Biomedical subjects

B Rocha

Publications and source records attributed to B Rocha.

At least 37 records · Page 2Linked to original sources

PubMed1997

T cell deletion induced by chronic infection with mouse mammary tumor virus spares a CD25-positive, IL-10-producing T cell population with infectious capacity.

We found that T cells recognizing viral superantigen (vSAG) can be subdivided into two distinct functional subsets based on IL-2R alpha (CD25) expression. CD4+Vbeta6+CD25- and CD4+Vbeta6+CD25+ T cells were sensitive to vSAG activation. When obtained from BALB/c(SW) mice, both subsets were infected and capable to induce the tolerance process when transferred into noninfected recipients. However, in contrast to CD4+Vbeta6+CD25- cells, which were gradually deleted in MMTV(SW)-infected mice, the pool of CD4+Vbeta6+CD25+ lymphocytes was constant even at the end of the deletion process, and maintained a limited reactivity to vSAG-induced activation. The constant number of Vbeta6+CD25+ observed in infected mice could not be explained by their rapid turnover (deletion and renewal), as their proliferative rate measured by BrdU incorporation was similar in infected and naive mice, as well as in virus-nonspecific (Vbeta8.2+) cells. Neither was the Vbeta6+CD25+ subset dependent on vSAG activation since it was also present in MMTV-free mice and was not generated from Vbeta6+CD25- cells upon in vivo vSAG stimulation. Vbeta6+CD25+ T cells constitutively expressed IL-4 and IL-10 mRNA. IL-10 has been shown to be associated with viral, bacterial, and parasitic infections. This permanent CD25+ subpopulation may play a role in the control of viral infection and tolerance induction via vSAG recognition and IL-10 production.

Animals↗

PubMed1997

Lymphocyte homeostasis.

B- and T-lymphocyte populations have an independent homeostatic regulation of resting (B and T) and activated (B) or memory (T) cell compartments. This organization may provide an efficient mechanism to ensure simultaneously a first natural barrier of protection against common pathogens, the maintenance of immunological T-cell memory and a reservoir of repertoire diversity capable of dealing with new antigenic challenges.

Animals↗

PubMed1995

Persistence of V beta 6+ T cells in Mls-1a mice. A role for the third complementarity-determining region (CDR3) of the T cell receptor beta chain in superantigen recognition.

We have studied V alpha 2 and J beta usage by V beta 6+CD4+ peripheral T cells isolated from the congenic mice strains BALB/c (Mls-1b) and BALB.D2 (Mls-1a). We found that the TCR beta-chain of V beta 6+CD4+ T cells present in adult Mls-1a mice differed from those in Mls-1b mice; the fraction of V beta 6+CD4+T cells using the J beta 2.7 segment was reduced, while the number of V beta 6+CD4+ T cells using J beta 1.2 was augmented. These results indicate that the CDR3 region of the TCR beta-chain participates in recognition of the Mls superantigen. We also found that in Mls-1a mice an increased fraction of V beta 6+CD4+ T cells expressed the V alpha 2 chain. The study of J beta usage by V beta 6+CD4+V alpha 2+ and V beta 6+CD4+V alpha 2- T cells indicates that both J beta segment and TCR V alpha 2 chain expression confer complementary protection against deletion by Mls-1a superantigen. These results suggest a novel view of Mls-1a-driven selection, where the CDR3 region of the V beta chain modulates superantigen recognition, and the affinity/avidity of the TCR-MHC-superantigen complex determine the fate of the T cell.

Animals↗

PubMed1995

Anergy and exhaustion are independent mechanisms of peripheral T cell tolerance.

We studied the interactions of male-specific T cell receptor (TCR)-alpha/beta-transgenic (TG) cells with different concentrations of male antigen in vivo. We constructed mouse chimeras expressing different amounts of male antigen by injecting thymectomized, lethally irradiated mice with various ratios of male (immunoglobulin [Ig] Ha) and female (IgHb) bone marrow. These chimeras were injected with male-specific TCR-alpha/beta-trangenic cells. These experiments allowed us to monitor antigen persistence and characterize antigen-specific T cells in terms of their frequency, reactivity, and effector functions (as tested by elimination of male B cells in vivo). In the absence of antigen, virgin TG cells persisted but did not expand. Transient exposure to antigen resulted in cell expansion, followed by the persistence of increased numbers of antigen-reactive T cells. In contrast, antigen persistence was followed by two independent mechanisms of tolerance induction: anergy (at high antigen concentrations), where T cells did not differentiate into effector functions but persisted in vivo as unresponsive T cells, and exhaustion (at lower antigen concentrations), where differentiation into effector functions (B cell elimination) occurred but was followed by the disappearance of antigen-specific T cells.

Animals↗

PubMed1995

Intra- and extra-thymic expression of the pre-T cell receptor alpha gene.

We have analyzed pre-T cell receptor alpha (pT alpha) gene expression in cells from various anatomical sites to investigate the lineage specificity of pT alpha RNA as well as its presence in pro-T cells and in sites of extrathymic T cell development. pT alpha RNA is found in precursors of alpha beta T cells but is absent from mature alpha beta T cells as well as T cells that express the gamma delta T cell receptor on the cell surface. pT alpha expression is exquisitely T lineage specific in that mature and immature B cells, myeloid cells, NK cells and pluripotent stem cells are pT alpha negative. On the other hand, pT alpha expression is found in pro-T cells outside the thymus as well as in intra- and extra-thymic sites of T cell development. The latter finding is consistent with the notion that early steps of T cell development within and outside the thymus may be similar.

Animals↗

PubMed1995

The peripheral T cell repertoire: independent homeostatic regulation of virgin and activated CD8+ T cell pools.

Mature T cells may be produced in the thymus, or by expansion in the periphery. While thymus output of virgin cells ensures repertoire diversity, peripheral expansion increases the size of rare clones, and thus the efficiency of immune responses. We studied the role of both phenomena in the generation of the CD8+ T cell pool using RAG-/- female mice expressing a transgenic T cell receptor specific for the male antigen; nude mice injected with peripheral T cells; and euthymic irradiated chimeras injected with bone marrow and mature T cells. Our results show that the total number of virgin and activated T cells, each constituting about half of the peripheral T cell pool, was regulated independently, revealing an efficient mechanism to maintain repertoire diversity while optimizing the immune response.

Allergy and Immunology↗

PubMed1995

Extrathymic T cell differentiation.

In the mouse, the gut mucosa is a major site of extrathymic differentiation of T cells. Recent data in this past year show that this process differs from the main thymic differentiation pathway not only in its location, but also in its use of costimulatory molecules, signal transduction modules, and mechanisms of repertoire selection. The thymus exerts an influence on the expansion of the extrathymically differentiated gut intraepithelial lymphocytes that appears to be varied in nature, including acting as a source of TCR- progenitors. All gut intraepithelial lymphocytes, whatever their extrathymic or thymic site of differentiation, have common features of activated and specialized cytotoxic cells. Other T cells may differentiate extrathymically, in particular in the liver; these later cells appear to have a very restricted, probably autoreactive repertoire, and also display natural killer cell features.

Animals↗

PubMed1994

Chronic mianserin or eltoprazine treatment in rats: effects on the elevated plus-maze test and on limbic 5-HT2C receptor levels.

Rats were chronically treated with mianserin (10 mg/kg i.p.) or eltoprazine (1 mg/kg i.p.) and were tested in the elevated plus-maze test for anxiety. 5-HT2C (previously 5-HT1C, see Humphrey et al., 1993, Trends Pharmacol. Sci. 14, 223) binding sites and their mRNA were evaluated in limbic structures (i.e., amygdala, hippocampus, septum) of a sample of these rats by autoradiographic binding studies and in situ hybridization histochemistry. Mianserin and eltoprazine displayed opposite effects in the elevated plus-maze: mianserin induced anxiolytic-like effects, while eltoprazine showed anxiogenic-like ones. Within the amygdala, but not in other structures, the quantitative autoradiographic analysis of the 5-HT2C binding sites showed a differential effect: mianserin treatment induced a decrease in the number of these sites, while eltoprazine treatment resulted in an increase. In spite of this, neither mianserin- nor eltoprazine-treated rats displayed an alteration in the 5-HT2C receptor mRNA levels in the brain regions examined. Our results are suggestive of a relation between anxiolytic/anxiogenic-like effects and the level of 5-HT2C binding sites in the amygdala.

Acetylcholinesterase↗

PubMed1994

Different use of T cell receptor transducing modules in two populations of gut intraepithelial lymphocytes are related to distinct pathways of T cell differentiation.

Most gut intraepithelial cells (IEL) of the mouse are T cells that bear CD8 molecules, present either as alpha-beta chain heterodimers (CD8 beta+) or as alpha chain homodimers (CD8 beta-). All CD8 beta+ IEL bear alpha/beta T cell receptors (TCR); CD8 beta- IEL bear either alpha/beta or gamma/delta TCR and are considered to be a thymus-independent (TI) population, probably arising locally from a small fraction of CD3- IEL containing the recombinant activating gene RAG proteins. Here we report that TI CD8 beta- IEL, whether bearing alpha/beta or gamma/delta TCR, contain, in normal mice, mRNAs for both zeta and Fc epsilon RI gamma chains. These chains are present in their CD3-TCR complexes as homo- or heterodimers. In contrast, only zeta chain mRNA and homodimers are found in gut CD8 alpha/beta+ IEL and in peripheral T lymphocytes. Intestinal CD3- precursor cells contain only gamma chain, and CD3- IL-2R+ thymocyte precursors only zeta chain mRNAs. Only very primitive thymocyte precursors contain detectable gamma chain mRNA, and it thus appears that Fc epsilon RI gamma chain use is switched off at a very early stage during thymocyte differentiation. Thus, T cell differentiation in the gut epithelium differs from that occurring in the thymus, from which CD8 beta+ IEL appear to derive. Use of different TCR transducing modules and CD8 accessory molecules between the TI and the thymus-derived T cell populations provides an explanation for their difference in reactivity to antigenic stimulations and thus in selection of repertoires.

Animals↗

PubMed1994

Thymic and extrathymic origins of gut intraepithelial lymphocyte populations in mice.

We have investigated the origin of intraepithelial lymphocytes (IEL) populations in the murine gut, using reconstitution experiments in which the presence of thymus-derived cells of host or donor origin is rigorously controlled: RAG-/- mutant mice which have no T cells, were injected either with the bone marrow (BM) cells of nude mice or with selected peripheral lymph node (LN) T cells of euthymic mice. In thymectomized RAG-/- mice, injection of BM cells from nude mice led, after 2 mo, to the development of a peripheral B cell compartment and to the appearance, in the gut, of IEL bearing homodimeric CD8 alpha chains and either gamma/delta or alpha/beta TCR. In RAG-/- mice with a thymus, a similar injection led to complete lymphoid reconstitution, with the additional appearance in the gut of CD4+, CD8 alpha/beta+ or CD4+CD8 alpha/alpha+ IEL, all bearing alpha/beta TCR. In contrast, injection of LN T cells into these mice reconstituted a gut IEL population made of CD4+, CD8 alpha/beta+, or CD4+ CD8 alpha/alpha+ cells, all bearing alpha/beta TCR; CD8 alpha/alpha+ TCR-gamma/delta+ or alpha/beta+ IEL were not observed. These results demonstrate that the thymus and/or thymic-derived peripheral T cells are absolutely required for the generation of CD4+, CD8 alpha/beta+, and CD4+CD8 alpha/alpha+ IEL, which are thus thymus dependent. In contrast, TCR+ CD8 alpha/alpha+ IEL appear in the absence of the thymus, and thus are thymus independent.

Animals↗

PubMed1994

Generation of mature T cell populations in the thymus: CD4 or CD8 down-regulation occurs at different stages of thymocyte differentiation.

We have studied the differentiation and repertoire selection during the maturation of CD4+CD8+ (DP) thymocytes into CD4+CD8- (CD4SP) and CD8+CD4- (CD8SP) T cells, in normal mice, mice transgenic for T cell receptor (TcR)-alpha beta restricted by either class I or class II major histocompatibility (MHC), and in mice deficient in class I or class II MHC expression. Our data suggest that mature CD4 and CD8 T cells derive from different pathways of T cell differentiation in the thymus. Thus, interaction of DP thymocytes with MHC class II leads to the immediate down-regulation of CD8, which occurs simultaneously with an increase in TcR expression; DPTcR(lo)HSA(hi) thymocytes mature into a CD4+CD8(lo) TcR(hi)HSA(hi) intermediate population. This cell population generates CD4SP thymocytes, the majority of which are still HSA(hi). In contrast, interaction with MHC class I induces the up-regulation of TcR, which precedes the down-regulation of CD4; DPTcR(lo) generate DPTcR(hi) thymocytes, the majority of which are the committed precursors of CD8SP cells. Further differentiation results in CD4 down-regulation and the transition from DPTcR(hi) into CD8+CD4(lo) TcR(hi)HSA(lo) and +D8SPTcR(hi)HSA- T cells. Since down-regulation of CD4 and CD8 occurs at different stages of thymocyte differentiation, our results do not support a stochastic/selective model of lineage commitment in the thymus.

Animals↗

PubMed1994

Activation and 'deletion' of self-reactive mature and immature T cells during ontogeny of Mls-1a mice: implications for neonatal tolerance induction.

We assessed the kinetics of V beta 6+ T cell elimination in the lymph nodes and thymus during Mls-1a mouse ontogeny. Our results suggest that induction of tolerance to Mls-1a antigens involves mechanisms other than clonal deletion of immature T cells in the thymus. Mature CD4+CD8- (CD4SP) T cells were affected by Mls-1a antigens earlier than immature thymocyte populations. Up to 2 weeks after birth, reduced frequencies of V beta 6+ T cells were detected only in CD4SP cells from the thymus and lymph nodes, and generation of CD4SP cells in the thymus was blocked at least 1 week earlier than that of their CD4+CD8loTCRhi immature precursors. The number of V beta 6+CD4SP T cells increased during the first 2 weeks of life and remained constant thereafter. We thus found no evidence of deletion of mature V beta 6+CD4SP T cells, as the reduced frequencies in adult mice can be attributed to the dilution of previously generated cells in lymphoid organs of growing mice, which increase in cellularity after birth. V beta 6+CD4+ T cells were activated in vivo shortly after birth, as shown by a selective increase in IL-2 receptor alpha chain expression in the thymus and lymph nodes from day 0 to day 2 after birth. It is therefore likely that endogenous expression of Mls-1a antigen shortly after birth activates V beta 6+CD4SP T cells and renders them anergic. This process of tolerance induction may precede the clonal deletion of immature T cells in the thymus, described in the adult mouse.

Animals↗

PubMed1993

What brain structures are active during emotions? Effects of brain stimulation elicited aversion on c-fos immunoreactivity and behavior.

Aversive behavior is produced by stimulating some brain structures, such as the dorsal periaqueductal gray and the medial hypothalamus. We have used c-fos immunoreactivity to map brain areas which are influenced by stimulation of these two structures. Stimulation was produced in freely moving rats by electrical stimulation or by microinjections of either excitatory amino acids or GABA blocking drugs. Behavior was monitored to detect emotional changes. The effects on labeling induced by the stimulation of either structure were then compared. Structures labeled include the amygdala, the stria terminalis, the supramamillary area, the hypothalamus, the periaqueductal gray, the superior colliculus, the nucleus cuneiformis, and the locus coeruleus. Regardless whether chemical or electrical stimulation was used or the structure stimulated, there was a large overlap among the brain areas labeled. We then compared our results with data from the literature where other methods of inducing aversion have been used, including pain and stress. There was remarkable similarity in the patterning of labeling irrespective of the type of stimulation (central-peripheral, chemical-electrical). There was, however, one interesting difference produced by central vs. peripheral stimulation. Labeling was unilateral in the former case and bilateral in the latter case. Our results suggest that there is a neural substrate that mediates aversive behavior, no matter how it is produced. Nevertheless, that peripheral stimulation produces mainly bilateral activation of this substrate whereas central stimulation produces mainly unilateral activation suggests that natural peripheral stimuli are also integrated at a higher functional level. Future work could be directed toward explicit comparisons of central versus peripheral stimulation to identify the structures involved in higher level integration of aversive behavior.

Animals↗

PubMed1993

Clonal anergy blocks in vivo growth of mature T cells and can be reversed in the absence of antigen.

Experiments in various models have indicated that immunological tolerance can result from the physical elimination (deletion) of reactive lymphocytes as well as from anergy. We have previously reported that mature CD4-CD8+ T cells when confronted with their antigen can proliferate extensively but are finally eliminated or become intrinsically anergic such that remaining cells are refractory to stimulation by any T cell receptor ligands, even in the presence of exogenous interleukin 2. Here we show that in vivo the anergy can be reversed in the absence of antigen, such that the cells are then able to proliferate extensively in vivo to a new challenge with the antigen in question.

Animals↗

PubMed1993

T cell development in mice lacking the CD3-zeta/eta gene.

The CD3-zeta and CD3-eta polypeptides are two of the components of the T cell antigen receptor (TCR) which contribute to its efficient cell surface expression and account for part of its transducing capability. CD3-zeta and CD3-eta result from the alternative splicing of a single gene designated CD3-zeta/eta. To evaluate the role of these subunits during T cell development, we have produced mice with a disrupted CD3-zeta/eta gene. The analysis of thymocyte populations from the CD3-zeta/eta-/- homozygous mutant mice revealed that they have a profound reduction in the surface levels of TCR complexes and that the products of the CD3-zeta/eta gene appear to be needed for the efficient generation and/or survival of CD4+CD8+ thymocytes. Despite the almost total absence of mature single positive thymocytes, the lymph nodes from zeta/eta-/- mice were found to contain unusual CD4+CD8- and CD4-CD8+ single positive cells which were CD3-. In contrast to the situation observed in the thymus, the thymus-independent gut intraepithelial lymphocytes present in zeta/eta-/- mice do express TCR complexes on their surface and these are associated with Fc epsilon RI gamma homodimers. These results establish an essential role for the CD3-zeta/eta gene products during intrathymic T cell differentiation and further emphasize the difference between conventional T cells and thymus-independent gut intraepithelial lymphocytes.

Animals↗

PubMed1993

Evidence for state-dependent retrieval in conditioned place aversion.

State-dependent retrieval (SDR) in conditioned place aversion (CPA) was observed using Long-Evans male rats, with three different aversive drugs injected ip: lithium chloride (31.8 mg/kg), FG 7142 (10 mg/kg), and naloxone (5 mg/kg). Experiment 1 showed that state-dependent dissociation was complete with lithium chloride following two conditioning trials, but disappeared when the number of learning sessions was increased. Experiment 2 showed that following two conditioning trials, such complete state-dependent dissociations were also observed when FG 7142 or naloxone were used during conditioning. The results of this study showed that the expression of SDR is not monotonic in CPA paradigm but depends on the stage of learning. In addition, SDR can be observed following administration of a variety of drugs known to produce CPA, but differing in their neurochemical targets and the nature of the aversive effects they induce. Finally, these results are discussed with regard to what is known about SDR in other paradigms.

Animals↗

PubMed1993

Effect of 5,7-dihydroxytryptamine lesion on mianserin-induced conditioned place aversion and on 5-hydroxytryptamine1C receptors in the rat brain.

The lesion of serotonergic neurons (by an intraventricular injection of 5,7-dihydroxytryptamine) potentiated the conditioned place aversion induced by the 5-hydroxytryptamine1C/5-hydroxytryptamine2 antagonist mianserin in rats. This effect was selective for mianserin as the same lesion suppressed the conditioned place aversion induced by the benzodiazepine inverse agonist FG-7142. Previous results had shown the involvement of the 5-hydroxytryptamine1C receptors in the conditioned place aversion induced by mianserin [Rocha et al. (1993) Behav. Pharmac. 4, 101-106]. It was thus of interest to investigate the effect of the lesion on these receptor binding sites. Autoradiographic binding studies showed that the lesion significantly increased the concentration of the 5-hydroxytryptamine1C binding sites in various brain regions, including the amygdala, the hippocampus and the nucleus accumbens. Contrastingly, in these same brain regions, in situ hybridization histochemistry did not reveal an alteration of the level of messenger RNA coding for these receptors. On the one hand, correlating potentiation of the aversive effects of mianserin and increase of 5-hydroxytryptamine1C binding sites in the limbic system represent an interesting step in the comprehension of the molecular and motivational effects of serotonergic drugs. On the other hand, showing a dissociation between the expression of 5-hydroxytryptamine1C receptors and their corresponding messenger RNA, suggest that post-transcriptional mechanisms are involved in the regulation of these receptors.

5,7-Dihydroxytryptamine↗

PubMed1993

Tissue distribution of Mtv-7-like exogenous retroviral transcripts and clonal deletion of V beta 6+ T cells in Mls-1b BALB/c mice.

We have previously described an Mls-1a-like clonal deletion of mature CD4+ T cells which express V beta 6 and V beta 8.1 chains of the TCR in half of the mice of a BALB/c, Mls-1b colony (BALB/c IC). This occurs in the absence of the Mtv-7 provirus which is responsible for the clonal deletion in Mls-1a mice. We developed a polymerase chain reaction assay in order to study the presence of retroviral transcripts homologous to the viral superantigen gene (vSAG) of Mtv-7 and Mtv-6 in various tissues. Mtv-7 homologous transcripts were present in the mammary glands of lactating BALB/c IC mice and in the thymuses and/or spleens of BALB/c IC virgin mice with deletion of V beta 6+ lymph node T cells, and not in BALB/c IC with normal V beta 6 expression. These results indicate that this BALB/c colony is infected with an exogenous mouse mammary tumour virus (MMTV) whose vSAG is similar to Mtv-7, as recently reported. Thymectomies performed at 4-5 weeks of age (at least 4 weeks before detection of clonal deletion), did not affect the occurrence of clonal deletion in peripheral lymph nodes when tested 20 weeks later. This suggests that clonal deletion can be achieved without further intrathymic contact with the antigen. Since MMTV is transmitted through milk and is likely to be present in the gut, we evaluated the percentage of V beta 6+CD4+ T cells within the gut intraepithelial lymphocyte (IEL) population. Mice with normal V beta 6 expression in lymph nodes may show partial deletion of V beta 6+CD4+ IEL.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗