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1.
Biol Res ; 43(4): 393-402, 2010.
Article in English | MEDLINE | ID: mdl-21526265

ABSTRACT

We have already demonstrated (Stojanovic et al., 2009) a connection between tetanus toxoid (TTd) hyperimmunization and the induction of anti-phospholipid syndrome (APS) in BALB/c mice. Here we show that C57BL/6 mice subjected to an identical procedure do not exhibit any like pathology attributable to anti-phospholipid antibodies; we explain that this absence results from idiotypic connectivity. Six groups of C57BL/6 mice were hyperimmunized with TTd in aluminum hydroxide or glycerol, with or without pretreatments. Pretreated mice had been injected with polyclonal or nonspecific immune stimulators, such as complete Freund's adjuvant (CFA) or glycerol. The epitope specificity of induced antibodies was tested by indirect ELISA using a tetanus toxoid immunogen and these autoantigens: phospholipids, gangliosides, laminin. Idiotypic connectivity was tested by competitive ELISA and gauged from the degree to which the interaction of idiotypic/anti-idiotypic complementary antibodies was inhibited in the presence of immunized sera antibodies. Higher idiotypic connectivity was noted amongst pretreated mice. There was a positive correlation between higher connectivity and autoantibody levels that acted to favor the participation of natural autoantibodies in the inhibitory process. We conclude that idiotypic connectivity plays a protective role in immunization-induced autoimmunity.


Subject(s)
Antibodies, Anti-Idiotypic/immunology , Antibodies, Antiphospholipid/immunology , Immunoglobulin G/immunology , Mice, Inbred C57BL/immunology , Tetanus Toxoid/immunology , Animals , Dose-Response Relationship, Immunologic , Enzyme-Linked Immunosorbent Assay , Female , Immunization/methods , Mice , Time Factors
2.
Biol. Res ; 43(4): 393-402, 2010. ilus
Article in English | LILACS | ID: lil-582853

ABSTRACT

We have already demonstrated (Stojanovic et al., 2009) a connection between tetanus toxoid (TTd) hyperimmunization and the induction of anti-phospholipid syndrome (APS) in BALB/c mice. Here we show that C57BL/6 mice subjected to an identical procedure do not exhibit any like pathology attributable to anti-phospholipid antibodies; we explain that this absence results from idiotypic connectivity. Six groups of C57BL/6 mice were hyperimmunized with TTd in aluminum hydroxide or glycerol, with or without pretreatments. Pretreated mice had been injected with polyclonal or nonspecific immune stimulators, such as complete Freund's adjuvant (CFA) or glycerol. The epitope specificity of induced antibodies was tested by indirect ELISA using a tetanus toxoid immunogen and these autoantigens: phospholipids, gangliosides, laminin. Idiotypic connectivity was tested by competitive ELISA and gauged from the degree to which the interaction of idiotypic/anti-idiotypic complementary antibodies was inhibited in the presence of immunized sera antibodies. Higher idiotypic connectivity was noted amongst pretreated mice. There was a positive correlation between higher connectivity and autoantibody levels that acted to favor the participation of natural autoantibodies in the inhibitory process. We conclude that idiotypic connectivity plays a protective role in immunization-induced autoimmunity.


Subject(s)
Animals , Female , Mice , Antibodies, Anti-Idiotypic/immunology , Antibodies, Antiphospholipid/immunology , Immunoglobulin G/immunology , /immunology , Tetanus Toxoid/immunology , Dose-Response Relationship, Immunologic , Enzyme-Linked Immunosorbent Assay , Immunization/methods , Time Factors
3.
Neuropeptides ; 41(6): 485-93, 2007 Dec.
Article in English | MEDLINE | ID: mdl-17761280

ABSTRACT

The role of somatostatin on inhibition of both normal and tumor cell cycle, secretion of endocrine and exocrine cells, as well as induction apoptosis is well documented. However, its effect on T cell development and thymic structure is not fully clarified. In order to investigate the influence of somatostatin in vivo on the thymus structure and T cell development, the young adult Albino Oxford male rats were intracerebroventriculary treated with somatostatin-14. We examined the thymus compartments and its cellularity, through assessment of morphometric parameters by stereological method, and the relation between thymocytes subpopulations, over expression of CD4, CD8 and T-cell receptor (TCR) alpha beta by flow cytometry. Additionally, we also determined the body and thymus weight of the rats, during the first three months of life, to define the time of SRIH-14 application. A decrease of relative thymus weight from the fourth weeks of postnatal life, and an unchanged relative thymus weight obtained in treated group indicates that SRIH-14 in young adult rats inhibits growth of whole organism, not only thymus. The changes in the absolute number and numerical density of cortical thymocytes indicate that SRIH-14 alters the true lymphoid tissue. SRIH-14 changes relation between thymocyte subsets, increase number of CD4(-)CD8(-)TCR alpha beta(-) and CD4(-)CD8(+)TCR alpha beta(hi) thymocyte subsets as well as the CD4(-)CD8(-)TCR alpha beta(low/hi) thymocytes, while decrease number of CD4(+)CD8(+) TCR alpha beta(-/low/hi) thymocyte subsets. These results indicate that somatostatin-14 is not involved in the control of the physiologic involution of the thymus, although induces thymic weight loss through the reduction of true lymphoid tissue. In addition, changes in frequency of thymocyte subpopulations, especially immature cells, indicate that SRIH-14 modulates thymocytes development and maturation.


Subject(s)
Somatostatin/pharmacology , T-Lymphocyte Subsets/cytology , Thymus Gland/cytology , Animals , Cell Differentiation , Flow Cytometry , Lymphocyte Count , Male , Organ Size/drug effects , Rats , Rats, Inbred Strains , T-Lymphocyte Subsets/drug effects , Thymus Gland/drug effects
4.
J Neuroimmunol ; 160(1-2): 77-86, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15710460

ABSTRACT

The aim of this study was to investigate whether chronic stress, induced by repeated daily swimming during 21 days, alters the morphofunctional parameters in the thymus of adult rats. Our results showed that chronic stress reduced thymus mass, total number of thymocytes, volume of the thymus compartments and numerical density of thymocytes within thymus inner cortex and medulla. However, the percentage of apoptotic cells and the level of corticosterone were significantly increased. The percentages of CD4-CD8-TCRalphabeta(low/high) and CD4-CD8+TCRalphabeta(-)thymocytes were significantly increased, while the percentage of the least mature CD4+CD8-SP TCRalphabeta(-) thymocytes was significantly decreased. These results show that recurred swimming procedure induces thymus hypotrophy and elevated percentage of DN TCRalphabeta(+) cells.


Subject(s)
Stress, Physiological/immunology , Stress, Physiological/physiopathology , Swimming/physiology , Thymus Gland/physiopathology , Animals , Apoptosis/immunology , CD4-CD8 Ratio , Corticosterone/blood , Flow Cytometry , Lymphopenia/immunology , Lymphopenia/pathology , Lymphopenia/physiopathology , Male , Organ Size/immunology , Rats , Rats, Inbred Strains , Receptors, Antigen, T-Cell, alpha-beta/biosynthesis , Stress, Physiological/pathology , T-Lymphocytes/metabolism , T-Lymphocytes/pathology , Thymus Gland/metabolism , Thymus Gland/pathology
5.
Histochem J ; 34(11-12): 573-82, 2002.
Article in English | MEDLINE | ID: mdl-14626348

ABSTRACT

The aim of this study was to investigate the effects of centrally applied somatostatin-28 on morphometric characteristics of the thymus, the thymocyte subpopulations, as well as, on apoptosis and phases of cell cycle in thymocytes. For this purpose, peripubertal male rats were cannulated intracerebroventriculary and treated with repeated, nanomolar concentrations of somatostatin-28 (experimental group) or saline (control group). Animals were sacrificed and their thymuses were used for the analysis of thymocyte subpopulations, cell cycle and apoptosis by flow cytometry and for the evaluation of morphometric parameters by stereological analysis. Our results showed that somatostatin-28 caused decrease of the thymic mass and volume, as well as total thymocytes number. Stereological analysis revealed volume decrease of thymic cortex and medulla accompanied with cellularity decrease. Somatostatin in the deeper cortex decreased the number of thymocytes, per volume unit, while in outer cortex raised their number. A significant increase in the percentage of double-negative and both single-positive thymocyte subpopulations, in parallel with a diminished percentage of double-positive cells was found. The cellularity of double-positive and single-positive thymocyte subpopulations was decreased. Somatostatin-28 treatment augmented the percentage of apoptotic cells, while the percentage of the cells represented in phases of cell cycle was reduced. These results suggest that somatostatin-28 induce thymus hypotrophy as result of decreasing cortex and medulla volume and cellularity. Changes in the percentage and cellularity of thymocyte subpopulations and numerical density of thymocytes in outer and deeper cortex, indicate that somatostatin-28 evoked disturbance in transition of double-negative to double-positive thymocytes.


Subject(s)
Protein Precursors/pharmacology , Somatostatin/pharmacology , Thymus Gland/drug effects , Animals , Apoptosis/drug effects , Body Weight/drug effects , Cell Count , Cell Cycle , DNA/analysis , Flow Cytometry , Injections, Intraventricular , Male , Organ Size/drug effects , Protein Precursors/administration & dosage , Rats , Rats, Inbred Strains , Sexual Maturation/physiology , Somatostatin/administration & dosage , Somatostatin-28 , Thymus Gland/growth & development , Thymus Gland/pathology
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