Ig Concentrations Research Articles

Rapid depletion of CD20+ B and T cells following ofatumumab therapy onset

BackgroundThe humanized monoclonal anti-CD20-antibody ofatumumab is highly effective in treating relapsing multiple sclerosis (MS). ObjectiveThis study aimed to investigate the immanent effect of ofatumumab on the peripheral immune system, particularly targeting B and T cells expressing CD20. MethodsBlood samples of 53 MS patients receiving ofatumumab were collected prior to first application and after one week, two weeks and three months. Multicolor flow cytometry was used to phenotype peripheral blood mononuclear cells, and immunoglobulin (Ig) concentrations were measured by nephelometry. ResultsAmong CD20+ lymphocytes, 13 % co-expressed CD3 (identifying them as CD3+CD20+ T lymphocytes), with a noticeable shift in the CD4/CD8-ratio towards CD8+ T cells. One week after administering ofatumumab, a significant reduction of CD20+ lymphocytes with complete depletion of CD3+CD20+ T lymphocytes was observed, persisting during the investigation period. During the treatment, IgM levels showed a slight but significant decrease, whereas IgA and IgG levels remained stable. ConclusionOfatumumab effectively depletes CD20+ lymphocytes already after the first administration. This depletion affects not only B cells, but also a small proportion of T cells (CD3+CD20+), affirming the hypothesis that the anti-inflammatory effects of CD20+ cell depletion might extend to the reduction of CD3+CD20+ T lymphocytes.

Serum immunoglobulin levels in group E of chronic obstructive pulmonary disease: insights for clinical management and immunoglobulin therapy strategies

ObjectiveThe study aimed to characterize serum immunoglobulin (Ig) concentrations and their relationship with clinical and paraclinical features in patients with COPD group E in the stable stage. Additionally, the study focused on evaluating the relationship between serum Ig levels and the risk of exacerbations over the next 12 months, thereby clarifying the role of serum Ig deficiency in affecting the future risk for these patients.MethodsA prospective observational study assessed IgG, IgA, IgM, and IgE levels in 67 COPD patients and 30 healthy controls at Military Hospital 103 from October 2017 to August 2020. Primary outcomes included Ig isotype levels in COPD patients, with secondary outcomes exploring differences compared to controls and associations with clinical variables.ResultsCOPD patients showed significantly lower IgG concentrations and higher IgA levels than controls. IgM and IgE levels did not differ significantly. Subgroup analysis revealed notable decreases in IgG1 and IgG3 concentrations, with 10.4% of patients exhibiting reduced IgG levels and 0.3% diagnosed with common variable immunodeficiency. No significant associations were found between Ig levels and exacerbation risk or clinical variables.ConclusionsSerum IgG and IgM concentrations were significantly reduced in COPD patients compared to normal individuals, with IgG1 and IgG3 concentrations notably low. Serum IgA levels were significantly higher in COPD patients compared with normal controls. However, no significant association was found between Ig concentrations, particularly serum IgG deficiency and its subclasses, with the frequency and risk of exacerbations during 12 months of longitudinal follow-up. Caution is warranted in the use of immunoglobulin therapy in the treatment of COPD patients.Trial registrationAn independent ethics committee approved the study (Ethics Committee of Military Hospital 103 (No. 57/2014/VMMU-IRB), which was performed in accordance with the Declaration of Helsinki, Guidelines for Good Clinical Practice.

Survey on colostrum management by German dairy farmers focusing on frozen colostrum storage

Because calves are born with low levels of antibodies, effective colostrum management is one of the most critical factors for successful calf rearing. A timely and adequate supply of sufficiently high-quality colostrum immediately after birth is essential to ensure the passive immunization of calves. Frozen colostrum reserves are recommended to fulfill the immunological and nutrient requirements of newborn calves, even in exceptional situations; however, the implementation rates on German dairy farms and challenges of realization remain unclear. A 33-question online survey, focused on frozen colostrum reserves, was developed to obtain an overview of colostrum management practices on German dairy farms. The questionnaire was divided into 3 sections: 1. personal data; 2. farm characteristics; 3. colostrum management. Of the 155 responses we received, 63.9% were from female farmers, and 35.5% were from male farmers. Conventional farming was practiced on 89.0% of farms, and organic farming was practiced on 7.1% of farms. Of the respondents, 89.0% froze colostrum. The main reasons for freezing colostrum were: 1. the dam does not produce enough colostrum; 2. the dam cannot be milked; or 3. in cases where the dam dies during birth. Farmers primarily froze colostrum from cows during their third to fifth lactation. Before freezing, 33.1% of the respondents measured indicators in the colostrum to estimate Ig concentrations, whereas 2.3% determined the colostrum quality after freezing. Reusable and disposable PET deposits (23.1%, 22.3%) and ColostroBags (20.0%) were the primary containers used to freeze colostrum. The main reasons for not freezing colostrum were the high labor intensity and the availability of fresh colostrum from other cows. Thawing methods included buckets (47.7%) and professional water baths (13.8%). The survey identified areas in which improved knowledge transfer could enhance colostrum management. Furthermore, there appeared to be a lack of specific, feasible instructions for employees concerning the practical implementation of colostrum management. Most importantly, the regular determination and documentation of Ig concentrations should be emphasized. The added value of stored colostrum, relative to a greater workload, should also be promoted, particularly on smaller farms.

Exploring the Impact of Extra Virgin Olive Oil on Maternal Immune System and Breast Milk Composition in Rats.

Maternal breast milk plays a key role in providing newborns with passive immunity and stimulating the maturation of an infant's immune system, protecting them from many diseases. It is known that diet can influence the immune system of lactating mothers and the composition of their breast milk. The aim of this study was to establish if a supplementation during the gestation and lactation of Lewis rats with extra virgin olive oil (EVOO), due to the high proportion of antioxidant components in its composition, has an impact on the mother's immune system and on the breast milk's immune composition. For this, 10 mL/kg of either EVOO, refined oil (control oil) or water (REF group) were orally administered once a day to rats during gestation and lactation periods. Immunoglobulin (Ig) concentrations and gene expressions of immune molecules were quantified in several compartments of the mothers. The EVOO group showed higher IgA levels in both the breast milk and the mammary glands than the REF group. In addition, the gene expression of IgA in mammary glands was also boosted by EVOO consumption. Overall, EVOO supplementation during gestation and lactation is safe and does not negatively affect the mother's immune system while improving breast milk immune composition by increasing the presence of IgA, which could be critical for an offspring's immune health.

#2684 Free light chains levels in ANCA associated vasculitis: a new potential inflammatory biomarker

Abstract Background and Aims Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis is a systemic disease characterized by an immune-mediated process where inflammation plays a central role. The widely available serum biomarkers of inflammation are not always able to detect ongoing disease activity in ANCA-associated vasculitis (AAV) patients. Free light chains (FLCs) expression levels have been proposed as new promising biomarker in some immune-mediated and inflammatory diseases. The aim of this study is to evaluate the expression levels of FLCs in patients affected by AAV and investigate the potential effect of rituximab treatment on FLCs levels. Method We conducted a retrospective multicenter study enrolling AAV patients in complete remission of AAV (defined as BVAS/GPA = 0); those with clinical suspicion of underlying hematologic malignancy were excluded. We analyzed κ and λ FLCs serum concentrations and evaluated their correlation with biomarkers of immune system activation, such as erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), serum immunoglobulin (Ig) levels, serum C3 and C4 complement and albumin levels (which has a negative correlation with acute inflammatory phase). We assessed impact of rituximab (RTX) therapy within the last 6 months on the same laboratory parameters. Results A total of 137 patients with AAV were recruited, who were gender balanced, had an average age of 68 years and an estimated glomerular filtration rate of 68 (43-89) mL/min/1.73 m2. Fifty-one patients received rituximab treatment (RTX group), while 86 patients were on other immunosuppressive maintenance therapies (non-RTX group). In comparison to non-RTX group, patients treated with RTX experienced higher complement C3 levels, lower ESR levels and lower FLCs κ (p = 0.063) and λ (p = 0.013) levels. The differences between RTX and non-RTX groups in terms of FLCs were even more prominent after correction to total Ig concentration (p = 0.004 for κ /Ig and p = 0.003 for λ /Ig, respectively). Both, serum free κ and λ levels were found to correlate significantly with ESR (R = 0.55 and p < 0.001 for κ; R = 0.53 and p < 0.001 for λ) and albumin (R = ‒0.38 and p < 0.001 for κ; R = ‒0.33 and p < 0.001 for λ). Additionally, serum free κ levels showed an inverse association with C3 (R = ‒0.18 and p = 0.041). Conclusion In subjects with AAV, FLC levels were found to be associated with markers of inflammation and complement activation; hence FLCs levels may represent a promising biomarker for detection of subclinical inflammation in AAV patients on maintenance therapy. Rituximab therapy showed a potential to reduce the levels of markers of subclinical inflammation, including FLCs.

Immunobiological status of the organism of young pigs during their triphasic period cultivation and actions of corrective factors

The problem of the impact and consequences of technological stress is accompanied by several reactions that do not always have a positive effect on the animal's body. The work aimed to investigate the dynamics of indicators of the immunobiological status of piglets during their three-phase rearing and the use of immunostimulating agents Globigen® Pig Doser and Globigen® Jump Start. Parallel studies were conducted based on the private farm “Meat Resources” LLC of the Lviv region of the Starosambir district, which practices three-phase pig breeding. The experiment involved 36 analog piglets and hybrids of Landrace and Great White breeds, which were formed into 3 groups of 12 animals each. Piglets of the control group were fed the standard ration provided by the technology. During the first three days of life, newborn piglets of the first experimental group were given 2 ml/gal of liquid Globigen®Pig Doser. The piglets of the second experimental group were given a powdered dry product Globigen®Jump Start, which was added to the main diet at 0.2 % per ton of feed. It was established that the use of means with an immunostimulating effect was accompanied by a decrease in the content of Ig M in the blood samples of experimental groups of piglets. Thus, in the blood serum of 30-day-old piglets under the influence of Globigen® Pig Doser with a high level of probability (Р < 0.02), a decrease in the content of this class of antibodies was found to the level of 1.59 mg/ml, and under the influence of Globigen® Jump Start the content in the blood serum of 60-day-old piglets, having reached the lowest value, was 2.08 mg/ml. The positive effect of immunostimulating agents, which was characterized by an increase in the amount of Ig G, was found in experimental groups of pigs at the age of 60 days: 17.10 mg/ml for the effects of Globigen® Pig Doser and 17.08 mg/ml for the impact of Globigen® Jump Start. The highest content of Ig A with high probability (P < 0.001) was found in blood samples of animals that were given Globigen® Jump Start – 3.45 mg/ml and 3.86 mg/ml under the influence of Globigen® Pig Doser. The practicality and effectiveness of the use of means with immunostimulating properties, the main active ingredient of which is the immunoglobulin of a chicken egg, is explained by the ability to influence the pathogenesis of diseases of the gastrointestinal tract, which arise as a result of weakening the resistance of the animal body under the influence of a large number of technological stresses. Research data prove that the specificity of the action of Ig Y has a positive effect on the preservation of the integrity of the villi of the intestinal mucosa, which is reflected by the strengthening of its protective and barrier function, which in general contributes to the increase in the percentage of animal survival: using Globigen®Pig Doser 80.0 % and 86.7 % using Globigen®Jump Start. Based on such results, it can be asserted that the use of Globigen® Pig Doser and Globigen® Jump Start for three-phase pig farming is scientifically justified and economically feasible.

Effect of Salvia sclarea L. extract on growth performance, antioxidant capacity, and immune function in lambs.

This study aimed to explore the effects of Salvia sclarea extract on the growth performance, apparent nutrient digestibility, antioxidant capacity, and immune function of the lambs. Sixty female lambs (Chinese Merino sheep) aged 2 months and weighing 20 ± 2 kg were selected and randomly divided into five groups of 12 lambs each. The control group (CK) received only basal feed, whereas the experimental group was supplemented with different concentrations of salvia sclarea extract in the basal feed at 0.04, 0.08, 0.12, and 0.16 mL/kg (CL1, CL2, CL3, and CL4, respectively). The feeding period was 85 days, including 15 days of pre-feeding and 70 days of regular feeding. Body weight and feed intake were recorded during the test period, and blood was collected at the end of the test to determine immune and antioxidant indices. The results showed that the average daily weight gain and feed intake of the lambs were significantly higher in the CL3 group than in the CK group (p < 0.05). In addition, the apparent nutrient digestibility of crude protein and neutral detergent fiber increased significantly (p < 0.05). The dry matter, acid detergent fiber, and ether extract were not significantly different (p > 0.05). Serum levels of superoxide dismutase, catalase, and glutathione peroxidase and antioxidant capacity were significantly higher in the CL2, CL3, and CL4 groups than in the CK group, whereas malondialdehyde levels were significantly lower (p < 0.05). The serum levels of immune globulin immune globulin A, immune globulin G, immune globulin M, interferon-γ, and interleukin-10 were significantly higher and the levels of tumor necrosis factor-α and interleukin-1β were significantly lower in the CL2, CL3, and CL4 groups (p < 0.05). In conclusion, the addition of the S. sclarea extract to the diet promoted growth performance and nutrient digestion in lambs. Immune response was improved by increasing Ig and cytokine concentrations. It enhances antioxidant status by increasing antioxidant enzyme activity in lambs.

Immunoglobulins in COVID-19 pneumonia: from the acute phase to the recovery phase

BackgroundCOVID-19 pneumonia causes hyperinflammatory response that culminates in acute respiratory syndrome (ARDS) related to increased multiorgan dysfunction and mortality risk. Antiviral-neutralizing immunoglobulins production reflect the host humoral status and illness severity, and thus, immunoglobulin (Ig) circulating levels could be evidence of COVID-19 prognosis.MethodsThe relationship among circulating immunoglobulins (IgA, IgG, IgM) and COVID-19 pneumonia was evaluated using clinical information and blood samples in a COVID-19 cohort composed by 320 individuals recruited during the acute phase and followed up to 4 to 8 weeks (n = 252) from the Spanish first to fourth waves.ResultsCOVID-19 pneumonia development depended on baseline Ig concentrations. Circulating IgA levels together with clinical features at acute phase was highly associated with COVID-19 pneumonia development. IgM was positively correlated with obesity (ρb = 0.156, P = 0.020), dyslipemia (ρb = 0.140, P = 0.029), COPD (ρb = 0.133, P = 0.037), cancer (ρb = 0.173, P = 0.007) and hypertension (ρb = 0.148, P = 0.020). Ig concentrations at recovery phase were related to COVID-19 treatments.ConclusionsOur results provide valuable information on the dynamics of immunoglobulins upon SARS-CoV-2 infection or other similar viruses.

The ability of a handheld near-infrared spectrometer to do a rapid quality assessment of bovine colostrum, including the immunoglobulin G concentration

Portable infrared-based instruments have made significant contributions in different research fields. Within the dairy supply chain, for example, most of portable devices are based on near-infrared spectroscopy (NIRS) and are nowadays an important support for farmers and operators of the dairy sector, allowing fast decision-making, particularly for feed and milk quality evaluation and animal health and welfare monitoring. The affordability, portability, and ease of use of these innovative devices have been pivotal factors for the implementation in dairy farms. In fact, pocket-sized devices enable non-expert users to perform quick, low cost and non-destructive analysis on various samples without complex preparation. As bovine colostrum (BC) quality is mostly given by the Ig G (IgG) level, evaluating the ability of portable NIRS tools to measure antibodies concentration is advisable. In this study we used the wireless device SCiO manufactured by Consumer Physics Inc. (Tel Aviv, Israel) to collect BC spectra and then attempt to predict IgG concentration and gross and fine composition in individual samples collected as soon as possible after calving (<6 h) in primiparous and pluriparous Holstein cows farmed in 9 Italian farms. Chemometric analyses revealed that SCiO has promising predictive performance for colostral IgG concentration, total Ig concentration, fat, and AA (R2CV ≥ 0.75). Excellent accuracy was observed for dry matter, protein, and S prediction in cross-validation and good prediction ability in external validation (R2CV ≥ 0.93; R2V ≥ 0.82). Nonetheless, SCiO's ability to discriminate between good- and low-quality samples was satisfactory. The affordable cost, the accurate predictions, and the user-friendly design coupled with the increased interest in colostrum quality within the dairy sector may boost the collection of extensive BC data for management and genetic purposes in the near future.

Administration of Red Macroalgae (Galaxaura oblongata) in the Diet of the Rainbow Trout (Oncorhynchus mykiss) Improved Immunity and Hepatic Gene Expression

This study is designed to evaluate the effects of dietary red macroalgae (Galaxaura oblongata) on growth performance, serum, and skin mucus immunological and antioxidant responses in rainbow trout (Oncorhynchus mykiss). For this, rainbow trout were fed diets containing different levels of G. oblongata (0 (ctrl), 0.5 (G1), and 1 (G2) %) for 8 weeks. Following the feeding trial, there were no significant differences in growth performance between the experimental treatments (p &gt; 0.05). Total immunoglobulin (Ig) content and lysozyme (LYZ) activity in serum were increased in fish fed G. oblongata (p &lt; 0.05), with the highest value at (0.5%). Regardless of the inclusion level, mucus total Ig levels were significantly increased in the G. oblongata groups (p &lt; 0.05), and mucus LYZ activity was not changed (p &gt; 0.05). All groups fed G. oblongata showed higher serum catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activities than the control group (p &lt; 0.05). However, skin mucus SOD activity increased more in the group fed 1% of G. oblongata than the other groups (p &lt; 0.05). Additionally, the skin mucus GPx activity showed higher values in the group fed 0.5 and 1% G. oblongata than in the control (p &lt; 0.05). No significant differences were recognized between the experimental treatments in terms of CAT activity and malondialdehyde (MDA) concentration (p &gt; 0.05). G. oblongata up-regulated gpx gene expression with the maximum value at the group fed 1% G. oblongata (p &lt; 0.05). Additionally, interleukin 6 (il-6) and tumor necrosis factor-alpha (tnf-α) gene expressions were significantly up-regulated in fish fed 1% compared with the control and 0.5% groups. Based on the results, 0.5–1% G. oblongata can be used in the fish diet and enhance immunity without causing impairment in growth.

P338 The dependence of the level of Glycoprotein type 2 on the level of Zonulin in Crohn's disease

Abstract Background Introduction. Type 2 glycoprotein (GP2) is a quantitatively predominant membrane protein of excretory granules of pancreatic acinar cells. In a number of studies, it has been demonstrated that this protein is the main antigen for pancreatic antibodies associated with Crohn's disease Zonulin – a protein that can reversibly increase the permeability of the intestinal wall by changing the structure of tight junctions of the lateral surfaces of intestinal epithelial cells. The determination of fecal zonulin is used for noninvasive assessment of increased intestinal permeability. Normal values of zonulin in stool (≤ 110 ng/ml) indicate the absence of damage to the villous surface of the intestinal mucosa and normal density of intercellular contacts The aim of the study was to establish the dependence of the level of type 2 glycoprotein on the level of fecal zonulin (FZ) in the feces of CD patients with exacerbation of the disease. Methods 122 patients with CD in the form of ilekolitis with exacerbation of the disease (Me age — 32 years) were examined. FZ was evaluated by ELISA (IDK ® Zonulin ELISA Kit, Immunodiagnostik AG, Germany) in ng/ml. Reference values: &amp;lt; 83.15 ng/ml is a variant of the norm, 83.15-110 ng/ml is an elevated level, 110 ng/ml is a high level. The level of IgG and IgA to GP2 (antibodies to glycoprotein) antigen of pancreatic centroacinar cells (Anti-GP2, IgG, IgA) was determined by ELISA. Reference values: IgG GP-2 less than 10 units/ml, Ig and GP-2 less than 5 units/ml – positive values. Results In the stool samples of CD patients during the period of exacerbation in 92 patients, an increase in FZ was detected, the average value was 333.4 ±16.9 ng/ml. In the blood samples of 87 patients with CD during the period of exacerbation, an increase in Ig A GP-2 was detected, the average value was 43.4±6.9 units/ml. In the blood samples of 107 CD patients during the period of exacerbation, an increase in Ig G GP-2 was detected, the average value was 17.1 ± 1.3 units/ml. There is a high correlation force between the concentration of FZ and the concentration of Ig A GP-2 (r-0.760) (p=0.010). There is a moderate correlation force (r-0.490) (p=0.023) between the concentration of FZ and the concentration of Ig G GP-2. Conclusion An increase in the concentration of FZ in the feces of CD patients during exacerbation significantly correlates with an increase in the concentration of Ig A GP-2 and Ig G GP-2.

Dose-response of inactivated yeast in diets of late gestating and lactating gilts on immunoglobulin transfer and offspring preweaning growth performance.

Fifty gilts (initial body weight [BW] 190.7 ± 4.2kg) were recruited on day 85 of gestation and were used until day 19 of lactation to assess the dose-response of inactivated yeast via hydrolyzation (HY) inclusion on offspring growth and immunoglobulin (Ig) transfer prior to weaning. Gilts were assigned to one of the 5 experimental diets: a control with no HY (HY0) or inclusion of 0.25% (HY0.25), 0.5% (HY0.5), 1.0% (HY1.0), or 1.2% (HY1.2) HY. Gilts were weighed on days 85 and 110 of gestation and days 1 and 19 (weaning) after farrowing. Offspring were weighed on days 1 and 19 of age. On lactation day 1 (approximately 24h after farrowing), colostrum, gilt plasma, and plasma from 2 median BW piglets were collected and on day 19, plasma from each gilt and 2 median BW piglets per litter were collected for determination of Ig concentrations. Contrast statements were used to assess the linear, quadratic, cubic, and quartic effects of HY inclusion. The inclusion of HY had minimal effects on gilt BW or litter characteristics at birth (total number born and born alive, piglet birth weight). Lactation average daily feed intake of the gilts tended to increase then decrease with increasing HY inclusion (quadratic; P = 0.085). Piglet preweaning average daily gain (linear, quadratic, and quartic; P < 0.05) and BW at weaning (quadratic and quartic; P < 0.05) increased then decreased with increasing HY inclusion. On lactation day 1, colostrum and gilt plasma Ig concentrations were not affected by dietary treatment (P > 0.10) but piglet IgA and IgM decreased then increased with HY inclusion level (cubic; P < 0.05). On lactation day 19, piglet plasma IgG tended to increase with HY inclusion (linear; P = 0.099). In summary, increasing HY inclusion in late gestating and lactating gilt diets improved immune transfer in the first 24h after birth and piglet preweaning growth rates and BW at weaning. Therefore, maternal feeding of HY could be used as a strategy to improve offspring immunocompetence and BW at weaning, with possible carryover benefits for the postweaning phase.

Impact of the mare colostral immunoglobulins on the expression of TLR3, TLR4, and TLR7 in foals.

Colostrum contains substances such as hormones, enzymes, polyamides, nucleic acid derivatives, and amino acid derivatives and also includes immunomodulating substances, proline-rich polypeptides, immunoglobulins (Ig) as well as bacteriostatic compounds. The aim of the study was to evaluate the impact of colostral Ig on the level of expression of selected Toll-like receptor genes (TLR3, TLR4, and TLR7). The experiments were conducted on 25 Polish Pony foals. Blood samples were collected according to the following scheme: before first suckling, at the 1st, 3rd, 5th, 10th, 20th, and 30th, day of age. Colostrum was obtained before the first suckling, 24 hr, and 3 days after the delivery. Gene expression analyses were performed on Illumina Eco using a commercial kit TaqMan®MGB probes. The quality of colostrum was assessed via refractometer and BRIX value. The total Ig level was calculated from the spectrophotometric method. IgG levels were measured using ELISA assay. The quality of colostrum significantly correlated with TLR4 and TLR7 expression. Foals that ingested colostrum with low Ig content displayed a higher level of TLR4 gene expression, while in the case of TLR7 the opposite trend was shown. This result indicates that colostrum may play an important role in shaping the mechanisms of building the immune system's response to bacterial pathogens.

HUMORAL IMMUNITY IN PATIENTS WITH MULTIPLE SCLEROSIS

Introduction. More than 1 million people in the world suffer from multiple sclerosis (MS). The etiology of MS remains not fully understood. The main role in the pathogenesis of MS belongs to the combination of chronic inflammation and neurodegenerative processes, in the genesis of which autoimmune processes play a significant role. Progress in studying the mechanisms of the pathological process of demyelinating diseases has been achieved mainly thanks to immunological research. One of the manifestations of activation of humoral immunity in MS is increased synthesis of immunoglobulins in brain tissue and cerebrospinal fluid&#x0D; The aim of the study. To study the state of humoral immunity in multiple sclerosis patients with different types of disease course&#x0D; Research methods. 68 patients with a verified diagnosis of multiple sclerosis (MS) based on the MacDonald et al criteria aged 19 to 65 years (37 women, 29 men) were examined. The primary progressive type of the course was established in 19 patients, the secondary-progressive - in 3, the remitting-relapsing type of the course - in 26 patients. The control group consisted of 80 practically healthy male volunteers aged 25-45 years.&#x0D; The concentration of immunoglobulins A, G, M in blood serum was determined by the Mancini method (1965). The content of circulating immune complexes (CIC) in blood serum was determined by the method of precipitation in polyethylene glycol.&#x0D; Results and discussion. The highest content of Ig A was observed in the group of patients with primary progressive form of MS, which was 3.7 times higher than normal values and 1.2 times higher than in the general group of MS patients. In the group of patients with the secondary progressive form of MS, the content of Ig A was 3.1 times higher than in the control group and 1.2 times lower than in the group with the primary progressive form. In the group of patients with relapsing-remitting form of MS, there was an increase in the level of Ig A three times relative to the normal indicator, and a decrease (by 1.2 times) relative to the level in the group with a primary progressive form of MS.&#x0D; The content of Ig M in the blood serum of patients with MS exceeded the norm by 5 times. In the group with the primary progressive form of MS, the level of Ig M was 3.5 times higher than the level in the control group and 1.5 times lower than the level in the group of patients with MS. In patients with secondary progressive form of MS, the content of Ig M was 2.6 times higher than the norm, but 1.3 times lower than the level in the group with primary progressive form of MS and 1.9 times lower than in the group of patients with MS. In patients with the relapsing-remitting form of MS, the level of Ig M was: 4.6 times higher than the level in the control group and 1.7 times higher than the content in patients with the secondary-progressive form of MS, and also 1.3 times higher, than in the group with the primary progressive form.&#x0D; In all groups of patients with MS with different types of the course of the disease, the most pronounced increase in the level of Ig G was noted.&#x0D; The highest level of CIC was recorded in the general group of patients with MS and in the group with secondary progressive form of MS.&#x0D; Conclusions. An imbalance of the humoral link of immunity was observed in patients with multiple sclerosis. A probable increase in the concentration of Ig A in the blood serum of patients with multiple sclerosis indicates the presence of infection of the mucous membranes, which is especially pronounced in patients with the primary progressive form of multiple sclerosis. A significant increase in the content of Ig G in the blood serum of patients with multiple sclerosis indicates a chronic course of the process, which is especially evident in the relapsing-relapsing form of multiple sclerosis. An increased concentration of IgM in the blood serum of patients with multiple sclerosis may indicate an exacerbation of the course of chronic inflammation, especially in the case of relapsing-remitting multiple sclerosis. The detected fluctuations of the average content of CIC in the blood serum of patients with multiple sclerosis did not go beyond the physiological norm.

Synergistic Suppression of NF1 Malignant Peripheral Nerve Sheath Tumor Cell Growth in Culture and Orthotopic Xenografts by Combinational Treatment with Statin and Prodrug Farnesyltransferase Inhibitor PAMAM G4 Dendrimers.

Neurofibromatosis type 1 (NF1) is a disorder in which RAS is constitutively activated due to the loss of the Ras-GTPase-activating activity of neurofibromin. RAS must be prenylated (i.e., farnesylated or geranylgeranylated) to traffic and function properly. Previous studies showed that the anti-growth properties of farnesyl monophosphate prodrug farnesyltransferase inhibitors (FTIs) on human NF1 malignant peripheral nerve sheath tumor (MPNST) cells are potentiated by co-treatment with lovastatin. Unfortunately, such prodrug FTIs have poor aqueous solubility. In this study, we synthesized a series of prodrug FTI polyamidoamine generation 4 (PAMAM G4) dendrimers that compete with farnesyl pyrophosphate for farnesyltransferase (Ftase) and assessed their effects on human NF1 MPNST S462TY cells. The prodrug 3-tert-butylfarnesyl monophosphate FTI-dendrimer (i.e., IG 2) exhibited improved aqueous solubility. Concentrations of IG 2 and lovastatin (as low as 0.1 μM) having little to no effect when used singularly synergistically suppressed cell proliferation, colony formation, and induced N-RAS, RAP1A, and RAB5A deprenylation when used in combination. Combinational treatment had no additive or synergistic effects on the proliferation/viability of immortalized normal rat Schwann cells, primary rat hepatocytes, or normal human mammary epithelial MCF10A cells. Combinational, but not singular, in vivo treatment markedly suppressed the growth of S462TY xenografts established in the sciatic nerves of immune-deficient mice. Hence, prodrug farnesyl monophosphate FTIs can be rendered water-soluble by conjugation to PAMAM G4 dendrimers and exhibit potent anti-tumor activity when combined with clinically achievable statin concentrations.

The immunomodulating activity of trimodulin (polyvalent IgM, IgA, IgG solution): a post hoc analysis of the phase II CIGMA trial

BackgroundThe phase II CIGMA trial performed in 160 patients with severe community-acquired pneumonia (sCAP) found treatment with trimodulin (human polyvalent immunoglobulin [Ig]: ~ 23% IgM, ~ 21% IgA, ~ 56% IgG) was associated with a lower mortality in those patients with elevated baseline serum levels of C-reactive protein (CRP) and/or subnormal IgM.MethodsIn this post hoc analysis, the pharmacodynamic effects of trimodulin treatment (182.6 mg/kg/day for 5 days) were investigated on Ig replenishment, cellular markers of inflammation (absolute neutrophil [ANC] and lymphocyte [ALC] count, neutrophil-to-lymphocyte ratio [NLR]), and soluble markers of inflammation (procalcitonin [PCT] and CRP). The impact of these pharmacodynamic effects on mortality was also evaluated.ResultsCompared with healthy subjects, baseline serum levels of IgM, IgG, and ALC were significantly lower, and ANC, NLR, PCT and CRP significantly higher in sCAP patients (p < 0.0001). Low Ig concentrations increased with trimodulin. Normalization of ANC (analysis of variance [ANOVA] p = 0.016) and PCT (ANOVA p = 0.027) was more rapid with trimodulin compared with placebo. These and other effects were more evident in patients with low baseline IgM levels. Normalization of PCT and CRP levels was both steadier and faster with trimodulin treatment. In patients with low baseline ALC, trimodulin was associated with a lower 28-day all-cause mortality rate (14.5% vs 32.1% in placebo, p = 0.043) and more ventilator-free days ([VFD]; median VFD: 3.5 vs 11 in placebo, p = 0.043). These numerical differences were greater if baseline IgM was also low (low ALC, low IgM: 8.1% mortality vs 34.1% placebo, p = 0.006; 3 VFD vs 15 VFD, p = 0.009, respectively). Results were consistent in patients with high baseline CRP (low ALC, high CRP: 10.9% mortality vs 34.1% placebo, p = 0.011).ConclusionsThis post hoc pharmacodynamic analysis of a blinded phase II trial suggests that trimodulin compensates for, and more rapidly modifies, the dysregulated inflammatory response seen in sCAP patients. Trimodulin was associated with significantly lower mortality and more VFD in subgroups with high CRP and low ALC. This effect was particularly marked in patients who also had low baseline IgM values. These findings require confirmation in prospective trials.

PSXIV-30 Effects of Vitamin/Mineral Supplementation During Gestation in Beef Heifers on Immunoglobulin Concentrations in Colostrum and Immune Responses in Naturally and Artificially Reared Calves

Abstract Two experiments were conducted to evaluate the impacts of feeding a vitamin/mineral supplement to beef heifers throughout gestation on concentrations of immunoglobulin (Ig) in colostrum and in calf serum 24 h after consumption of maternal colostrum (Exp. 1) or a colostrum replacement product (Exp. 2). Angus-based beef heifers [n = 72, initial body weight (BW) = 380.4 ± 50.56 kg] were managed in an individual feeding system and randomly assigned to a basal diet targeting gain of 0.45 kg·heifer-1·d-1 (CON; n = 36) or the basal diet plus 113 g·heifer-1·d-1 of a vitamin/mineral supplement (VTM; n = 36). For Exp. 1 treatments began at the time of artificial insemination (AI) to sexed semen, and heifers becoming pregnant after first service AI (CON, n = 14; VTM, n = 17) continued treatments throughout gestation. At calving, heifer calves were allowed to nurse their dams and remained alongside their dams until weaning. For Exp 2. non-pregnant heifers were rebred 60 d after initial dietary treatments began and continued treatments throughout gestation, with resultant heifer calves (CON, n = 7; VTM, n = 7) being removed from their dams at birth and receiving 1.5 L of colostrum replacer containing 150 g globulin protein via an esophageal feeder, followed every 12 h by 1.9 L of milk replacer. In both experiments samples of colostrum were collected after completely milking the rear-right quarter of the udder. Blood samples were collected from calves before (within 2 h of birth) and 24 h after colostrum consumption. Concentrations of IgG, IgM, and IgA were quantified in colostrum and serum via bovine radial immunodiffusion plates. Data for both experiments were analyzed for the effect of treatment using the MIXED procedure in SAS, with additional analysis of data from Exp. 1 using the CORR procedure to determine correlations among dam colostrum and calf serum Ig concentrations at 24 h. In both experiments, maternal treatment did not affect (P ≥ 0.21) concentrations of IgG, IgM, or IgA in colostrum at calving or in calf serum at 24 h. All calves from both experiments had undetectable concentrations of these respective immunoglobulins at birth. Concentrations of IgG at 24 h in calves that received maternal colostrum were 2,595.8 ± 535.38 mg/dL (Exp. 1), whereas those that received colostrum replacer were 1,610.6 ± 335.09 mg/dL (Exp. 2). Concentrations of IgG, IgM, and IgA were positively correlated (r ≥ 0.34; P ≤ 0.05) with concentrations of calf serum IgG, IgM, and IgA at 24 h after suckling. Our results suggest that maternal vitamin/mineral supplementation throughout gestation did not impact Ig concentrations in colostrum or the resultant serum Ig concentrations in calves either 24 h after suckling or after delivery of a commercial colostrum replacer.

106 Using Postbiotic Saccharomyces Cerevisiae Yeast in Gestation and Lactation Diets of Gilts to Improve Immunoglobulin Transfer and Offspring Growth Pre-Weaning

Abstract Gilts [n = 50; initial body weight (BW) 190.7 ± 4.2 kg] were used to determine the optional inclusion level of postbiotic Saccharomyces cerevisiae in late gestation and lactation diets to improve immunoglobulin transfer and piglet growth performance during the suckling period. Gilts were assigned to one of five (n = 10) diets: standard gestation and lactation diets (Y0) or diets with postbiotic yeast included at either 0.25% (Y0.25), 0.5% (Y0.5), 1.0% (Y1), or 1.2% (Y1.2). Diets were fed from d 85 of gestation until weaning (d 21 after farrowing). During gestation gilts were fed 2.6kg/d and feed was offered ad libitum during lactation. Gilt BW was recorded on d 85 and 110 of gestation, and gilt and piglet BW were recorded on d 1 and 21 of lactation. Twenty-four hours after farrowing, plasma was collected from each gilt and two median BW piglets per litter via orbital sinus. Additionally, 50 mL of colostrum was collected in equal parts from all functional teats on the right side. On d 21, plasma samples were collected from each gilt and two median BW piglets per litter. Concentrations of immunoglobulin (Ig) A, G, and M were quantified in plasma and colostrum. Data were analyzed in SAS with treatment as the main effect. Linear, quadratic, and yeast vs. no yeast contrast statements were used to assess responses to dietary inclusion of yeast. Time (minutes) between farrowing (defined as birth of the first piglet, confirmed by video recording) and sampling was used as a covariate for d 1 Ig concentrations. There were no differences in number of piglets born alive (12.5 ± 0.9), birth weight (1.40 ± 0.45kg), or gilt BW change during lactation (-7.98 ± 6.68kg). Lactation average daily feed intake tended to be 14.6% greater for gilts provided yeast versus no yeast (5.11 vs. 4.46 ± 0.31kg, respectively; P = 0.069). At weaning, offspring from Y0.25 fed gilts had greater BW than piglets from Y0- or Y1.2-fed gilts (6.48 vs. 5.66 and 5.82 kg ± 0.16, respectively; P &amp;lt; 0.001), with all other treatments intermediate. On d 1, colostrum and gilt plasma Ig concentrations were unaffected by diets. However, yeast increased piglet plasma concentrations of IgM (linear; P = 0.034) and tended to increase IgA (linear; P=0.093), and total Ig (linear; P = 0.074). On d 21, gilts fed yeast-containing diets tended to have greater plasma IgA (yeast vs. no yeast and quadratic; P = 0.063 and P = 0.076, respectively), but IgM was greater for gilts fed Y0 (yeast vs. no yeast; P &amp;lt; 0.05). On d 21, piglet plasma IgA tended to be greater when gilts were provided yeast-containing diets (yeast vs. no yeast; P = 0.069). Therefore, postbiotic Saccharomyces cerevisiae was effective in improving maternal transfer, with piglet IgA and IgM concentrations increasing linearly with increasing yeast inclusion, while Y0.25 also improved piglet growth performance during the suckling phase.

STATE OF HUMORAL IMMUNITY IN DIABETIC RETINOPATHY AGAINST THE BACKGROUND OF THE METABOLIC SYNDROME

Introduction. The most common cause of vision loss in case of diabetes is retinal pathology (diabetic retinopathy). The study of blood pressure levels in blood serum in patients with nonproliferative diabetic retinopathy in type 2 diabetes is a promising direction of research with the aim of studying the links of the pathogenesis of the development of this pathology, diagnosis, and possible prognosis of the disease&#x0D; The aim of the study: to find out the peculiarities of humoral immune dysfunction in patients with diabetic retinopathy against the background of metabolic syndrome&#x0D; Results and discussion. PSA increases in patients with decompensated diabetes (by 1.2 times compared to the control group). We found a statistically significant increase in the level of Ig A in patients of group 2 by 2.5 times compared to the control group and by 1.5 times compared to group 1. The content of Ig M exceeds the control indicator by 2.3 times and in group 1 by 1.7 times. The level of Ig G increased compared to the control indicator by 1.4 times, and did not differ from the indicator in group 1. The content of CIC in group 2 exceeded the indicators in the control group by 1.4 times and by 2 times the indicators in group 1, which indicates a higher probability of occurrence of III-type hypersensitivity reactions in the group of non-insulin-dependent patients with DR.&#x0D; Probable strong correlations of the concentration of immunoglobulins with other indicators in group 2 were revealed, which may indicate the preservation of relationships between immune indicators in compensated diabetes.&#x0D; The analysis of the ratio of the indicators implies the predominant activation of a specific link of humoral immunity and suggests the chronicity of the process, in the group of non-insulin-dependent patients there is a pronounced activation of humoral protection on the mucous membranes, as well as a predominant activation of early humoral immune mechanisms.&#x0D; Conclusion. Determination of the content of humoral factors of immunity in blood serum often does not reveal a probable difference. Therefore, it is advisable to determine the ratio of indicators. The obtained data indicate more pronounced changes in indicators of specific humoral immunity - immunoglobulins in patients with diabetic retinopathy, compared to the indicator of systemic inflammation - SRP.

Minerals and essential amino acids of bovine colostrum: Phenotypic variability and predictive ability of mid- and near-infrared spectroscopy

Colostrum quality and volume are fundamental for calves because it is the primary supplier of antibodies and the first source of energy, carbohydrates, lipids, proteins, minerals, and vitamins for the newborn. Assessing the detailed composition (i.e., AA and mineral content) of bovine colostrum (BC) on-line and at a reasonable cost would help dairy stakeholders such as farmers or veterinarians for precision feeding purposes and industries producing preparations containing BC such as foodstuff, supplements, and medicaments. In the present study we evaluated mid- (MIRS) and near-infrared spectroscopy (NIRS) prediction ability for AA and mineral composition of individual BC. Second, we the investigated the major factors affecting the phenotypic variability of such traits also evaluating the correlations with the Ig concentration. Results demonstrated that MIRS and NIRS were able to provide sufficiently accurate predictions for all the AA. The coefficient of determination in external validation (R2V) fell, in fact, within the range of 0.70 to 0.86, with the exception of Ile, His, and Met. Only some minerals reached a sufficient accuracy (i.e., Ca, P, S, and Mg; R2V ≥ 0.66) using MIRS, and also S (R2V = 0.87) using NIRS. Phenotypically, both parity and calving season affected the variability of these BC composition traits. Heifers' colostrum was the one with the greatest concentration of Ca and P, the 2 most abundant minerals. These minerals were however very low in cows calving in summer compared with the rest of the year. The pattern of AA across parities and calving season was not linear, likely because their variability was scarcely (or not) affected by these effects. Finally, samples characterized by high IgG concentration were those presenting on average greater concentration of AA. Findings suggest that infrared spectroscopy has the potential to be used to predict certain AA and minerals, outlining the possibility of implementing on-site analyses for the evaluation of the broad-sense BC quality.