scholarly journals Systemic Administration of Proteoglycan Protects BALB/c Retired Breeder Mice from Experimental Arthritis

2016 ◽  
Vol 2016 ◽  
pp. 1-11
Author(s):  
Larissa Lumi Watanabe Ishikawa ◽  
Priscila Maria Colavite ◽  
Thais Fernanda de Campos Fraga-Silva ◽  
Luiza Ayumi Nishiyama Mimura ◽  
Thais Graziela Donegá França ◽  
...  
Keyword(s):  
Tolerance Induction ◽  
Experimental Arthritis ◽  
Delayed Type Hypersensitivity ◽  
Spleen Cells ◽  
The Third ◽  
Dimethyldioctadecylammonium Bromide ◽  
Specific Tolerance ◽  
Lower Production ◽  
Score Evaluation

This study was undertaken to evaluate the prophylactic potential of proteoglycan (PG) administration in experimental arthritis. Female BALB/c retired breeder mice received two (2xPG50 and 2xPG100 groups) or three (3xPG50 group) intraperitoneal doses of bovine PG (50 μg or 100 μg) every three days. A week later the animals were submitted to arthritis induction by immunization with three i.p. doses of bovine PG associated with dimethyldioctadecylammonium bromide adjuvant at intervals of 21 days. Disease severity was daily assessed after the third dose by score evaluation. The 3xPG50 group showed significant reduction in prevalence and clinical scores. This protective effect was associated with lower production of IFN-γand IL-17 and increased production of IL-5 and IL-10 by spleen cells restimulatedin vitrowith PG. Even though previous PG administration restrained dendritic cells maturation this procedure did not alter the frequency of regulatory Foxp3+T cells. Lower TNF-αand IL-6 levels and higher expression of ROR-γand GATA-3 were detected in the paws of protected animals. A delayed-type hypersensitivity reaction confirmed specific tolerance induction. Taken together, these results indicate that previous PG inoculation determines a specific tolerogenic effect that is able to decrease severity of subsequently induced arthritis.

scholarly journals In vitro tolerance induction of neonatal murine B cells.

1976 ◽  
Vol 143 (6) ◽  
pp. 1327-1340 ◽  
Author(s):  
E S Metcalf ◽  
N R Klinman
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocytes ◽  
Specific Interaction ◽  
Tolerance Induction ◽  
Antigen Receptors ◽  
Spleen Cells ◽  
Protein Conjugates ◽  
Immature B Cells

The susceptibility of neonatal and adult B lymphocytes to tolerance induction was analyzed by a modification of the in vitro splenic focus technique. This technique permits stimulation of individual hapten-specific clonal precursor cells from both neonatal and adult donors. Neonatal or adult BALB/c spleen cells were adoptively transferred into irradiated, syngeneic, adult recipients which had been carrier-primed to hemocyanin (Hy), thus maximizing stimulation to the hapten 2,4-dinitrophenyl coupled by Hy (DNP-Hy). Cultures were initially treated with DNP on several heterologous (non-Hy) carriers and subsequently stimulated with DNP-Hy. Whereas the responsiveness of adult B cells was not diminished by pretreatment with any DNP conjugate, the majority of the neonatal B-cell response was abolished by in vitro culture with all of the DNP-protein conjugates. During the 1st wk of life, the ability to tolerize neonatal splenic B cells progressively decreased. Thus, tolerance in this system is: (a) restricted to B cells early in development; (b) established by both tolerogens and immunogens; (c) achieved at low (10(-9) M determinant) antigen concentrations; and (d) highly specific, discriminating between DNP- and TNP-specific B cells. We conclude that: (a) B lymphocytes, during their development, mature through a stage in which they are extremely susceptible to tolerogenesis; (b) the specific interaction of B-cell antigen receptors with multivalent antigens, while irrelevant to mature B cells, is tolerogenic to neonatal (immature) B cells unless antigen is concomitantly recognized by primed T cells; and (c) differences in the susceptibility of immature and mature B lymphocytes to tolerance induction suggest intrinsic differences between neonatal and adult B cells and may provide a physiologically relevant model for the study of tolerance to self-antigens.

scholarly journals Role of IgD in the immune response and tolerance. I. Anti-delta pretreatment facilitates tolerance induction in adult B cells in vitro.

1977 ◽  
Vol 146 (6) ◽  
pp. 1473-1483 ◽  
Author(s):  
D W Scott ◽  
J E Layton ◽  
G J Nossal
Keyword(s):  
Immune Response ◽  
B Cells ◽  
B Cell ◽  
Tolerance Induction ◽  
Spleen Cells ◽  
Cell Compartment ◽  
Immature B Cells

Adult spleen cells from C57BL.Ige mice, which generally are resistant to in vitro tolerance induction in the B-cell compartment, became hyporesponsive (tolerant) when cultured with antigen in the presence of an anti-allotype serum. Both antigen and anti-delta had to be present for this effect, which was hapten-specific and did not occur in C57BL/L mice, which lack the Ig5-1 allotype of the delta-chain detected in this system. Preculture with anti-mu serum plus antigen, in contrast, did not cause tolerance induction in adult spleen B cells of either strain. These results suggest that the surface IgD may act as a failsafe receptor to prevent tolerance induction in adult B cells. Tolerance studies with spleen cells from mice with markedly reduced numbers of IgD+ve cells, because of regimen of repeated injections of anti-delta serum beginning at birth (delta-suppressed mice), confirmed the importance of membrane IgD in preventing tolerance, because such delta-suppressed mice were hypersusceptible to tolerance by antigen alone. Inasmuch as immature B cells lack IgD on their surface, these studies suggest that acquisition of IgD is an important maturational step in the ability of murine B cells to discriminate tolerogenic and immunogenic signals.

scholarly journals Induction of immunological tolerance requires that the B cells can respond to the polyclonal B-cell-activating properties of the thymus-independent antigens.

1977 ◽  
Vol 146 (1) ◽  
pp. 308-312 ◽  
Author(s):  
C Fernandez ◽  
G Möller
Keyword(s):  
B Cells ◽  
B Cell ◽  
Tolerance Induction ◽  
Immunological Tolerance ◽  
Spleen Cells ◽  
Normal Cells ◽  
Purified Protein Derivative ◽  
High Affinity

Mice were rendered specifically tolerant to the fluorescein isothiocyanatedextran (FITC) epitope by injection of FITC-dextran B512. Their spleen cells were removed at various times and cultivated in vitro with different polyclonal B-cell activators, such as lipopolysaccharide (LPS), purified protein derivative of tuberculin, and native dextran. LPS caused the appearance of high affinity anti-FITC plaque-forming cells to an equal extent with cells from untreated and tolerant animals, whereas native dextran failed to activate cells from tolerant mice, although it was a potent activator of normal cells. It was concluded that tolerance induction only affects those B cells that could respond to the polyclonal B-cell-activating properties of the tolerogen, but not other B cells having an identical set of Ig receptors directed against the tolerogen.

scholarly journals In vitro tolerance induction of primed, IgD-negative murine spleen cells.

1981 ◽  
Vol 153 (3) ◽  
pp. 653-664 ◽  
Author(s):  
S M Walker ◽  
W O Weigle
Keyword(s):  
B Cells ◽  
Tolerance Induction ◽  
Keyhole Limpet Hemocyanin ◽  
Immunological Tolerance ◽  
Target Cells ◽  
Spleen Cells ◽  
Murine Spleen ◽  
In Vitro Response

The above observations demonstrated induction of immunological tolerance in vitro in primed IgD-, IgG+ B cells. In these studies, addition of trinitrophenylated (TNP) turkey gammaglobulin (TGG) or TNP ovalbumin conjugates suppressed the secondary in vitro response in mice primed with TNP keyhole limpet hemocyanin (TNP-KLH). Suppression was not a reflection of a shift in kinetics of the antibody response, was not dependent on suppressor T cells, and could only be eliciate when conjugate was added within 4 h of addition of TNP-KLH moreover, preincubation of the primed spleen cells with TNP-TGG for 20 h at 37 degrees C, followed by extensive washing, was as effective in inhibiting the response to TNP-KLH as when TNP-TGG was present throughout the 5 d of culture, reflecting induction of a tolerant state. Amounts of conjugate in the concentration range that have been shown by others to tolerize immature or neonatal B cells or mature B cells that have been stripped of surface IgD were sufficient to induce tolerance. The target cells being tolerized did not bear IgD, as determined by B cell depletion and blocking procedures with anti IgD. Whether the lack of surface IgD on the primed cells contributed to the relative ease of tolerance induction was not established by these studies, but the advantages of using primed B cells to examine further the role of surface IgD in tolerance susceptibility was discussed.

scholarly journals Genetic Vaccination against Coccidioides immitis: Comparison of Vaccine Efficacy of Recombinant Antigen 2 and Antigen 2 cDNA

1999 ◽  
Vol 67 (2) ◽  
pp. 630-635 ◽  
Author(s):  
Chengyong Jiang ◽  
D. Mitchell Magee ◽  
Teresa N. Quitugua ◽  
Rebecca A. Cox
Keyword(s):  
Fusion Protein ◽  
Vaccine Efficacy ◽  
Water Soluble ◽  
Delayed Type Hypersensitivity ◽  
Coccidioides Immitis ◽  
Genetic Immunization ◽  
Spleen Cells ◽  
Lethal Challenge ◽  
Water Soluble Extract

ABSTRACT Previous studies from our laboratory established that C-ASWS, an alkali-soluble, water-soluble extract from cell walls ofCoccidioides immitis, protects mice against lethal challenge with this fungus. The C-ASWS extract contains a glycosylated protein, designated antigen 2 (Ag2), and a polysaccharide antigen. We recently cloned Ag2 cDNA and showed that the recombinant fusion protein elicited strong delayed-type hypersensitivity responses in immunized mice. This investigation was undertaken to determine if the recombinant Ag2 protein, expressed as an Ag2-glutathione S-transferase (GST) fusion protein, or Ag2 cDNA would protect mice against lethal challenge with C. immitis. The recombinant Ag2-GST protein protected BALB/c mice against intraperitoneal challenge with 250 arthroconidia, as assessed by a decrease in fungal CFU in tissues. The Ag2-GST-immunized mice did not show, however, an increased survival during a 30-day period postinfection. By contrast, immunization of mice with Ag2 cDNA ligated into the pVR1012 plasmid engendered protection against intraperitoneal challenge with 2,500 arthroconidia and against pulmonary challenge with 50 arthroconidia. Vaccine efficacy paralleled the development of delayed-type hypersensitivity responses to C. immitis antigen. Whereas mice vaccinated with the recombinant Ag2-GST protein did not mount footpad hypersensitivity to C-ASWS or the recombinant Ag2-GST protein, mice vaccinated with the pVR1012-Ag2 construct mounted a strong footpad hypersensitivity and their spleen cells secreted gamma interferon upon in vitro stimulation with the Ag2-containing C-ASWS extract. This is the first investigation to show that genetic immunization can protect against lethal challenge withC. immitis.

scholarly journals GENETIC CONTROL OF IMMUNE RESPONSES IN VITRO

1974 ◽  
Vol 140 (1) ◽  
pp. 172-184 ◽  
Author(s):  
Judith A. Kapp ◽  
Carl W. Pierce ◽  
Baruj Benacerraf
Keyword(s):  
Immune Response ◽  
Gene Regulation ◽  
B Cells ◽  
Bovine Serum Albumin ◽  
Immune Responses ◽  
Bovine Serum ◽  
Tolerance Induction ◽  
Spleen Cells

Although nonresponder, H-2s and H-2q, mice fail to develop GAT-specific PFC responses to GAT, they do develop GAT-specific PFC responses when stimulated by GAT complexed to an immunogenic carrier such as methylated bovine serum albumin. The studies described in this paper show that injection of nonresponder mice with GAT specifically decreases their ability to develop anti-GAT PFC responses to a subsequent challenge with GAT-MBSA. Addition of GAT to cultures of spleen cells from nonresponder mice also prevents development of the GAT-specific PFC responses stimulated by GAT-MBSA. Thus, interaction of nonresponder spleen cells with GAT leads to the induction of unresponsiveness in vivo and in vitro. Various parameters of the tolerance induction have been investigated and described. A comparison of the effects of GAT on B cells indicates that nonresponder B cells are more readily rendered unresponsive by soluble GAT than are responder B cells. The significance of these data for our understanding of Ir gene regulation of the immune response is discussed.

scholarly journals IMMUNOLOGICAL TOLERANCE IN BONE MARROW-DERIVED LYMPHOCYTES

1972 ◽  
Vol 136 (6) ◽  
pp. 1404-1429 ◽  
Author(s):  
David H. Katz ◽  
Toshiyuki Hamaoka ◽  
Baruj Benacerraf
Keyword(s):  
Bone Marrow ◽  
B Lymphocytes ◽  
Adoptive Transfer ◽  
Tolerance Induction ◽  
Adoptive Cell Transfer ◽  
Secondary Response ◽  
Specific Tolerance ◽  
Tolerant State

Administration of the 2,4-dinitrophenyl (DNP) derivative of the copolymer of D-glutamic acid and D-lysine (D-GL) to inbred mice induces a state of DNP-specific tolerance in such animals irrespective of their immune status at the time of treatment. Taking advantage of the relative ease with which DNP-D-GL can induce tolerance in an animal previously primed with an immunogenic DNP-carrier conjugate, we have established conditions for tolerance induction in an adoptive cell transfer system. Thus, the adoptive secondary anti-DNP antibody response of DNP-keyhole limpet hemocyanin (KLH)-primed spleen cells was completely, or almost completely, abolished by exposure of such cells to DMP-D-GL either in vivo or in vitro. Tolerance induction in vivo occurred irrespective of whether the DNP-primed cells were exposed to DNP-D-GL in the donor animal before adoptive transfer or in recipient mice after transfer. In the latter situation, it was possible to show that tolerance induction in this model occurs very rapidly (1 hr) and with relatively low doses of tolerogen (50 µg). Incubation of DNP-KLH-primed cells with DNP-D-GL in vitro under varying culture conditions also resulted in depression of the adoptive secondary response of such cells, although the kinetics and degree of tolerance induction in this way were slightly different from that obtained by in vivo tolerization. Utilizing the adoptive transfer tolerance system, it was possible to approach certain questions concerning the mechanism of tolerance induction and fate of tolerant bone marrow-derived (B) lymphocytes in the DNP-D-GL model. The possibility that suppression of anti-DNP antibody from the DNP-D-GL reflects blocking of surface receptor molecules on B lymphocytes has been ruled out by several experimental observations. The most conclusive evidence on this point derives from the failure of enzymatic treatment with trypsin to reverse the tolerant state induced by in vitro exposure of primed cells to DNP-D-GL, whereas trypsinization completely restored the immunocompetence of DNP-KLH-primed cells rendered unresponsive by exposure to DNP-ovalbumin in vitro. The present studies also demonstrate that the tolerant state induced by DNP-D-GL represents a predominantly irreversible inactivation of specific B lymphocytes. This conclusion is derived from experiments in which it was found that tolerance was maintained through as many as two serial adoptive transfers performed over a period of time of at least 24 days from the single exposure of such cells to the tolerogen. Moreover, the possibility that maintenance of tolerance through such serial transfers was due to inadvertent transfer of tolerogenic doses of DNP-D-GL was definitively ruled out. It appears, therefore, that DNP-specific tolerance induced by DNP-D-GL is an example of irreversible inhibition of cell reactivity to antigen reflecting yet-to-be-determined events at the intra- and subcellular levels.

scholarly journals The mechanism of tolerance induction in thymus-derived lymphocytes; I. intracellular inactivation of hapten-reactive helper T lymphocytes by hapten-nonimmunogenic copolymer of D-amino acids.

1975 ◽  
Vol 141 (6) ◽  
pp. 1308-1328 ◽  
Author(s):  
T Hamaoka ◽  
U Yamaskita ◽  
T Takami ◽  
M Kitagawa
Keyword(s):  
T Cell ◽  
T Lymphocytes ◽  
Cell Activity ◽  
Tolerance Induction ◽  
Adoptive Cell Transfer ◽  
Helper T Cell ◽  
Cell Reactivity ◽  
Helper T Lymphocytes ◽  
Specific Tolerance

Treatment of a p-azobenzoate (PAB) derivative of a copolymer of D-glutamic acid and D-lysine (D-GL) induced a profound state of unresponsiveness to PAB-reactive helper T lymphocytes generated in PAB-mouse gamma globulin (MGG)-primed mice. This unresponsiveness in T lymphocytes was specific for PAB-reactive cells, since the bacterial alpha-amylase-, keyhole limpet hemocyanin-, or ovalbumin-primed helper T lymphocytes were not suppressed by PAB-D-GL treatment. Taking advantage of the relative ease with which PAB-D-GL can induce specific unresponsiveness to helper T lymphocytes in an animal previously primed with PAB-MGG, it was possible to approach certain questions concerning the mechanisms of tolerance-induction and the fate of tolerant helper T lymphocytes in the PAB-D-GL model by utilizing a classical adoptive cell transfer systemmelimination of the possibility of carry-over of the tolerogen with cells or of the generation of suppressor cells as the result of PAB-D-GL treatment as an explanation of the suppression of helper T-cell activity strongly inplicates the existence of a central intracellular mechanism of specific tolerance on the helper T-cell level. The possibility that suppression of the activity of PAB-reactive helper T lymphocytes by PAB-D-GL reflects simple blocking of surface receptor molecules on T lymphocytes was ruled out as it was found that the helper activity of PAB-reactive cells was minimally suppressed even when PAB-D-GL was directly exposed in vitro to helper T lymphocytesmmoreover, the most conclusive evidence on te the tolerant state induced by in vivo exposure of primed T cells to PAB-D-GL. It appears, therefore, that specific tolerance induced by PAB-D-GL' TO PAB-reactive helper T lymphocytes is an example of irreversible inhibition of T-cell reactivity to antigen, reflecting yet to be determined events at the intra- and subcellular levels.

scholarly journals Antigen recognition. IV. Discrimination by antigen-binding immunocompetent B cells between immunity and tolerance is determined by adherent cells.

1976 ◽  
Vol 143 (4) ◽  
pp. 805-821 ◽  
Author(s):  
E Diener ◽  
N Kraft ◽  
K C Lee ◽  
C Shiozawa
Keyword(s):  
B Cell ◽  
Short Pulse ◽  
Tolerance Induction ◽  
Mouse Spleen ◽  
Adherent Cells ◽  
Antigen Receptors ◽  
Spleen Cells ◽  
A Cells ◽  
Tolerant State

Mouse spleen cells capable of specifically binding intrinsically tritium-labeled polymerized flagellin (POL) (labeling by biosynthesis of flagellar protein) via IgM receptors were found to comprise a distinct population of about 20-50 cells per 10(6) lymphocytes. Evidence is presented that the majority of mouse spleen cells binding tritium-labeled POL undergoes blastogenesis after antigen capping, antigen shedding, and receptor reformation. Under conditions of tolerance induction in vitro, however, loss of antigen from the cell surface was inhibited. Such inhibition of antigen redistribution and shedding was reversed by a short pulse of colchicine and new antigen receptors were formed. In spite of this, colchicine had no effect on the tolerant state. However, tolerance could be broken, regardless of presence or absence of the alkaloid, with radioresistant theta-negative accessory (A) cells (adherent cells) from normal but not from tolerant spleen cell populations. "Tolerant" A cells, although they were incapable of cooperating in a response to POL, were capable of participating in a response to a second unrelated antigen. It is concluded that tolerance to POL in vitro is induced by mechanisms other than the physical blocking of bone marrow-derived (B) cell receptors by antigen. Most likely, the discrimination by the B cell between a tolerogenic and immunogenic signal is mediated by A cells.

Sign in / Sign up

Export Citation Format

Share Document