Search PubMed⌕ Search

Biomedical subjects

K Bottomly

Publications and source records attributed to K Bottomly.

At least 73 records · Page 4Linked to original sources
PubMed1991

Differential expression of the alternatively spliced exons of murine CD45 in Th1 and Th2 cell clones.

Antigen-specific murine CD4+ T cell clones can be divided into functionally distinct subsets known as Th1 and Th2. To date these cells have been indistinguishable by surface phenotype. This report identifies two anti-CD45R monoclonal antibodies (14.8 and C363.16A) that bind preferentially to Th2 cells. Further analysis of the CD45-specific mRNA in Th1 and Th2 cells shows clear differences between these two cell types. Th1 cell clones express mRNA for the two smallest forms of CD45 containing none or only one of the alternatively splices exons. In contrast, Th2 cell clones express predominantly the high molecular weight isoforms of CD45 containing two or three of the alternatively spliced exons.

Animals↗
PubMed1991

Regulated expression and structure of T cell receptor gamma/delta transcripts in human thymic ontogeny.

Gamma delta (gamma delta) T cells have been found in all vertebrates examined, yet their function in vivo remains unknown. Because gamma delta T cell receptors are related to immunoglobulin, and because they are encoded by rearranging, multi-gene families, the receptors are thought to be antigen recognition molecules. However, a capacity to recognize naturally diverse antigens has not yet been shown. In this work, the expression and structure of human gamma delta transcripts have been examined in the fetal and early post-natal thymus. The data indicate that many gamma and delta genes are rearranged and expressed throughout ontogeny, but that as ontogeny proceeds, quite dramatic changes occur in the patterns of gene expression and rearrangement. In particular, receptors encoded by early to mid-gestation fetal thymic transcripts would be of quite restricted diversity. Only later in ontogeny can receptors of substantial diversity be generated. These properties are very similar to the patterns of gamma delta gene activation in the mouse, and they serve to reiterate similarities both in gene rearrangement and in gamma delta across vertebrate species.

Base Sequence↗
PubMed1991

Cell proliferation and cytokine production by CD4+ cells from old mice.

Splenocytes from young adult or old C57BL/6NNia mice were stimulated in vitro with the anti-CD3 epsilon mAb, 145-2C11, in either soluble (2C11s) or plate-bound (2C11i) form. In the young group, each mode of cell activation resulted in peak DNA synthesis at approximately 48 h of culture; at this time point, the old group exhibited response levels to 2C11s or 2C11i that were approximately 40% of those in the young group. However, in the presence of 2C11i, splenocytes from old donors showed a delayed peak response which approached the peak levels attained in the young group. To analyze the responsiveness of the CD4+ T cell subpopulation, this cell type was isolated from spleens of young or old mice and was stimulated in vitro with 2C11s or 2C11i, in the presence or absence of added accessory cells (T cell-depleted, irradiated splenocytes). The induction of DNA synthesis by 2C11s was accessory cell dependent, and the response in the old group were markedly reduced in comparison to those in the young group. In contrast, stimulation of DNA synthesis with 2C11i was relatively accessory cell independent, resulted in higher response levels in both age groups, and lessened the disparity between age groups. The analysis of IL-2 and IL-4 secretion by stimulated CD4+ cells revealed that, in response to 2C11s and accessory cells, only IL-2 accumulation was detectable and the levels in the young group were approximately 10-fold higher than the IL-2 levels in the old group. However, stimulation of CD4+ cells with 2C11i and accessory cells yielded improved IL-2 production and a detectable IL-4 response in the old group, whereas the young group exhibited a response profile similar to that induced by 2C11s. Further analysis of the IL-2, IL-4, and IFN gamma mRNA levels in 2C11i-stimulated CD4+ cells revealed that old donor cells accumulated similar levels of IL-2 transcripts, but higher levels of IL-4 and IFN gamma transcripts, than young donor CD4+ cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1991

The role of gamma interferon in infection of susceptible mice with murine coronavirus, MHV-JHM.

Infection of BALB/c mice with mouse hepatitis virus, strain JHM (MHV-JHM), at any of several intervals relative to ovalbumin (OVA) administration resulted in elevated OVA-specific IgG 2 a titers. Since gamma interferon (IFN) has been implicated as an up-regulator of IgG 2 a production, attempts were made to determine whether levels of this cytokine were modified in sera of infected mice. Serum IFN-gamma was not detected, but treatment of MHV-JHM-infected mice with monoclonal anti-IFN-gamma antibody resulted in high mortality with decreased survival times, enhanced virus titers in liver and spleen, and more severe virus-associated pathology, compared to mock-treated, infected mice. Immunotherapy with recombinant IFN-gamma ameliorated disease as reflected by mortality rates and virus titers in target organs.

Animals↗
PubMed1991

Cytokines in T-cell development.

The thymus provides a unique environment for the development of T cells, supporting both precursor cell proliferation and differentiation. The control of these processes is unknown but they may be mediated by cytokines, or other soluble factors, or by interactions with specific elements of the thymic stroma. Here, Simon Carding, Adrian Hayday and Kim Bottomly describe cellular, immunochemical and molecular studies of the production and action of cytokines within the human and mouse thymus and demonstrate their essential role in T-cell development.

Animals↗
PubMed1991

Selective activation of Th1- and Th2-like cells in vivo--response to human collagen IV.

Mice immunized with human collagen IV develop either antibody responses or T-cell proliferative responses as a function of the MHC genotype of the immunized mice. CD4+ T cells, similar to Th1 and Th2 cells, participate in these two types of responses, with CD4+ T-cell proliferative responses associating with IL-2 and IFN gamma release, and antibody production associating with CD4+ T-cell IL-4 and IL-5 release. Thus it would appear that the same antigen can induce responses consistent with either cell-mediated or humoral immunity depending on MHC class II genotype. In attempting to understand how MHC genotype controls the class of immunity observed several models are discussed. It was proposed, based on results obtained upon priming with human collagen IV, that the activation of Th1 and Th2 responses may be regulated at several levels (presentation by different APCs, presentation of different densities of the T-cell receptor ligand and presentation of different T-cell epitopes). With the identification of the peptide recognized by the CD4+ T cells, it was clear that the inability to induce Th1 responses in H-2b and the inability to induce Th2 responses in H-2s could not be accounted for by the failure to generate an immunodominant peptide during processing or the failure of the peptides to bind to the MHC class II molecules. Furthermore, the difference in the type of response generated could not be explained by the use of different peptides of the human collagen IV molecule in the two mouse strains, as a single peptide will induce both types of CD4+ T-cell response. However, it cannot be ruled out that the a2 peptide-class II interaction forms different T-cell ligands in the two strains either because the two class II MHC molecules are different or that the peptide is processed and reveals a different antigenic activity (Fox et al. 1988). Perhaps the most important finding from the peptide studies is that the lack of proliferative response in H-2b mice is not absolute, but can be overcome either by priming with high doses of the a2 peptide or by increasing the amount of peptide needed to elicit a recall response. It seems reasonable to speculate that changes in the dose required for priming Th1 or Th2 responses may reflect differences in the activation requirements of the two types of cells with Th1 cells requiring a high ligand density and Th2 cells a low ligand density.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1991

Infection of BALB/cByJ mice with the JHM strain of mouse hepatitis virus alters in vitro splenic T cell proliferation and cytokine production.

Earlier studies from this laboratory showed that infection of BALB/cByJ mice by a natural route with mouse hepatitis virus, strain JHM (MHV-JHM), results in functional splenic T cell suppression in vitro. This was evidenced by reduced concanavalin A-driven spleen cell proliferation and interleukin (IL)2 production measured after conventional intervals of cell culture (72 and 24 h, respectively). The purpose of the present work was to determine whether MHV-induced T cell dysfunction is kinetic or absolute and whether production of other T-cell derived cytokines is defective. Bioassays revealed that production of IL2, gamma interferon, and IL4 by spleen cells from acutely infected mice is suppressed and that some of the defects are kinetic as well as absolute. Proliferative responses of both CD4+ and CD8+ T cells were depressed, but neither cell type contained infectious virus. Cells that proliferated poorly in response to concanavalin A were fully capable of responding to specific virus stimulation. These results further emphasize the potential complications that MHV infection may pose to immunologic research using mice.

Animals↗
PubMed1990

Late dominance of the inflammatory process in murine influenza by gamma/delta + T cells.

The inflammatory response in the lungs of mice infected with an influenza A virus consists largely of macrophages and CD3+ T cells. Most T lymphocytes recovered before day 7 after infection express mRNA for the T cell receptor alpha/beta (TCR-alpha/beta), while TCR-gamma/delta mRNA+ cells are found at much higher frequency over the next 7 d. The predominant surface phenotype for the TCR-gamma/delta mRNA+ population is CD3+4-8-TCR-alpha/beta-. Some lymphocytes expressing all the known V gamma genes are found in the inflammatory exudate, but V gamma 2+/V gamma 1+ and V gamma 4+ T cells are present at highest frequency. The response is staged, with maximal numbers of V gamma 4+ cells occurring on day 10 after infection, while the predominant phenotype on day 13 is V gamma 2/V gamma 1+. The emerging peak in numbers of V gamma 4+ lymphocytes is paralleled by increasing numbers of macrophages expressing hsp mRNA. The later maxima found for the V gamma 2+/V gamma 1+ T cells is consistent with the possibility that at least some of these lymphocytes are responding to the hsp+ cells and are functioning to resolve the inflammatory process.

Animals↗
PubMed1990

Differences in the expression profiles of CD45RB, Pgp-1, and 3G11 membrane antigens and in the patterns of lymphokine secretion by splenic CD4+ T cells from young and aged mice.

Previous studies indicate that the 3G11, CD45RB, and Pgp-1 determinants are differentially expressed on CD4+ T cell subsets in the mouse. We used multicolor immunofluorescence staining and flow cytofluorometric analysis to examine the expression of each of these determinants on splenic CD4+ cells from young (age 3 to 6 mo) and aged (age 24 to 26 mo) C57BL/6 mice. The CD4+ pool from aged mice contained significantly reduced numbers of 3G11+ and CD45RBhi cells, but increased numbers of Pgp-1hi cells, in comparison with the young group. Analysis of the simultaneous expression of all three subset determinants on CD4+ cells revealed that, in young mice, the major fraction (greater than 50%) was 3G11+CD45RBhiPgp-1lo. Among the less prevalent cell phenotypes, reductions in 3G11 expression correlated with decreases in CD45RB levels and increases in Pgp-1 levels. The phenotype that dominated the young group (3G11+CD45RBhiPgp-1lo) was approximately fivefold less represented in the aged group. The CD4+ pool from aged mice was characterized by increases in the 3G11-CD45RBvariablePgp-1hi and the 3G11+CD45RBloPgp-1hi phenotypes. To evaluate possible age-associated differences in cytokine secretion patterns by splenic CD4+ cells, purified CD4+ cells from each age group were stimulated in vitro with immobilized anti-CD3 epsilon mAb and accessory cells. At various times thereafter, supernatants from cultures were tested for IL-2 and IL-4 content by using the CTLL.6 and 11.6 bioassays, respectively, and the CD4+ cells were assayed for [3H]TdR uptake. Cell cultures from the aged group exhibited similar peak IL-2 accumulation and lower peak [3H]TdR uptake, but greatly increased peak IL-4 accumulation, as compared with cell cultures from the young group. The expression patterns of subset determinants, in conjunction with cytokine secretion profiles, indicate that, in aged mice, marked alterations occur in the subset composition of the splenic CD4+ cell pool. These findings are discussed in the context of previous findings on changes in T cell reactivity with advancing donor age.

Aging↗
PubMed1990

Characterization of gamma/delta T cell clones isolated from human fetal liver and thymus.

The origin and development of T cells bearing gamma/delta T cells receptors (TcR) has been extensively studied in the mouse. By contrast, little is known about development patterns and diversity of the human gamma/delta T cell lineage. To study the repertoire of human gamma/delta+ T cells during T cell ontogeny, we have isolated clonal populations of gamma/delta+ T cells from 14-week fetal thymus and liver and characterized the molecular composition of their TcR. The technique of in situ hybridization was used to identify cells expressing TcR genes in fetal liver and thymus. A panel of clones representative of developing T cell populations found in vivo was subsequently isolated from both tissues and clones expressing cell surface gamma/delta receptors were identified. Although both the liver-derived gamma/delta+ T cell clone, L2, and the thymus-derived gamma/delta+ T cell clone, T6, had similar cell surface phenotypes, namely CD3+, CD7+, CD45+ and CD8-, their reactivity with anti-CD2 and -CD4 antibodies was different. L2 was CD2high, CD4- whereas T6 was CD2low, CD4low. Both clones possessed effector functions similar to those of adult T cells as demonstrated by the synthesis and secretion of cytokines in response to stimulation through the CD3/TcR complex. Analysis of the TcR composition of the fetal clones showed both clones to possess similar or identical gamma chain components, C gamma 1, J gamma 1/2, V gamma 8, and both utilize V delta gene segments other than V delta 1. This TcR genotype has not been previously reported in the analysis of adult gamma/delta+ T cells. Our studies have identified a unique population of human gamma/delta+ T cells that may be derived extrathymically and appear to be preferentially and perhaps transiently expressed during fetal life.

Biological Factors↗
PubMed1990

Molecular associations on the T cell surface correlate with immunological memory.

Different isoforms of CD45 are expressed on naive and memory CD4 T cells in the mouse, as revealed by an antibody to a set of isoforms of CD45 that utilize exon B, called CD45RB. Cloned TH1 and TH2 lines also differ for expression of isoforms detected by this antibody. Differential expression of CD45 isoforms correlates with different behavior of cell surface molecules involved in transmembrane signal transduction. On naive T cells, CD4, CD45 and the CD3/T cell receptor complex behave as independent entities. On memory T cells, these three molecules are stably associated on the T cell surface. Furthermore, on TH2 cells, which express intermediate levels of CD45RB, CD4 is stably associated with CD45 isoforms other than CD45RB, but this complex is not associated with the CD3/T cell receptor. These results lead us to propose that immunological memory in CD4 T cells consists of an altered structure of the T cell's specific signal transduction apparatus controlled by low-molecular weight CD45 isoforms. This altered receptor structure would allow the more sensitive triggering of the T cell characteristic of memory cells. The organization of multimolecular signal transduction systems may be a general means by which cells alter their physiological behavior, allowing the acquisition of new phenotypic characteristics.

Animals↗
PubMed1990

Developmentally regulated fetal thymic and extrathymic T-cell receptor gamma delta gene expression.

The gamma delta T-cell receptor (TCR) is the first TCR to be expressed in ontogeny in all vertebrates in which it has been examined thoroughly. Murine gamma delta cell-surface protein is detected by the fourteenth day of gestation. In this work, the activation of gamma delta RNA has been studied. Data indicate that the first TCR protein to appear in the thymus is encoded by gamma genes that are activated after cells colonize the thymus. However, the sequential appearance of different gamma delta TCR proteins during thymic ontogeny cannot be readily explained by differential temporal activation of V gamma genes in the thymus. There are distinct patterns of gamma and delta gene expression during fetal liver development and in the fetal gut (or tissue associated with it). Cells apparent in the liver of mice at birth express gamma delta cell-surface protein, but they disappear from the liver very soon afterward. One V gamma gene is rearranged and expressed prethymically. In addition, gamma gene expression is detectable in the livers of newborn athymic mice. Together, these observations indicate a thymic-independent pathway of activation of TCR genes.

Animals↗
PubMed1990

Cultured human luteal peripheral monocytes secrete increased levels of interleukin-1.

Interleukin-1 (IL-1), an immune monokine secreted by activated macrophages and monocytes, appears to be intimately involved in endocrine phenomena. It is well established that IL-1 mediates a number of immune and inflammatory responses. In addition, the production of IL-1 appears to be modulated by the gonadal estradiol and progesterone, leading to this study of IL-1 secretion by cultured peripheral monocytes isolated at varying times during the menstrual cycle and pregnancy. Circulating peripheral monocytes were isolated from eight women undergoing human menopausal gonadotropin-stimulated ovulation induction during the late follicular phase just before hCG administration and from eight similarly stimulated women on the 12th day after hCG administration. Peripheral monocytes were also isolated from six women during both second and third trimester pregnancy. After 48 h in culture, conditioned media were assayed for IL-1 bioactivity using the D1O lymphocyte proliferation assay. Significantly more IL-1 bioactivity was secreted by monocytes isolated in the luteal phase of the cycle (52.4 +/- 17.5 IU/mL) compared to that in late follicular phase (5.2 +/- 0.9 IU/mL; P = 0.001) or third trimester of pregnancy (7.0 +/- 1.5 IU/mL; P = 0.006). Thus, peripheral monocyte IL-1 secretion appears to be increased by luteal levels of progesterone, although further elevation of progesterone during pregnancy returns IL-1 levels to the preovulatory baseline. In addition, basal body temperatures obtained in four women during third trimester pregnancy were 98 F or less. Thus, IL-1 secretion from cultured monocytes appears to increase with luteal concentrations of progesterone and decrease to preovulatory levels at higher concentrations of the steroid during pregnancy, which may account for the dissociation of high progesterone levels and elevated basal body temperature during late pregnancy.

Adult↗
PubMed1989

MHC control of CD4+ T cell subset activation.

The present results demonstrate that CD4+ T cells activated in the primary in vivo response to antigen produce distinct patterns of cytokines depending upon the MHC class II haplotype of the responding mice. I-As mice were found to selectively activate IL-2/IFN-gamma-producing CD4+ T cells, whereas I-Ab mice exhibited selective activation of IL-4-producing CD4+ T cells in response to collagen IV. The effector response phenotype was found to correlate with the cytokine phenotype of CD4+ T cells activated in vivo; IL-2/IFN-gamma-producing cells giving rise to proliferative (cell-mediated) responses, IL-4-producing cells leading to secondary IgG (humoral) responses. Together the data support the notion that the outcome of a given immune response (e.g., protection vs. onset, tolerance vs. autoimmunity) may be determined in part by the type of CD4+ T cells initially activated by antigen. Moreover, the present experiments demonstrate for the first time that polymorphism in class II MHC can determine such selective activation of different cytokine-producing CD4+ T cell phenotypes.

Animals↗
PubMed1989

Monoclonal antibodies to murine CD3 epsilon define distinct epitopes, one of which may interact with CD4 during T cell activation.

The TCR is comprised of two variable chains that confer specificity, called alpha:beta or gamma:delta, physically associated with five different molecules that comprise the complex known as CD3. Antibodies to this complex are very useful, as they react with all T lymphocytes. A rat mAb to mouse CD3 has been prepared. It reacts with 100% of T cells in all mouse strains tested but with no other cell type. It binds to the CD3 epsilon chain. This antibody activates cloned T cell lines and normal T cells, provided suitable accessory cells and signals are present. This antibody detects a determinant similar to but not identical with those detected by two previously reported hamster anti-CD3 epsilon antibodies. This antibody fixes C efficiently, and it is thus useful for depletion of T cells from bulk populations. Activation of T cells by one of the three different anti-CD3 epsilon antibodies was inhibited by the Fab fragment of anti-CD4, similar to the effects of anti-CD4 Fab on two previously reported anti-TCR V region antibodies that bind a CD3 epsilon-associated epitope. This further defines a site involving TCR V regions and CD3 epsilon with which CD4 appears to associate during T cell activation.

Animals↗