Immunoglobulins IgM Research Articles

Analysis of IgM, IgA, and IgG isotype antibodies Directed against SARS-CoV-2 spike glycoprotein and ORF8 in the course of COVID-19

Immunoassays are a standard diagnostic tool that assesses immunity in severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) infection. However, immunoassays do not provide information about contaminating antigens or cross-reactions and might exhibit inaccurately high sensitivity and low specificity. We aimed to gain insight into the serological immune response of SARS-CoV-2 patients by immunoblot analysis. We analyzed serum immunoglobulins IgM, -A, and -G directed against SARS-CoV-2 proteins by immunoblot analysis from 12 infected patients. We determined IgG isotype antibodies by commercially available ELISA and assessed the clinical parameters of inflammation status and kidney and liver injury. Unexpectedly, we found no correlation between the presence of antibodies and the future course of the disease. However, attention should be paid to the parameters CRP, IL-6, and LDH. We found evidence of antibody cross-reactivity, which questions the reliability of results for serum samples that tested negative for anti-SARS-CoV-2 antibodies when assessed by immunoassays. Nevertheless, for the detection of IgG anti-SARS-CoV-2 antibodies, our data suggest that the use of the spike glycoprotein in immunoassays should be sufficient to identify positive patients. Using a combination of the spike glycoprotein and the open reading frame 8 protein could prove to be the best way of detecting anti-SARS-CoV-2 IgM antibodies.

Does the Course of Disease Influence the Development of Fatigue in Rheumatoid Arthritis Patients?

Abstract Patients with rheumatoid arthritis (RA) typically have many permanently inflamed joints. The inflammation inside the body can lead to general physical weakness, exhaustion, and drowsiness. This feeling of extreme tiredness is also called “fatigue”. Some people find this to be the worst symptom of the disease. However, the clinical significance of fatigue and its pathogenesis have not been recognised. This study aimed to determine the development of fatigue depending on activity and aggressiveness of RA. To achieve the goal, patients were interviewed and indicators of disease activity and aggressiveness were determined: rheumatoid factor (RF), anti-cyclic citrullinated peptide antibodies (anti-CCP), erythrocyte sedimentation rate (ESR), C reactive protein (CRP), immunoglobulins IgA RF, IgM RF, IgG RF and anti-carbamylated protein antibodies (anti-CarP). Based on the results of the survey, RA patients were divided into two groups — with and without fatigue. In the group of RA patients with fatigue, statistically more often an increase in IgA RF, IgM RF, and IgG RF levels was observed in those with elevated RF level, higher IgM RF and IgG RF levels were associated with increase in IgA RF level, and increase in the IgG RF and anti-CarP levels with elevation in the IgM RF level. A higher IgG RF level contributed to a higher anti-CarP level increase. Significant differences in the levels of clinical and laboratory inflammatory markers were not observed between the RA patients with and without fatigue. The obtained data suggest that the aggressive course of RA, more than inflammation, may contribute to the development of fatigue in RA patients.

Synergistic effect of Spirulina platensis and selenium nanoparticles on growth performance, serum metabolites, immune responses, and antioxidant capacity of heat-stressed broiler chickens

This study examined the effects of dietary Spirulina platensis (SP) at levels of 0, 5, and 10 g.kg-1 and selenium nanoparticles (SeNPs) at 0, 0.1, and 0.2 mg.kg-1, individually and in combination, on heat-stressed broiler chickens for 5 weeks. Four hundred fifty one-day-old Ross-308 chicks were allocated to 9 dietary groups with 5 replicates (10 chicks each). The control diet was consisted of corn-soybean-based basal diet. The obtained results displayed a significant increase in final body weight (p = 0.005) and weight gain during the periods from 22 to 35 days (p = 0.002) and 1 to 35 days (p = 0.005) in birds fed supplemented diets compared to those fed control diet, with the highest being in birds fed with both 10 g SP and 0.1 mg SeNPs. Feed conversion ratio was also improved in birds fed supplemented compared to control group. Dietary supplements significantly improved carcass dressing (p < 0.001), carcass yield (p = 0.001) percentages, and blood lipid profile. Blood triiodothyronine was higher (p = 0.005) with all treated diets except that contain 5 g SP compared to the control, with the highest being in birds fed diet contains 5 g SP + 0.2 mg SeNPs. Immunoglobulin subclasses IgG, IgM, and IgA were higher in birds fed supplemented diets compared to the control group. Antibody titers to Newcastle disease, avian influenza, and infectious bursal disease were numerically increased with dietary supplementation compared to the control group. Dietary treatments increased (p < 0.001) glutathione peroxidase and superoxide dismutase (SOD) levels, except diet contains 5 g SP for SOD level and decreased (p < 0.001) malondialdehyde level. It is concluded that dietary inclusion of SP and SeNPs, particularly their combination at levels 5 g SP plus 0.2 mg SeNPs kg-1 and 10 g SP plus 0.1 mg SeNPs kg-1, improved growth performance, carcass yield, immunity, and antioxidant capacity of heat-stressed broilers.

Seroprevalence and risk factors of Trichomonas vaginalis among couples in Al-Hamza city-Iraq.

Background: Trichomoniasis is a sexually transmitted disease. It is a public health risk factor. This disease associated with many sexual diseases and is likely to be a cause of infertility, abortion, and the birth of low-weight babies. The purpose of the study is to establish a database of parasite infection in the city, especially with no previous research on the rate of infection among men. Materials and method: Two hundred forty serum samples were collected from 120 couples between 18-43 years old, for the period from 2 Jan. 2020 to 25 jun. 2020, all of which were examined by ELISA test to detect immunoglobulin IgM and IgG. All results were analyzed by SPSS 20.

Traffic generated emissions alter the lung microbiota by promoting the expansion of Proteobacteria in C57Bl/6 mice placed on a high-fat diet

Air pollution has been documented to contribute to severe respiratory diseases like asthma and chronic obstructive pulmonary disorder (COPD). Although these diseases demonstrate a shift in the lung microbiota towards Proteobacteria, the effects of traffic generated emissions on lung microbiota profiles have not been well-characterized. Thus, we investigated the hypothesis that exposure to traffic-generated emissions can alter lung microbiota and immune defenses. Since a large population of the Western world consumes a diet rich in fats, we sought to investigate the synergistic effects of mixed vehicle emissions and high-fat diet consumption. We exposed 3-month-old male C57Bl/6 mice placed either on regular chow (LF) or a high-fat (HF: 45% kcal fat) diet to mixed emissions (ME: 30 µg PM/m3 gasoline engine emissions+70 µg PM/m3 diesel engine emissions) or filtered air (FA) for 6 h/d, 7 d/wk for 30 days. Levels of pulmonary immunoglobulins IgA, IgG, and IgM were analyzed by ELISA, and lung microbial profiling was done using qPCR and Illumina 16 S sequencing. We observed a significant decrease in lung IgA in the ME-exposed animals, compared to the FA-exposed animals, both fed a HF diet. Our results also revealed a significant decrease in lung IgG in the ME-exposed animals both on the LF diet and HF diet, in comparison to the FA-exposed animals. We also observed an expansion of Enterobacteriaceae belonging to the Proteobacteria phylum in the ME-exposed groups on the HF diet. Collectively, we show that the combined effects of ME and HF diet result in decreased immune surveillance and lung bacterial dysbiosis, which is of significance in lung diseases.

Immunoglobulins in teleosts.

Immunoglobulins are glycoproteins which are produced as membrane-bound receptors on B-cells or in a secreted form, known as antibodies. In teleosts, three immunoglobulin isotypes, IgM, IgT, and IgD, are present, each comprising two identical heavy and two identical light polypeptide chains. The basic mechanisms for generation of immunoglobulin diversity are similar in teleosts and higher vertebrates. The B-cell pre-immune repertoire is diversified by VDJ recombination, junctional flexibility, addition of nucleotides, and combinatorial association of light and heavy chains, while the post-immune repertoire undergoes somatic hypermutation during clonal expansion. Typically, the teleost immunoglobulin heavy chain gene complex has a modified translocon arrangement where the Dτ-Jτ-Cτ cluster of IgT is generally located between the variable heavy chain (VH) region and the Dμ/δ-Jμ/δ-Cμ-Cδ gene segments, or within the set of VH gene segments. However, multiple genome duplication and deletion events and loss of some individual genes through evolution has complicated the IgH gene organization. The IgH gene arrangement allows the expression of either IgT or IgM/IgD. Alternative splicing is responsible for the regulation of IgM/IgD expression and the secreted versus transmembrane forms of IgT, IgD, and IgM. The overall structure of IgM and IgT is usually conserved across species, whereas IgD has a large variety of structures. IgM is the main effector molecule in both systemic and mucosal immunity and shows a broad range of concentrations in different teleost species. Although IgM is usually present in higher concentrations under normal conditions, IgT is considered the main mucosal Ig.

ОЦІНКА ВПЛИВУ ДОНОРА МОНООКСИДУ КАРБОНУ (CORM-2) НА ШВИДКІСТЬ ЗГОРТАННЯ КРОВІ В УМОВАХ ІМУННОЇ ВІДПОВІДІ

The balance between the liquid state support of the blood and the thrombus formation process depends on the activity of plasma factors and thrombocytes. In a vascular injury condition, thrombocytes form a thrombocyte plug, vasoconstriction and, further, take part in the restoration of the tissues. Reducing the clotting function of thrombocytes is a current problem. The search for agents - modulators of thrombocyte activity - is a promising area of research. Compounds - gas-transmitters can be used to model the activity of thrombocytes hemostasis. Carbon monoxide (CO) gas belongs to this group as well. CO in very low concentrations affects in a different way the process of apoptosis, stimulates calcium-dependent potassium channels, and changes the activity of mitochondria. To investigate the effect of CO on blood clotting rate, laboratory mice were injected with the compound CO donor (CORM-2). This substance was administered separately during the inductive and productive phases of the immune response. The dynamics of the immune response were measured by IgA, IgM and IgG immunoglobulin levels. It was found that blood clotting was intensified under the influence of CORM-2 (20 mg/kg). Especially, the increase was during the productive phase. At the end of the experiment, the percentage of megakaryoblasts in the total population of thrombocyte precursors decreased in the bone marrow of animals (to which CORM-2 was administered). Against this background, an increased content of megakaryocytes was found in the group that received CORM-2 during the productive phase. In the group that received CORM-2 during the inductive phase, in addition to an increase in megakaryocyte levels, there was an increase in thrombocyte levels. The group that received CORM-2 during the productive phase also had a predominance of metamegakaryocytes and reducing of thrombocyte amount. Key words: thrombocytes, carbon monoxide donor, CORM-2, immune response phases.

Evaluation of immunoglobulins IgG, IgM, IgA and Complement Components C3 and C4 Levels in Iraqi full term Neonates with Severe Hyperbilirubinemia

Background and aim: Hyperbilirubinemia is an elevation of the bilirubin levels in blood of newborn babiesthat presented as jaundice. This study aimed to evaluate the levels of important of immunological systemproteins (IgG, IgM, IgA, C3 and C4) in both newborn babies with severe hyperbilirubinemia and healthycontrols.Methods: Thirty (30) full-term neonates with severe hyperbilirubinemia (serum bilirubin &gt;20 mg/dl) andtwenty five (25) healthy control full-term neonates were included in the study. Blood samples taken and Hb,total serum bilirubin, total protein measured immediately by spectrophotometer. IgG, IgM, IgA, C3 and C4measured by immune-nephelometric method.Results: In hyperbilirubinemia group (Male/ Female percentages) were (63% / 37%), (IgG 373.4±218 mg/dl),( IgM 20.5 ± 24.03 mg/dl), and (C3 209.4±17.81 mg/d)l values were significantly lower than controlgroup ( 588.2±298.5 mg/dl IgG, 26.44 ± 2.92 mg/dl IgM and 627.5±221.1 mg/dl for C3). Correlationanalysis revealed that in control group IgG, IgM, and C3 shown to be significantly (P value &lt;0.05) andpositively correlated (r-value &gt; 3) with both age and weight.Conclusions: jaundice is a risk factor for sepsis that could be due to lower levels of immunoglobulins andcomplements components. The normal development of immune system that observed with increasing ageand weight was impaired in patients with neonatal hyperbilirubinemia.

Seroprevalence and Risk factors of Toxoplasma gondiiamong children in Al-Qadisiyah Province –Iraq

Background: Toxoplasmosis is a disease associated with the nature of the population, domestic quality and general health culture. Toxoplasma gondii causes many healthy and psychological problems with the possibility of transmission from the mother to her fetus. The primary objective of this study is to establish a database of infection rates among the governorate's children, with the absence of a database of the parasite prevalence rate among children.&#x0D; Materials and method: Four hundred sixty-three serum samples from children 13 old and under were collected during the period from 6 Jan. 2019 to 12 Dec. 2019, all of which were examined by ELISA test to detect immunoglobulin IgM and IgG. All results were analyzed by SPSS 20.&#x0D; Results: The current study referred to high prevalence rates of T. gondii (23.3%), where the ratios of IgM and IgG were (4.1% and 17.9%) respectively. Compared with the present rates of immunoglobulin with gender, age, environment and animal contact.&#x0D; Conclusion: Our current study registered high prevalence rates among children in Al-Qadisiyah governorate, this result represents a database of researchers and workers of health. It's the first study concerning the prevalence of toxoplasmosis in children in Al-Qadisiyah Province.

Seroprevalence of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) infection in an Italian cohort in Marche Region, Italy.

Background and aim:The COVID-19 is an infectious disease caused by the novel coronavirus SARS-CoV-2, declared a public health emergency by the World Health Organization. In this study, we evaluated the seroconversion of SARS-CoV-2 antibodies to find predictors of infection in terms of symptoms, health status, and professions.Methods:Serological samples of 341 volunteers in a cohort in Marche Region, Italy, were analyzed for the presence of IgM and/or IgG immunoglobulins specific for the SARS-CoV-2. Contextually, an anamnestic questionnaire was administered. The binary logistic regression analysis was used to find the predictors of seroconversion.Results:Forty-nine subjects (14.4 %) were found positive, without significant differences between gender and age groups. The predictors identified inside the variable categories “symptoms,” “risk factors” (smoking habit and established pathologies), and “professions” were the loss of taste and smell (OR, 8.563), cardiovascular diseases (OR, 2.912), and police officers profession (OR, 3.875), respectively.Conclusions:Although the limited number of subjects recruited in this study, our results could give important findings to be considered for planning preventive strategies in the view of the next COVID-19 waves. (www.actabiomedica.it)

Exposure to Antibodies Anti-Chlamydophila Pneumoniae Associated to Respiratory Symptoms of Asthma among Adolescents.

Previous report shows relationship between C. pneumonie and Asthma. In this study, we assessed the association between exposure to C. pneumoniae infection and the risk of bronchial asthma in adolescents in the State of Morelos, Mexico. An analysis was performed on a population-based cohort study of 80 adolescents with respiratory symptoms of asthma defined by ISAAC and 202 healthy adolescents between 12 and 17 years old. The information was collected twice from questionnaires, anthropometry, and sampling. Excessive weight gain was determined by calculating body mass index, and exposure to the specific antibodies IgM, IgG, and IgA was detected by microimmunofluorescence. The geometric means were calculated for titers of C. pneumoniae. The odds ratio was used across multiple models. The results showed that the exposure to C. pneumoniae was very high in the study population (67.2%). All immunoglobulins were significantly increased in patients with asthma symptoms compared with the healthy population (17% for IgG, 34% for IgM, and 52% for IgA). In multiple models, the IgM and IgA immunoglobulins were found to be associated with asthma (OR, 2.4; 95% CI, 1.4–4.2 and OR, 2.4; 95% CI, 1.2–4.8, respectively). Our study reflects a high seroprevalence of C. pneumoniae in the population; this seroprevalence is higher in young people with asthma. Specific immunoglobulins to C. pneumoniae are associated with IgM and IgA. The epidemiological significance of our results influences the timely monitoring and management of infections acquired at an early age that persist, or recur, for much of the juvenile life. Additional studies are needed to validate our findings.

Evaluation of Concurrent Malaria and Dengue Infections among Febrile Patients- A Cross-sectional Study

Introduction: There are several tropical mosquito borne infections, such as Malaria and Dengue, these are the two major and common arthropod borne infections that cause high morbidity and mortality in many patients and are major public health concern worldwide. Concurrent malaria and dengue infection is an important condition that is infrequently reported. Aim: To find out the prevalence rate of co-infection for both dengue and malaria and also to associate the severity of such co-infections with symptoms and haematological parameters. Materials and Methods: In observational cross-sectional study, samples were collected from 604 febrile patients clinically suspected for malaria and dengue attending the Medicine Out Patient Department (OPD) of Dhiraj Hospital, Gujarat, India. The samples were diagnosed for malaria by using rapid malaria antigen test kit and peripheral blood smear microscopy for the identification of Plasmodium spp. The dengue NS-1 antigen, (Immunoglobulin) IgM and IgG antibodies rapid kit were use to rule out dengue infection. The statistical analysis done by software EPI info. Results: In the present study, out of 604, 58 patients (9.6%) were positive for malaria and 80 patients (13.24%) were positive for dengue, while 21 patients (3.47%) were having concomitant infection with both dengue and malaria. The most affected age group was 31-60 years, 45.53%. The study revealed male preponderance (61.59%), hepatomegaly and jaundice were seen in 52.38% patients, haemorrhagic manifestations in 23.80%, kidney failure in 4.76% (1), and thrombocytopenia (platelet count &lt;150,000/cubic millimeter (cmm)) were noted in 95.23% patients. Conclusion: It is mandatory to test any febrile patient for both malaria and dengue so that the diagnosis of the patient is not held back by missing any one. The clinicians who treat febrile patients in or from endemic areas must thoroughly examine and diagnose for both malaria and dengue, still one or the other is positive. Vector control, health education and good hygiene are community based preventive measures that are needed to control both the diseases.

Effects of hypoxic preconditioning combined with altitude training on CD55, CD59 and the immune function of swimmers

Hypoxic preconditioning refers to a certain intensity and time of hypoxic exposure before hypoxic stress, which makes the body produce endogenous protection to enhance the body's tolerance to subsequent more severe hypoxia. However, there are few studies on the effects of hypoxic preconditioning combined with altitude training on the immune system of athletes. Nine swimmers from Shanghai underwent 3-week hypoxic preconditioning [living high-training low (HiLo)] combined with 3-week altitude training. CD55 and CD59 expression in red blood cells (RBCs), CD55 and CD59 expression in white blood cells (WBCs), RBC count, WBC count, T lymphocyte CD3, CD4, CD8 expression, and immunoglobulins IgG, IgM, and IgA were measured 4 times: before the start of hypoxic preconditioning, in the first week of hypoxic preconditioning, at the end of hypoxic preconditioning (i.e., before the start of altitude training), and at the end of altitude training. CD55 and CD59 expression in RBCs significantly increased in the first week of hypoxic preconditioning (P<0.05), returned to baseline levels at the end of preconditioning, and significantly increased again during altitude training (P&lt0.05). CD55 and CD59 expression in WBCs decreased significantly during hypoxic preconditioning (P&lt0.05) and increased significantly during altitude training (P&lt0.05). CD3 expression first decreased and then increased in the hypoxic preconditioning phase, then decreased again in the altitude training phase. However, there was no significant difference in each phase. CD4/CD8 expression after altitude training was significantly lower than that before altitude training (P&lt0.05), but was not significantly different from that before the start of hypoxic preconditioning. IgG, IgM, and IgA did not fluctuate significantly throughout the experimental phase. After hypoxic preconditioning combined with altitude training, the expression of CD55 and CD59 on the surface of RBCs and WBCs increased significantly, and T lymphocyte CD4/CD8 expression also increased. These results suggest an improvement in the complement regulation system and RBC immune function. Hypoxic preconditioning can therefore improve immunity and enhance the physical function of athletes during altitude training.

Effect of inhaled anesthetic gases on immune status alterations in health care workers

The objective of this research was to evaluate consequences to the immune system of long-term exposure to waste anesthetic gases (WAG) by medical theater personnel. Two groups were recruited: (i) 60 healthy male controls; (ii) 120 medical professionals exposed to WAG, subdivided according to theater role, i.e. surgeons, surgical assistants (SA), anesthetists, anesthetic assistants (AA), nurses, and workers. Serum levels of fluoride, hexafluoroisopropanol (HFIP), total lymphocyte counts, as well as of CD3, CD4, and CD8 cells, CD4/CD8 ratios, and immunoglobulins IgA, IgG, IgM, and IgE were assayed. The results showed that fluoride and HFIP titers were significantly increased in anesthetists and AA compared with the other exposed groups. All exposed groups demonstrated significant elevation in lymphocyte count, CD4+ cell levels, CD4/CD8 ratios, as well as levels of IgE, IgM and IgG compared with the controls. With regard to the latter outcomes, a significant increase in IgE was seen in the surgeon, nurse, and worker groups compared with the other professions. Surgeons, anesthetists and AA exhibited higher IgM titers compared with their colleagues. Significantly higher IgG levels were identified in the SA, anesthetists, AA, and workers than in their nurses and surgeon coworkers. Of the six sub-groups, only the anesthetists and their assistants (AA) displayed a significant increase in CD4+ cells and CD4/CD8 ratios and a decrease of CD8+ cells compared with the controls. This spectrum of results suggests that variation exists in immunomodulatory responses to WAG exposure amongst hospital personnel.

Genetic and acquired blistering disorders of pediatric age group: An experience from Eastern India.

Blistering or vesiculobullous disorders in pediatric population are either immunobullous or mechanobullous. Spectrum was analyzed using demographic details, clinical features, histopathology, direct immunofluorescence (DIF) and Immunofluorescence mapping (IFM). This was a single institution based observational study in children below 18 years. The demographic details were collected using proforma containing particulars of the patient, history, complaints, and other parameters. Punch biopsy of the skin lesion was done. Biopsy samples were examined under light microscope followed by DIF using fluorescent conjugated polyclonal antibody against immunoglobulins IgG, IgM, IgA, and complement C3. The salt-split technique was also used in particular cases. IFM was done using anticytokeratin (CK) 5 & 14, antilaminin 332, anticollagen VII, and anticollagen IV antibodies. Out of total 50 cases, linear IgA bullous dermatosis (LABD) was the commonest. The average concordance between clinical and final diagnosis (histopathological examination + DIF) was 87.5% and discordance was 12.5%. The agreement between histopathological examination and DIF was found to be substantially significant (κ = 0.6892). IFM depicted epidermolysis bullosa simplex with reduced CK 14 expression, dystrophic epidermolysis bullosa with reduced Collagen VII expression and junctional epidermolysis bullosa with absent laminin 5 expression. The spectrum of bullous lesions in childhood was properly delineated and subcategorization of EB was done. Histopathological examination showed the hallmarks that were conclusive in most of the cases except in LABD and EB. DIF and IFM proved indispensable in those cases. Thus, DIF is not a substitute for histopathology but complementary to it.

EFFECTІVENESS OF LASER THERAPY ІN COMPLEX TREATMENT OF HERPETIC STOMATITIS

The aim: The purpose of research is to improve the effectiveness of treatment by applying laser in complex treatment in patients with herpetic stomatitis. Materials and methods: Were examined 47 people (22 men and 25 women) with acute herpetic stomatitis. The patіents' age was from 20 to 72 years. All patients underwent immunological tests such as (T-lymphocytes (CD3 +), T-helpers (CD4 +), cytotoxic T-lymphocytes (CD8 +), the content of immunoglobulins IgM, IgG, IgA and slgA in saliva to assess the іmmunological reactivity of the organism. Results: Clinical observations have shown that when using laser therapy, the results of treatment were better than in the control group. Positive dynamics was observed as early as 1-2 days after the start of laser use and was more pronounced when used in the early stages of the disease. The use of the laser promoted a more rapid suspension of new lesions, a decrease in pain syndrome, and a more rapid regression of herpetic lesions. We have found significant improvements in the arameters of cellular and humoral іmmunity in patients of the main group. Conclusions: Treatment of herpetic stomatitis is relevant and requires further improvement and the search for new methods. The use of a laser in the complex treatment of herpetic stomatitis has a positive effect on the course and іmmunological status of patients with herpetic stomatіtis. The manifestations of the disease healed faster, and the pain syndrome was less severe. Duration between іllness and treatment decreased on 19.7% and 26%. It is advіsable to include the method of treatment using a laser in the complex therapy of herpetic stomatitis.

Monoclonal Antibodies to Myeloid Differentiation Antigens: In Vivo Studies of Three Patients With Acute Myelogenous Leukemia

Three patients with acute myelogenous leukemia (AML) in relapse were treated with intravenous infusions of one or more purified murine monoclonal antibodies (MoAbs) specific for differentiation antigens on normal and malignant myeloid cells. Three of the MoAbs used were IgM immunoglobulins that react with glycolipids, while the fourth, an IgG2b, reacts with a protein antigen. Peripheral blood leukemia cell counts decreased significantly, but transiently, during treatment. Evidence of in vivo binding of each MoAb to leukemia cells was obtained, although two of the four MoAbs could not be detected in the plasma following infusion, perhaps due to circulating blocking factors. Antigenic modulation was not encountered in these studies. However, the induction of human antibody to murine MoAb was observed in one patient who was treated over a 70-day period. Toxicities encountered were minimal and included fever (3 patients), back pain (1 patient), and arthralgias and myalgias (1 patient). This is the first reported clinical trial of (1) IgM MoAbs, (2) MoAb therapy in patients with AML, (3) combinations of MoAbs directed toward different myeloid differentiation antigens, and (4) MoAbs directed to glycolipids. The relative lack of toxicity and the positive effects of MoAb treatment in the reduction of leukemia cell counts permit the continued study of more innovative approaches to the treatment of AML with MoAbs.

Isolation and identification in human blood serum of the proteins possessing the ability to bind with 48 kDa form of unconventional myosin 1c and their possible diagnostic and prognostic value.

We firstly identified 48 kDa molecular form of the unconventional myosin 1c (p48/Myo1C), and isolated it from blood serum of multiple sclerosis patients. The amount of p48/Myo1C in human blood serum correlated with some autoimmune, hemato-oncological and neurodegenerative diseases and thus may serve as a potential molecular biomarker. The biological functions of this protein in human blood remain unknown. Previously, we used the monodisperse magnetic poly (glycidyl methacrylate)(mag-PGMA-NH2 ) microspheres with immobilized 48/Myo1C and western-blot analysis, which allowed us to identify IgM and IgG immunoglobulins presenting an affinity to this protein. Here, we used mass spectrometry followed by the western blotting in order to identify other blood serum proteins with affinity to 48/Myo1C. The obtained data demonstrate that 48/Myo1C binds to component 3 of the complement and the antithrombin-III proteins. A combination of magnetic microparticle-based affinity chromatography with MALDI-TOF mass spectrometry and an in silico analysis provided an opportunity to identify the partners of interaction of 48/Myo1C with other proteins, in particular those participating in complement and coagulation cascades.

An accelerated method for determining «self/non-self» microorganisms in the agglutination reaction

A simple accelerated method for determining «self/non-self» microorganisms by using the agglutination reaction (RA) and therapeutic/prophylactic serum (Immunoglobulin complex preparation, lyophilized IgG, IgA, IgM immunoglobulins, developed by CSC Immuno-Gem, Moscow) is proposed to test for pathogenic, opportunistic and dominant probiotic Bifidobacteria spp. In parallel, all the microbial cell cultures examined were registered in the databases of Russia-wide and international collections and tested by the intermicrobial “self/non-self” recognition method, previously developed by us. 16 collection strains of various microorganisms were assessed by the RA with relevant therapeutic and prophylactic serum. Biological samples were obtained from the collection bacterial strains of Bifidobacterium bifidum 791, Escherichia coli LEGM-18, Klebsiella pneumoniae 278, Lactobacillus fermentum 90T-C4, Bifidobacterium longum MC-42, Escherichia coli M-17, Shigella sonnei 177b, Shigella flexneri 170, Escherichia coli 157, Staphylococcus aureus 209, Candida albicans 10231 and Salmonella serovar Enteritidis ATCC 10708. In addition, cell cultures obtained from the Museum of the Institute of Cellular and Intracellular Symbiosis UB RAS such as Bifidobacterium longum ICIS-505, Lactobacillus acidophilus ICIS-1127, Bifidobacterium bifidum ICIS-202, Bifidobacterium bifidum ICIS-310 were also included into the study. To assess microbial peptidoglycan foreignness, the intermicrobial “self/non-self” recognition method was also used based on inducing metabolites produced by the “dominant” test strain Bifidobacterium longum MC-42 after pre-incubation with metabolites collected from the studied cell cultures (“associates”) followed by established “dominant-associate” feedback loop. The data were evaluated by assessing change-fold in reproduction (growth/replication) and adaptation (biofilm formation and anti-lysozyme test) of microbial cultures in accordance with the described technique followed by comparing these two methods for intermicrobial “self/non-self” recognition. All the RA data were found to fully agree with those obtained after previous studies by using intermicrobial “self/non-self” recognition method coupled to “dominant-associate” system. Moreover, compared to analogous “intermicrobial recognition” method (5 days), ease of use and test timeframe (24 hours) allow to consider RA attractive for screening studies to select strains for scientific and industrial purposes.

Restricted Immunoglobulin Joining Chain (IgJ) Protein Expression in B Lymphoblastic Leukemia (B-ALL) Based on B-ALL Subtype

Background Philadelphia-like (Ph-like) B Lymphoblastic Leukemia (B-ALL) is a high-risk subtype of B-ALL that lacks the BCR-ABL1 fusion but has a gene expression profile similar to Philadelphia positive (Ph+) B-ALL. Gene expression profiling has previously identified Immunoglobulin Joining chain (IgJ) overexpression at the mRNA level in Ph-like B-ALL. This is surprising since IgJ is normally expressed by mature or maturing B cells. The normal function of IgJ protein is to concatenate monomers of immunoglobulin IgM or IgA into the mature pentameric and dimeric forms of these molecules respectively. IgJ also plays a crucial role in transport of IgA protein across mucosal epithelium to facilitate mucosal humoral immunity. In hematopathology, J chain immunohistochemistry (IHC) has been used to identify neoplastic cells in nodular lymphocyte predominate Hodgkin lymphoma (NLPHL) and can be used in distinguishing this disease from morphologic mimickers. It does not have known diagnostic utility outside of this context. Lymphoblasts do not typically express immunoglobulins at the protein level. Therefore, we sought to determine the protein expression of IgJ in B-ALL and to determine whether IgJ immunohistochemistry may be employed in identifying particular subtypes of B-ALL. Methods We selected a total of 46 B-ALL cases diagnosed from a bone marrow sample at our institution from 2016-2019 with adequate diagnostic material for IHC. This included 5 cases of Ph-like B-ALL, all with a CRLF2 rearrangement and overexpression, 7 de novo Ph+ B-ALL and 34 cases representing the other most commonly recognized WHO subtypes of B-ALL, determined based on cytogenetic studies performed at the time of diagnosis. No cases of B-ALL with t(5;14) and B-ALL with iAMP21 was represented. Our cohort included 23 pediatric cases and 24 adult cases and the patients ranged from 1 to 82 years old at the time of initial diagnosis. A total of 8 normal bone marrow cases (negative staging bone marrow biopsies for diffuse large B cell lymphoma, neuroblastoma or classic Hodgkin lymphoma) were used as controls. IgJ IHC was performed on B-plus fixed paraffin embedded bone marrow biopsy specimens using a commercially available and validated anti-IgJ monoclonal antibody (clone OTI3B3). Staining of bone marrow samples was performed at 2 different dilutions; tonsil secondary follicles and neoplastic cells from NLPHL were used as technical controls. Cellular staining in the lymphoblasts was scored in a blinded manner by a board certified hematopathologist and a pathologist in training as diffusely positive, partially positive, or negative. Results Cellular staining was distinguishable from background staining due to circulating immunoglobulins and there was almost perfect inter-observer concordance in identifying positive and negative cases (agreement of 98%, kappa test 0.94). All normal bone marrow controls cases were negative for IgJ cellular staining. A total of 11/46 (23%) B-ALL cases demonstrated partial or diffuse cellular staining for IgJ in the lymphoblasts. This included 4/5 (80%) Ph-like cases, 5/7 (71%) Ph+ cases, 1/3 MLL rearranged cases and 1/6 ETV6-RUNX1. All TCF3-PBX1 (0/4), hyperdiploid B-ALL (0/10), hypodiploid B-ALL (0/2), and B-ALL, NOS cases (0/9) were negative for IgJ. Diffuse IgJ staining was restricted to Ph-like (2/4) or Ph+ (2/5) B-ALL subtypes; the positive MLL rearranged and ETV6-RUNX1 B-ALL cases only showed weak partial staining. IgJ protein was significantly expressed in Ph+/Ph-like B-ALL (p&amp;lt;0.0001) and in our cohort, detected these cases with a 75% sensitivity, 95% specificity, 82% positive predictive value and 92% negative predictive value. Conclusion We conclude that IgJ protein expression occurs in a subset of B-ALL, predominantly restricted to Ph+ and Ph-like cases. Although, these findings will need to be validated in larger studies, our results suggest that IgJ IHC, in concert with routine standard cytogenetics studies may be a rapid and cost effective method in identifying Ph-like B-ALL. Disclosures No relevant conflicts of interest to declare.