disorder to Disruptive Mood Dysregulation.

(1) Background: Internet gaming disorder (IGD) in youths likely leads to disruptive mood dysregulation, especially among those with attention-deficit/hyperactivity disorder (ADHD). Whether IGD mediates the pathways leading ADHD to disruptive emotional dysfunction remains unclear. This study aims to elucidate the direct or indirect influence of IGD on ADHD; (2) Method: The Swanson, Nolan, and Pelham Version IV questionnaire was used to evaluate symptoms of ADHD and oppositional defiant disorder, and the Chen gaming disorder scale was used to measure IGD. A psychiatrist diagnosed ADHD, IGD, and disruptive mood dysregulation disorder (DMDD)-like symptoms. Structural equation modeling was applied to evaluate the role of IGD in mediating ADHD progression to disruptive mood dysregulation; (3) Results: Among a total of 102 ADHD youths, 53 (52%) of them with IGD were significantly more likely to have poor interpersonal relationships (p < 0.01) and DMDD-like symptoms (p < 0.01) than ADHD youths without IGD. IGD played a mediating role in increasing the risk of disruptive mood dysregulation in ADHD youths; (4) Conclusions: The findings suggest that IGD mediates ADHD's progression to disruptive mood dysregulation. Intensive biopsychosocial interventions are warranted for ADHD youths with IGD. More children and adolescents became mood-dysregulated after excessive gaming during the COVID-19 pandemic; this study's results suggest that child mental health experts develop earlier detection and prevention strategies for children and adolescents hidden behind internet addiction.

Daratumumab for Refractory IgD Multiple Myeloma with Lung Cancer and Persistent Thrombocytopenia: a Case Report.

BACKGROUND: Immunoglobulin D multiple myeloma (IgD-MM) is a rare but aggressive disease. The safety and effectiveness of anti-CD38 monoclonal antibody (daratumumab) have not been known in either IgD-MM or MM complicated with secondary neoplasm. METHODS: A fragile IgD-MM patient had an aggressively relapsed disease concurrent with lung cancer and severe thrombocytopenia, which led to a dilemma for management. After a failure of ixazomib-based chemotherapy, a salvage therapy with daratumumab unexpectedly induced complete remission and platelet recovery, and the patient successfully proceeded to lung cancer surgery. CONCLUSIONS: Our case indicates daratumumab is both safe and effective for refractory IgD-MM with severe complications.

Novel approaches for treating Internet Gaming Disorder: A review of technology-based interventions.

Internet Gaming Disorder (IGD) has become a significant issue in mental healthcare over the past decades as the number of people engaging in excessive and unhealthy gaming increases with each year. Despite its inclusion in the 5th Edition of Diagnostic Statistical Manual and the development of a number of treatment methods that have been designed and tested for IGD, treatment remains a challenge. This review attempts to give an overview of the current state of IGD and its treatment with a specific focus on the potential of technology-based solutions, such as web-based programs, mobile applications, and virtual reality. The review also highlights the need for additional work in the area of treatment development for IGD and the preliminary evidence for the usefulness and importance of technology-based treatment methods which offer unique advantages, such as accessibility, scalability, and cost-effectiveness, over other existing treatment options.

The Comparative Efficacy of Treatments for Children and Young Adults with Internet Addiction/Internet Gaming Disorder: An Updated Meta-Analysis.

Internet gaming disorder (IGD) is a formal mental disorder leading to bad outcomes for children and adolescents. This study comprehensively compared the estimated effect of various pharmacotherapy and psychosocial interventions for IGD from randomized controlled trials (RCT) through updated meta-analysis, using meta-regression. A search of PubMed/MEDLINE, Cochrane Library, and Airiti Library between 2000 and 2017 was conducted for various IA/IGD intervention modalities. A total of 124 studies from 29 selected papers involving 5601 children and young adults with IA/IGD were found. Meta-analyzing the pooled standardized mean difference (SMD) revealed a preliminary random effect of 1.399 with a 95% confidence interval of 1.272-1.527, suggesting highly effective treatment of IA/IGD. After adjusting for the confounding risks of age, publication year, type of subjects, and type of study, this study revealed that combining pharmacotherapy with cognitive behavioral therapy (CBT) or multi-level counseling (MLC) was the most effective treatment option. Using a scale of time spent online or a severity of IA symptoms scale was a more effective measurement, with p-values = 0.006 and 0.002, respectively. IA/IGD patients with comorbid depression showed worse outcomes than youth with another comorbidity. The corresponding model goodness-of-fit indices were τ2 = 1.188; I2-Residual = 89.74%; and Adjusted-R2 = 16.10%. This systematic review indicates that pharmacotherapy combined with CBT or MLC might be an effective therapeutic strategy for youth with gaming disorder.

Severity of SARS-CoV-2 infection is linked to double-negative (CD27- IgD-) B cell subset numbers.

OBJECTIVES: The role of B cells in COVID-19, beyond the production of specific antibodies against SARS-CoV-2, is still not well understood. Here, we describe the novel landscape of circulating double-negative (DN) CD27- IgD- B cells in COVID-19 patients, representing a group of atypical and neglected subpopulations of this cell lineage. METHODS: Using multiparametric flow cytometry, we determined DN B cell subset amounts from 91 COVID-19 patients, correlated those with cytokines, clinical and laboratory parameters, and segregated them by principal components analysis. RESULTS: We detected significant increments in the DN2 and DN3 B cell subsets, while we found a relevant decrease in the DN1 B cell subpopulation, according to disease severity and patient outcomes. These DN cell numbers also appeared to correlate with pro- or anti-inflammatory signatures, respectively, and contributed to the segregation of the patients into disease severity groups. CONCLUSION: This study provides insights into DN B cell subsets' potential role in immune responses against SARS-CoV-2, particularly linked to the severity of COVID-19.

Imbalanced Immune Response of T-Cell and B-Cell Subsets in Patients with Moderate and Severe COVID-19.

BACKGROUND: The immunological changes associated with COVID-19 are largely unknown. METHODS: Patients with COVID-19 showing moderate (n = 18; SpO2 > 93%, respiratory rate > 22 per minute, CRP > 10 mg/L) and severe (n = 23; SpO2 < 93%, respiratory rate >30 per minute, PaO2/FiO2 ≤ 300 mmHg, permanent oxygen therapy, qSOFA > 2) infection, and 37 healthy donors (HD) were enrolled. Circulating T- and B-cell subsets were analyzed by flow cytometry. RESULTS: CD4+Th cells were skewed toward Th2-like phenotypes within CD45RA+CD62L- (CM) and CD45RA-CD62L- (EM) cells in patients with severe COVID-19, while CM CCR6+ Th17-like cells were decreased if compared with HD. Within CM Th17-like cells "classical" Th17-like cells were increased and Th17.1-like cells were decreased in severe COVID-19 cases. Circulating CM follicular Th-like (Tfh) cells were decreased in all COVID-19 patients, and Tfh17-like cells represented the most predominant subset in severe COVID-19 cases. Both groups of patients showed increased levels of IgD-CD38++ B cells, while the levels of IgD+CD38- and IgD-CD38- were decreased. The frequency of IgD+CD27+ and IgD-CD27+ B cells was significantly reduced in severe COVID-19 cases. CONCLUSIONS: We showed an imbalance within almost all circulating memory Th subsets during acute COVID-19 and showed that altered Tfh polarization led to a dysregulated humoral immune response.

Shared B cell memory to coronaviruses and other pathogens varies in human age groups and tissues.

Vaccination and infection promote the formation, tissue distribution, and clonal evolution of B cells, which encode humoral immune memory. We evaluated pediatric and adult blood and deceased adult organ donor tissues to determine convergent antigen-specific antibody genes of similar sequences shared between individuals. B cell memory varied for different pathogens. Polysaccharide antigen-specific clones were not exclusive to the spleen. Adults had higher clone frequencies and greater class switching in lymphoid tissues than blood, while pediatric blood had abundant class-switched convergent clones. Consistent with reported serology, prepandemic children had class-switched convergent clones to severe acute respiratory syndrome coronavirus 2 with weak cross-reactivity to other coronaviruses, while adult blood or tissues showed few such clones. These results highlight the prominence of early childhood B cell clonal expansions and cross-reactivity for future responses to novel pathogens.