Vulnerability to Meningococcal Disease in Immunodeficiency Due to a Novel Pathogenic Missense Variant in NFKB1

NF-κB1 deficiency is suggested to be the most common cause of common variable immunodeficiency (CVID). NFKB1 encodes for the p105 precursor protein of NF-κB1, which is converted into the active transcriptional subunit p50 through proteasomal processing of its C-terminal half upon stimulation and is implicated in the canonical NF-kB pathway. Rare monoallelic NFKB1 variants have been shown to cause (haplo) insufficiency. Our report describes a novel NFKB1 missense variant (c.691C>T, p.R230C; allele frequency 0.00004953) in a family vulnerable to meningitis, sepsis, and late-onset hypogammaglobulinemia. We investigated the pathogenic relevance of this variant by lymphocyte stimulation, immunophenotyping, overexpression study and immunoblotting. The ectopic expression of p50 for c.691 C>T restricted transcriptionally active p50 in the cytoplasm, and immunoblotting revealed reduced p105/50 expression. This study shows that the deleterious missense variant in NFKB1 adversely affects the transcriptional and translational activity of NFκB1, impairing its function. Patients immunological parameters show a progressive course of hypogammaglobulinemia, which may partially account for the incomplete disease penetrance and suggest the need for closer immunological monitoring of those mutation carriers.

Seroprevalence of Anti-SARS-CoV-2 Antibodies in COVID-19 Patients in Hyderabad, Sindh

Aim: The world is affected by the severe acute respiratory syndrome coronavirus2 (SARS-CoV-2) pandemic. This virus has emerged as a human pathogen that can cause symptoms ranging from fever to Pneumonia, but it remains asymptomatic or mild. To better understand the virus's ongoing spread, identify those who have been infected, and track the immune response, accurate and robust immunological monitoring and SARS-CoV-2 detection assays are needed. Methods: The estimation of serology tests to assess the presence of antibodies to SARS-CoV-2 in COVID- 19 patients at Asian Institute of Medical Sciences (AIMS) and Isra University & hospital. 1229 patients were selected including males and females with the age being 25 to 65 years living in the territories from 1st August to 30th November 2020. The anti-SARS-CoV-2 test was performed by an electrochemiluminescence immunoassay analyzer. Results: Out of 1229 participants 206 (17%) were positive with COVID-19, and 1023 (83%) were negative. The results further revealed that a higher percentage of positive COVID-19 were detected in males in all age groups as compared to females, and most of them are affected at age of 46-65 years male 40 (24.69%) and female 14(17.5%). Conclusion: The seroprevalence of SARS-COV-2 antibodies has increased in the old age population, which may aid in determining the true number of infected cases. Although the current study is based on a small sample of participants, the findings suggest a study with a larger population to implement stronger and targeted interventions.

Everolimus-Induced Immune Effects after Heart Transplantation: A Possible Tool for Clinicians to Monitor Patients at Risk for Transplant Rejection

Background: Patients treated with an inhibitor of the mechanistic target of rapamycin (mTORI) in a calcineurin inhibitor (CNI)-free immunosuppressive regimen after heart transplantation (HTx) show a higher risk for transplant rejection. We developed an immunological monitoring tool that may improve the identification of mTORI-treated patients at risk for rejection. Methods: Circulating dendritic cells (DCs) and regulatory T cells (Tregs) were analysed in 19 mTORI- and 20 CNI-treated HTx patients by flow cytometry. Principal component and cluster analysis were used to identify patients at risk for transplant rejection. Results: The percentages of total Tregs (p = 0.02) and CD39+ Tregs (p = 0.05) were higher in mTORI-treated patients than in CNI-treated patients. The principal component analysis revealed that BDCA1+, BDCA2+ and BDCA4+ DCs as well as total Tregs could distinguish between non-rejecting and rejecting mTORI-treated patients. Most mTORI-treated rejectors showed higher levels of BDCA2+ and BDCA4+ plasmacytoid DCs and lower levels of BDCA1+ myeloid DCs and Tregs than mTORI non-rejectors. Conclusion: An mTORI-based immunosuppressive regimen induced a sufficient, tolerance-promoting reaction in Tregs, but an insufficient, adverse effect in DCs. On the basis of patient-specific immunological profiles, we established a flow cytometry-based monitoring tool that may be helpful in identifying patients at risk for rejection.

Efficiency of human plague vaccination in tuvinian natural plague focus. Message 2: dynamics of immune status indicators after revaccination

Relevance. In Russia, the live plague vaccine (LPV) is used for specific prophylaxis of plague. Immunological monitoring of humans vaccinated by LPV in order to search for informative diagnostic markers, as well as to improve the tactics of epidemiological surveillance of plague enzootic territories is an urgent area of research.The aim is to assess the parameters of cellular and humoral immunity in humans revaccinated by LPV who permanently reside on territory of the Tuvinian natural plague focus.Materials and methods. The study involved 76 volunteers from the Republic of Tuva, revaccinated by LPV. Blood sampling was performed before vaccination and 1, 3, and 6 months after revaccination. The study included the determination of cytokine production (IFN-γ, IL-4, TNF-α), specific antibodies, immunoglobulins (IgM, IgG, IgA and IgE) and lymphocyte subpopulation composition (CD3, CD4, CD8, CD16, CD19, immunoregulatory index).Results. A decrease in IgG and IgM and an increase in IgA were found after vaccination with LPV and their increase after revaccination. Correlation relationships were revealed between immunoglobulins, B cells and IL-4. Revaccination leads to an increase seroconversion. The activation of humoral immunity in humans vaccinated against plague is also evidenced by dynamics of changes in the subpopulation composition: an increase in B-lymphocytes and natural killer cells, a decrease in T-helpers and immunoregulatory index, and cellular immunity stimulation is an increase in spontaneous and induced production of pro- and anti-inflammatory cytokines.Conclusion. It has been shown that LPV is capable of causing the body’s immune restructuring and activating the cellular and humoral mechanisms of immunological protection. For a complete understanding of the development and preservation of antiplague immunity, it is necessary to continue the annual immunological monitoring of the population living on the territory of the Tuvinian natural plague focus, using additional modern research methods.

Comprehensive Immune Profiling of a Kidney Transplant Recipient With Peri-Operative SARS-CoV-2 Infection: A Case Report

To date there is limited data on the immune profile and outcomes of solid organ transplant recipients who encounter COVID-19 infection early post-transplant. Here we present a unique case where the kidney recipient’s transplant surgery coincided with a positive SARS-CoV-2 test and the patient subsequently developed symptomatic COVID-19 perioperatively. We performed comprehensive immunological monitoring of cellular, proteomic, and serological changes during the first 4 critical months post-infection. We showed that continuation of basiliximab induction and maintenance of triple immunosuppression did not significantly impair the host’s ability to mount a robust immune response against symptomatic COVID-19 infection diagnosed within the first week post-transplant.

Laboratory strategies for the diagnosis of COVID 19

To prevent further spread of the COVID 19 pandemic, rapid and accurate diagnosis of the infection is essential. The standard method for detecting coronavirus is polymerase chain reaction. Serological methods and tests for detecting antibodies against SARS-CoV-2 and determining protective immunity in the population are usually used to retrospectively identify those patients who have already suffered an asymptomatic and mild form of infection. Immunological monitoring is necessary to control the situation of infection in patients at the hospital and pre-hospital stages, in order to track contacts and conduct epidemiological surveillance of the situation with the spread of the SARS-CoV-2 virus

Experience of Using a Complex Antigenic Preparation of the Plague Microbe to Assess the Severity of a Specific Anti-Plague Response

Background. Improving the methodology of immunological monitoring in natural foci of plague in the Russian Federation and adjacent territories to increase the effectiveness of epidemiological surveillance of plague is an urgent line of research. The lack of correlation between the production of specific antibodies to the capsular antigen (F1) ofthe plague microbe with other indicators of the state of cellular defense reactivity indicates the need to search for new informative and accessible markers for assessing anti-plague immunity.Objective: to evaluate possibility of using the complex preparation (F1 and cell membranes) evaluate the possibilities of using an artificial antigenic complex based on F1 and cell membranes (CM) of the plague microbe in antigen-specific tests in vitro in people vaccinated against plague.resu. The study involved 153 volunteers living in the territory enzootic for plague (the village of Khandagayty ofthe Ovyur kozhuun of the Tyva Republic and the village of Kosh-Agach of the Kosh-Agach district of the Altai Republic). The study included the determination of spontaneous and mitogen-induced production of cytokines (IFN-γ, IL-4, TNF-α) by blood cells, titers of specific IgG antibodies to the capsular antigen F1 of the plague microbe and concentrations ofthe main classes of immunoglobulins (IgM, IgG, IgA and IgE) in blood serum, as well as immunophenotyping of blood lymphocytes (CD3, CD4, CD8, CD16, CD19).Results. Comparative assessment of the level of cytokines (TNF-α, IFN-γ and IL-4) in spontaneous/induced F1+CM Y. pestis tests revealed a statistically significant increase in the production of cytokines TNF-α and IFN-γ in the antigeninduced tests compared with spontaneous (p < 0.01).Conclusion. Thus, the effectiveness of the use of artificial antigenic complex based on F1 and cell membranes ofthe plague microbe has been shown to assess the production of cytokines in antigen-specific cell tests in vitro, which justifies the need for further research.

MO438STUDY OF IMMUNOLOGICAL STATUS IN PATIENT OF CKD IN PREDIALYSIS STAGES

Background and Aims It is known that inflammation is an important factor associated with the variability of the clinical picture in patients with chronic diseases. However, the question remains not completely clear: are immunological changes predictors of less stable control of the course of CKD. The aim of this study was a comprehensive assessment of the immunological status in patients with pre-dialysis stages of CKD. Method 80 patients (46 men and 36 women) with CKD in the outcome of nephropathies of diabetic and nondiabetic etiology, admitted to the Republican Medical Scientific and Practical Center for Therapy and Medical Rehabilitation, were examined. Chronic pyelonephritis without exacerbation was diagnosed in 31 patients, chronic glomerulonephritis - in 29, diabetic nephropathy - in 25 polycystic kidney disease - in 6 patients. Glomerular filtration was calculated based on serum creatinine concentration using the CKD-EPI formula (2011). A complex of immunological studies was performed. Immunotyping of lymphocytes was performed by flow cytofluorometry with the characterization of the main markers of immunocompetent cells. Results In patients with CKD, violations were observed in almost all links of immunological reactivity, first of all, a significant decrease in indicators of natural resistance (CD16 +, PS, PI) (p &lt;0.05). The results of immunological monitoring showed a predominant violation of cellular defense mechanisms (CD3 +, CD4 +, CD8 +) (p &lt;0.05), a multiple increase in inflammatory cytokines Il-4, TNF-α, IL-6 (p &lt;0.05), in the absence of activation of humoral link and inadequacy of elimination mechanisms (CEC, p &lt;0.05). Conclusion Our study showed that the immunological parameters were characterized by insufficient activation of the first line of defense of natural resistance, as well as specific mechanisms - an imbalance of the cellular and humoral links of immunity, hyperactivation of cytokine regulation in the absence of effector functions.