Search PubMed⌕ Search

Biomedical subjects

M D Cooper

Publications and source records attributed to M D Cooper.

At least 127 records · Page 7Linked to original sources
PubMed1993

In situ analyses of in ovo graft-vs.-host reaction induced by thymic nurse cell lymphocytes.

In both mammalian and avian systems, thymic nurse cells (TNC) have been shown to harbor a heterogeneous population of T lymphocytes (TNC-L) some of which exhibit a postselectional phenotype. By transplanting micromanipulated single chicken TNC onto the chorionallantoic membrane (CAM) of major histocompatibility complex (MHC)-disparate embryos, an experimental system which allows for the detection of lymphocytes with graft-vs.-host (GVH) reactivity, we demonstrate here that TNC enclose lymphocytes that can develop into both CD4+ single-positive (sp) and CD8+ sp, T cell receptor (TcR) alpha beta+, or TcR gamma delta+ cells. This finding was additionally confirmed by serial transfer of primary expanded alloreactive T cells onto the CAM of secondary hosts. All donor TNC-L expressed MHC class II molecules and the interleukin-2 receptor alpha chain in primary and secondary GVH reactions. Furthermore, we observed selective accumulation of CD8+ and TcR gamma delta+ host lymphocytes in the CAM upon the induction of a local GVH reaction, most probably as a consequence of the pathological alteration of the epithelium.

Animals↗
PubMed1993

Growth requirements for avian gamma delta T cells include exogenous cytokines, receptor ligation and in vivo priming.

These studies analyze growth requirements for the normal gamma delta T cell population in peripheral lymphoid tissues. Avian gamma delta T cells can respond well to T cell mitogens in the presence of alpha beta T cells, but our studies indicate that they do not grow well alone. Exogenous growth factors were required in order for gamma delta T cells to proliferate in response to receptor ligation by anti-T cell receptor antibodies or other T cell mitogens. Interleukin-2 was implicated as one of the necessary growth factors that the gamma delta cells cannot produce adequately on their own. The response to dual stimulation (receptor ligation plus exogenous T cell factors) was attributable to a discrete subpopulation of gamma delta T cells that could be identified by their cell surface CD8, major histocompatibility complex class II expression and relative increase in cell size. Conversely, non-responsive gamma delta T cells did not exhibit these activation markers. These observations suggest a physiological basis for the relatively late appearance of gamma delta T cells in inflammatory responses and their failure as a population to match the growth potential of alpha beta T cells. More importantly, the results imply that the biological role of gamma delta T cells must be understood within the context of their interaction with alpha beta T cells.

Animals↗
PubMed1993

Migration patterns of thymus-derived gamma delta T cells during chicken development.

Cell transfer experiments in congenic chick strains, one of which expresses the ov antigen marker, indicate that intestinal gamma delta T cells are derived from gamma delta+ thymocytes in embryos and newly hatched birds, and this early intestinal colonization occurs in two discrete waves. Here, we extend these studies to show that splenic colonization by gamma delta T cells occurs in essentially the same way. Following the engraftment of ov+ thymic lobes in thymectomized ov- recipients, gamma delta T cells migrate both to the spleen and intestine. By 1 week after hatching, a third generation of thymus-derived gamma delta T cells begins to migrate to both peripheral lymphoid organs, and this thymus-dependent seeding process is sustained over the first weeks of life. The survival time for splenic gamma delta migrants is significantly less than for the intestinal migrants. Tissue section analysis indicates that gamma delta T cells enter the intestinal epithelium at all villus levels. A shift in the gamma delta intraepithelial lymphocyte distribution toward the villus tip in thymectomized birds suggests the comigration of enterocytes and gamma delta intraepithelial lymphocytes. However, survival kinetics of the donor gamma delta population and a relatively high division rate of intestinal gamma delta T cells indicate that founder thymic migrants produce relatively long-lived clones of intestinal gamma delta T cells.

Animals↗
PubMed1993

cDNA cloning and expression of human glutamyl aminopeptidase (aminopeptidase A).

The murine B-lymphocyte differentiation antigen BP-1/6C3 has been identified as glutamyl aminopeptidase (E-AP), formerly known as aminopeptidase A, the new gene symbol for which is ENPEP. In mice, the enzyme is found on early B-lineage cells and certain stromal cells of the bone marrow and thymus. This ectopeptidase is also expressed by capillary endothelial cells, placenta, and epithelial cells of the intestine and proximal renal tubules. Here we have used a mouse E-AP cDNA to identify the human counterpart in a kidney library. Sequence comparison of the human and mouse cDNAs reveals approximately 80% homology at both nucleotide and predicted amino acid levels. The nucleotide sequence of human E-AP predicts a type II integral membrane protein of 957 amino acids with an 18-amino-acid aminoterminal intracellular domain, and a 22-amino-acid transmembrane domain. The large extracellular carboxyterminal domain contains the zinc-binding motif typical of zinc-dependent metallohydrolases. When the human E-AP cDNA was placed downstream of the SR alpha promoter in an expression vector and transfected into COS-7 cells, the transfected cells exhibited cell surface E-AP activity. A 4.1-kb transcript could be detected in a variety of human tissues, including heart, brain, placenta, lung, liver, skeletal muscle, kidney, and pancreas. However, in representative lymphoid leukemias, E-AP transcripts were restricted to pre-B leukemia and were not found in T- and B-cell leukemias. The cDNA cloning and successful expression of human E-AP will allow more precise analysis of its physiological role(s).

Amino Acid Sequence↗
PubMed1993

B-cell development in man.

The development of B-lineage cells requires a series of complex interactions with hemopoietic stromal cell elements during the generative phase, and with antigen and T lymphocytes during the subsequent proliferative/differentiative phases in lymphoid tissues. Recent advances have been made in defining developmental changes in structure and assembly of the antigen receptors and in identifying protein kinases involved in signal transduction via these receptors. The mechanism of T-cell help has also come into much clearer focus through elucidation of the interaction between CD40 on B cells and the CD40 ligand on activated T cells. Finally, progress has been made with the recent identification of defects in a cytoplasmic protein tyrosine kinase and in the CD40 ligand as causes of two B-cell immunodeficiencies in man.

Antigens, CD↗
PubMed1993

Normal B lymphocyte differentiation.

Normal differentiation of B lineage cells has been the subject of intensive investigation over the past three decades. Current models of this process in humans are melded from the results of studies in a variety of organisms, including humans, mice and birds. Several recent developments have significantly reshaped and refined these models. The technique of homologous recombination in embryonic stem cells has allowed the production of mice with selectively disrupted genes that are important for B cell development in mice. At the same time, functional studies of human B cell differentiation, together with analysis of naturally occurring mutations that disrupt this process, have progressed rapidly. This has provided insight into the pathogenesis of lymphoproliferative and immunodeficiency diseases as well as a clearer view of normal developmental events. In this chapter we have reviewed human B cell differentiation with particular emphasis on newly emerging concepts. We also discussed CD5, a pan-T cell antigen that is expressed in low levels on a subpopulation of B cells implicated in the pathogenesis of chronic lymphocytic leukaemia (CLL). Finally, we discussed the issue of restricted variable region gene usage during B cell ontogeny and in CLL.

Animals↗
PubMed1993

Shared idiotypes in mesangial deposits in IgA nephropathy are not disease-specific.

The antigenic specificity of the mesangial IgA in IgA nephropathy (IgAN) remains unknown. Because shared antigenic specificities may be reflected in the usage of shared idiotypes, we prepared five monoclonal anti-idiotypic antibodies (MoAbs) specific for the mesangial IgA eluted from the kidney of an IgAN patient. All five MoAbs reacted with the same idiotype, which proved to be of a public nature. Although the idiotype could be identified in the mesangial deposits of the majority of IgAN patients studied, it was not specific for the disease because it was also found in the glomerular deposits of other types of glomerulonephritis. The idiotype was also expressed in polyethylene glycol precipitates of sera and in pokeweed mitogen-induced plasma cells from both IgAN patients and healthy controls. The conclusion that no disease-specific idiotypes are present in the renal eluate was further supported by the failure to produce polyclonal anti-idiotypic antibodies by immunizing a rabbit with the eluted mesangial IgA. Our results support the concept that mesangial IgA deposits in IgAN are of a polyclonal nature.

Antibodies, Monoclonal↗
PubMed1993

Signal transduction in human B cells initiated via Ig beta ligation.

Ig alpha and Ig beta heterodimers are non-covalently associated with Ig to compose the antigen receptor complexes on B cells. The demonstration that different sets of tyrosine kinases bind to the cytoplasmic tails of Ig alpha and Ig beta suggests that Ig alpha and Ig beta may activate distinct second messenger pathways. In this study, we examined the effects of mAbs against an exposed epitope of human Ig beta on pre-B and B cell triggering. Cross-linkage of Ig beta on B cells leads to activation of tyrosine kinases, hydrolysis of phosphatidylinositides, and elevation of intracellular Ca2+, effects qualitatively identical to those of anti-mu mAbs. Our observations thus indicate that cross-linking of Ig beta does not segregate signal transduction pathways connected with the cytoplasmic tails of Ig alpha and Ig beta. Ig alpha ligation has been reported to be more effective in triggering pre-B than B cells, whereas our results indicated that Ig beta ligation is more efficient in triggering B than pre-B cells. In addition to their activation properties, the anti-Ig beta mAbs effectively modulated B cell receptor complexes and blocked terminal differentiation of all plasma cell isotypes. The findings support the idea that anti-Ig beta could serve as a universal B cell immunosuppressant.

Animals↗
PubMed1993

Differential regulation of V(D)J recombination during development of avian B and T cells.

The lymphoid immune system is comprised of two major cell types, B cells and T cells, originally identified in avian species. Although both lineages arise from hematopoietic stem cells, avian B cells require a period of development in the bursa of Fabricius while T cells undergo development in the thymus. Each cell type expresses a lineage-specific antigen receptor encoded by genes created by the rearrangement of individual members of variable (V), diversity (D), and joining (J) gene segment families during embryonic development. In this report, we demonstrate that productive rearrangement of the TCR beta gene occurs exclusively in the thymus during normal development. TCR beta rearrangements involving gene segments from the V beta 1 gene family can be detected beginning on day 12 of development, while rearrangements involving the other family of V beta gene segments, V beta 2, were first detected on day 14 of embryogenesis. In contrast, productive rearrangements of Ig light (IgL) and heavy (IgH) chain genes were not restricted to the bursa of Fabricius. Instead, VH-DJH heavy chain rearrangements and VL-JL light chain rearrangements were detected primarily in the embryonic spleen, beginning as early as embryonic day 10, even in birds bursectomized at 60 h of development. Within the spleen, Ig rearrangement was confined to the subset of cells that express the chB6 surface protein. Unlike bursal lymphocytes, which express the recombinase activating gene (RAG)-2 but not RAG-1, splenic B cell precursors also express RAG-1. The data indicate that, while B cell precursors initiate recombination prior to migration of the bursa of Fabricius, T cell precursors undergo V(D)J recombination following migration to the thymus. Thus, distinct developmental mechanisms appear to regulate the process of receptor rearrangement during avian B and T cell development.

Animals↗
PubMed1993

In vivo depletion of chicken T-cell subsets.

In the chicken three types of T-cell receptors can be defined by monoclonal antibodies TCR1, TCR2 and TCR3, which recognize gamma delta T cells, and V beta 1- and V beta 2-expressing alpha beta T cells, respectively. In the present report we have analysed means of selectively depleting the gamma delta T cells and the V beta 1+ alpha beta T cells. gamma delta T cells, which represent up to 66% of all T cells in blood of a 6-month-old chicken, can be effectively depleted by neonatal thymectomy (Tx) to levels as low as 1%. Immunohistology demonstrates a similar depletion in lymphoid organs while intestinal epithelium-associated gamma delta T cells are affected by Tx to a lesser extent. V beta 1-bearing alpha beta T cells, which comprise about 80% of the alpha beta T cells, were depleted by embryonic and neonatal injection of the TCR2 antibody. In the thymus such treatment depleted only the V beta 1+ alpha beta T cells with high density expression of T-cell receptor. Therefore, we thymectomized TCR2-treated animals in order to prevent development of mature V beta 1+ alpha beta T cells from the low density immature thymocytes. Treatment of chickens with a total of 22 mg of TCR2 antibody plus Tx reduced V beta 1+ alpha beta T cells from an average of 65% to 10% of all T cells. In these TCR2 antibody-treated animals the V beta 2-expressing alpha beta T cells become the predominant type of T cell (average 85%).

Age Factors↗
PubMed1993

Widespread tissue distribution of aminopeptidase A, an evolutionarily conserved ectoenzyme recognized by the BP-1 antibody.

Early B-lineage cells in mice express a cell surface glycoprotein, recognized by the BP-1 and 6C3 monoclonal antibodies, that has been identified as aminopeptidase A (APA E.C.3.4.11.7). In the present studies we obtained evidence by DNA "zoo-blot" analysis that the APA gene is highly conserved. This ectoenzyme catalyzes the removal of N-terminal Glu- and Asp-residues to convert angiotensin II to angiotensin III, a degradation step important in local regulation of blood pressure in mammals. To gain further insight into the physiology of this molecule, which is shared between immune and vascular systems, we examined the tissue distribution of BP-1 mRNA using a cDNA probe and of the protein antigen using the BP-1 antibody for immunohistology. APA transcripts were present in all tissues examined. Abundant BP-1/APA was found in the intestinal brush border of the small intestine, renal glomeruli, proximal renal tubules, pulmonary alveolar walls and vascular endothelium in many organs. Other tissues containing the BP-1 antigen included stromal cells in the thymus cortex, bile canaliculi in liver, gall bladder epithelium, interlobular ducts in pancreas, the ovarian theca interna, basement membrane of the epididymis and the splanchnopleure in placenta. APA enzyme activities have been identified in most of these locations, in keeping with identification of the BP-1/6C3 antigen as APA. The data suggest this ectopeptidase may serve diverse physiologic roles in a broad spectrum of tissues.

Aminopeptidases↗
PubMed1993

Definition of immunoglobulin A receptors on eosinophils and their enhanced expression in allergic individuals.

Fc alpha receptors (Fc alpha R), detected by the binding of IgA and by anti-Fc alpha R antibodies, were found to be differentially expressed on eosinophils and neutrophils. Neutrophils were the major granulocyte population expressing Fc alpha R, and they expressed much higher levels of Fc alpha R than eosinophils. The expression of Fc alpha R by eosinophils could be upregulated approximately threefold by Ca2+ ionophore treatment in a dose- and time-dependent manner. This effect, which was blocked by a chelating agent, was not duplicated by other cellular stimuli. Eosinophils in allergic individuals displayed enhanced Fc alpha R expression, whereas neutrophils did not. The Fc alpha R on eosinophils had a higher molecular mass (70-100 kD) than those identified on neutrophils (55-75 kD). However, removal of N-linked carbohydrates from Fc alpha R of eosinophils and neutrophils revealed a major protein core of 32 kD for both cell types. The data indicate that expression of Fc alpha R molecules with a characteristic glycosylation pattern is upregulated on eosinophils in allergic individuals.

Adult↗
PubMed1992

Heterogeneity of immunoglobulin-associated molecules on human B cells identified by monoclonal antibodies.

Two covalently linked transmembrane molecules, encoded in mice by the mb-1 and B29 genes, have been defined as integral components of the antibody receptor units expressed on B cells. We have produced monoclonal antibodies against an exposed extracellular epitope on the putative human equivalent of the mouse B29 product. These antibodies, CB3-1 and -2, were used to show that cytoplasmic expression of this molecule begins in human pro-B cells (terminal deoxynucleotidyltransferase-positive, mu chain-negative), whereas surface expression coincides strictly with surface immunoglobulin expression of all isotypes. Immunochemical analysis of the human immunoglobulin-associated molecules revealed greater molecular heterogeneity than has been noted for the murine analogues. This molecular heterogeneity of immunoglobulin-associated molecules varied as a function of differentiation stage and the immunoglobulin isotypes expressed by B-lineage cells. Our data support the hypothesis that biochemical heterogeneity of the surface immunoglobulin-associated molecules may contribute to the variability in biological effects of antigen receptor crosslinkage on B cells of different maturational stages. Because the CB3 antibodies are capable of down-modulating the antigen receptors on all B cells, they may prove therapeutically useful as universal B-cell suppressants.

Antibodies, Monoclonal↗
PubMed1992

Molecular heterogeneity of Fc alpha receptors detected by receptor-specific monoclonal antibodies.

Fc alpha receptors (Fc alpha R) were isolated from a human monocytic cell line and used to raise four mAb with receptor specificity. The antibodies were used to identify the types of white blood cells that express Fc alpha R and the molecular heterogeneity of the receptor molecules. Nonpolymorphic epitopes, outside of the Fc alpha-binding site, were recognized only on blood cells of granulocyte and monocyte/macrophage lineages. The molecules identified, both by the antibodies and by the IgA ligand, were glycoproteins ranging in relative molecular mass from 55 to 75 kDa. However, one antibody detected a subpopulation of Fc alpha R molecules characterized by relatively restricted size heterogeneity. A complex glycosylation pattern was revealed by the resolution of discrete 32- and 36-kDa molecular species after removal of N-linked oligosaccharides and by evidence for O-linked carbohydrate moieties on at least a portion of the Fc alpha R molecules. In biosynthetic studies, all four anti-Fc alpha R antibodies and the IgA ligand bound a single 32-kDa core protein present in tunicamycin-treated cells, and the exceptional antibody again recognized molecules with relatively restricted glycosylation in the nontreated cells. These antibodies and native IgA ligands thus provide complementary reagents for definition of the complex structure and function of Fc alpha R in systemic IgA antibody responses.

Antibodies, Monoclonal↗
PubMed1992

Genetic and immunologic analysis of a family containing five patients with common-variable immune deficiency or selective IgA deficiency.

A family with 13 members included 2 subjects with selective IgA deficiency (IgA-D) and 3 subjects with common-variable immune deficiency (CVID), diseases which usually occur sporadically. Reciprocal combinations of B and T cells in vitro between one normal and two immune-deficient family members and normal subjects revealed that defective Ig synthesis was determined by the B cells, while the patient T cells functioned normally. Normal T helper and suppressor function was demonstrated even in one patient with CVID who developed a T-cell lymphoproliferative disorder associated with elevated IgM; this patient's B cells made only IgM in vitro. Immune deficiencies were inherited in this family in a pattern consistent with an autosomal dominant trait with incomplete penetrance. All the immune-deficient patients in this family possessed at least one copy of an MHC haplotype previously shown to be abnormally frequent in IgA-D and CVID: HLA-DQB1*0201, HLA-DR3, C4B-Sf, C4A-deleted, G11-15, Bf-0.4, C2-a, HSP70-7.5, TNF alpha-5, HLA-B8, and HLA-A1. The patient who developed the lymphoproliferative disorder was homozygous for this haplotype. Four immunologically normal members, one of whom was 80 years old, also possessed this MHC haplotype, indicating that its presence is not sufficient for disease expression. A small segment of another MHC haplotype associated with Ig deficiency in the population also occurred in this family, but it was not associated with immune deficiency.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1992

Immunization-induced decrease of the CD4+:CD8+ ratio in cats experimentally infected with feline immunodeficiency virus.

In a previous experiment a group of 15 specified pathogen free (SPF) cats were experimentally infected with a Swiss isolate of feline immunodeficiency virus (FIV). A group of 15 SPF cats served as FIV negative controls. Nine cats of each group were vaccinated with a recombinant feline leukemia virus (FeLV) vaccine, six cats in each group with a placebo vaccine. All vaccinated cats developed high antibody titers to FeLV and were protected against subsequent FeLV challenge infection. In both control groups five of six cats became persistently infected with FeLV. Unexpectedly, the primary immune response to the vaccine antigen was significantly higher in the FIV positive group than in the FIV negative. The secondary response was stronger in the FIV negative cats. The goal of the present investigation was to further study the immune response in these 30 cats. They were immunized twice with the synthetic peptide L-tyrosine-L-glutamic acid-poly(DL-alanine)-poly(L-lysine) (TGAL) 21 days apart. Blood samples were collected on four occasions during the immunization process. They were tested for antibodies to TGAL, complete blood cell counts and CD4+, CD8+ and pan-T-lymphocyte counts. The following observations were made: (1) in contrast to the FeLV vaccine experiment, the primary immune response to TGAL was not significantly stronger in the FIV positive cats when tested by enzyme-linked immunosorbent assay (2). The absolute size of the CD4+ lymphocyte population was distinctly smaller in the FIV positive than in the FIV negative cats. The lowest CD4+ values were found in the dually FIV/FeLV infected cats. (3) A population of CD8+ lymphocytes was identified that was characterized by a distinctly weaker fluorescence. The size of this population increased in FIV positive and decreased in FIV negative cats during the TGAL immunization experiment. (4) The CD4+:CD8+ ratio increased in FIV negative cats during TGAL immunization from 1.9 to 2.3. In contrast, in FIV positive animals the CD4+:CD8+ ratio decreased significantly from 1.9 to 1.3 during the same period. From these and earlier data it was concluded that in short-term FIV infection the immune response to T-cell dependent antigens may be increased over that of the controls. Immune suppression develops gradually with duration of the infection. The significant drop of the CD4+:CD8+ ratio over a 5 week immunization period suggests that antigenic stimulation may accelerate the development of immune suppression in FIV positive cats. If this is a general feature, FIV infection may provide a particularly interesting model for studying the pathogenesis of AIDS.

Amino Acid Sequence↗