Tag Archives: antibodies
Comparison of adjuvant properties of chitosan during oral and subcutaneous immunization of mice with BSA
M. R. Kozak1*, I. M. Petruh1, V. V. Vlizlo2
1Institute of Animal Biology NAAS, Lviv, Ukraine;
2Stepan Gzhytskyi National University of Veterinary Medicine and Biotechnologies Lviv, Ukraine;
*e-mail: mariyarkozak@gmail.com
Received: 15 December 2021; Accepted: 01 July 2022
Vaccination is the best method to prevent the spread of infectious diseases, its disadvantages are side effects. Potentially safe DNA, RNA or protein molecules possess antigenic properties, but are low-immunogenic and therefore require conjugation with an adjuvant. The aim of the research was to evaluate Chitosan (CS) potency as an adjuvant and compare its effectiveness depending on the route of drug administration. The experiments were carried out on 3 groups of BALB/c mice. Mice of the first group were injected subcutaneously with 20 µl of a mixture of CS (3.3 mg/kg) and BSA (1.7 mg/kg). The mixture of CS and BSA at the same doses and volume was administered orally to mice of the second experimental group. The third group – control – unvaccinated mice. Anti-BSA antibody levels were measured by ELISA. Aspartate aminotransferase, alanine aminotransferase activity and cholesterol, creatinine and urea levels were determined in the serum. It was found that both subcutaneous and mucosal immunizations provided a 2-fold increase in anti-BSA antibody titers against the background of maintaining all biochemical blood parameters at the level of the physiological norm. However, AST activity in the serum of oral-immunized mice was elevated as compared to subcutaneous-immunized mice. Serum cholesterol level in the group of subcutaneously immunized mice and creatinine and urea levels in both experimental groups were reduced compared to the control. It is concluded that oral immunization with CS is the optimal route for antigen-specific IgG antibody response induction.
Avidity of bivalent antibodies. Problems of its experimental determination and theoretical evaluation
S. A. Bobrovnik, M. A.Demchenko, S. V. Komisarenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kiyv;
e-mail: s-bobrov@bk.ru
Some problems of experimental determination or theoretical evaluation of antibody avidity are considered. It was shown that in order to determine the fraction of nonoccupied antibodies in their mixture with the excess of the corresponding antigen which is required to estimate avidity the methods should be used which are more sensitive than ELISA. The available methods did not allow determining the avidity of bivalent antibodies because of many reasons. However, in the recent years new methods were suggested that make it possible to evaluate the avidity of bivalent antibodies and that of the receptors which consist of two binding sites connected by a flexible linker of the known length. Thus, there are all possibilities now for determining the avidity of bivalent antibodies in experiments or by theoretical methods.
Analysis of titration curves of serum antibodies obtained by ELISA
S. A. Bobrovnik
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: s-bobrov@bk.ru
Different forms of titration curves obtained by ELISA for serum antibodies, monoclonal antibodies and polyreactive immunoglobulins were considered. A new interpretation of dose-dependent titration curves was suggested. It was shown that our interpretation of dose-dependent titration curves for antibodies which are presented in some biological liquids allow obtaining additional information about properties of the samples. This information was not obtained earlier because of the wrong understanding of the considered problem.
Nicotinic acetylcholine receptors: specific antibodies and functions in humoral immunity
M. V. Skok, L. M. Koval, O. Yu. Lykhmus, O. M. Kalashnyk,
G. L. Gergalova, S. V. Komisarenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: skok@biochem.kiev.ua
Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels initially discovered in muscles and neurons and further found in many non-excitable cells. The present review summarizes the results of studies performed in the Department of Molecular Immunology during the last decade and concerning the structure and functions of nAChRs in B lymphocytes and in mitochondria, as well as the role of nAChR-specific antibodies in the development of neurodegenerative disorders like Alzheimer disease.
Correlation between base concentration of C-reactive protein in the blood, levels of serum antiendotoxin antibodies and endotoxin-binding capacity of monocytes and granulocytes of healthy people
A. I. Gordienko
State Institution S. I. Georgievsky Crimea State Medical University, Simferopol, Ukraine;
e-mail: uu4jey@csmu.strace.net
The associative links between base concentration of C-reactive protein (hsCRP), levels of serum antiendotoxin antibodies of different classes and endotoxin-binding capacity of monocytes and granulocytes of healthy volunteers were investigated by cluster analysis. In the group of healthy volunteers with increased base concentration of hsCRP in blood the levels of serum antiendotoxin antibodies of different classes and endotoxin-binding capacity of monocytes and granulocytes were reduced. Thus, the disbalance of detoxification and clearance of endotoxin by humoral and cellular mechanisms can be one of the possible causes of development of low intensity inflammation and increase of hsCRP concentration in blood.
Avidity of polyreactive immunoglobulins
S. A. Bobrovnik
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: s-bobrov@bk.ru
An analysis of the mechanism of interaction between polyreactive immunoglobulins (PRIG) and antigen was conducted and it was shown that most of the traditional methods of antibody affinity evaluation are not applicable for PRIG affinity. The comparative assessment of the mouse and human PRIG avidity against ovalbumin and horse myoglobin and the avidity of specific monoclonal antibodies against ovalbumin have shown that the avidity of PRIG not only is much less than the avidity of monoclonal antibodies but even exceeds it.
Proteolytic activity of IgGs from blood serum of wistar rats at experimental rheumatoid arthritis
Yu. Ya. Kit1, S. L. Myronovsky3, I. I. Kril’2,
A. M. Havrylyuk2, V. V. Chop’yak2, R. S. Stoika1
1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Danylo Halytsky Lviv National Medical University, Ukraine;
3Ivan Franko National University of Lviv, Ukraine;
e-mail: kit@cellbiol.lviv.ua
The aim of this work was to study the proteolytic activity of IgGs purified from blood serum of Wistar rats at experimental rheumatoid arthritis (ERA) induced by an injection of bovine collagen of type II. Twenty rats were immunized with a preparation of bovine collagen II (Sigma-Aldrich, USA) in the presence of complete Freund’s adjuvant. ERA development was determined by inflammation in limbs of treated animals. IgG preparations were isolated from blood serum of immunized and non-immunized animals by precipitation of antibodies with 33% ammonium sulfate followed by chromatography on the Protein G-Sepharose column. Human histone H1, bovine collagen II, calf thymus histones, myelin basic protein (MBP), bovine serum albumin (BSA), and bovine casein were used as substrates of the proteolytic activity of IgGs. It was found that IgG preparations from blood serum of rats with ERA were capable of cleaving histone H1 and MBP, however, they were catalytically inactive towards collagen II, casein, BSA, and core histones. IgGs from blood serum of non-immunized rats were proteolytically inactive towards all used protein substrates. Thus, we demonstrated that immunization of rats with bovine collagen II induced IgG-antibodies possessing the proteolytic activity towards histone H1 and MBP. This activity might be associated with the development of inflammatory processes in the immunized rats.
Proteolytic activity of IgG-antibodies of mice, immunized by calf thymus histones
Yu. Kit1, N. Korniy1,3, I. Kril’2, I. Magorivska1, V. Tkachenko1, R. Bilyy1,2, R. Stoika1
1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
e-mail: kit@cellbiol.lviv.ua;
2Danylo Galytsky Lviv National Medical University, Ukraine;
3Ivan Franko L’viv National University, Ukraine
The main goal of the study was to determine the ability of histones to induce production of the proteolytically active IgG-antibodies in BALB/c mice. In order to perform this study 8 mice were immunized with the fraction of total calf thymus histones. IgGs were isolated from the serum of the immunized and not immunized animals by means of precipitation with 33% ammonium sulfate, followed by affinity chromatography on protein G-Sepharose column. Histones, myelin basic protein (MBP), lysozyme, BSA, ovalbumin, macroglobulin, casein and cytochrome c served as substrates for determining the proteolytic activity. It was found that IgGs from the blood serum of immunized mice are capable of hydrolyzing histone H1, core histone and MBP. On the contrary, the proteolytic activity of IgGs from the blood serum of not immunized mice was not detected. The absence of proteolytical enzymes in the fraction of IgGs was proven by HPLC chromatography. High levels of proteolytic activity toward histones have been also detected in affinity purified IgGs from blood serum of patients with rheumatoid arthritis, but not in healthy donors. These data indicate that eukaryotic histones may induce production of protabzymes in mammals. The possible origin of these protabzymes and their potential biological role in mammalians is discussed.