Search NEL Online

Journal Article 2015; 36(Suppl 1): 38-45 PubMed PMID: 26757124 Citation  Keywords:  Animals, Antibodies:immunology, Antibody Formation, Blotting, Western, Cell Line, Tumor, Chickens, Cytochrome P-450 Enzyme System:immunology, Cytochrome P450 Family 2, Eggs, Electrophoresis, Gel, Two-Dimensional, Enzyme-Linked Immunosorbent Assay, Humans,.   

OBJECTIVES: Cytochromes P450 (CYP) are monooxygenases, which metabolize mostly hydrophobic endogenous and exogenous compounds. CYPs without any clear connection to metabolism are called "orphans". Interestingly, these "orphan" CYPs are over-expressed in tumor tissues. Thus, the main aim of the paper is the development of antibodies for immunodetection of these CYPs as potential malignancy markers.

METHODS: Unique sequences of CYP2S1 and 2W1 were selected and peptides synthesized. Chickens were immunized with peptides bound to hemocyanin (KLH). The antibodies were isolated from egg yolks and their reactivity was tested by ELISA. Antibodies were further affinity purified on immobilized peptides. Western blots containing CYP2S1 and 2W1 standards were developed with purified antibodies.

RESULTS: Using unique peptide immunogens of CYP2S1 and 2W1 the antibodies were developed. As judged from ELISA all chickens produced specific antibodies against the respective peptides. Both affinity purified antibodies against CYP2S1 peptide recognized the CYP2S1 standard on Western blots, but only one of four anti-peptide antibodies against CYP2W1 reacted with CYP2W1 standard. The antibodies were used for the detection of CYPs in cancer cell lines and human tissues samples. Although both CYPs were frequently co-expressed in cancer cells, CYP2S1 was solely induced in the cell line BxPC3, while CYP2W1 was predominantly present in cell lines MCF7 and HeLa. Our data show that anti-peptide antibodies are an indispensable tool for detection of homologous CYPs.

CONCLUSIONS: The anti-peptide antibodies successfully recognized CYP2S1 and 2W1 in the cancer cell lines and tissue samples....

Read abstract  Full text PDF

Journal Article 2015; 36(5): 458-464 PubMed PMID: 26707047 Citation  Keywords:  Adolescent, Autoantibodies:immunology, Candida albicans, Candidiasis:immunology, Carrier State:immunology, Child, Child, Preschool, Cross Reactions, Female, Ghrelin:immunology, Growth Disorders:immunology, Helicobacter Infections:immunology, Helicobacter .   

OBJECTIVES: Many of peptides synthesized in gastrointestinal tract (GI) and adipose tissues, regulate growth and food intake. The GI microflora is an antigenic source. Based on the molecular mimicry hypothesis, intestinal microbe-derived antigens may trigger the production of autoantibodies cross-reacting with some neuropeptides.

DESIGN: The aim of the study was to assess whether in idiopathic short stature (ISS) children with Candida albicans (C.albicans) colonisation and/or Helicobacter pylori (H.pylori) infection the autoantibodies (in positive levels) against selected neuropeptides [anti-NP Abs(+)]: ghrelin, leptin, orexin A, αMSH are more prevalent than in Controls.

SETTING: The study group comprised 64 children with ISS and 36 children with normal height (Controls). In each child, IgG antibodies against H.pylori, ghrelin, leptin, orexin A and αMSH were assessed in serum, while presence of C.albicans - in stool samples.

RESULTS: The higher prevalence of anti-NP Abs(+) in ISS children with C.albicans and/or H.pylori than in normal height children with the colonization in question (34.4% vs 21.1%, p<0.01) was found. The prevalence of anti-NP Abs(+) in groups of children without C.albicans and H.pylori were low, anti-NP Abs(+) were detected in 9.4% of ISS children only, while in Controls they were not found.

CONCLUSIONS: In short children with C.albicans and/or H.pylori the incidence of autoantibodies against selected neuropeptides is high. It probably is connected with molecular mimicry between antigens of these microbiota and the mentioned peptides. It is tempting to speculate that presence of cross-reacting autoantibodies against regulatory neuropeptides may results in worse growth velocity. However, further studies are necessary to elucidate this issue....

Read abstract  Full text PDF

Journal Article 2014; 35(Suppl 2): 99-104 PubMed PMID: 25638373 Citation  Keywords:  Animals, Antibodies:immunology, Bacterial Infections:prevention & control, Cell Line, Chickens, Egg Yolk:immunology, Epithelial Cells:immunology, Humans, Immunoglobulin Fab Fragments:immunology, Immunoglobulins:immunology, Inflammation:immunology, Lung:im.   

OBJECTIVES: As an alternative therapeutic approach for the prevention and treatment of bacterial infections with P. aeruginosa of cystic fibrosis (CF) patients, chicken yolk antibodies (IgY) could be used. The most significant advantage of IgY, in contrast to mammalian IgG, consists in the fact, that when bound to the antigen, they usually do not induce inflammatory reaction. In addition, the simplicity of egg production and the ease of IgY preparation makes this kind of antibody an excellent tool for passive immunization. Thus, the aim of our project was to study the effect of IgY and its Fab fragment on the potential induction of pro-inflammatory reactions in lung epithelial cells.

METHODS: Chicken IgY were prepared from pooled egg yolks. Fab fragmens of IgY were purified from the papain digest of IgY using DEAE-Sephacel ion exchange chromatography. Their purity was verified by SDS electrophoresis in polyacrylamide gel. Immortalized human cell lines, CuFi (CF patient) and NuLi (healthy subject), and A549 (human adenocarcinoma cells) were exposed to IgY, Fab, OVA, LPS (positive control), PBS (negative control), and human and goat IgG for 24 hours. The concentration of pro-inflammatory cytokines TNF-α, IL-1β, IL-6 and GM-CSF were determined in cell media using the BioPlex method, which enables the quantification of multiple analytes simultaneously in one sample.

RESULTS: Our results show that i) the Fab fragment induced levels of some proinflammatory cytokines, when compared to the PBS control, whereas ii) chicken IgYs did not induce any notable production of pro-inflammatory cytokines in contrast to intense effect of LPS on TNF-α and GM-CSF. In summary, our data show that levels of all cytokines are comparable with physiological values in human serum except of IL-1β, which concentration in cell medium was markedly elevated by Fab fragment.

CONCLUSIONS: The present data indicate that IgY are not inflammatory for lung cells and thus they are possibly applicable for prevention of airway bacterial infections.

...

Read abstract  Full text PDF

Case Reports 2014; 35(5): 335-341 PubMed PMID: 25275265 Citation  Keywords:  Adolescent, Antithyroid Agents:administration & dosage, Autoantibodies:immunology, Child, Child, Preschool, Female, Graves Disease:drug therapy, Hashimoto Disease:drug therapy, Humans, Hypothyroidism:chemically induced, Male, Methimazole:administration & .   

BACKGROUND: The main clinical manifestations of autoimmune thyroid diseases are Graves' disease (GD) and Hashimoto's thyroiditis (HT). Graves' disease is the cause of most cases of hyperthyroidism in childhood. Indications for radical therapy (surgery or 131I treatment) in children are still a matter of discussion, as sustained (sometimes very long) remission of GD is possible, while the radical therapy almost always leads to hypothyroidism. Spontaneous evolution from GD with hyperthyroidism to HT with hypothyroidism may also be observed.

OBJECTIVE: The aim of the study was to analyze the clinical course of 6 cases of hyperthyroid girls with GD in whom a normalization of previously increased autoantibodies against thyrotropin (TSH) receptor (anti-TSHR) was observed together with a significant increase in autoantibodies against thyroid peroxidase (anti-TPO) and thyroglobulin (anti-Tg), with concomitant hypo- or euthyroidism but no recurrence of hyperthyroidism.

SUBJECTS: Patients' age at diagnosis ranged from 5.0 to 16.5 years. Two (2) patients had Turner syndrome, another one (1), diabetic, was on insulin therapy.

RESULTS: In all the girls, antithyroid drugs were administered and euthyroid state was achieved during the first 2.0-3.5 months of the treatment. Mild side effects were observed in only one case. The therapy was continued up to 1.5-4.0 years. Relapses during the therapy were observed in 2 cases. Up to now, no relapses have been observed for 0.5-7.5 years since the therapy withdrawal in 5 patients (1 patient was lost to follow-up), 2 patients are currently treated with levothyroxine due to hypothyroidism.

CONCLUSIONS: It seems that the prolonged pharmacotherapy with antithyroid drugs, followed by observation after remission of hyperthyroidism, may be an appropriate therapeutic option at least in some children with GD as they can be cured without radical therapy and the potential risks of such treatment....

Read abstract  Full text PDF

Comparative Study 2014; 35(1): 20-27 PubMed PMID: 24625913 Citation  Keywords:  Adult, Antibodies:immunology, Blood Glucose:analysis, Cholesterol:blood, Cholesterol, LDL:blood, Comorbidity, Cytomegalovirus:pathogenicity, Cytomegalovirus Infections:blood, Dehydroepiandrosterone:blood, Female, Humans, Immunoglobulin G:immunology, Male,.   

OBJECTIVES: Toxoplasma gondii, the protozoan parasite infecting about 30% population worldwide, is suspected to be the etiological agent of certain form of schizophrenia disease. Toxoplasma is known to change levels of certain neurotransmitters, cytokines and several hormones in both infected animals and humans. A common feature of toxoplasmosis and schizophrenia is a disorder of immune system.

METHODS: Here we studied the levels of five neuro- and immunomodulatory steroids, selected hormones and lipids in sera of 173 schizophrenia patients.

RESULTS: Toxoplasma infected schizophrenia patients expressed only insignificantly lower concentration of neuro- and immunomodulatory DHEA metabolites. Infected women had higher concentration of glucose while infected men had higher concentration of cholesterol and LDL cholesterol. No significant effect of human cytomegalovirus infection on the concentration of the above parameters was observed. The difference in the concentration of DHEA metabolites faded with the decrease of the concentration of anti-Toxoplasma IgG antibodies (i.e. with the duration of Toxoplasma infection) while the difference in the concentration of cholesterol and LDL-cholesterol increased with the decrease of the concentration of anti-Toxoplasma IgG antibodies. The prevalence of toxoplasmosis in male (53.2%) but not female (29.8%) schizophrenia patients was unusually high in comparison with prevalence of toxoplasmosis in a general population.

CONCLUSION: Our results provided an explanation for seemingly decreasing prevalence of toxoplasmosis in schizophrenia patients observed in current studies (increased concerns about the rights of patients resulting in absence of non-cooperative Toxoplasma-positive patients in the study population) and suggest possible explanation for reported positive correlation between prevalence of toxoplasmosis and incidence of cardiovascular diseases (accelerated atherosclerotic development due to increased level of cholesterol and LDL in Toxoplasma infected humans)....

Read abstract  Full text PDF

Case Reports 2013; 34(7): 601-605 PubMed PMID: 24463996 Citation  Keywords:  Autoantibodies:immunology, Brain Stem:immunology, Encephalitis:complications, Gangliosides:immunology, Guillain-Barre Syndrome:complications, Humans, Magnetic Resonance Imaging, Male, Tomography, X-Ray Computed, Young Adult,.   

: We report a case of a 23-year-old man diagnosed with overlapping Bickerstaff brainstem encephalitis (BBE) and Guillain-Barre syndrome (GBS). The patient initially presented with fever and headache and gradually developed ataxia, disturbance of consciousness, respiratory muscle paralysis, bilateral facial paralysis and quadriplegia accompanied by significant atrophy of limb, temporalis and masseter muscles. Brain MRI revealed abnormality in the left basal ganglia, thalamus, and rightside posterior limb of the internal capsule. Electromyogram indicated neurogenic damage (mainly axonal damage) in the upper and lower limbs and bilateral facial nerve damage. Cerebrospinal fluid (CSF) collected via lumbar puncture was colorless and transparent with a pressure of 330 mm H2O. The white blood cell count in CSF was 200×106/L, the protein concentration was 1.25 g/L, and Pandy's reaction was positive. Both the blood and CSF were negative for GQ1b antibody. The patient was clinically diagnosed with overlapping BBE and GBS. After treatment with ventilator assisted breathing, hormone therapy, neurotrophic and anti-infection therapies, and symptomatic and supportive care for more than three months, spontaneous breathing was restored. By the 5-month follow-up examination, the patient had completely recovered and returned to work. Like GBS and Fisher syndrome, BBE might be an anti-GQlb IgG antibody syndrome. Although the serum GQlb IgG antibody-positive rate for BBE is only 66%, a normal brainstem MRI or GQlb lgG antibody-negative finding cannot completely rule out BBE. Therefore, identifying critical illness polyneuropathy for patients with respiratory muscle paralysis and tracheal extubation difficulties at early stages is clinically important....

Read abstract  Full text PDF

Journal Article 2007; 28(4): 422-426 PubMed PMID: 17693974 Citation  Keywords:  Adolescent, Anorexia Nervosa:immunology, Antibodies:immunology, Antigens, CD28:immunology, Antigens, CD3:immunology, CD4-Positive T-Lymphocytes:metabolism, Calcium:metabolism, Case-Control Studies, Child, Female, Humans, Interleukin-2:metabolism, Lymphocy.   

OBJECTIVES: Anorexia nervosa (AN), a disease of chronic human starvation has a deep impact on the function of several organ systems. We hypothesized that disturbed cellular activation may contribute to complications in AN. We tested our assumption on short-term activation kinetics of lymphocytes.

PATIENTS AND METHODS: Blood was taken from 11 AN and 10 healthy adolescents. Peripheral blood mononuclear cells were isolated and CD4+ lymphocytes were then activated with phythohemagglutinin for the determination of calcium-influx and membrane potential. Moreover, cells were also activated by anti-CD3/anti-CD28 coated beads and three days after the prevalence of interleukin-2 positive CD4+ cells were determined.

RESULTS: After activation, more time was required to reach maximal calcium content in CD4+ cells of AN patients than in those of controls (control vs. AN (median, range): 86 [45-232] vs. 215 [59-235] second, p<0.05), but the rate of membrane potential alteration was similar. The number of interleukin 2 positive CD4+ cells was lower in AN (11.50 [7.60-15.30] vs. 13.50 [12.00-22.00] %, p<0.05). No association was detected between cell activation and any of clinical or anthropometric data of AN patients.

CONCLUSIONS: These results suggest that AN may have an impact on calcium handling of the cells and, hence, cell activation characteristics. We assume that altered calcium flux kinetics may contribute to complications present in AN....

Read abstract  Full text PDF

Journal Article 2005; 26(4): 277-282 PubMed PMID: 16136024 Citation  Keywords:  Adult, Antibodies:immunology, Dental Amalgam:adverse effects, Female, Humans, In Vitro Techniques, Infertility:epidemiology, Interferon-gamma:metabolism, Lymphocyte Activation:drug effects, Lymphocytes:drug effects, Male, Mercury Poisoning, Nervous System.   

INTRODUCTION: Heavy metals can negatively influence the reproduction due to the fact that they are able to impair the immune reactions including autoantibody production in susceptible individuals. In such a way the infertility could be also caused by altered pathologic immune reaction.

AIM OF THE STUDY: To investigate the in vitro lymphocyte reaction after stimulation with metals and production of gamma interferon and antisperm antibodies in supernatants after lymphocyte stimulation in patients with infertility and with proven antisperm antibodies in their serum. The cause of antisperm antibodies presence was not determined.

METHODS: The diagnosis of metal allergy was performed by the lymphocyte proliferation method modified for metals (MELISA) in supernatants of tissue cultures of lymphocytes without the antigen stimulation and after stimulation with mercury chloride, the in vitro production of gamma interferon and antisperm antibodies was studied by ELISA.

RESULTS: More than 50% of patients were reacting to mercury, iron, aluminium and silver as mean by lymphocyte reactivity. When compared the lymphocyte reaction in patients with and without mercury allergy we found that the lymphocytes of patients with mercury intolerance produced less gamma interferon and more antisperm antibodies in supernatants after mercury stimulation of their lymphocytes.

CONCLUSION: In patients with metal intolerance diagnosed by the MELISA test the release of metal ions from dental materials can be one of the stimulating factors which may adversely affect fertility....

Read abstract  Full text PDF