Immunoglobulin Concentrations Research Articles

Effects of moringa polysaccharides on growth performance, immune function, rumen morphology, and microbial community structure in early-weaned goat kids

The aim of this research was to investigate the effects of adding moringa polysaccharides (MOP) on the growth performance, immune function, rumen tissue morphology, and rumen microbial community in early-weaned goat kids. Twenty-one 7-day-old Leizhou male goat kids weighing (3.05 ± 0.63) kg, were randomly divided into a control group (CON group), a low-dose group (LOW group), and a high-dose group (HIG group). MOP was added to the goat kids’ milk replacer (MR) at 0, 0.15, and 0.3% (on dry matter basis),fed until 60 days of age, and four goat kids in each group with body weights close to the mean of each group were selected for slaughter. The results showed that, compared to the CON group, the MOP groups significantly improved final body weight, body measurements, daily weight gain, and feed intake of the early weaned goat kids; significantly reduced the content of propionic acid, butyric acid, valeric acid, and ammoniacal nitrogen; and in addition, the addition of MOP could significantly increase the height of rumen nipple, the content of immunoglobulin G (IgG) in the serum. The HIG group significantly increased rumen pH, rumen muscularis layer thickness, rumen wall thickness, and serum immunoglobulin A (IgA), and immunoglobulin M (IgM). In conclusion, the addition of MOP positively impacted the growth performance, serum immune function, and rumen tissue morphology in early-weaned goat kids.

Enhanced activity of split trehalase biosensors by interspecies domain combineering.

The split trehalase biosensor has potential as a versatile diagnostic technology. Split enzymes are engineered proteins, divided into inactive fragments, which can reassemble and regain activity when brought together by an analyte. The split TreA biosensor requires no sample processing and produces stable signals (in the form of glucose). Split trehalase reagents can function in blood, but periplasmic trehalase of E.coli requires blood acidification for maximal activity. The objective of this study was to obtain split trehalase with near physiological pH optimum. For this purpose, periplasmic trehalases of Cellvibrio spp. with higher activity at neutral pH, were split in analogy with the E. coli TreA into hood and catalytic domains. However, these split trehalases displayed self-complementation due to spontaneous reassembly. In contrast, when catalytic domains of Cellvibrio trehalases were combined with E.coli hood domains, these hybrids displayed conditional complementation capacity when split trehalase fragments fused to immunoglobulin-binding protein G (STIGA) were used to quantify immunoglobulin concentrations. Other hybrid combinations of Cellvibrio spp. had increased activity compared to the cognate pairs, albeit with strong self-complementation. A mutagenesis analysis of residues responsible for self-complementation led to uncoupling of self-complementation from allostery. The Michaelis-Menten kinetics of Cellvibrio enzymes and fragment pairs confirmed improved activity of a mutated hybrid pair of Cellvibrio hood and catalytic domains at physiological pH. In conclusion, the improvements in pH optimum and activity, demonstrated with STIGA, can now be leveraged to enhance other variations of the split trehalase biosensor platform, broadening its utility for testing clinical samples.

Warning values of serum total kappa/lambda ratio for M-proteinemia

BackgroundTo introduce the serum total kappa/lambda ratio (K/L) in humoral immunity testing reports to improve the detection rate of M-proteinemia.Methods156 M protein-positive and 5464 M protein-negative samples confirmed by serum immunofixation electrophoresis(sIFE) were accumulated from January 2021 to December 2023 in the First Affiliated Hospital of Soochow University and the contents of immunoglobulins (IgG, IgA, IgM, kappa and lambda) were tested by Beckman IMMAGE800. All the samples were divided into two groups by time: the modeling group and the validation group. The K/L values in the modeling group were analyzed by SPSS 27.0 to get the receiver operating characteristic curve (ROC). Furthermore, a more in-depth analysis was conducted to verify the reliability of the optimal cutoff values in the validation group. In addition, the levels of immunoglobulins of another group including 106 patients with definite diagnosis of monoclonal gammopathy ranging from January 2021 to June 2024 were traced back to improve the diagnostic efficiency.ResultsThe optimal cutoff values of K/L were 2.31 and 1.43 corresponding to K-type and L-type M-proteinemia respectively by ROC analysis. The sensitivity and specificity were validated as 76.14% and 77.42%. False positives were mainly found in samples with systemic sclerosis (36.84%), hypohepatia (28.71%) and sicca syndrome (27.27%). While false negatives were mainly found in IgA monoclonal gammopathy (38.39%) and IgM monoclonal gammopathy (28.57%). Combining with the detection rules of IgG, IgA and IgM, the sensitivities for the diagnosis of immunoglobulin light chain amyloidosis(AL) and monoclonal gammopathy of undetermined significance(MGUS) can be increased to 83.33% and 85%.ConclusionsK/L > 2.31 and K/L < 1.43 can be used as warning values for M-proteinemia. In addition, the content of the heavy chain in IgA- or IgM-type M-proteinemia may be considered to improve the detection rate.

Comparative Analysis of Skim Milk and Milk Fat Globule Membrane Proteomes between Human and Farm Animal Milk for Infant Formula Production.

Human and animal milk contain a rich variety of milk proteins that meet the needs of their newborns. In total, 1263 skim milk proteins and 1754 MFGM proteins were identified in human milk and six types of animal milk, respectively. Both similarities and differences were observed among the species. Human milk contained more immunoglobulins involved in the adaptive immune response, playing a crucial role in mucosal defense in newborn babies. In contrast, ruminant milk contained higher amounts of antimicrobial proteins, which protect newborns from bacterial infections. The most dominant difference in MFGM proteins between human and animal milk was related to protein processing in the endoplasmic reticulum. Goat milk and camel milk were more similar to human milk in terms of skim milk and MFGM proteins compared to the other five types of animal milk. Moreover, immunoglobulins and complement and coagulation cascade proteins in goat milk were most similar to those in human milk. A higher content of immunoglobulin A was observed in donkey milk, which could be considered as a source of IgA in infant formula. These results provide more comprehensive and novel insights into protein variation among animal milk, which may support improving dairy products such as infant formula.

Immune system dysfunction in personnel of chemical hazard facilities: purpose and effectiveness of immunotherapeutic drugs

Clinical signs of the presence of immune dysfunction/insufficiency, identified by questioning workers at the landfill for the storage and destruction of industrial toxic waste, were the basis for performing a screening study of immune status. This included a clinical blood test with determination of the leukocyte formula, assessment of the phenotypic status of lymphocytes for the main subpopulations, determination of the concentration in serum immunoglobulins of various classes, assessment of the migration activity of leukocytes, assessment of the microbicidal capacity of neutrophils and their ability to phagocytosis. When the examined employees were found to have more than 20% changes in the normative range of immune status parameters and the presence of combined disorders in different parts of immunoreactivity, in particular combined disorders in the cellular part of the immune system, it was the basis for conducting an in-depth stage of immunoepidemiological examination. This included determination of functional activity cellular elements of the adaptive component of immunity, assessment of the severity of autosensitization, assessment of the severity and specificity of atopic hypersensitization, assessment of cytokine status in normal conditions and in response to mitogen. When assessing the phenotypic status of immunoreactivity cells, it turned out that the absolute number of T lymphocytes (CD3+ phenotype), helper T lymphocytes (CD3+CD4+ phenotype) and killer T lymphocytes (CD3+CD8+ phenotype) was significantly reduced in the peripheral blood of test site employees (p ≤ 0.05, compared with similar indicators in the control group). On the contrary, the content of NK cells (CD3-CD56+ phenotype) tended to increase (both the absolute number and the proportion among other lymphocyte populations). Dysfunction of the cellular component of humoral immunoreactivity (a significant decrease in the absolute number of lymphocytes with the CD3-CD20+ phenotype [B lymphocytes] in the peripheral blood), as well as a significant increase in the concentration of serum IgA-class immunoglobulin and pronounced disimmunoglobulinemia in the serum of the examined individuals (compared to individuals in the control group ) indicated a predominantly inhalation method of exposure to immunotoxic industrial toxicants from the landfill on the immune system. Pathogenetically targeted immune-oriented therapy for the personnel of the test site was carried out with drugs that have thymomimetic activity (cytovir-3, thymogen) and are a means of replacement immunocorrection with a cytokine drug (genetically engineered recombinant yeast IL-2 [roncoleukin]). The established effectiveness of the immunotherapy, in particular the reduction in the incidence rate of various nosological forms of immune-related pathology among the site employees who received immunotherapy and the presence of an immunocorrective effect in the immunoactive drugs used for therapy, indicate the advisability of carrying out preventive medical measures in persons professionally exposed to xenobiotic influences as a component of improving medical provision.

Dissecting Secondary Immunodeficiency: Identification of Primary Immunodeficiency within B-Cell Lymphoproliferative Disorders

Distinguishing between primary (PID) and secondary (SID) immunodeficiencies, particularly in relation to hematological B-cell lymphoproliferative disorders (B-CLPD), poses a major clinical challenge. We aimed to analyze and define the clinical and laboratory variables in SID patients associated with B-CLPD, identifying overlaps with late-onset PIDs, which could potentially improve diagnostic precision and prognostic assessment. We studied 37 clinical/laboratory variables in 151 SID patients with B-CLPD. Patients were classified as “Suspected PID Group” when having recurrent-severe infections prior to the B-CLPD and/or hypogammaglobulinemia according to key ESID criteria for PID. Bivariate association analyses showed significant statistical differences between “Suspected PID”- and “SID”-groups in 10 out of 37 variables analyzed, with “Suspected PID” showing higher frequencies of childhood recurrent-severe infections, family history of B-CLPD, significantly lower serum Free Light Chain (sFLC), immunoglobulin concentrations, lower total leukocyte, and switch-memory B-cell counts at baseline. Rpart machine learning algorithm was performed to potentially create a model to differentiate both groups. The model developed a decision tree with two major variables in order of relevance: sum κ + λ and history of severe-recurrent infections in childhood, with high sensitivity 89.5%, specificity 100%, and accuracy 91.8% for PID prediction. Identifying significant clinical and immunological variables can aid in the difficult task of recognizing late-onset PIDs among SID patients, emphasizing the value of a comprehensive immunological evaluation. The differences between “Suspected PID” and SID groups, highlight the need of early, tailored diagnostic and treatment strategies for personalized patient management and follow up.

The influence of professional oral hygiene on the indicators of local immunity

Professional oral hygiene is an important part of measures aimed at preventing and maintaining the patient’s health. Local immune response is the first barrier of immune protection capable of preventing infection diseases. The aim of the study - the influence of professional hygiene on the indicators of local immunity has been studied. The study involved two groups of patients: the first group included 19 individuals with chronic caries of varying depths (class I), and the second group comprised 15 individuals with moderate periodontal disease. To evaluate nonspecific oral protection, the index of natural colonization of the buccal epithelium and the activity of microbial adsorption by oral mucosal cells were assessed. Specific immunity was evaluated by measuring the concentration of immunoglobulins in unstimulated saliva. All indicators were recorded both before and after professional oral hygiene procedures. When studying the index of natural colonization of the buccal epithelium, it was found that professional cleaning in both groups led to normalization of the spectrum of bacteria adhered to the epithelium: there was a decrease in the amount of gram-negative microflora, atypical for this biotope, while maintaining a high adhesion index of gram-positive cocci. In persons with caries, before sanation, “unsatisfactory” resistance of oral mucosa was revealed in a third of the examined, after sanation, the number of such patients decreased by 3 times, with “satisfactory” resistance increased by 2 times. In the group with periodontal disease, all patients exhibited “unsatisfactory” mucosal resistance before the hygiene procedures. After sanitation, 20 % “unsatisfactory” resistance persisted. In the majority of patients (80 %), the resistance of the oral mucosa improved to “satisfactory”. The concentration of all classes of immunoglobulins in both groups before sanitation was significantly below the norm, after the measures taken, the indicators increased significantly and reached the level typical for healthy individuals. Carrying out professional cleaning of the teeth leads to the normalization of local immune status even without additional medical immunocorrection.

Colostrum conductivity, pH and Brix index as predictors of passive immunity transfer in foals.

Foal immunity relies heavily on the absorption of colostrum immunoglobulins; thus, colostrum evaluation is used to predict the transfer of passive immunity (TPI), and its conductivity is associated with TPI in cattle. Leading up to and at parturition, a reduction in colostrum pH and conductivity is thought to be necessary for TPI; however, this remains to be determined. To assess conductivity, pH, and Brix refractometer index of colostrum to predict the TPI in foals. Field study. The colostrum of 241 mares was assessed for conductivity, pH, and Brix index using handheld devices immediately after parturition. Twenty-hour postpartum foals had complete blood cell count and plasma IgG concentrations assessed. Foals were split in complete versus incomplete TPI (i.e., IgG ≥8 g/L vs. <8 g/L). Mare (breed, duration of pregnancy, parity, and age), foal (sex), and colostrum (conductivity, pH, and Brix index) data were computed to assess factors affecting TPI. Multivariate regression and receiver operating characteristic (ROC) curves were employed for analysis. Complete TPI was achieved in 89.4% of foals. The ROC TPI values were conductivity ≤5 mS/cm (AUC = 0.68), Brix ≥23.4% (AUC = 0.70), and pH ≤6.4 units (AUC = 0.73). The odds ratio for TPI in 24 h post-partum was 6.1 (2.1-17.1, 95% CI) for conductivity, 3.2 (1.2-9.3, 95% CI) for pH, and 4.5 (1.5-12.7, 95% CI) for Brix index. Sensitivity and specificity of 93% (88-96 95% CI), 56% (44-7, 95% CI), 92% (87-96, 95% CI), 28% (14-47, 95% CI), 94% (88-97, 95% CI) and 18% (14-47, 95% CI) for conductivity, Brix and pH, respectively. Low incidence of incomplete TPI. Conductivity, pH and Brix are strong predictors of foals that succeed in the transfer of passive immunity; however, all three presented low to moderate specificity. The established cutoff values obtained herein can be used on-field by other foaling programs. All three handheld devices used in the study are inexpensive and ready to use on-farm.

Effect of Fermented Mulberry Leaves on Gut Health of Finishing Pigs

This study was conducted to investigate the effects of supplementing fermented mulberry leaves (FML) on intestinal morphology, antioxidant capacity, and immune function in the gut of finishing pigs. Eighteen 132-day-old healthy crossbred (Duroc × Landrace × Yorkshire) male castrated pigs were randomly divided into two treatment groups with nine replicates per group. The control (CON) group was fed the basal diet, and the FML group was fed the basal diet supplemented with 10% FML. The experiment lasted 69 days. The results showed that 10% FML improved gut health. The apparent total tract digestibility in dry matter, crude protein, crude fiber, neutral detergent fiber, acidic detergent fiber, ether extract, and crude ash increased in the 10% FML group of finishing pigs compared to the CON group (p &lt; 0.05). Duodenal, jejunal, and ileal intestinal morphology, such as villus height and villus-height-to-crypt-depth ratio, increased in the 10% FML group compared to the CON group, whereas crypt depth decreased in the duodenum, jejunum, and ileum (p &lt; 0.05). Total antioxidant capacity increased in the ileum of the 10% FML group compared with the CON group (p &lt; 0.05). The FML supplementation improved the contents of duodenal immunoglobulin A, jejunal interleukin-1β, interleukin-8, ileal interleukin-1β, interleukin-6, interferon-γ, and immunoglobulins A and M compared to the control group (p &lt; 0.05). Moreover, FML downregulated the mRNA expression levels of tumor necrosis factor-α in the duodenum, Toll-like receptor 4, nuclear factor-κ B-P65, and myeloid differentiation factor 88 in the jejunum, and Toll-like receptor 4 and nuclear factor-κ B-P65 in the ileum (p &lt; 0.05). The FML also upregulated Montrose uniting church 1 in the duodenum and claudin 2 in the ileum (p &lt; 0.05). In conclusion, dietary supplementation with 10% FML improved the gut health of finishing pigs and FML is a potential feed ingredient for pig breeding.

Dietary supplementation with pterostilbene activates the PI3K-AKT-mTOR signalling pathway to alleviate progressive oxidative stress and promote placental nutrient transport

BackgroundProgressive oxidative stress easily occurs as a result of a gradual increase in the intensity of maternal metabolism due to rapid foetal development and increased intensity of lactation. However, studies on the effects of processive oxidative stress on nutrient transport in the placenta have received little attention. The present study was conducted on sows at 85 days of gestation to study the effects of pterostilbene (PTE) on maternal oxidative stress status and placental nutrient transport.ResultsPTE increased the antioxidant capacity and immunoglobulin content in mothers’ blood and milk, reduced the level of inflammatory factors, and improved the nutrient content of milk. PTE also reduced sow backfat loss and the number of weak sons, and increased piglet weaning weight and total weaning litter weight. We subsequently found that PTE enhanced placental glucose and fatty acid transport and further affected glycolipid metabolism by increasing the expression of LAL, PYGM, and Gbe-1, which activated the PI3K phosphorylation pathway. Moreover, PTE addition altered the relative abundance of the Firmicutes, Proteobacteria, Parabacillus, and Bacteroidetes-like RF16 groups in sow faeces. PTE increased the levels of acetate, propionate, butyrate and isovalerate in the faeces.ConclusionsThese findings reveal that the addition of PTE during pregnancy and lactation mitigates the effects of processive oxidative stress on offspring development by altering maternal microbial and placental nutrient transport capacity.Graphical

A-111 “Activation” Of macro-ast by pyridoxal phosphate supplementation

Abstract Background Type 1 macro-AST is a high molecular weight AST (Aspartate Aminotransferase) formed between an AST monomer and immunoglobulins in plasma, which can cause elevated AST values. It is believed that reduced clearance or excretion of macro-AST can cause elevated AST results. Here we report a case of macro-AST that produced high AST values only by the AST assay that includes pyridoxal phosphate (PLP), not by the one without PLP. Case description A 54 year-old male patient diagnosed with disseminated Bartonellosis was monitored with ‘liver function” tests on a monthly basis, with AST varying between 24 to 73 U/L (Reference interval, 13-47 U/L). The patient’s AST value suddenly jumped to 378 U/L when the laboratory switched to an AST assay supplemented with PLP (named ASTPM) on Roche Cobas c702 analyzer (c702). When the same patient sample was tested using the previous AST assay which does not include PLP, a result of 14 U/L was obtained. Macro-AST was suspected, as all other “liver function” tests were within reference interval. Methods Protein-A and Protein-G were used to remove immunoglobulins from the patient sample and another sample with matching AST as a control, respectively. Three hundred microliters of washed Protein-A or Protein-G Sepharose bead slurry was mixed with 300 microliters of each sample, respectively. Each mixture was incubated at 37oC for 3 hours on a rocker. After centrifugation, the supernatant was transferred to a new tube. AST, ASTPM, and all other tests were performed on c702, and results corrected for dilution. The recovery of enzyme activity or immunoglobulin concentration after the removal of immunoglobulins was calculated, respectively. Results In the original sample, the ratio of ASTPM to AST was 28 (389 U/L vs. 14 U/L) for the patient, and 1.1 (304 U/L vs. 275 U/L) for the control. After the complete removal of immunoglobulin G (IgG), the recovery of ASTPM for the patient was 3% and for the control was 74%. The recovery of ALTPM (supplemented with PLP) was 89% for the patient and 85% for the control, showing the expected loss of enzymatic activity by Protein-A or Protein-G treatment. Conclusions The significant reduction of ASTPM activity (recovery of 3%) after removal of IgG in the patient sample proved that the AST enzyme was associated with IgG. The slight increase of ASTPM activity compared to AST (ratio of 1.1) in the control sample was an expected effect of supplementing PLP in the AST assay, however, the huge increase of ASTPM activity (ASTPM/AST ratio of 28) in the patient sample suggests that PLP “activated” the macro-AST and produced high AST result. This is a concerning effect of supplementing PLP in AST assays because it can cause spurious AST elevations in cases of macro-AST. As more laboratories begin to employ the PLP supplemented aminotransferase assays, it is important to acknowledge the interactions between PLP and macro-AST and the role of the laboratory to identify macro-AST in order to avoid unnecessary, costly, and time-consuming testing and procedures for these patients.

B-080 Development and Validation of a GQ1b IgG ELISA as a Diagnostic Aid for Subacute Demyelinating Polyneuropathies

Abstract Background Autoimmunity targeting GQ1b is associated with group of disorders that includes Miller-Fisher Syndrome (MFS), Bickerstaff Brainstem Encephalitis (BBE) and classic Guillain Barre Syndrome (GBS) with ophthalmoplegia. Collectively these are referred to as GQ1B-IgG related syndromes. The prevalence of GQ1b-IgG in this population of patients is high and has been reported to be &amp;gt;80% in well-defined clinical cohorts. Clinical phenotypic presentation is diverse and requires highly accurate antibody testing. The objective of this study was to evaluate a newly developed custom GQ1b-IgG assay, using a semi-quantitative ELISA-based method. Methods A GQ1b-IgG ELISA was developed and validated. Patient sera along with assay controls and a single-point calibrator are diluted and incubated in duplicate wells in a 96-well plate coated with GQ1b antigen. The plate is washed and incubated with anti-human-IgG secondary antibody conjugated with HRP. After a colorimetric reaction, optical densities (OD) are read. The OD of patient samples are compared against the calibrator, generating a cut-off index (COI). Samples ≥1.0 COI are considered positive. During the validation phase, analytical and clinical performance characteristics were evaluated. Studies included assessment of imprecision at multiple concentrations of GQ1b-IgG, accuracy using method comparison, analytical sensitivity and specificity, reference range, common interferences, plate drift, and clinical performance in relevant disease and control cohorts. Results Intra-assay imprecision (coefficient of variation; %CV) ranged from 1.0 - 16% across all concentrations of GQ1b-IgG. Inter-assay imprecision (%CV) ranged from 12.4 - 44.5% but was &amp;lt;20% at or above the assay cut-off. We found perfect qualitative agreement with an independent ELISA reference method (20 positive and 20 negative samples selected based on reference method results [reference interval &amp;lt;1:100]). Limit of detection (LOD) was calculated at a COI of 0.22. To assess analytical specificity, 40 samples with high concentrations of immunoglobulins (hypergammaglobulinemia [N=15], IgG M-protein [N=15] and systemic lupus erythematosus [N=10] were tested and all were negative. A reference interval study was performed on 140 healthy donor serums, all were negative. Interference studies were conducted on 2 positive and 2 negative serum samples, spiked for highly hemolytic (1000 mg/dL), lipemic (2000 mg/dL) and icteric (60 mg/dL) conditions. There was no impact on the assay even with high levels of a given interferent. Plate drift on 2 distinct patient pools tested across the plate (COI range 0.7 - 1.3) had overall plate CV’s ≤10%. ROC analysis was performed on a cohort of 278 patient samples (15 MFS cases and 263 controls [suspected demyelinating neuropathy or disease mimics]). A cut-off value of 1.0 (COI) was selected based on the Youden index. At this cut-off, clinical sensitivity for MFS was 87% while clinical specificity was 99.2%. Conclusions The observed performance of the GQ1b-IgG ELISA was acceptable for clinical laboratory implementation. A COI of ≥ 1.0 was selected for clinical use.

B-337 Mass Spectrometry to evaluate treatment response in patients with Multiple Myeloma. A comparative study versus Immunofixation

Abstract Background The International Myeloma Working Group (IMWG) defines treatment response categories based on monoclonal protein (MP) changes by serum protein electrophoresis (SPE) and immunofixation (sIFE). Patients that after treatment remain positive by sIFE while SPE is negative, are categorized as very good partial response (VGPR) and, if sIFE becomes negative, patients would be considered in complete remission (CR). However, the interpretation of sIFE is qualitative and subjective. In this study we evaluate the discrepancies between sIFE and Mass Spectrometry (MS) to evaluate tumor burden and the accuracy of treatment response and it´s clinical impact in terms of progression free survival (PFS). Methods Patient samples were automatically prepared on the EXENT-iP®500 liquid handler, and sample spots were analysed using the EXENT-iX®500 MALDI-TOF mass spectrometer. Light chain spectral peaks were identified by EXENT-iQ® software and then quantified indirectly in combination with total immunoglobulins concentrations by immunoturbidimetry in the Optilite® platform (The Binding Site, part of Thermo Fisher Scientific). MPs were also identified and quantified by SPE and sIFE. Results From the 430 samples from the GEM12, GEM14 and GEM-CESAR clinical trials analyzed at pre-determined time points to evaluate treatment response (post-Induction, post-ASCT, post-cons, 1st and 2nd year after maintenance), MS identified the M protein in 45% of cases (193/430) and sIFE was positive in 32% (137/430). Combining the results of both methods, we found that MS and sIFE were concordant in 76% of cases (326/430), 26.28% IFE+/ MS+ and 49.53% IFE-/ MS-. Considering sIFE as a reference, the negative predictive value (NPV) of MS was 0.8987, with a sensitivity and a specificity of 0.8248 and 0.7270, respectively (p&amp;lt;0.0001). Nevertheless, the positive predictive value (PPV) was 0.5855 which could be explained by the discordances observed between IFE and MS techniques (5.58% of patients IFE+/MS- and 18.6% of patients IFE-/MS+). Indeed, when the 137 patients in VGPR (IFE+ patients) were analyzed according to MS status we found that patients IFE+/ MS+ had a trend to an inferior mPFS in comparison to IFE+/MS- (5.96 years vs not reached). These discordances were even more noticeable when comparing the 293 patients in CR (IFE- patients) with MS status, since IFE-/ MS+ patients had a mPFS of 4.65 years vs mPFS not reached for IFE-/ MS- patients (p=0.0001). Conclusions Our data confirms that IFE sensitivity limitations can affect the evaluation of treatment response in patients with MM, being more noticeable when the presence of MP band in sIFE is not clear because is very fuzzy or oligoclonal bands appear. Consequently, heterogeneity in interpretation practices across labs is a major cause of misattributed response categories, especially on those patients achieving treatment responses &amp;gt;VGPR. In an era of highly effective MM treatments that induce quick and deep responses, we believe IMWG response criteria should be updated with an MS response category.

B-336 Analytical verification of the EXENT® solution for the identification an quantitation of monoclonal immunoglobulins by mass spectrometry

Abstract Background Recently a new mass spectrometry (MS) technology, EXENT® solution (The Binding Site, part of Thermo Fisher Scientific), for the identification and quantification of IgG, IgA and IgM intact monoclonal immunoglobulins (M-Ig) has been launched in the European markets. In this study we present the results of onsite verification study performed in our hospital. Methods Study design, using patients serum samples, including data analysis were based on the Clinical &amp; Laboratory Standards Institute (CLSI). Relevant standards used were: Precision-CLSI EP15-A3:2014, Linearity-EP06 ED2:2020 and Comparison - EP09C ED3:2018. Samples were automatically prepared on the EXENT-iP®500 liquid handler, sample spots were analysed using the EXENT-iX®500 MALDI-TOF mass spectrometer. Light chain spectral peaks were identified by EXENT-iQ® software and then quantified indirectly in combination with total immunoglobulins concentrations by immunoturbidimetry in the Optilite® platform (The Binding Site, part of Thermo Fisher Scientific). We compared the results of MS with capillary electrophoresis (CZE) (IgG: n=82; IgA n=19; IgM n=19) and immunofixation (sIFE) (n=541). Results Precision was, for IgG with concentrations between 50.77 g/L and 3.51 g/L of 4.14% and 4.47%, for IgA between 61.88g/L and 5.02 g/L of 1.42% and 7.17%, and for IgM between 68.32 g/L and 7.30 g/L of 5.32% and 8.44%, respectively. Deviation from linearity of this assay, using serially diluted serum samples over the following ranges for IgG (65.18 g/L - 0.23 g/L), IgA (48.91 g/L - 0.26 g/L) and IgM (49.65 g/L -0.23 g/L) was &amp;lt;10% for all isotypes. M-Ig concentration comparison for specimens with detectable IgG, IgA or IgM M-Ig with CZE, assessed by Passing Bablok regression was IgG g/L MS = -0.9756 + 1.523x CZE g/L (slope -2.058 to -0.3322 and intercept 1.331 to 1.679), IgA g/L MS = -2.114 + 1.646x CZE g/L (slope -12.50 to -0.5059 and intercept 1.105 to 3.296), IgM g/L MS = -1.886 + 1.935x CZE g/L (slope -5.205 to -0.3571 and intercept 1.239 to 2.462). All samples MS g/L = -1.182 + 1.556x CZE g/L (n=120). Agreement between EXENT/IFE was of 99.1% for IgGK(209/211), 97.2% IgGL(105/108), 100% AK(97/97), 100% AL(51/51), 100% MK(6/6), 100% ML(3/3), 89.2% free K(33/37), 89.3% free L(25/28), 66,7% non-secretory(4/6). Discordances intact immunoglobulin was due to samples with two M-Ig, where IFE wasn’t able to identify the major clone, for free K, free L was due to EXENT’s ability to identify the heavy chain that matched with the light chain, and for non-secretory patients due to EXENT was able to identify one M-Ig in 2/6 samples. Conclusions These on-site verification results demonstrate that the EXENT solution has a wide measuring interval, to detect M proteins with very low concentrations and to provide stable and reproducible performance with a high level of accuracy for the detection and typing of monoclonal immunoglobulins. However, the bias observed against CZE to quantify M-Ig would be justified for the lowest CZE sensitivity, CZE variability and also due to the CZE quantification uses the total protein concentration to quantify M-Ig.

Efficacy and safety of telitacicept in patients with lupus nephritis.

Although telitacicept is a promising drug for treating systemic lupus erythematosus, there are limited studies on its efficacy and safety in patients with lupus nephritis in China. This lack of research data restricts its potential for broader application and acceptance on a global scale. The present study aimed to determine the efficacy and safety of telitacicept in patients with lupus nephritis (LN) in China. Using a self-controlled before-after comparison method, patients with LN were recruited at Lishui Central Hospital between February 2022 and April 2023, who received telitacicept weekly as part of the standard treatment. Data on the systemic lupus erythematosus disease activity index 2000 (SLEDAI-2K), glucocorticoid dosing and the quantity of immunosuppressive medicines prescribed was collected. Additionally, serum complements, erythrocyte sedimentation rate (ESR), urinary protein levels, immunoglobulin concentrations, serum creatinine levels, plasma albumin concentrations, platelet counts and renal function parameters were documented throughout the study. A total of 13 patients were enrolled in the trial, comprising 11 women and two men. Following 12-48 weeks of treatment with telitacicept (80 or 160 mg per week), 84.6% (n=11) of all patients experienced symptom relief and their SLEDAI-2K score was reduced by more than four points. By the observation endpoint, the median glucocorticoid dosage of the 13 patients was decreased from 15 to 2.5 mg/d, and six patients discontinued their glucocorticoids. Furthermore, 46.1% of patients (n=6) reduced their dose and number of immunosuppressive medicines, while 15.4% (n=2) stopped their immunosuppressive medicines. Minimal changes were observed in serum creatinine, platelet count, C3 levels and C4 levels among patients. Immunoglobulin levels (IgG, IgA and IgM) remained stable or showed an upward trend. Plasma albumin levels remained within the normal range in three patients and increased in ten patients. It increased to the normal range in three of these ten patients. At the endpoint, ESR levels decreased in all patients. Additionally, three patients displayed varying degrees of renal function improvement, and their estimated glomerular filtration rate (ml/min/l.73 m2) increased from 127.8 to 134.2, 95.1 to 123.1 and 61.5 to 67.3, respectively. Urinary protein levels decreased in all patients. It decreased >0.5 g/l in seven patients and reached the normal levels in three patients. The adverse events of telitacicept were manageable. Among the patients infected with COVID-19, three patients had fever, 10 patients remained asymptomatic and none of them exhibited severe respiratory syndromes. In this study, telitacicept effectively stabilized LN activity and alleviated the clinical symptoms of most patients. Furthermore, it reduced the dose of glucocorticoid and immunosuppressive medicines. Therefore, telitacicept may be a promising treatment option for individuals with lupus nephritis.

Effects of including different levels of equal mix of soybean and flaxseed oils in Japanese quail diets on the growth, carcass quality, and blood biomarkers

Corn, the primary ingredient in modern poultry feeds, contains high levels of ω-6 fatty acids but lacks sufficient ω-3 fatty acids, creating an imbalance. Maintaining a balance between ω-6 and ω-3 fatty acids in poultry diets is crucial due to their competition. This study aimed to evaluate the effects of incorporating different concentrations of an equal mix of soybean oil (SO) and flaxseed oil (FO) into quail diets on growth performance, carcass quality, and blood biochemistry. One-week-old Japanese quail birds (n=200) were randomly assigned to four dietary groups, each comprising five replicates with 10 chicks per replicate. Four isonitrogenous/isocaloric basal diets were formulated. Group 1 (control) received a basal diet without SO or FO, while Groups 2-4 received basal diets supplemented with an equal mix of SO+FO at levels of 1.0%, 1.5%, and 2.0%, respectively. The inclusion of oil mixes significantly increased body weight (BW) at five weeks and daily weight gain (DWG) during weeks 3-5 and 1-5. The feed conversion ratio (FCR) improved with the addition of oil mixes throughout the trial period. Supplementing quail diets with oil mixes resulted in reduced serum total cholesterol (TC) and LDL cholesterol, elevated serum HDL cholesterol, and no significant effect on triglycerides (TG) and VLDL cholesterol levels. Quails fed oil-supplemented diets showed lower serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and creatinine levels, while urea and uric acid were significantly affected. Birds fed diets with oil mixes also had increased serum concentrations of immunoglobulin G (IgY), immunoglobulin M (IgM), and superoxide dismutase (SOD). However, serum immunoglobulin A (IgA), malondialdehyde (MDA), and total antioxidant capacity (TAC) levels did not significantly change across experimental groups. Overall, adding up to 2% of the SO and FO mix in growing quail feeds improved growth performance, blood lipid profile, liver and kidney function markers, immune response, and antioxidant defense. The highest level of oil mix (2%) yielded the most beneficial effects.

Interactive Effects of Dietary Probiotic and Succinic Acid on the Growth Performance, Digestive Enzyme Activities, Immunomodulation, Antioxidative Capacity, and Disease Resistance in Rainbow Trout (Oncorhynchus mykiss) Juveniles

Abstract The current study was carried out to explore the effects of lactofeed (LA) as a multi-strain probiotic and succinic acid (SA) on growth variables, gut lactic acid bacteria count, digestive enzymes, innate immune responses, antioxidant capacity, and resistance against yersinia ruckeri in rainbow trout juveniles (Oncorhynchus mykiss). Three hundred sixty healthy rainbow trout juveniles (13.21±0.41 g) were randomly divided into twelve tanks (300 L) as four experimental groups. They were fed with basal diet (Control; BD), FT1: BD + 1 g/kg LA, FT2: BD + 5 g/kg SA, and FT3: BD + 1 g/kg LA + 5 g/kg SA for eight weeks. According to the results, final weight (FW), weight gain (WG), protein efficiency rate (PER), and feed conversion rate (FCR) revealed a remarkable discrepancy compared to the control group. In addition, dietary inclusion of LA (FT1 and FT3) significantly increased the specific growth rate (SGR). Dietary supplementation of LA+SA (FT3) remarkably improved total bacteria count (TBC) and amylase activity compared to the unsupplemented group. Gut lactic acid bacteria (LAB) count and digestive protease activity in all supplemented fish were remarkably higher than in unsupplemented group. Blood immunological factors including white blood cell (WBC) count, total immunoglobulin content (Ig), and the activity of lysozyme (LYZ), alternative complement pathway (ACH50), and acid phosphatase (ACP) were significantly enhanced in the supplemented groups. Neutrophil (NEU) count, skin mucus Ig and hepatic glutathione peroxidase (GPX) increased in SA (FT2 and FT3) treatments. In addition, monocyte (MON) count and skin mucus LYZ activity were significantly elevated following feeding with the FT3 diet. Other immunological parameters of skin mucus including protease activity, alkaline phosphatase (ALP) activity, and ACH50 activity, as well as hepatic superoxide dismutase (SOD) and catalase (CAT) activities increased in fish fed with diets containing LA and/or SA. Malondialdehyde (MDA) value was remarkably decreased in all supplemented rainbow trout compared to the fish fed with BD. Disease resistance against Y. ruckeri in fish fed with supplemented diets significantly improved with respect to the results obtained in the control specimens. Overall, dietary LA+SA supplementation was beneficial to improve growth performance, gut LAB count, digestive enzyme activities, innate immune responses, antioxidant capacity, and disease resistance in rainbow trout. According to these findings, 1 g/kg LA + 5 g/kg SA is suggested for adding to rainbow trout diet.

Immunoglobulin E and Serum Interleukin-13 and Bacteria in Patients with Alopecia Areata

Alopecia areata (AA) is a form of hair loss that does not result in permanent scars. The etiopathology of it is not completely understood. To evaluate the serum concentration of immunoglobulin E (IgE) and interleukin-13 (IL-13) in patients diagnosed with AA and determine the bacterial strain present in individuals with infections. The study comprised a sample of 60 women afflicted with alopecia areata and 30 women who were in good health, serving as controls.The levels of IgE and IL-13 in the serum were evaluated using an enzyme-linked immunosorbent assay test(ELISA). The microorganisms were identified by conventional techniques and verified using Vitek technology. for IgE showed highly significant differences (p value &lt; 0.1) Between patients and the control group, Regarding IL-13, the results showed significant differences (p value &lt; 0.05), Between patients and the control group, Regarding age As for IgE, there is no significant difference between patients in the three groups (p value &gt; 0.05), but it is different from the control (p value &lt; 0.05).IL-13 There are significant differences (p value &lt; 0.05). Among adults, the level of interleukin is higher than that of middle and young people. The weight-related results demonstrated significant variability across patients (p value &lt; 0.05), with obese individuals exhibiting higher levels of IGE and IL-13 compared to those of medium and underweight. Regarding, In relation to bacteria, the percentages were as stated S.epedermidis (50.98%) , (S.aureus33.33%) mixed , S.aureus + S.epedermidis , (9.8%) E.coli (1.96%) Pseudomonas aeruginosa(1.96 %) , Granulicatella elegans (1.96%) .Total is IgE and IL-13 increased in patients with alopecia areata, Regarding bacteria, we need other studies to prove their role in alopecia areata.

The Platform Trial In COVID-19 Priming and BOOsting (PICOBOO): The immunogenicity, reactogenicity, and safety of different COVID-19 vaccinations administered as a second booster (fourth dose) in AZD1222 primed individuals aged 50-

PICOBOO is a randomised, adaptive trial evaluating the immunogenicity, reactogenicity, and safety of COVID-19 booster strategies. We report data for second boosters among individuals 50-<70 years old primed with AZD1222 (50-<70y-AZD1222) until Day 84. Immunocompetent adults who received any first booster ≥three months prior were eligible. Participants were randomly allocated to BNT162b2, mRNA-1273 or NVX-CoV2373 1:1:1. The concentrations of ancestral anti-spike immunoglobulin were summarised as the geometric mean concentrations (GMC). Reactogenicity and safety outcomes were captured. Additional analyses including neutralising antibodies were performed on a subset. ACTRN12622000238774. Between Mar 2022 and Aug 2023, 743 participants were recruited and had D28 samples; 155 belonged to the 50-<70y-AZD1222 stratum. The mean adjusted GMCs (95% credible intervals) were 20,690 (17 555-23 883), 23,867 (20 144-27 604) and 8654 (7267-9962) U/mL at D28 following boosting with BNT162b2, mRNA-1273 and NVX-CoV2372, respectively, and 10,976 (8826-13 196), 15,779 (12 512-19 070) and 6559 (5220-7937) U/mL by D84. IgG against Omicron BA.5 was 2.7-2.9 times lower than the ancestral strain. Limited neutralisation against Omicron subvariants was found following all vaccines. Severe reactogenicity events were <4%. All vaccines were immunogenic with more rapid waning after mRNA vaccines. These data support boosting with vaccines with greater specificity for circulating Omicron subvariants.

Long-Term Survival and Immune Reconstitution of Donor-Derived Chimeric Antigen Receptor T-Cell Therapy for Childhood Molecular Relapse of B-Cell Acute Lymphoblastic Leukemia After Allogeneic Hematopoietic Stem Cell Transplantation

Measurable residual disease (MRD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an independent risk factor for relapse in patients with acute lymphoblastic leukemia (ALL). This study aimed to assess the efficacy, safety, and immune reconstitution of chimeric antigen receptor T-cell (CAR-T) therapy in patients with molecular relapse after allo-HSCT. Eleven patients with molecular relapse of B-cell-ALL who underwent CAR-T therapy after allo-HSCT were enrolled. The rate of MRD negativity after a month of CAR-T infusion was 81.8%. Patients who bridged to second-HSCT after CAR-T therapy (n = 3) showed a trend of higher 3-year leukemia-free survival and 3-year overall survival than those who did not (n = 8; 100% vs. 75.0%; 95% CI, 45.0–104.9%; p = 0.370). No treatment-related mortalities were observed. Among patients who did not bridge to second-HSCT and remained in complete remission until the last follow-up (n = 6), five of them had not recovered normal immunoglobulin concentrations with a median follow-up of 43 months. CAR-T therapy may be a safe and effective treatment strategy to improve survival after allo-HSCT; however, the problem of prolonged hypogammaglobulinemia in patients who do not bridge to second-HSCT is worth noting.