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Biomedical subjects

A A Freitas

Publications and source records attributed to A A Freitas.

At least 19 recordsLinked to original sources
PubMed2005

Predicting hydration free energies of neutral compounds by a parametrization of the polarizable continuum model.

A parametrization of the polarizable continuum model (PCM) is presented having the experimental hydration free energies of 215 neutral molecules as target. The cavitation and dispersion contributions were based on the Tuñon-Silla-Pascual-Ahuir (Tuñon; et al. Chem. Phys. Lett. 1993, 203, 289) and Floris-Tomasi (Floris, F.; Tomasi, J. J. Comput. Chem. 1989, 10, 616) expressions, respectively. Both the polar and nonpolar contributions were evaluated on the same solvent-excluding molecular surface that used unscaled Bondi atomic radii. The parametrization was provided for the HF, Xalpha, LSDA, B3LYP, and mPW1PW91 methods at the 6-31G(d) basis set, and the results are in fair agreement with the experimental data. For the sake of comparison, the PCM(UAHF) and our parametrization (PCM2), both at HF level, have produced DeltaG(PCM(UAHF)) = aDeltaGexp (a = 1.02 +/- 0.02, r = 0.945, sd = 0.987, Ftest = 1778) and DeltaG(PCM2) = aDeltaGexp (a = 0.95 +/- 0.02, r = 0.952, sd = 0.843, Ftest = 2070), respectively. The mean absolute deviations from experimental data were 0.67 and 0.68 kcal/mol for PCM(UAHF) and PCM2, respectively.

Journal Article↗
PubMed2001

CD8+ T lymphocytes in double alpha beta TCR transgenic mice. I. TCR expression and thymus selection in the absence or in the presence of self-antigen.

We derived Rag2-deficient mice bearing two rearranged alphabeta TCR transgenes, one specific for the HY male Ag and the second specific for the gp33-41 peptide of lymphocytic choriomeningitis virus, both restricted to the MHC H-2D(b) class I molecule. We found that, in female double transgenic (DTg) mice, most CD8 T cells express only the TCRbeta chain from the aHY transgene. By comparing the mRNA species for both beta-chains, we observed that in T cells from DTg mice the aHY TCRbeta chain transcripts are abundant, whereas the anti-lymphocytic choriomeningitis virus TCRbeta chain transcripts are rare. In contrast to TCRbeta chain expression, most of the T cells from DTg mice express two TCRalpha chains. We examined the thymus selection of the dual-receptor CD8 T cells in the presence of self-Ag. We found that the presence of a second TCRalpha chain allows a significant number of CD8 T cells expressing a self-reactive receptor to escape central deletion and migrate to the peripheral pools of male mice. Differences in TCR and coreceptor expression between female and male MoaHY and DTg mice suggest that peripheral T cell survival requires an optimal level of signaling, which implies a process of "adaptation" of lymphocyte populations to the host environment.

Animals↗
PubMed2001

CD8+ T lymphocytes in double alpha beta TCR transgenic mice. II. Competitive fitness of dual alpha beta TCR CD8+ T lymphocytes in the peripheral pools.

We studied Rag2-deficient mice bearing two rearranged alphabeta TCR transgenes, both restricted to the MHC H-2D(b) class I molecule. We have previously shown that, in these DTg mice, most peripheral CD8 T cells express one TCRbeta chain associated with two TCRalpha chains, as in one-third of the mature T cells from normal mice. We examined the functional behavior of the dual-receptor CD8 T cells developing either in the absence or in the presence of self-Ag. The dual-receptor CD8 T cells, which develop in absence of self-Ag, show efficient responses to immunization and remain sensitive to induction of peripheral tolerance. In contrast to single TCR T cells, the dual-TCR cells, when tolerized upon exposure to high levels of self-Ag, are not deleted and therefore may exert important regulatory functions. When developing in the presence of self-Ag, the dual-receptor-expressing CD8 T cells escape central deletion, but are not fully competent to respond to cognate stimuli. Overall, we found that the dual-TCR CD8 T cells show a poor competitive value and can be out-competed by single-TCR cells, both in the course of immune responses and in reconstitution experiments. The decreased fitness of the dual-receptor cells may contribute to diminishing the autoimmune hazard that they could represent.

Animals↗
PubMed2001

Resource competition determines selection of B cell repertoires.

Previous experiments with mouse chimeras demonstrated that cellular competition for antigen-specific survival signals plays a crucial role in the maintenance of the naive B cell repertoire. Transgenic (Tg) B cell populations in these chimeras have a shortened lifespan and poor competitive abilities as compared to more diverse non-Tg populations in the same mice. We develop a mathematical model to investigate the mechanism of B cell competition. The model allows for various B cell clones, generated in the bone marrow, to go into the peripheral circulation, where they compete specifically for various ligands providing survival signals. In the model we also find the observed poor competitive abilities of the Tg repertoire. Investigating the nature of the competition in the model, we find that most of the competition is "intraspecific" occurring largely within the clone of truly Tg B cells, and within the repertoire of leaky Tg and non-Tg B cells. This is confirmed by analysing a simplified version of the model, which only allows for intraspecific competition, and resembles a simple ecological model with density-dependent death. The fact that our model accounts for the data, casts doubt on a previous interpretation of the same data arguing that more diverse repertoires outcompete repertoires of lower diversity. Here, we conclude that most of the data can be explained with intraspecific competition, and formulate an experimental prediction that allows one to distinguish between the previous interpretation of inter-specific competition between repertoires, and the current interpretation of intraspecific competition.

Animals↗
PubMed2001

T cell homeostasis: thymus regeneration and peripheral T cell restoration in mice with a reduced fraction of competent precursors.

We developed a novel experimental strategy to study T cell regeneration after bone marrow transplantation. We assessed the fraction of competent precursors required to repopulate the thymus and quantified the relationship between the size of the different T cell compartments during T cell maturation in the thymus. The contribution of the thymus to the establishment and maintenance of the peripheral T cell pools was also quantified. We found that the degree of thymus restoration is determined by the availability of competent precursors and that the number of double-positive thymus cells is not under homeostatic control. In contrast, the sizes of the peripheral CD4 and CD8 T cell pools are largely independent of the number of precursors and of the number of thymus cells. Peripheral "homeostatic" proliferation and increased export and/or survival of recent thymus emigrants compensate for reduced T cell production in the thymus. In spite of these reparatory processes, mice with a reduced number of mature T cells in the thymus have an increased probability of peripheral T cell deficiency, mainly in the naive compartment.

Animals↗
PubMed2000

B cell positive selection by self antigens and counter-selection of dual B cell receptor cells in the peripheral B cell pools.

The presence of B cells expressing two B cell receptors (BCR), described in BCR-transgenic, gene-targeted and normal mice, may represent an autoimmune hazard. We generated RAG-2-deficient mice bearing two complete rearranged immunoglobulin transgenes. In these mice most mature resting B cells express chains from the two transgenes. We studied selection of these dual receptor B cells in the presence of self antigens. In spite of the reduced surface density of the anti-self receptor, self-reactive B cells are deleted in the presence of membrane-bound self antigens. In contrast, the presence of soluble self antigen positively selects single receptor B cells expressing the self-reactive receptor. At the periphery these positively selected B cells down-regulate surface IgM expression and become unresponsive. A few dual receptor cells, however, escape tolerance induction. We examined the peripheral fate of the dual receptor B cells and showed that they are poorly selected into the activated B cell compartment and show a poor competitive capacity when in presence of populations of single receptor B cells. These results indicate that peripheral selection contributes to the very low frequencies of dual receptor B cells in normal mice and that multiple safeguard mechanisms operate to minimize the autoimmune hazard that allelically included B cells could represent.

Animals↗
PubMed2000

Peripheral B cell survival.

Recent findings suggest that lymphocyte survival is a continuous active process and support the role of B cell receptor engagement in B cell survival. In this context the conflict of survival interests between the diverse B cells gives rise to a pattern of interactions which mimics the behavior of complex ecological systems. In response to competition lymphocytes modify their survival requirements and diverge to occupy different immunological niches through differentiation. Thus naive and memory-activated B cell populations show independent homeostatic regulation. We discuss how niche differentiation allows the coexistence of different cell types and guarantees both repertoire diversity and efficient immune responses.

Animals↗
PubMed2000

Population biology of lymphocytes: the flight for survival.

In this essay we suggest that the primary goal of the cells of the immune system is to ensure their own growth and survival. In adults, in steady-state conditions, the number and distribution of lymphocyte populations is under homeostatic control. New lymphocytes that are continuously produced in primary and secondary lymphoid organs must compete with resident cells for survival. We discuss recent findings supporting lymphocyte survival as a continuous active process and implicating cognate receptor engagement as fundamental survival signals for both T and B lymphocytes. The conflict of survival interests between different cell types gives rise to a pattern of interactions that mimics the behavior of complex ecological systems. In their flight for survival and in response to competition, lymphocytes use different survival signals within different ecological niches during cell differentiation. This is the case for T and B lymphocytes and also for naive and memory/activated T and B cells. We discuss how niche differentiation allows the co-existence of different cell types and guarantees both repertoire diversity and efficient immune responses.

Adult↗
PubMed1999

Transfer of small resting B cells into immunodeficient hosts results in the selection of a self-renewing activated B cell population.

We studied the role of bone marrow B cell production in the renewal of peripheral B cells and the feedback mechanisms that control the entry of newly formed B cells into the peripheral B cell pools. When resting lymph node B cells are injected into B cell-deficient hosts, a fraction of the transferred cells expands and constitutes a highly selected population that survives for prolonged periods of time by continuous cell renewal at the periphery. Although the number of donor B cells recovered is low, a significant fraction shows an activated phenotype, and the serum immunoglobulin (Ig)M levels are as in normal mice. This population of activated B cells is resistant to replacement by a new cohort of B cells and is able to feedback regulate both the entry of newly formed B cells into the peripheral pool and terminal differentiation. These findings suggest that peripheral B cell selection follows the first come, first served rule and that IgM-secreting cells are generated from a pool of stable activated B cells with an independent homeostasis.

Animals↗
PubMed1999

Peripheral T cell survival.

T cell survival in the periphery is an active process, depending on continuous TCR engagement by peptide-MHC complexes and/or response to environmental cytokines. Naive T cells require interactions with the MHC restricting element. The survival requirements of memory T cells are as yet insufficiently characterized, but MHC-restricted interactions are not necessary.

Animals↗
PubMed1998

The role of the B cell receptor V region in peripheral B cell survival.

We investigate the role of the antigen-specific B cell receptor (BCR) in the establishment and maintenance of the peripheral B cell pools. We studied the fate of a population of transgenic B cells expressing a BCR without V region (Tg(deltaVmu)). We found that the Tg(deltaVmu) B cells can populate the peripheral B cell pools in the absence of other B cells, but when in the presence of a second population of non-transgenic B cells, they are virtually absent from the mature B cell compartments. By studying the rate of accumulation of 5-bromo-2'-deoxyuridine we show that the peripheral Tg(deltaVmu) B cells have a shorter life-span compared to non-transgenic B cells. By directly comparing the fate of two populations of transgenic B cells, either lacking or expressing a V region, we were able to assign the poorest competitive ability and the short peripheral survival of the Tg(delatVmu) B cells to the lack of an antigen-binding site. The results obtained support the involvement of the V region in the persistence of peripheral B cell populations.

Animals↗
PubMed1997

Differential requirements for survival and proliferation of CD8 naïve or memory T cells.

The requisite molecular interactions for CD8 T cell memory were determined by comparison of monoclonal naïve and memory CD8(+) T cells bearing the T cell receptor (TCR) for the HY antigen. Naïve T cells required only the right major histocompatibility complex (MHC) class I-restricting molecule to survive; to expand, they also needed antigen. In contrast, for survival, memory cells did not require the restricting MHC allele, but needed only a nonspecific class I; for expansion the correct class I, but not antigen, was required. Thus, maintenance of CD8 T cell memory still required TCR-MHC class I interactions, but memory T cells may have a lower functional activation threshold that facilitates secondary responses.

Adoptive Transfer↗
PubMed1997

Resource competition as a mechanism for B cell homeostasis.

Cellular competition for survival signals offers a cogent and appealing mechanism for the maintenance of cellular homeostasis [Raff, M. C. (1992) Nature (London) 356, 397-400]. We present a theoretical and experimental investigation of the role of competition for resources in the regulation of peripheral B cell numbers. We use formal ecological competition theory, mathematical models of interspecific competition, and competitive repopulation experiments to show that B cells must compete to persist in the periphery and that antigen forms a part of the resources over which B cells compete.

Animals↗
PubMed1997

Independent homeostatic regulation of B cell compartments.

In the present study we used mice with a developmental arrest of B cell production to study the ability of a limited number of normal B cell precursors to populate peripheral B cell pools. In chimeras reconstituted with mixtures of bone marrow (BM) cells from normal and B cell-deficient donors, we show that the rate of BM B cell production is a constant function of the number of BM pre-B cells and is not modified by the peripheral B cell pool size, i.e. there is no feedback regulation of the central pre-B cell compartment by the number of mature B cells. We also show that the physiological number of peripheral B cells requires a minimum continuous input of newly formed cells, but is not determined by the number of B cell precursors. Chimeras with a threefold reduced rate of BM B cell production have normal numbers of peripheral B cells. Parabiosis between normal and B cell-deficient mice showed that the BM B cell production of one mouse suffices to replenish the B cell pool of three mice. Finally, we show that the compartment of activated IgM-secreting B cells is homeostatically autonomous since the number of cells it comprises is regulated independently of the size of the mature B cell pool. The results presented here support a model of the immune system in which the size of the different B cell compartments, i.e. pre-B, resting B and IgM-secreting, is autonomously regulated.

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↗
PubMed1996

Cellular competition modulates survival and selection of CD8+ T cells.

In this investigation we compare the repopulation of the CD8+ T cell compartments of bone marrow (BM) chimeras by either normal nontransgenic or T cell receptor (TcR) alpha beta-transgenic (TG) CD8+ T cells, the fate of TG and non-TG CD8+ T cells in different parabionts and the survival of TG and non-TG peripheral CD8+ T cells after transfer into athymic hosts. We found that cellular competition among CD8 T cells occurs at several steps of T cell differentiation including a) during the DN to DP transition, b) positive selection in the thymus, c) export from the thymus and d) in the periphery. Comparison of the results obtained in the BM chimeras and in the parabionts shows that an important step of T cell selection occurs during seeding of peripheral lymphoid tissues. Once established, peripheral T cells resist replacement by recent thymus migrants, i.e. in the periphery, selection of T cell repertoires follows the rule "first come, first served". Peripheral dominance correlates with T cell activation and division. Cell cycling and CD44 expression are more frequent among non-TG CD8 T cells than TG CD8 T cells and within the latter, more frequent among P14 TG CD8 T cells than anti-HYTG CD8 T cells. Thus, in the absence of intentional immunization, the frequencies of CD8+ T cells follow a hierarchy of selection in which non-TG > or = P14 TG > anti-HY TG. We also show that the equilibrium size and the fate of one CD8 T cell population differs according to the presence or absence of other CD8 T cell populations. Under these circumstances, selection of T cell repertoires and T cell survival and memory rely not only on the interactions of each T cell with their respective ligands, but also on the nature and number of other competing cells.

Animals↗