Circulating Immune Complexes Levels Research Articles

Clinical validation of circulating immune complexes for use as a diagnostic marker of canine leishmaniosis.

Canine leishmaniosis (CanL) is a systemic disease that affects dogs. When multiplication of the parasite cannot be controlled, dogs consistently show high levels of antigen and IgG antibodies, which lead to the formation of circulating immune complexes (CIC). Timely intervention to reduce the parasite load and CIC levels is crucial for preventing irreversible organ damage. However, a diagnostic test to quantify CIC levels is currently lacking. In this real-world study, we aimed to examine the performance of a new ELISA to measure CIC levels in dogs naturally infected with Leishmania infantum. Thirty-four dogs were treated according to their clinical condition and followed for 360 days. Before (day 0) and after treatment (days 30, 90, 180, 270, and 360), all dogs underwent a physical examination, and blood samples were obtained for CBC, biochemical profile, serum protein electrophoresis and IFAT. Serum PEG-precipitated CIC were determined by ELISA. Our results indicate higher CIC levels in dogs in advanced disease stages showing higher antibody titres (p < 0.0001, r = 0.735), anemia (p < 0.0001), dysproteinemia (p < 0.0001), and proteinuria (p = 0.004). Importantly, dogs responding well to treatment exhibited declining CIC levels (p < 0.0001), while in poor responders and those experiencing relapses, CIC were consistently elevated. CIC emerged as a robust discriminator of relapse, with an area under the curve (AUC) of 0.808. The optimal cut-off to accurately identify relapse was an optical density of 1.539. Our findings suggest that declining CIC levels should be expected in dogs showing a favorable treatment response. Conversely, in dogs displaying a poor response and recurrent clinical relapses, CIC levels will be high, emphasizing the need for vigilant monitoring. These findings suggest that CIC could serve as a valuable biomarker for disease progression, treatment efficacy, and relapse detection in CanL. Our study contributes to enhancing diagnostic approaches for CanL and underscores the potential of CIC as a complementary tool in veterinary practice. As we move forward, larger studies will be essential to confirm these findings and establish definitive cut-offs for clinical application.

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.

POSSIBILITIES OF CORRECTING CELLULAR-HUMORAL IMMUNE DISORDERS AND CYTOKINE STATUS IN HIGH-RISK PREGNANT WOMEN FOR THE DEVELOPMENT OF PREECLAMPSIA

POSSIBILITIES OF CORRECTING CELLULAR-HUMORAL IMMUNE DISORDERS AND CYTOKINE STATUS IN HIGH-RISK PREGNANT WOMEN FOR THE DEVELOPMENT OF PREECLAMPSIA

Mechanisms of Long-Term Persistence of Mycoplasmas in Children with Asthma.

Improving the management of children with asthma associated with mycoplasma infection is important. Aim: To study the duration of the persistence of antigens, and DNA in a free state, in the structures of circulating immune complexes (CICs) and living cells of Mycoplasma pneumoniae (Mpn) and Mycoplasma hominis (Mh) in children with asthma. In total, 205 children with asthma from 1 to 14 years were observed. The reaction of aggregate-hemagglutination (AHAA), the direct immunofluorescence reaction (DIF), the reaction of the polymerase chain reaction (PCR), and the culture method were used. In addition, 47 children were re-examined 1.5 months after the treatment of mycoplasma infection with azithromycin. The number of samples positive for antigens and DNA in the free state and in the structures of CICs significantly decreased. Then, 50 blood serum samples containing Mh antigens, and 50 samples containing Mpn antigens were analyzed by culture method. Mh was isolated in 21 (65.5%) of 32 samples containing DNA. Mpn was isolated from antigen-positive samples in nine cases. The presented data indicate the long-term persistence of antigens, and DNA of mycoplasma cells in the free state, in the structure of CICs, as well as in the form of "microcolonies". A high level of CICs can be used to predict the course of the disease and the response to therapy.

Impact of nitric oxide synthesis modulators on the state of humoral immune system in experimental antiphospholipid syndrome

Background: Antiphospholipid syndrome is an autoimmune disease of multiple venous and/or arterial thrombosis and/or pregnancy loss. Oxidative stress only enhances the body’s immune response. In pathological conditions, the formation of nitric oxide is disrupted, which can be manifested by vasoconstriction, increased coagulation, and endothelial dysfunction. Objective: The aim of the research was to study the level of immunoglobulins and circulating immune complexes (CICs) in experimental antiphospholipid syndrome and its correction with L-arginine and aminoguanidine. Materials and methods: Antiphospholipid syndrome was modeled on white female BALB/c mice. L-arginine (25 mg/kg) and aminoguanidine (10 mg/kg) were used for its correction. The content of immunoglobulins and CICs was studied. Results: It was established that the level of immunoglobulins (Ig) and circulating immune complexes increased in the group of animals with antiphospholipid syndrome compare to the control. The levels of IgA and CICs decreased significantly, and the levels of IgM and IgG did not change in the mice with antiphospholipid syndrome and L-arginine correction. In cases of aminoguanidine administration, decreased IgM and IgG levels and no significant decrease in IgA and CICs was evidenced compare to the animals with antiphospholipid syndrome. In cases of using a combination of L-arginine and aminoguanidine agents, only IgM did not change, all other parameters decreased compare to the animals with APS. Conclusion: The parameters of the humoral immunity in female mice with experimental antiphospholipid syndrome increase. The level of immunoglobulins and circulating immune complexes decrease depending on the chosen correction agents or their complex administration. Thus, L-arginine and aminoguanidine has a positive effect on various immunity responses by decreasing the negative impact of pathobiochemical alterations.

Иммунный статус в развитии рестеноза у пациентов с окклюзионно-стенотическим поражением бедренно-подколенного артериального сегмента после бедренно-подколенного шунтирования

Objective: to assess the immune status of systemic and local blood flow after femoral-popliteal bypass surgery in patients with and without restenosis. Materials and methods. The analysis of the treatment of 82 patients who underwent femoral-popliteal bypass surgery was carried out. Depending on the condition of the reconstruction zone after 12 months, the patients were divided into two groups: group I - 21 patients without restenosis, group II - 61 patients with the development of restenosis. The analysis of IgA, IgM, IgG, circulating immune complexes (CIC), interleukin (IL)-6, IL-1 in the systemic and local blood flow before and after surgery was performed. Results. In patients with the development of restenosis of the reconstruction zone, the level of IgA in the systemic and IgA, IgM in the local bloodstream was initially lower, the level of CIC, IL-1, IL-6 in the systemic and IgG, IL-1, IL-6 in the local bloodstream was higher, after surgery, the level of IgA in the local bloodstream was lower, higher - IgM in systemic and IL-6 in local blood flow in comparison with patients without restenosis. The correlation between the immune status and the development of restenosis was established: before surgery in the systemic circulation - IgA (r=-0.31, p&lt;0.01), CIC (r=0.39, p&lt;0.001), IL-6 (r=0.25, p&lt;0.05), IL-1 (r=0.26, p&lt;0.05), in the local bloodstream - IgA (r=-0.26, p&lt;0.05), IgM (r=-0.30, p&lt;0.01), IgG (r=0.26, p&lt;0.05), IL-6 (r=0.30, p&lt;0.01), IL-1 (r=0.29, p&lt;0.01); after surgery in the systemic circulation - IgM (r=0.33, p&lt;0.01), in the local - IgA (r=-0.24, p&lt;0.05), IL-6 (r=0.40, p&lt;0.001). The relationship between the degree of chronic arterial insufficiency (r=0.22, p&lt;0.05), the ankle- brachial index after surgery (r=-0.27, p&lt;0.05) with the development of restenosis was revealed. Conclusion. The interrelation of the immune status of the systemic and local blood flow before and after surgery and restenosis in the anastomosis zone after femur-popliteal bypass surgery has been shown. The study of the immune status will make it possible to develop an algorithm for restenosis prevention.

Regional peculiarities of specific immunity in inhabitants of different mountain heights of the Kyrgyz Republic

Kyrgyzstan’s challenging natural conditions and fragile mountain ecosystems can weaken the immune system of individuals residing permanently in high-altitude areas. Our study aimed to explore specific immune protection indicators in healthy populations from the Chui and Naryn regions at varying mountain altitudes. We assessed percentages of CD3+ (T-lymphocytes), CD20+ (B-lymphocytes), CD4+ (T-helper lymphocytes), CD8+ (cytotoxic T-lymphocytes), IRI (immunoregulatory index), CIC (circulating immune complexes), and quantified immunoglobulins of classes A, M, and G. Using immunofluorescent methods and monoclonal antibodies; we determined the percentages of CD3+, CD20+, CD4+, and CD8+. Radial immunodiffusion was used to quantify immunoglobulins of classes A, M, and G, while Falk’s method helped study CIC levels in blood sera. The participant count was based on statistical expert recommendations and G power analysis. The analysis revealed a decrease in the total number of T-lymphocytes (CD3+), primarily due to reduced T-helper and cytotoxic T-lymphocyte subpopulations. However, a significant increase in cytotoxic T-lymphocytes (CD8+) was observed among high-altitude residents. Mountain residents exhibited wide-ranging individual fluctuations for each immunoglobulin class. Our findings suggest varying degrees of immune mechanism disruptions, which maintain natural protection and internal environment constancy at different altitudes, resulting in a mixed type of cellular and humoral immunity suppression. These results emphasize the need for further research in the region and consideration of altitude when implementing disease prevention strategies related to the immune system for the local population.

Assessment of circulating immune complexes in canine leishmaniosis and dirofilariosis.

Canine leishmaniosis (CanL) by Leishmania infantum (L.i.) and heartworm disease by Dirofilaria immitis (D.i.) are common zoonotic vector-borne diseases (VBDs) characterized by a variety of pathological and clinical signs. The immunopathology in both VBDs is extremely complex, and their clinical manifestations are strongly dependent on the type of immune response elicited by the parasites. In particular, the formation of circulating immune complexes (CICs) plays an important role in the pathogenesis of these VBDs. Based on the international guidelines, dogs with high anti-L. infantum antibody titres and one or more clinical and/or laboratory signs related to CanL require anti-Leishmania treatment. Consequently, the CICs measurement could be used for improving the clinical staging process of CanL. The aim of the study was to assess the CICs level by a competitive inhibition enzyme immunoassay, in healthy or sick dogs seropositive to L.i. and in healthy dogs positive to D.i.. Out of 51 enrolled dogs, 11 were included in Group A (seronegative to L.i., D.i. negative and healthy), 15 in Group B (exposed to L.i., D.i. negative and healthy), 12 in Group C (seropositive to L.i., D.i. negative and sick) and 13 in Group D (seronegative to L.i, D.i. positive and healthy). The comparison of CIC level in canine sera revealed a significant difference among groups (P < 0.001), with the highest concentration (i.e., median = 104.6µg/mL) in dogs with CanL. The findings of the study highlight the CICs measurement as a useful tool in the clinical staging of CanL for avoiding misclassification of dogs as leishmaniotic, thus not requiring anti-Leishmania therapy, as well as the possibility of results misuse in geographical areas where both leishmaniosis and heart-worm disease are endemic.

The effect of the combination of thiotriazoline and corvitin on the values of some parameters of the cellular and humoral immune system during the experimental reproduction of allergic alveolitis under immobilization stress

The purpose of the study was to determine the effectiveness of the combination of corvitin and thiotriazoline for the correction of levels of T- and B-lymphocytes, circulating immune complexes (CIC) of the blood of male guinea pigs (GP) if experimental allergic alveolitis (EAA) and immobilization stress (IS) are combined. The study was performed on 50 male GP weighing 180-210 g. EAA was reproduced by the method of O.O. Orekhov, Yu.A. Kyrylov. The production of IS was carried out by the method of P.D. Horizontov. The content of T- and B-lymphocytes in the blood was determined by the method of E.F. Chernushenko, L.S. Kogosova. CIC were detected by the method of Haskova V., Kaslik J., Math J., Matejckava M. The withdrawal from the experiment was carried out by the method of decapitation after previous administration of nalbuphine hydrochloride in the experimental group (EG) of intact GP; in the EG of EAA and IS during the study days corresponding to the stages of development of IS: 1st – anxiety, 14th – resistance, 24th – exhaustion; EG of EAA with IS on the 24th day after pharmacocorrection with the above-mentioned combination of drugs. There was a progressive decrease in the level of T-lymphocytes by 33,47% (p&lt;0,01), 36,23% (p&lt;0,01) and 39,62% (p&lt;0,01), an increase in B-lymphocytes by 27,82% (p&lt;0,01), 131,13% (p&lt;0,01) and 44,37% (p&lt;0,01), an increase in CIC by 41,15 % (p&lt;0,01), 46,62% (p&lt;0,01), and 50,78% (p&lt;0,01) on the 1st, 14th and 24th days respectively, compared to the indicators of the control group under the combination of EAA and IS. An increase in T-lymphocytes by 40% (p&lt;0,01), a decrease in B-lymphocytes by 24,31% (p&lt;0,01), and a decrease in the level of CIC by 26,08% (p&lt;0,01) were observed in the EG using corvitin and tiotriazoline relative to the corresponding indicators of the EG with a combination of EAA and IS without pharmacological interventions on 24th day of the experiment. The results indicate an immunocorrective effect of the combination of corvitin and thiotriazoline which is manifested by activation of the cellular and suppression of the humoral link of immunity of GP exposed to EAA and IS.

Circulating immune-complexes and complement activation through the classical pathway in myeloperoxidase-ANCA-associated glomerulonephritis

Background Anti-neutrophil cytoplasmic antibody (ANCA)-associated glomerulonephritis (AAGN) is the fulminant glomerular diseases with poor renal prognosis. Activation of the complement system has recently been reported in the pathogenesis of AAGN, but it remains to be clarified as to which complement pathway is mainly involved. Methods 20 patients with myeloperoxidase (MPO)-AAGN were retrospectively evaluated. Using serum samples, circulating immune-complexes (CICs) were assessed by the monoclonal rheumatoid factor assay, and C5a and C5b-9 were assessed by ELISA. Complement activation through the classical pathway was further evaluated by the WIESLAB® Complement System Classical Pathway kit. The affinities of ANCAs were evaluated by a competitive inhibition method using ELISA, and were classified into the high, and low-affinity group. Deposition of complement components, such as C3, C5, C4d, C5b-9, factor Bb, mannan-binding lectin serine peptidase (MASP)-1, MASP-2, and mannose/mannan-binding lectin (MBL), in frozen renal sections were analyzed by immunofluorescence staining. Results CICs were found to be positive in 65% of the patients. All CIC-positive patients belonged to the high-affinity group. Furthermore, serum C5a and C5b-9 were significantly increased in MPO-AAGN patients, and these levels positively correlated with CIC levels. A significant negative correlation was also found between levels of WIESLAB® classical pathway kit and CICs. By immunofluorescence staining, glomerular deposition of C4d, C5, and C5b-9 were observed in similar distributions in MPO-AAGN patients, whereas the deposition of MASP-1, MASP-2, MBL, and factor Bb were minimal. Conclusions These results suggest the involvement of immune-complex induced complement activation through the classical pathway in the pathogenesis of MPO-AAGN.

Microbial landscape and immune status in maxillary sinusitis of stomatogenic origin

Oroanthral fistula (anastomosis) is an element preventing the restoration of homeostasis in the maxillary sinus due to the constant flow of microbes from the oral cavity. It is also contributes to frequent exacerbations of maxillary sinusitis. Saprophytic gram-positive cocci and fungi of the oral cavity are dominating representatives of the microbial flora in the maxillary sinus. As the result of research, we found that in the case of maxillary sinusitis with oroantral fistula fungi made up 25.0% of microbiota, gram-positive bacteria – 41.7%, gram-negative bacteria – 33.3%. Gram-positive cocci from the Staphy­lococcus genus (Staphylococcus aureus and Staphylococcus epidermidis) and fungi (Candida albicans) comprised the biggest proportion of microbial flora that 33.3% and 16.7%, respectively. Slightly decreased levels of monocytes in venous blood was noted in 69.2% of patients. The average value of total serum IgE in group with oroantal fistula was 226.2 (70.4) IU/ml, the result exceeded normal limits almost in 2.26 times. Large circulating immune complexes (CICs) were normal in all patients in the group with iatrogenic maxillary sinusitis. The average lavels of small size CICs was 170.2 (4.23) ОU, which is in 1.06 times higher the upper limit of the norm (160 ОU). Elevated levels of total Ig E in serum of patients with oroanthral fistula indicates allergic sensitization. The detection of increased levels of CICs with small and medium sizes in serum may indicate a susceptibility of this category of patients to the development of immunopathological reactions.

Markers of progression of liver fibrotic changes in patients with chronic toxic drug-induced hepatitis

Background. In recent years, as a result of the growing expansion of the pharmaceutical market, there has been a clear trend towards an increase in the incidence of chronic toxic drug-induced hepatitis of drug genesis (TDIH). The appearance of fibrosis is considered the most important histological change that determines the further course of the disease. Therefore, the search for non-invasive or minimally invasive markers for assessing fibrotic changes in the liver remains an urgent issue in clinical practice. The purpose was to determine the diagnostic value of immunological parameters for stratification of the severity of liver fibrosis in patients with TDIH. Materials and methods. The study included 41 patients with TDIH, who were divided into three groups: group I consisted of 12 people without liver fibrosis (F0), group II— 22 patients with moderate fibrosis (F1-F2), group III— 7 individuals with severe liver fibrosis (F3-F4). Shear wave elastography was performed using a Soneus P7 system (Ukraine-Switzerland). All patients underwent a biochemical blood test with the determination of alanine aminotransferase (ALT), aspartate aminotransferase (AST). The subpopulation composition of lymphocytes, circulating immune complexes (CIC), the level of interleukins (IL-6, IL-10) and tumor necrosis factor α were assessed. Results.The progression of liver fibrosis is accompanied by an increase in cytolytic syndrome: patients with severe fibrosis have a 3.3-fold increase in the ALT (p&lt;0.05) compared to the controls and a 2.1-fold (p&lt;0.05) compared to that in patients with moderate fibrosis. The AST level is significantly higher— by 4.6 times (p=0.023) in patients with severe fibrosis than in those with moderate fibrosis. With the progression of liver fibrosis, there is a significant decrease in cellular immunity, an increase in the level of CIC and pro-inflammatory cytokines with a simultaneous decrease in the content of anti-inflammatory cytokines, which is confirmed by correlations between the liver stiffness index according to shear wave elastography data and the level of T-helpers (r=–0.466; p=0.03), IL-6 (r=0.364; p=0.01), IL-10 (r=–0.331; p=0.039) and CIC (r=0.381; p=0.017). Conclusions. Markers of the diagnosis of severe liver fibrosis in patients with TDIH are indicators such as the ratio of IL-6/IL-10 higher than 0.83 (sensitivity 81.8%, specificity 78.9%), CIC level more than 4.3 optical density units (sensitivity 77.3%, specificity 72.2%), the ratio of T-helpers/T-suppressors is less than or equal to 1.6 (sensitivity 72.7%, specificity 57.9%).

The Dynamics of Circulating Immune Complexes in Horses with Severe Equine Asthma.

Simple SummaryEquine asthma syndrome is a cost-consuming equine respiratory disease of the lower airways in horses. Non-invasive biomarkers from blood or urine are sought. The aim of this study was to assess the circulating immune complexes (CICs) during the exacerbation and remission of an asthma episode—with and without additional treatment and the potential usefulness of CIC levels in the diagnosis, monitoring, and treatment progression. The control group, asthma group, and treated asthma group each contained six horses. The horses were kept in a dusty environment for seven days and then moved to an asthma-friendly environment for over three weeks (the treated group received injections of glucocorticoids). Blood was collected at baseline and on the 1st, 2nd, 3rd, 7th, 14th and 30th days. CICs measured in the time points did not show statistical differences. When CICs were analysed within the groups, there was a significant decrease in CIC in the treated group and a significant increase in CIC in the non-treated group on day 30. CIC did not support the diagnosis procedure of equine asthma syndrome, although it may help in monitoring patients with and without treatment. To the best of the authors’ knowledge, this is the first study to analyse the dynamics of CIC in equine asthma patients during an environmental challenge, remission, and treatment.Non-invasive diagnostic biomarkers of equine asthma syndrome (EAS) from blood or urine are sought. The aim of this study was to assess the absorbance of circulating immune complexes (CICs) during the exacerbation, remission, and treatment of an asthma episode and assess the potential usefulness of CIC levels in the diagnosis and monitoring of the disease. The control group, asthma group, and treated asthma group each contained six horses. Following an initial examination and group classification, the horses were kept in a dusty environment for seven days and then moved to an asthma-friendly environment for three weeks (the treated group received injections of glucocorticoids). Blood was collected at baseline and on the 1st, 2nd, 3rd, 7th, 14th and 30th days. CIC was measured using the modified Haskova method. The time points did not show significant statistical differences. There was a significant decrease in CIC in the treated group, and a significant increase in CIC in the non-treated group on day 30. CIC did not support the EAS diagnosis, although it may help in monitoring patients. To the best of the authors’ knowledge, this is the first study to analyse the dynamics of CIC during environmental challenge, remission, and treatment.

Evaluation of Serum Immunoglobulins (IgG, IgA, IgM) and Circulating Immune Complexes in Oral Precancer and Cancer Patients.

Objectives The aim of the present study is to evaluate the role of immunoglobulins (IgA, IgG, and IgM) and circulating immune complexes (CIC) as tumor marker in oral cancer and precancer patients. Materials and Methods The present study was performed on 45 individuals subdivided into three groups, that is, oral precancer, oral cancer and healthy individuals, and levels of immunoglobulins, and CIC was estimated by turbidometry and ELISA method. Results In the present study, the mean serum IgA levels in oral precancer were 161.00 ( ± 118.02) mg/dL, oral cancers were 270.67 ( ± 171.44) mg/dL, and controls were 133.73 ( ± 101.31) mg/dL. Mean serum levels of IgG in oral precancer were 1,430.87 ( ± 316) mg/dL, oral cancers were 1,234.27 ( ± 365.42) mg/dL, and controls were 593.87 ( ± 323.06) mg/dL. Conclusion We found that the levels of serum IgG and IgA were elevated consistently in precancer and cancer group, and Serum IgM levels were increased only in precancer. Also, significant increase in serum CIC levels were seen in oral precancer and cancer group on comparison with control.

Role of Circulating Immune Complexes in the Pathogenesis of Canine Leishmaniasis: New Players in Vaccine Development.

During canine visceral leishmaniasis (CanL), due to Leishmania infantum (L. infantum), uncontrolled infection leads to a strong humoral immune response. As a consequence of the production of high antibody levels and the prolonged presence of parasite antigens, circulating immune complexes (CIC) are formed, which can be deposited in certain organs and tissues, inducing vasculitis, uveitis, dermatitis and especially glomerulonephritis and renal failure. A method to detect CIC and quantify their levels in serum samples from dogs infected with L. infantum has been recently described. It allowed demonstration of a correlation between CIC levels and disease severity. Thus, CIC measurement may be useful for diagnosis, assessment of disease progression and monitoring response to treatment. This is an interesting finding, considering that there remains an urgent need for identification of novel biomarkers to achieve a correct diagnosis and for optimal disease staging of dogs suffering from Leishmania infection. The objective of the present review is to shed light on the role of CIC in CanL, as well as to highlight their potential use not only as diagnostic and prognostic biomarkers but also as a valuable tool in vaccine development and new immunotherapy strategies to prevent or control disease outcome.

Assessing the values of circulating immune complexes in multiple sclerosis patients following immunomodulator or corticosteroid treatment

Multiple sclerosis is defined as an immune-mediated disease that affects the central nervous system, and also is characterized by the presence of immune cells and mediators which contribute to the subsidiary neuroinflammation associated with multiple sclerosis. Throughout the evolution of multiple sclerosis, it has been observed that circulating immune complexes (CICs) have higher values in these patients, especially in the acute phase of the disease. Thus, the aim of the present study was to observe, if in acute attack, relapsing-remitting multiple sclerosis patients still present high values of CICs after treatment with glatiramer and prednisone. We divided 70 patients with multiple sclerosis with high values of CICs into two treatment groups, one treated with glatiramer (Copaxone) (immunomodulatory treatment) and the other with prednisone (corticosteroid treatment). After three months of treatment, we assessed the levels of CICs of the two multiple sclerosis groups and we observed that the patients that followed the immunomodulatory treatment had lower values of CICs than the group that followed the corticosteroid treatment. In addition, another observation established was that the glatiramer treatment group had higher levels of vitamin D in the serum than the prednisone group of multiple sclerosis patients. To conclude, better outcomes, from the point of view of the results obtained from the comparative analysis of the values of CICs and vitamin D, were demonstrated by following immunomodulatory treatment.

The vaginal bacterial dysbiosis severity predicting model according to the normobiota index

The local and systemic immunodeficiency is the main mechanism for vaginal bacterial dysbiosis and its extreme manifestation – bacterial vaginosis (BV) development. The complex immune response study and the establishment of the main mechanisms and factors, reflecting it and corresponding to the microbiocenosis severity disorder are relevant. Aim – to develop a neural network model of the severity of vaginal bacterial dysbiosis based on the assessment of normobiota. Divided into the following groups according to the Conditionally pathogenic microflora index (CPMI) and normobiota index (NBI): normocenosis (n=53), dysbiosis I (n=128) and II degree (n=117) among the latter 83 patients with PNB&gt;1 lg GE/sample were identified, in whom BV was established. Molecular genetic studies of the epithelium scraping from the vagina posterolateral wall were carried out by Polymerase chain reaction (“DNK-Technologiia” LLC, RF). Facultative and obligate anaerobes, myco- and ureplasmas, and yeast-like fungi were quantified. The content of immunoglobulins, lysozyme, cytokines, complement, phagocytosis activity of leukocytes, hormones, the number of lymphocytes and their fractions, as well as the vaginal discharge pH (a total of 58 indexes) were identified in blood and vaginal discharge. For statistical and mathematical analysis, the Statistica 10 software (StatSoft, Inc., USA) was used. Using neural network modeling, it was revealed that among all the factorial signs for determining the bacterial dysbiosis degree, the complement component C4 and γ-INF content in the vaginal discharge and circulating immune complexes (CIC) and TNFα in the blood were important. A linear neural network model was built on the selected set of factor signs (the Cohen’s kappa coefficient consent index on the training set was k=0.87 (95% CI 0.82-0.91) for confirming plurality – k=0.89 (95% CI 0.77-1.00). With normocenosis, the complement activation PNB was decreased, and γ-INF and TNFα content was increased. The CIC levels blood increase corresponded to the opportunistic microflora growth and reflected the humoral immune response activation, which suggests that this indicator is an early dysbiosis marker. With I degree dysbiosis all factors had positive relationship with NBI, which reflected the immune system stress state. In case of II degree dysbiosis, NBI had a negative relationship with γ-INF content in the vaginal discharge, and CIC in the blood, while positive – with C4 content in vaginal discharge, and TNFα in the blood, which proved the immune system dysregulation and caused its further suppression with the BV-association immunodeficiency development. The immune system reaction during the BV development evolved from non-specific resistance reactions to cytokine-induced reactions of specific humoral immunity in response to the BV-associated microbiota growth, which subsequently experienced depletion and loss of immune control.

Assessment of Circulating Immune Complexes During Natural and Experimental Canine Leishmaniasis.

Canine leishmaniasis (CanL) is a disease characterized by a large variety of clinical alterations, the majority of which being due to immune mediated mechanisms. Sick dogs usually produce high levels of Leishmania-specific immunoglobulins which may give rise to circulating immune complexes (CICs) whose defective clearance by scavenging macrophages induces vasculitis and their deposition in specific organs. The aim of this study was to assess the serum level of CICs in dogs exposed to natural and experimental infection. Fifty-two sera were examined, belonging to untreated groups of naïve beagles previously studied to assess the performance of anti-leishmanial vaccines under natural (no. 22 dogs) or experimental (no. 30 dogs) transmission. Sera were classified in five groups according to the dog's health condition, IFAT titer, and the bone marrow (BM) nested (n)-PCR result. A: no.10 healthy dogs before the experimental infection; B: no.10 clinically healthy dogs infected experimentally, IFAT negative (= reciprocal titer <160) and n-PCR positive; C: no.10 clinically healthy dogs naturally infected, IFAT positive at titers 160–320 and n-PCR negative; D: no.10 sick dogs experimentally infected, IFAT positive at titer >320 and n-PCR positive; E: no.12 sick dogs naturally infected, IFAT positive at titer >320 and n-PCR positive. CICs levels were assessed by ELISA method (canine CIC assay—Cloude-Clone Corporation, USA). The two groups characterized by negative IFAT (A and B) had the lowest median level of CICs (16.09 and 12.78 μg/ml, respectively). CICs value increased progressively in the group C and reached the highest levels in the groups D and E, both characterized by high antibodies titer and severe disease, independently from the mode of infection. Significant differences in CICs concentration (p < 0.0001) were demonstrated between A, B, and C groups when compared with D or E groups of dogs. No differences were found inside the first three groups, while differences were recorded between the last two groups of sick dogs. CICs serum concentration increased with the progress of leishmaniasis, being significantly correlated with the increase of specific antibodies over time. High CICs levels detectable by commercial ELISA proved specific to an established Leishmania infection in dogs in the absence of other concomitant infections, as demonstrated by the similar trend assessed in experimentally and naturally infected dogs.

Immune-Complexome Analysis Identifies Immunoglobulin-Bound Biomarkers That Predict the Response to Chemotherapy of Pancreatic Cancer Patients.

Pancreatic Ductal Adenocarcinoma (PDA) is an aggressive malignancy with a very poor outcome. Although chemotherapy (CT) treatment has poor efficacy, it can enhance tumor immunogenicity. Tumor-Associated Antigens (TAA) are self-proteins that are overexpressed in tumors that may induce antibody production and can be PDA theranostic targets. However, the prognostic value of TAA-antibody association as Circulating Immune Complexes (CIC) has not yet been elucidated, mainly due to the lack of techniques that lead to their identification. In this study, we show a novel method to separate IgG, IgM, and IgA CIC from sera to use them as prognostic biomarkers of CT response. The PDA Immune-Complexome (IC) was identified using a LTQ-Orbitrap mass spectrometer followed by computational analysis. The analysis of the IC of 37 PDA patients before and after CT revealed differential associated antigens (DAA) for each immunoglobulin class. Our method identified different PDA-specific CIC in patients that were associated with poor prognosis patients. Finally, CIC levels were significantly modified by CT suggesting that they can be used as effective prognostic biomarkers to follow CT response in PDA patients.

Antiviral treatment perspective against Borna disease virus 1 infection in major depression: a double-blind placebo-controlled randomized clinical trial

BackgroundWhether Borna disease virus (BDV-1) is a human pathogen remained controversial until recent encephalitis cases showed BDV-1 infection could even be deadly. This called to mind previous evidence for an infectious contribution of BDV-1 to mental disorders. Pilot open trials suggested that BDV-1 infected depressed patients benefitted from antiviral therapy with a licensed drug (amantadine) which also tested sensitive in vitro. Here, we designed a double-blind placebo-controlled randomized clinical trial (RCT) which cross-linked depression and BDV-1 infection, addressing both the antidepressant and antiviral efficacy of amantadine.MethodsThe interventional phase II RCT (two 7-weeks-treatment periods and a 12-months follow-up) at the Hannover Medical School (MHH), Germany, assigned currently depressed BDV-1 infected patients with either major depression (MD; N = 23) or bipolar disorder (BD; N = 13) to amantadine sulphate (PK-Merz®; twice 100 mg orally daily) or placebo treatment, and contrariwise, respectively. Clinical changes were assessed every 2–3 weeks by the 21-item Hamilton rating scale for depression (HAMD) (total, single, and combined scores). BDV-1 activity was determined accordingly in blood plasma by enzyme immune assays for antigens (PAG), antibodies (AB) and circulating immune complexes (CIC).ResultsPrimary outcomes (≥25% HAMD reduction, week 7) were 81.3% amantadine vs. 35.3% placebo responder (p = 0.003), a large clinical effect size (ES; Cohen’s d) of 1.046, and excellent drug tolerance. Amantadine was safe reducing suicidal behaviour in the first 2 weeks. Pre-treatment maximum infection levels were predictive of clinical improvement (AB, p = 0.001; PAG, p = 0.026; HAMD week 7). Respective PAG and CIC levels correlated with AB reduction (p = 0,001 and p = 0.034, respectively). Follow-up benefits (12 months) correlated with dropped cumulative infection measures over time (p < 0.001). In vitro, amantadine concentrations as low as 2.4–10 ng/mL (50% infection-inhibitory dose) prevented infection with human BDV Hu-H1, while closely related memantine failed up to 100,000-fold higher concentration (200 μg/mL).ConclusionsOur findings indicate profound antidepressant efficacy of safe oral amantadine treatment, paralleling antiviral effects at various infection levels. This not only supports the paradigm of a link of BDV-1 infection and depression. It provides a novel possibly practice-changing low cost mental health care perspective for depressed BDV-1-infected patients addressing global needs.Trial registrationThe trial was retrospectively registered in the German Clinical Trials Registry on 04th of March 2015. The trial ID is DRKS00007649; https://www.drks.de/drks_web/setLocale_EN.do