The preparation and study of anti-idiotypic (secondary) antibodies (Ab2) against monoclonal primary antibodies (Ab1) specific to biologically active molecules with a known structure is of great scientific and practical importance. Due to partial antigenic similarity of Ab2 and the initial antigen structures, these antibodies can be the basis of the vaccine, if the antigen usage is not possible, or is limited by law. In particular, one may create Ab2-based preparations, designed for immunization, in order to prevent and treat the drug addiction. The value of Ab2 properties increases even more if Ab1, used to obtain them, recognize different parts of the antigen molecule, which makes it possible to obtain second-generation antibodies with a wide range of specificity. In this work, the morphine-like polyclonal and monoclonal Ab2 were obtained. In each case, as the first-generation immunoglobulins for immunization, we used two murine monoclonal antibodies (mAbs) specific to different morphine derivatives: 3K11 antibodies to 3-0-carboxymethyl (CMM) and 2-p-carboxyphenylazomethyl (FAM) derivatives, as well as 6G1 antibodies to 6-hemisuccinyl derivative (GSM). After immunization of the horse with Ab1 and development of immune response, three pools of specific polyclonal antibodies were isolated from the animal blood serum: horse anti-species antibodies to the total mouse immunoglobulins (HAM); horse anti-idiotypic antibodies against 3K11 antibodies (HAM-K11), and against 6G1 antibodies (HAM-G1). In parallel, immunization of mice with 3K11 and 6G1 antibodies and fusion of obtained lymphocytes with Sp2/0 mouse myeloma cells by the Milstein-Köhler method resulted in three producers of anti-idiotypic antibodies: a clone producing mouse monoclonal Ab2 specific for mAb-6G1 (AIG1), as well as clones producing anti-mAb-3K11 antibodies (AI-K11A and AI-K11B). The physico-chemical and antigenic properties of all the Ab2 obtained were characterized. It was shown that the horse anti-idiotypic immunoglobulins not only belong to different classes, but are also polyvalent, while all monoclonal Ab2 obtained are represented by IgM immunoglobulins, being also strictly specific to the corresponding first-generation antibodies. Subsequently, the morphine-like properties of the first domestic polyclonal and monoclonal Ab2 obtained in the work will be investigated in a cellular model. Likewise, we shall study their ability to induce Ab3 as well as morphine-specific Ab1.
147 women aged 31-47 years with autoimmune thyroiditis were examined. The control group included 63 appar- ently healthy women of the same age without thyroid disease. Using enzyme-linked immunosorbent assay, serum levels of tumor necrosis factor α (TNF-α), interleukins (IL) -1β, -6, -8, -17A, receptor activator of nuclear factor-kappa β ligand (RANKL) and osteoprotegerin (OPG) were determined. The median, interquartile range and Spearman’s rank correlation coefficient were calculated. The Mann-Whitney test was used to compare two independent samples. Autoimmune damage of the thyroid gland was characterized by a significant (p<0,001) increase in the levels of IL-1β and IL-6 against the background of a trend towards an increase of serum IL-17A concentration (p=0,067). At the same time, the values of TNF-α and IL-8 did not differ significantly from those of women in the control group (p=0,166 and p=0,102, respectively). Also, autoimmune thyroiditis in women was accompanied by a significant increase of RANKL concentration (p=0.029), while the OPG content remained at control levels (p=0,988). A regular decrease in the OPG/RANKL ratio was also recorded (p=0,017). The presence of a significant (p<0,05) positive relationship of IL-6 concentrations with TNF-α, IL-8, IL-17A and RANKL values, as well as between RANKL and IL-8 values was established. In addition, the concentration of RANKL was characterized by a direct correlation with the content of the anti-inflammatory mediator OPG. Attention is drawn to the revealed negative correlation between the OPG/RANKL index and IL-6 values. Autoimmune thyroiditis in women is characterized by a significant increase of serum levels of IL-1β, IL-6, RANKL and a decrease in the OPG/RANKL index, while the indices of IL-8, TNF-α and OPG production do not change signifi- cantly. For IL-6 concentrations, a positive correlation was established with the values of TNF-α, IL-8, IL-17 and nega- tive - with the OPG/RANKL ratio. RANKL values have a direct relationship with IL-6, IL-8 and OPG levels.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.