Biomedical subjects

A Coutinho

Publications and source records attributed to A Coutinho.

At least 55 records · Page 3Linked to original sources

PubMed1996

A model of the immune network with B-T cell co-operation. II--The simulation of ontogenesis.

This paper is based on a new model of the immune network which explicitly incorporates B-T cell co-operation. A major feature of this model is the simplifying assumption that inhibition by anti-TCR soluble Ig is the only possible down-regulatory influence on activated T-cells. This model is capable of coupling with antigens in both an "immune response" mode and a "tolerant" mode. In the present paper, we simulate the ontogenesis of the immune system by metadynamical recruitment of T- and B-cell clones from the thymus and the bone marrow, seeking to identify the conditions under which each of these modes of antigen coupling occurs. Achieving the tolerant mode depends principally on four parameters: a high value of SB, the rate of bone-marrow production of B-cells; a relatively high efficiency of T-help through mIg-TCR recognition compared with (MHC + peptide)-TCR interaction; and a relatively high value of the product PR.NA, where PR is the average probability that an Ig recognizes another molecule and NA is the number of antigens which are present throughout ontogeny. Analysis of the conditions under which these two modes can coexist, shows that this is possible when a sufficiently numerous set of founder antigens couple in a tolerant mode, whereas isolated antigens first presented once development is completed couple in an immune response mode. The present model thus provides a possible mechanism for the distinction (hitherto purely descriptive) between a Central Immune System organized as a network and responsible for tolerance, and a Peripheral Immune System responsible for immune responses.

B-Lymphocytes↗

PubMed1996

Quantitative analysis of multiple V-region interactions among normal human IgG.

There is to date no quantitative method for scoring putative V-region interactions among serum antibodies, and the available qualitative techniques are not amenable to routine utilization. This deficit may explain the paucity of observations on the characteristics of the immune network, in contrast with the multiplicity of phenomenological descriptions on idiotype regulation. We describe here a novel methodology that uses isoelectric focusing (IEF) to resolve human F(ab')2 preparations from large pools of normal serum IgG into multiple bands, and computer-aided data processing to analyze interactions between the resulting blotted proteins and normal serum IgG from individual donors. Our results show that in all normal human sera tested, there are IgG-mediated interactions with a large number of IEF fractions of human F(ab')2. These interactions are V region specific, as assessed by inhibition experiments and by lack of binding of IgG monoclonal antibodies, and are characterized by average affinities that are in the micromolar range, as measured by surface plasmon resonance.

Antibody Affinity↗

PubMed1996

Establishment of tissue-specific tolerance is driven by regulatory T cells selected by thymic epithelium.

Grafts of thymic epithelium (TE) rudiments restore T cell development and function in allogeneic athymic mice. These TE chimeras are specifically tolerant to grafts of peripheral tissues (e.g. skin and heart) from the TE donor strain, although they harbor peripheral immunocompetent T cells capable of rejecting those grafts. Initial analysis has shown that TE chimeras also harbor TE-selected CD4 T lymphocytes that inhibit graft rejection by tissue-reactive T cells in immunocompetent recipients. Peripheral tolerance in TE chimeras is thus maintained by dominant mechanisms dependent on regulatory CD4 T lymphocytes. Here we show that TE-selected regulatory T cells recruit nontolerant tissue-reactive CD4 and CD8 T cells to express similar regulatory functions. Only recent thymic emigrants, but not peripheral resident mature T cells are susceptible to this process of functional education, which also requires exposure to specific antigens and occurs entirely in the periphery. We propose that these mechanisms play a major role in establishing and maintaining natural self tolerance to tissue-specific antigens.

Animals↗

PubMed1996

Murine acariasis: I. Pathological and clinical evidence suggesting cutaneous allergy and wasting syndrome in BALB/c mouse.

We describe here a disease related to mite-associated ulcerative dermatitis in BALB/c mice, a strain previously classified as resistant to this condition. The disease was recognized by pruritic cutaneous pathology and wasting. Pathologic studies showed a marked allergic-type inflammation in the skin. The dominant histologic feature was extensive mast cell infiltration in cutaneous lesions and in lymphoid tissues, associated with a greatly elevated serum IgE concentration. The disease was secondary to infestation with an acarian ectoparasite Myocoptes musculinus, and seemed to represent an allergic reaction to the parasite-derived substances, with an associated wasting syndrome. This condition may be a useful experimental model for allergic diseases.

Animals↗

PubMed1996

Murine acariasis. II. Immunological dysfunction and evidence for chronic activation of Th-2 lymphocytes.

The authors describe the immunological profile of BALB/c mice with Mite-Associated Ulcerative Dermatitis (MAUD)-like disease, due to Myocoptes musculinus (Koch 1844) infestation. The disease probably involves allergic mechanisms and is characterized by erythematous and pruritic skin lesions, widespread hair loss, lymphadenopathy, lymphocytopenia, granulocytosis and wasting. Affected individuals had much reduced numbers of pre-B and B cells in bone marrow and B cells in blood; decreased T-cell numbers in peripheral lymphoid organs and blood; hypergammaglobulinaemia with selective increases of IgG1, IgE and IgA, and depletion on IgM and IgG3, the same isotype distribution being detected in splenic plasmocytes; qualitative modifications of the serum antibody reactivity pattern; and increased production of IL-4 with decreased IL-2 production after in vitro polyclonal stimulation of T cells. Taken together, these results suggest that infestation by M. musculinus in BALB/c mice leads to a significant immunological disorder resulting in a T-helper-2 (Th-2) type response, with marked systemic consequences. This pathological condition may thus provide a useful model system for the immunobiological perturbation associated with chronic allergic disease.

Animals↗

PubMed1996

The self-reactive antibody repertoire of normal human serum IgM is acquired in early childhood and remains conserved throughout life.

The authors have used a quantitative immunoblotting technique to analyse the antibody repertoire of IgM in cord blood and in the serum of young children, young adult males and aged males directed towards antigens in homologous tissues utilized as sources of self antigens. The reactivities of IgM with self antigens exhibited striking homogeneity and invariance among newborns. Self-reactive IgM repertoires of children, young adults and aged males were markedly conserved among individuals and comprised most of the anti-self reactivities that prevailed in neonates. Reactivities of IgM with bacterial antigens showed a high degree of homogeneity among newborns but were more diverse in children, young adults and elderly individuals. Diversity of IgM reactivities with self and non-self antigens did not vary significantly with aging. Principal component analysis (PCA) and linear discriminant analysis (LDA) of the data discriminated between self-reactive IgM repertoires of newborns and children, but failed to discriminate between repertoires of children, young adults and aged males. The data indicate that the self-reactive antibody repertoire of IgM differentiates during the first years of life and remains relatively constant thereafter.

Adult↗

PubMed1996

The age-associated increase in autoreactive immunoglobulins reflects a quantitative increase in specificities detectable at lower concentrations in young mice.

Serum immunoglobulins reactive with several autoantigens have been reported to increase with age. The authors have studied the reactivity of serum immunoglobulins from mice between 2 and 24 months of age with antigens present in lysates of syngeneic tissue extracts from young mice. The profile of immunoglobulin binding with the immunoblots of spleen and brain tissue increased progressively with age, showing only minor differences from mouse to mouse and, with one exception, revealing that the age-associated increase in binding of immunoglobulins occurred with antigens with the same migratory position in the immunoblots detectable, at lower concentration, in sera from young mice. Not all sera from older mice had increased immunoglobulin binding when tested with extracts of skin, muscle and liver but those that did expressed increased binding with antigens in all three lysates and with the same profile shown by sera from young mice. These results are consistent with the hypothesis that the age-associated increase in autoreactive immunoglobulins represents a selective increase in autoreactive specificities expressed by serum immunoglobulins from young animals at lower levels.

Aging↗

PubMed1996

Ig-isotype patterns of primary and secondary B cell responses to Plasmodium chabaudi chabaudi correlate with IFN-gamma and IL-4 cytokine production with CD45RB expression by CD4+ spleen cells.

In this work, the authors analysed T and B lymphocyte subsets and cytokine production in the spleen of BALB/c mice during polyclonal lymphocyte activation (primary infection) and parasite-specific response to Plasmodium chabaudi chabaudi (secondary infection). The secondary response was evaluated in fully immunoprotected animals, 60 days after a chloroquine-cured infection. The authors observed that in polyclonal lymphocyte activation antibody-secreting cells of all isotypes increased, with predominance of IgG2a and IgG3 classes. At that time, IFN-gamma was largely produced, but IL-4/IL-5 were just slightly enhanced. In mice re-infected after 60 days, the Ig-isotype pattern was restricted to IgG1 and only IL-4/IL-5 were produced. In both responses, however, the levels of IL-2 were greatly reduced, while those of IL-10 were enhanced to similar levels. The different involvement of Th1 and Th2 cells in both responses was confirmed through analysis of CD45RB expression by CD4+ cells. The authors observed that CD45RBhigh cells were the major CD4+ subpopulation in primary infected mice, while CD45RBlow cells predominated in 60 days re-infected animals. Moreover, the great majority of activated (large) CD4+ cells in the primary infection belonged to the CD45RBhigh subset, while after reinfection most of the CD4+ large had a CD45RBlow phenotype.

Animals↗

PubMed1996

Ultrasonographic assessment of the adulticidal efficacy of repeat high-dose ivermectin in bancroftian filariasis.

Since diethylcarbamazine, the drug recommended for treatment of lymphatic filariasis, seems only partially effective against the adult worm, intense interest persists in identifying a macrofilaricidal drug for this infection. To evaluate directly in vivo the macrofilaricidal activity of repeat high-dose ivermectin, 15 men who had living adult Wuchereria bancrofti detected in the scrotal area by ultrasound were treated with 400 micrograms/kg of ivermectin at 2-week intervals for 6 months (total dose, 4.8 mg/kg). Serial ultrasound examinations were performed before, during, and for 6 months after treatment. Profound suppression of microfilaraemia followed the first dose of ivermectin, but movements characteristic of the adult worm on ultrasound remained unchanged both in location and pattern. Even when given in total doses of 4.8 mg/kg, ivermectin appears to have no observable activity against adult W. bancrofti, although its ability to suppress microfilaraemia makes it potentially useful for the control of lymphatic filariasis.

Adult↗

PubMed1996

Rearrangement and expression of Vzeta1, Vzeta2 and Vzeta3 TCR zeta genes in C57BL/6 mice.

We have recently described a mAb (2.11) that recognizes the Vgamma1-Jgamma4-Cgamma4 chain. With this mAb and an anti-delta mAb we separated gammadelta+ 2.11(+) and gammadelta+ 2.11(-) intraepithelial lymphocytes (i-IEL) by FACS. Transcripts of rearranged TCR Vgamma1 and Vdelta2 genes in both i-IEL populations were analyzed by PCR followed by sequence analysis of cDNA spanning the junction of variable (V) and joining (J) genes. Roughly the same number of Vgamma1 and Vgamma2 transcripts were found in the 2.11(+) population while >90% of the transcripts in the 2.11(-)population contained a Vgamma gene sequence. Furthermore, gamma transcripts in the 2.11+ population were functional, while only 30-40% of the Vgamma transcripts in either population contained an in-frame sequence. The observed frequency of transcripts is what would be expected from cell populations that have not gone through cellular selection mediated by the RCR. Expression of Vgamma mRNA in RCRalphabeta and TCRgammadelta thymocytes was studied by a technique that analyzed populations of transcripts of rearranged genes. In both T cell population similar levels of Vgamma transcripts were found and about two out of three transcripts were out-of-frame. During the cloning and sequence analysis, we indentified a clone that expresses the Vgamma segment rearranged to the Jgamma3-Cgamma region in C57BL/6 mice. Together with the PCR cloning and sequencing of the complete Cgamma region in C57BL/6 mice these data demonstrate that the Jgamma-Cgamma gene is functional in this strain. Taken together, these studies revealed that: (i) cells expressing the Vgamma1 chain are an important subset of the gammadelta i-IEL population and that they show extensive junctional diversity; (ii) there is no correlation between expression of in-frame Vgamma transcripts and expression of Vgamma2 chains at the cell surface; and (iii) cells expressing the Vgamma3 chain might be a minor subset of the gammadelta T cell population in C57BL/6 mice.

Amino Acid Sequence↗

PubMed1996

B-lineage cell deficits in bone marrow of lpr/lpr mice.

Analyses of bone marrow (BM) lymphocytes in C57BL/6 mice homozygous for the lpr mutation (B6.lpr) disclosed low numbers of pre-B and B cells, as compared with age-matched control B6 mice. BM depletion in B6.lpr mice was selective for B-lineage cells, appeared in young adults, and developed markedly with age and disease progression, contrasting with the peripheral lymphocyte hypercellularity. Normalization of pre-B and B cellularity in BM of B6.lpr mice was observed after administration of polyclonal Ig, that also markedly improved the clinical condition. Isolated pre-B (B220+ IgM-) cells from B6 or B6.lpr mice, however, showed essentially the same rates of IL-7-dependent proliferation and differentiation to B (IgM+) cells in culture, indicating that the BM B-lineage deficit is not the result of an intrinsic defect in B cell generation.

Age Factors↗

PubMed1996

Ultrasonographic evidence of abnormal lymphatic vessels in young men with adult Wuchereria bancrofti infection in the scrotal area.

PURPOSE: We determined the prevalence and magnitude of dilatation of the lymphatic vessels of the spermatic cord in men infected with Wuchereria bancrofti, which is known major cause of hydrocele in the tropics. MATERIALS AND METHODS: Scrotal ultrasound was performed with a 7.5 MHz, transducer in 78 men from Recife, Brazil (endemic for filariasis) and in 15 from a nonendemic area. RESULTS: Among men from Recife the lymphatic vessels were dilated (1.3 to 15.0 mm., mean 3.8) at the location of the adult worm. Vessel diameter was not associated with hydrocele. CONCLUSIONS: Lymphatic dilation was observed in all men with ultrasonographically detectable W. bancrofti infection, even those who were asymptomatic.

Adolescent↗

PubMed1996

Studies on the periodicity and intravascular distribution of Wuchereria bancrofti microfilariae in paired samples of capillary and venous blood from Recife, Brazil.

We examined the periodicity and intravascular distribution of Wuchereria bancrofti microfilariae (mf) and determined the effect of these parasite properties on the accuracy of blood filming and filtration methods for diagnosis of bancroftian filariasis in the endemic area of Recife, Brazil. Microfilariae in both venous and capillary blood exhibited a nocturnal periodicity pattern with a relatively high amplitude. Overall, capillary blood contained approximately 1.25 times the number of mf present at the same time in the same volume of venous blood. However, the ratio of mf present in capillary and venous blood varied over a 24-hour period, so that the fewest mf were present in the capillary bed of the skin at the time when biting activity of the local Culex vector is the lowest. Twenty or 60 microliters blood films did not reliably detect carriers with fewer than 100 or 60 mf/ml venous blood, respectively, and were thus inadequate for the identification of low density mf carriers. In contrast, all carriers with > 1 mf/20 or 60 microliters blood smear at night could be identified during daytime hours by filtration of 1 micromilligram venous blood.

Animals↗

PubMed1996

Evidence for a thymus-dependent form of tolerance that is not based on elimination or anergy of reactive T cells.

The avian embryo has provided an appropriate model to study the ontogeny of the primary lymphoid organs, thymus and bursa of Fabricius. By using the quail-chick marker system the embryonic origin of the highly intricate cell components which form these organs could be traced back to the initial endodermal, mesodermal and ectodermal germ layers. The timing and dynamics of the incoming and outcoming flows of hemopoietic cells which characterize their lymphopoietic activity could be revealed in both quail and chick embryos. This knowledge served as a basis for an investigation on the role of the epithelial component of the thymus (derived from the pharyngeal endoderm) on tolerance to tissue graft and, by extension, tolerance to self. When this work was undertaken, the prevailing view was that exposure of the developing immune system to foreign antigens in the embryo allows them to be assimilated to self components in the mature animal. In fact, this was found to be true for allogeneic grafts between MHC-distinct chickens, of certain tissues, such as for instance wing tissues. However, in heterospecific transplantations, i.e. when a limb bud was grafted from quail to chick embryos, the chick host acutely rejected the foreign limb soon after birth. In contrast, grafts of the quail thymic epithelial (TE) rudiment resulted in the development of a chimeric thymus in which the foreign epithelial component was not only tolerated but able to induce full tolerance of the grafted wing from the same donor. By monitoring the amount of quail TE implanted we showed in addition that only part of the peripheral T-cell population had to differentiate in the context of the quail epithelial cells to induce tolerance to quail tissues. This pointed to the generation in the thymus of regulatory T cells, coexisting with specific anti-quail reactive T cells, but able to inhibit them from reacting against the quail wing antigenic determinants. A mammalian model was then devised to further study this mechanism of tolerance that we have qualified as "dominant" by opposition to the current model based on either clonal elimination or anergy which can be considered as recessive or passive. Nude mice of MHC type A were grafted with TE of E10 type B embryos. They became reconstituted for T-cell function but tolerant for B skin allografts. Spleen cells from such tolerant animals injected to naive A nude mice reconstituted T cell function in the recipient and transferred the tolerance to B skin grafts. Reducing the number of donor cells resulted in the segregation of the two phenomena. For low numbers the recipients were restored but not tolerant, thus showing the coexistence in the tolerant donor of anti-B reactive T cells together with regulatory cells able to abolish their reactivity against B determinants. Other experiments demonstrated that TE-induced tolerance does not rely on clonal deletion or anergy. This was shown on systems where elimination of cells directed toward superantigens was screened. It turned out that tolerance to skin grafts and superantigen T-cell deletion are unrelated phenomena. These observations strongly suggest that tolerance to self results at least in part from the interplay between cells potentially harmful for self component and others which exert a strong control on their reactivity. The latter cell type depends upon interactions of thymocytes with the endodermal component of the thymus.

Animals↗

PubMed1996

A model for developmentally acquired thymus-dependent tolerance to central and peripheral antigens.

Current models of tolerance to peripheral, tissue-specific antigens contain some major caveats. First, they consider peripheral tolerance independently from intrathymic T cell selection, a dichotomy that is challenged by observations on TE-induced tolerance. Second, they do not account for the fact that vertebrates are more readily tolerised in development than in adult life. Third, they do not explain the fact that embryonic/neonatal tolerance to foreign tissues can only be induced by HC or TE. A model of thymic selection and peripheral tolerance is developed here that resolves those problems, by assuming two classes of T cell effector functions, one being regulatory and the other aggressive. Three postulates are required: (1) both epithelial and hemopoietic cellular compartments of the thymic stroma can support both positive and negative selection of T cells, but with vastly different avidity requirements and efficiency; (2) positively selected T cells with the highest avidity that escape deletion are activated intrathymically and irreversibly committed for regulatory effector functions; (3) the functional phenotype of all other thymic emigrants is determined in the periphery upon encounter with antigen. Functional commitment in the periphery depends on the maturity stage (RTE or PMR) of the immunocompetent cell, on the nature of the antigen-presenting cells, and on the effector classes of other T lymphocytes interacting on the same presenting cell. This model explains a number of observations on experimental autoimmune disease and transplantation tolerance, and it contains several readily testable predictions.

Animals↗

PubMed1995

Lymphocytes selected in allogeneic thymic epithelium mediate dominant tolerance toward tissue grafts of the thymic epithelium haplotype.

Athymic mice grafted at birth with allogeneic thymic epithelium (TE) from day 10 embryos before hematopoietic cell colonization reconstitute normal numbers of T cells and exhibit full life-long tolerance to skin grafts of the TE haplotype. Intravenous transfers of splenic cells, from these animals to adult syngeneic athymic recipients, reconstitute T-cell compartments and the ability to reject third-party skin grafts. The transfer of specific tolerance to skin grafts of the TE donor strain, however, is not observed in all reconstituted recipients, and the fraction of nontolerant recipients increases with decreasing numbers of cells transferred. Furthermore, transfers of high numbers of total or CD4+ T cells from TE chimeras to T-cell receptor-anti-H-Y antigen transgenic immunocompetent syngeneic hosts specifically hinder the rejection of skin grafts of the TE haplotype that normally occurs in such recipients. These observations demonstrate (i) that mice tolerized by allogeneic TE and bearing healthy skin grafts harbor peripheral immunocompetent T cells capable of rejecting this very same graft; and (ii) that TE selects for regulatory T cells that can inhibit effector activities of graft-reactive cells.

Animals↗

PubMed1995

Analysis of the normal human IgG antibody repertoire. Evidence that IgG autoantibodies of healthy adults recognize a limited and conserved set of protein antigens in homologous tissues.

We have used a quantitative immunoblot technique to analyze the Ab repertoire of IgG in the serum of healthy adults with a large panel of homologous and foreign Ags in tissue extracts. Densitometric patterns of reactivity of purified IgG with self-Ags exhibited striking homogeneity among individuals with regard to the protein bands that were recognized. Purified IgG showed higher levels of reactivity with self-Ags than IgG in whole serum. Reactivity with self-Ags of IgG in whole serum was restricted to a small number of protein bands (fewer than 10). There were significant inter-individual differences in the intensity and nature of immunoreactivities. Purified IgG of different individuals exhibited heterogeneous patterns of immunoreactivity with Ags in bacterial extracts. Comparative analysis of repertoires of IgG and IgM indicated that all protein Ags recognized by IgM were also reactive with purified IgG. Some proteins in homologous and in foreign tissue extracts reacted solely with IgG. Our observations provide direct evidence for the restricted and conserved character of the antiself-repertoire of IgG of healthy adults and suggest that natural IgG Ab repertoires in serum are specifically selected for reactivity with a limited set of self-Ags. In addition, IgG autoreactivity in whole serum is controlled by non-IgG factors that determine the unique reactivity pattern of each individual.

Adult↗

PubMed1995

Invariance and restriction toward a limited set of self-antigens characterize neonatal IgM antibody repertoires and prevail in autoreactive repertoires of healthy adults.

Analysis of the reactivity of IgM with self-antigens in tissues by a quantitative immunoblotting technique showed striking invariance among newborns in the human and in the mouse. The self-reactive repertoire of IgM of adults was also markedly conserved; it comprised most anti-self reactivities that prevailed among neonates. Multivariate analysis confirmed the homogeneity of IgM repertoires of neonates toward self- and non-self-antigens. Multivariate analysis discriminated between newborn and adult repertoires for reactivity with two of five sources of self-proteins and with non-self-antigens. Our observations support the concept that naturally activated B lymphocytes are selected early in development and throughout life for reactivity with a restricted set of self-antigens.

Adult↗