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Biomedical subjects

I L Weissman

Publications and source records attributed to I L Weissman.

At least 127 records · Page 7Linked to original sources
PubMed1995

Two pathways of exocytosis of cytoplasmic granule contents and target cell killing by cytokine-induced CD3+ CD56+ killer cells.

Cytokine-induced killer (CIK) cells are non-major histocompatibility complex-restricted cytotoxic cells generated by incubation of peripheral blood lymphocytes with anti-CD3 monoclonal antibody (MoAb), interleukin-2 (IL-2), IL-1, and interferon-gamma. Cells with the greatest effector function in CIK cultures coexpress CD3 and CD56 surface molecules. CIK cell cytotoxicity can be blocked by MoAbs directed against the cell surface protein leukocyte function associated antigen-1 but not by anti-CD3 MoAbs. CIK cells undergo release of cytoplasmic cytotoxic granule contents to the extracellular space upon stimulation with anti-CD3 MoAbs or susceptible target cells. Maximal granule release was observed from the CD3+ CD56+ subset of effector cells. The cytoplasmic granule contents are lytic to target cells. Treatment of the effector cells with a cell-permeable analog of cyclic adenosine monophosphate (cAMP) inhibited anti-CD3 MoAb and target cell-induced degranulation and cytotoxicity of CIK cells. The immunosuppressive drugs cyclosporin (CsA) and FK506 inhibited anti-CD3-mediated degranulation, but did not affect cytotoxicity of CIK cells against tumor target cells. In addition, degranulation induced by target cells was unaffected by CsA and FK506. Our results indicate that two mechanisms of cytoplasmic granule release are operative in the CD3+ CD56+ killer cells; however, cytotoxicity proceeds through a cAMP-sensitive, CsA- and FK506-insensitive pathway triggered by yet-to-be-identified target cell surface molecules.

Biomarkers↗
PubMed1995

Transgenic mice carrying the diphtheria toxin A chain gene under the control of the granzyme A promoter: expected depletion of cytotoxic cells and unexpected depletion of CD8 T cells.

We have generated transgenic mice bearing the diphtheria toxin A chain (DTA) gene under the control of granzyme A (GrA) promoter sequences (GrA-DTA). GrA is expressed in activated cytotoxic cells but not in their immediate progenitors. These GrA-DTA mice are deficient in cytotoxic functions, indicating that most cytotoxic cells express GrA in vivo. Surprisingly, one founder strain containing a multicopy GrA-DTA insert show a marked and selective deficiency in CD8+ cells in peripheral lymphoid organs. This depletion was not observed in thymus, where the distribution of CD4+ and CD8+ cells is normal. Moreover, the emigration of T cells from thymus is normal, indicating that the depletion occurs in the periphery. GrA-DTA mice should be useful as models to dissect the role of cytotoxic cells in immune responses and as recipients of normal and neoplastic hematopoietic cells. The selective depletion of CD8+ cells in one founder strain could have implications for postthymic T-cell development.

Animals↗
PubMed1995

The purification and characterization of fetal liver hematopoietic stem cells.

Thy-1loSca-1+Lin-Mac-1+CD4- cells have been isolated from the livers of C57BL-Thy-1.1 fetuses. This population appears to be an essentially pure population of hematopoietic stem cells (HSC), in that injection of only six cells into lethally irradiated adult recipients yields a limit dilution frequency of donor cell-reconstituted mice. Sixty-seven to 77% of clones in this population exhibit long-term multilineage progenitor activity. This population appears to include all long-term multilineage reconstituting progenitors in the fetal liver. A high proportion of cells are in cycle, and the absolute number of cells in this population doubles daily in the fetal liver until 14.5 days postcoitum. At 15.5 days postcoitum, the frequency of this population falls dramatically. Long-term reconstituting HSC clones from the fetal liver give rise to higher levels of reconstitution in lethally irradiated mice than long-term reconstituting HSC from the bone marrow. The precise phenotypic and functional characteristics of HSC vary according to tissue and time during ontogeny.

Animals↗
PubMed1995

Extrathymic maturation of alpha beta T cells from hemopoietic stem cells.

The object of the study was to determine whether alpha beta T cells can develop from hemopoietic stem cells in the absence of the thymus. C57BL/6 (Ly-5.1 and Thy-1.2) mice were thymectomized or sham-thymectomized at 4 wk of age, and received lethal whole body irradiation 2 wk later. These mice were reconstituted with an i.v. injection of 500 highly purified hemopoietic stem cells (Mac-1-, B220-, TER-119-, CD3-, CD4-, CD8-, Thy 1low, SCA-1+) obtained from the bone marrow of C57BL/6 (Ly-5.2 and Thy-1.1) donors. A similar percentage of Ly-5.2+ alpha beta T cells (donor) was found in the marrow of thymectomized recipients, sham-thymectomized recipients, and normal donor mice at least 3 mo after stem cell transplantation. The percentage of Ly-5.2+ alpha beta T cells in the spleens of sham-thymectomized and normal donor mice was similar. The percentage in the spleens of thymectomized recipients was reduced by about 50%, and approximately one-half of the latter T cells expressed the CD4-CD8- alpha beta+ phenotype. A purified population of Ly-5.2+ alpha beta- cells obtained from the marrow of thymectomized recipients was incubated in vitro for 48 h without exogenous growth factors. After the incubation procedure a proportion of the marrow cells acquired alpha beta TCR surface receptors. The results show that alpha beta T cells can develop from hemopoietic stem cells in the absence of the thymus.

Animals↗
PubMed1995

The biology of hematopoietic stem cells.

Hematopoietic stem cells (HSC) are the only cells in the blood-forming tissues that can give rise to all blood cell types and that can self-renew to produce more HSC. In mouse and human, HSC represent up to 0.05% of cells in the bone marrow. HSC are almost entirely responsible for the radioprotective and short- and long-term reconstituting effects observed after bone marrow transplantation. The subsets of HSC that give rise to short-term vs long-term multilineage reconstitution can be separated by phenotype, demonstrating that the fates of HSC are intrinsically determined. Here we review the ontogeny and biology of HSC, their expression of fate-determining genes, and the clinical importance of HSC for transplantation and gene therapy.

Adult↗
PubMed1995

Life histories and senescence of Botryllus schlosseri (Chordata, Ascidiacea) in Monterey Bay.

The colonial ascidian Botryllus schlosseri is a model organism for research on invertebrate histocompatibility, development, and evolutionary biology. Nonetheless, the basic life history of Pacific Ocean populations of the species remains unknown. We determined field rates of growth, reproduction, and senescence in four cohorts of B. schlosseri colonies in Monterey Bay, California. Colonies grew exponentially as juveniles and reached sizes of up to 1400 zooids within 69 days. After a juvenile phase lasting at least 49 days, the colonies began to reproduce sexually. Each zooid produced up to 10 clutches, each with a maximum of 5 eggs, resulting in very high fecundity of up to 8000 eggs per colony. Following a short period (maximum 70 days) of continuous sexual reproduction, colonies abruptly senesced and died while still bearing a full clutch of eggs. Senescence progressed through four distinct stages over 1-2 weeks, and inevitably led to the simultaneous death of all zooids in the colony. Although senescence was the main cause of mortality, some colonies died as a result of predation or undermined causes. Certain life history traits varied significantly between cohorts that settled at different times of year. For example, lifespans in the field varied from about 3 months for spring to 8 months for fall-born colonies, but the lifetime fecundity of colonies did not vary between cohorts. The morphologies and life histories of colonies monitored in the field and reported here differed from those of colonies cultured previously in the laboratory.

Animals↗
PubMed1994

Characterization of preneoplastic thymocytes and of their neoplastic progression in irradiated C57BL/Ka mice.

Mice that receive whole body split-dose irradiation develop thymic lymphomas after a long latent period. Before emergence of frank lymphomas, preneoplastic thymocytes, which are defined by their ability to progress to full malignancy on intrathymic transfer to congenic hosts, appear. A combination of mAb 1C11, which binds to a cell surface glycoprotein on lymphoma cells, and of Abs to the differentiation markers CD4 and CD8 (MHC co-receptors), and CD3 (TCR complex) was used to characterize the phenotypes of preneoplastic thymocytes and to place them within the scheme of normal T cell ontogeny. In the irradiated, preneoplastic thymus, the 1C11 molecule was found to be present on CD4-8-, CD4-8+, and CD4+8+, but not CD4+8-, cells. After intrathymic transfer to Thy-1 congenic recipients, 1C11highCD4-8- cells from irradiated mice showed the highest leukemogenic potential, followed by the 1C11highCD4-8+ and 1C11highCD4+8+ subsets. Within the 1C11highCD4-8- subset, CD3+ cells were more leukemogenic than CD3- cells. The resulting lymphomas were 1C11highCD3+4-8+ and 1C11highCD3+4+8+, phenotypes that are absent or very rare in the normal thymus, but similar to those of primary radiation-induced lymphomas. Examination of the TCR V beta repertoire in these lymphomas shows a highly significant bias, in that approximately 50% express the V beta 8 gene product. These results indicate, but do not prove, that the 1C11highCD3+4-8- cells, a very small subset of normal thymocytes, are either the target of neoplastic transformation after radiation or the earliest identifiable cell population after the transforming event. These results also suggest at least one possible pathway to define the process leading to overt lymphoma.

Animals↗
PubMed1994

Demonstration that Thy(lo) subsets of mouse bone marrow that express high levels of lineage markers are not significant hematopoietic progenitors.

We have been unable to reproduce experiments suggesting the existence of three lineage-restricted progenitor populations from mouse bone marrow. Thy1.1loMac-1+B220+ cells were reported to give rise to greatly expanded numbers of myeloid and lymphoid cells, while Thy1.1loMac-1+B220- and Thy1.1loMac-1-B220+ cells were reported to be highly proliferative myeloid and B-lineage-restricted progenitors, respectively. Both Mac-1+ cell types appear to be much less frequent than previously reported, and we observed no activity consistent with their characterization as committed progenitors of expanded numbers of cells. The original identification of these populations may have resulted from a failure to distinguish bonafide signals from autofluorescent background and nonspecific staining. The progenitor activities originally associated with these populations may have been due to hematopoietic stem cell contamination. This study shows that low levels of Mac-1 are expressed on cells with multipotent progenitor activity. Thy1.1loB220+Mac-1- cells can be purified from bone marrow, but in these experiments they do not give rise to detectable levels of progeny on injection into lethally irradiated mice. Thy1.1loB220+Mac-1- cells appear to be pro-B cells without significant proliferation potential in vivo. The finding that the described populations do not have the reported progenitor activities leaves the pathways of stem cell differentiation open to further study.

Animals↗
PubMed1994

Rapid and sustained hematopoietic recovery in lethally irradiated mice transplanted with purified Thy-1.1lo Lin-Sca-1+ hematopoietic stem cells.

Hematopoietic stem cells (HSCs) are believed to play a critical role in the sustained repopulation of all blood cells after bone marrow transplantation (BMT). However, understanding the role of HSCs versus other hematopoietic cells in the quantitative reconstitution of various blood cell types has awaited methods to isolate HSCs. A candidate population of mouse HSCs, Thy-1.1lo Lin-Sca-1+ cells, was isolated several years ago and, recently, this population has been shown to be the only population of BM cells that contains HSCs in C57BL/Ka-Thy-1.1 mice. As few as 100 of these cells can radioprotect 95% to 100% of irradiated mice, resulting long-term multilineage reconstitution. In this study, we examined the reconstitution potential of irradiated mice transplanted with purified Thy-1.1lo Lin-Sca-1+ BM cells. Donor-derived peripheral blood (PB) white blood cells were detected as early as day 9 or 10 when 100 to 1,000 Thy-1.1lo Lin-Sca-1+ cells were used, with minor dose-dependent differences. The reappearance of platelets by day 14 and thereafter was also seen at all HSC doses (100 to 1,000 cells), with a slight dose-dependence. All studied HSC doses also allowed RBC levels to recover, although at the 100 cell dose a delay in hematocrit recovery was observed at day 14. When irradiated mice were transplanted with 500 Thy-1.1lo Lin-Sca-1+ cells compared with 1 x 10(6) BM cells (the equivalent amount of cells that contain 500 Thy-1.1lo Lin-Sca-1+ cells as well as progenitor and mature cells), very little difference in the kinetics of recovery of PB, white blood cells, platelets, and hematocrit was observed. Surprisingly, even when 200 Thy1.1lo Lin-Sca-1+ cells were mixed with 4 x 10(5) Sca-1- BM cells in a competitive repopulation assay, most of the early (days 11 and 14) PB myeloid cells were derived from the HSC genotype, indicating the superiority of the Thy-1.1lo Lin-Sca-1+ cells over Sca-1- cells even in the early phases of myeloid reconstitution. Within the Thy-1.1lo Lin-Sca-1+ population, the Rhodamine 123 (Rh123)hi subset dominates in PB myeloid reconstitution at 10 to 14 days, only to be overtaken by the Rh123lo subset at 3 weeks and thereafter. These findings indicate that HSCs can account for the early phase of hematopoietic recovery, as well as sustained hematopoiesis, and raise questions about the role of non-HSC BM populations in the setting of BMT.

Animals↗
PubMed1994

Regulation of the avidity of integrin alpha 4 beta 7 by the beta 7 cytoplasmic domain.

Integrins are cell-surface heterodimeric receptors with adhesive and transmembrane signaling properties. Their cytoplasmic domains can affect receptor avidity, cytoskeletal association, and post-receptor occupancy events. The alpha 4 beta 7 integrin mediates cell adhesion to Peyer's patch high walled endothelial venules (HEV), VCAM, and CS-1/fibronectin. To determine the role of the beta 7 cytoplasmic domain in these interactions, wild-type and truncated beta 7 subunits were stably expressed in the alpha 4+/beta 1-/beta 7- B cell lymphoma 38C13. The cell line delta 727 lacks the entire beta 7 cytoplasmic domain, delta 753 lacks the 34 C-terminal residues, and LXSN is vector-transfected 38C13. Cells expressing wild-type beta 7 bound Peyer's patch HEV, fibronectin, and immobilized VCAM constitutively and did not require prior activation with phorbol esters. Interestingly, delta 753 displayed no ligand binding activity, while delta 727 was constitutively active for all ligands and displayed greater avidity for fibronectin and Peyer's patch HEV than the wild-type beta 7. beta 7, delta 753, delta 727, and LXSN were also tested for the ability to bind soluble VCAM in the presence of various divalent cations. In the presence of Ca2+, but not Mg2+, delta 727 constitutively bound soluble VCAM, whereas beta 7, delta 753, and LXSN did not. beta 7 and delta 753 could bind soluble VCAM if first activated with Mn2+. The results suggest that: (i) alpha 4 beta 7 can be expressed in a constitutively active state, (ii) the beta 7 cytoplasmic domain regulates the avidity of alpha 4 beta 7, and (iii) 38C13 cell lines expressing wild-type and truncated beta 7 subunits define three stable activation states of alpha 4 beta 7: inactive (delta 753), partially active (beta 7), and fully active (delta 727) receptors.

Amino Acid Sequence↗
PubMed1994

Expression of the integrin alpha 4 beta 1 on melanoma cells can inhibit the invasive stage of metastasis formation.

Among a series of adhesion molecules, expression of integrin alpha 4 beta 1 showed a unique inverse correlation with the invasive potential of B16 melanoma cell lines. When an alpha 4 cDNA was introduced into an alpha 4-beta 1+ highly invasive melanoma line, alpha 4 beta 1 heterodimers were expressed on the surface. Matrigel invasion by the alpha 4+ beta 1+ cells was reduced. Pulmonary metastasis was also suppressed when the transfectants were placed subcutaneously, but not when injected intravenously. Expression of alpha 4 beta 1 promoted homotypic intercellular adhesion. The homotypic adhesion was abrogated, and the alpha 4+ beta 1+ (less invasive cell lines) increased matrigel invasion following the anti-alpha 4 MAb treatment. These results suggest that integrin alpha 4 beta 1 could play a role in controlling melanoma cell metastasis at the invasive stage.

Animals↗
PubMed1994

bcl-2 inhibits apoptosis of neutrophils but not their engulfment by macrophages.

Neutrophils, the most common inflammatory leukocytes, have the most limited life span of all blood cells. After they undergo apoptosis, they are recognized and engulfed by macrophages. bcl-2, a proto-oncogene rearranged and deregulated in B cell lymphomas bearing the t(14;18) translocation, is known to inhibit programmed death. bcl-2 expression is localized in early myeloid cells of the bone marrow but is absent in mature neutrophils. Transgenic mice that expressed bcl-2 in mature neutrophils showed that bcl-2 blocked neutrophil apoptosis. Despite this, homeostasis of neutrophil population is essentially unaffected. In fact, macrophage uptake of neutrophils expressing bcl-2 still occurred. This transgenic model indicates that the mechanism that triggers phagocytosis of aging neutrophils operates independently of the process of apoptosis regulated by bcl-2.

Animals↗
PubMed1994

Isolation and characterization of major histocompatibility complex class IIB genes from the nurse shark.

The major histocompatibility complex (MHC) contains a set of linked genes which encode cell surface proteins involved in the binding of small peptide antigens for their subsequent recognition by T lymphocytes. MHC proteins share structural features and the presence and location of polymorphic residues which play a role in the binding of antigens. In order to compare the structure of these molecules and gain insights into their evolution, we have isolated two MHC class IIB genes from the nurse shark, Ginglymostoma cirratum. Two clones, most probably alleles, encode proteins which differ by 13 amino acids located in the putative antigen-binding cleft. The protein structure and the location of polymorphic residues are similar to their mammalian counterparts. Although these genes appear to encode a typical MHC protein, no T-cell-mediated responses have been demonstrated in cartilaginous fish. The nurse shark represents the most phylogenetically primitive organism in which both class IIA [Kasahara, M., Vazquez, M., Sato, K., McKinney, E.C. & Flajnik, M.F. (1992) Proc. Natl. Acad. Sci USA 89, 6688-6692] and class IIB genes, presumably encoding the alpha/beta heterodimer, have been isolated.

Amino Acid Sequence↗
PubMed1994

In vitro development of B cells and macrophages from early mouse fetal thymocytes.

The developmental potentialities of early mouse fetal thymocytes were analyzed by in vitro culturing cell suspensions obtained from 12-15-gestational day thymus in contact with a bone marrow stroma clonal cell line that supports pre-B and myeloid cell differentiation. B cell and macrophage development from fetal thymocytes was observed at all fetal stages tested, but limiting dilution analysis revealed that the frequency of cells forming colonies on bone marrow stroma is the highest in the fetal thymus at day 12, then dramatically decreases until day 15. These observations suggest that the thymus rudiment is seeded by multipotential precursor cells which are not immediately committed to T cell development in the thymic cellular environment.

Animals↗