scholarly journals BIOLOGICAL CHARACTERISTICS OF T AND B MEMORY LYMPHOCYTES IN THE RAT

1973 ◽  
Vol 137 (5) ◽  
pp. 1275-1292 ◽  
Author(s):  
Samuel Strober ◽  
Jeanette Dilley
Keyword(s):  
Thoracic Duct ◽  
Cell Lineage ◽  
Cell Types ◽  
Thoracic Duct Lymph ◽  
Slight Reduction ◽  
Memory Cells ◽  
Spleen Cells ◽  
Dividing Cells ◽  
Experimental Findings

The adoptive secondary antibody response of rats to the hapten-protein conjugate dinitrophenyl-diphtheria toxoid (DNP-DT) was used to investigate the migratory properties and rate of formation of T and B memory cells in the spleen. The experimental findings show that hapten (DNP-BSA)- and carrier (DT)-primed spleen cells act synergistically in the restoration of the adoptive anti-DNP response. Passage of both hapten- and carrier-primed spleen cells through an intermediate host (intravenous injection and subsequent collection in the thoracic duct lymph) showed that both cell types are able to recirculate from the blood to the lymph. In addition, memory to the hapten or carrier could be withdrawn from the spleen by prolonged thoracic duct drainage. The rate of formation of hapten- and carrier-primed spleen cells was studied by treating donors with [3H]thymidine for 48 h before cell transfer in an attempt to "suicide" rapidly dividing cells. Only a slight reduction in the adoptive response to the hapten or carrier was noted upon transfer of treated cells to irradiated hosts. In further experiments, the cell lineage of hapten- and carrier-primed cells was determined by treating each cell type in vitro with rabbit antirat B cell serum (RARBS) and complement. Although treatment with RARBS did not affect the adoptive response restored by carrier-primed cells, the same treatment abolished the response restored by hapten-primed cells. These findings indicate that T and B memory cells in the spleen of the rat are relatively long-lived, recirculating lymphocytes. The contribution of fixed or rapidly turning over cells to immunological memory is small or negligible as compared with the latter cells.

scholarly journals CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE

1968 ◽  
Vol 128 (4) ◽  
pp. 801-820 ◽  
Author(s):  
J. F. A. P. Miller ◽  
G. F. Mitchell
Keyword(s):  
Thoracic Duct ◽  
Bone Marrow Cells ◽  
Cell Types ◽  
Cell Interaction ◽  
Thoracic Duct Lymph ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
Duct Cells ◽  
Duct Lymph ◽  
Thymectomized Mice

An injection of viable thymus or thoracic duct lymphocytes was absolutely essential to enable a normal or near-normal 19S liemolysin-forming cell response in the spleens of neonatally thymectomized mice challenged with sheep erythrocytes. Syngeneic thymus lymphocytes were as effective as thoracic duct lymphocytes in this system and allogeneic or semiallogeneic cells could also reconstitute their hosts. No significant elevation of the response was achieved by giving either bone marrow cells, irradiated thymus or thoracic duct cells, thymus extracts or yeast. Spleen cells from reconstituted mice were exposed to anti-H2 sera directed against either the donor of the thymus or thoracic duct cells, or against the neonatally thymectomized host. Only isoantisera directed against the host could significantly reduce the number of hemolysin-forming cells present in the spleen cell suspensions. It is concluded that these antibody-forming cells are derived, not from the inoculated thymus or thoracic duct lymphocytes, but from the host. Thoracic duct cells from donors specifically immunologically tolerant of sheep erythrocytes had a markedly reduced restorative capacity in neonatally thymectomized recipients challenged with sheep erythrocytes. These results have suggested that there are cell types, in thymus or thoracic duct lymph, with capacities to react specifically with antigen and to induce the differentiation, to antibody-forming cells, of hemolysin-forming cell precursors derived from a separate cell line present in the neonatally thymectomized hosts.

Abstract 313: Twist Contributes to Cardiomyocyte Renewal and is Required for Maintenance of Adult Cardiac Function

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Yi-Li Min ◽  
Svetlana Bezprozvannaya ◽  
Drazen Šošic ◽  
Young-Jae Nam ◽  
Hesham Sadek ◽  
...  
Keyword(s):  
Cardiac Function ◽  
Cardiac Fibroblasts ◽  
Cell Lineage ◽  
Cardiac Regeneration ◽  
Cell Types ◽  
Bhlh Transcription Factor ◽  
Adult Heart ◽  
Twist 1 ◽  
Essential Contribution

Cardiomyocyte renewal occurs very slowly in adult mammals, and little is known of the genetic basis of cardiac regeneration. Twist is a highly conserved bHLH transcription factor responsible for Drosophila mesoderm formation during embryogenesis. Recent studies have shown that Twist protein is essential for muscle regeneration in adult Drosophila, but the potential role of Twist in the mammalian heart has not been explored. There are two Twist genes in vertebrates, Twist-1 and -2. We show that Twist-1 and -2 are expressed in epicardium and interstitial cells but not in differentiated cardiomyocytes in mice. To understand the potential function of Twist-dependent lineages in the adult heart, we generated inducible Twist2CreERT2; ROSA26-tdTomato reporter mice. By treating these mice with tamoxifen at 8 weeks of age, we observed progressive labeling of various cell types, such as epithelial cells, cardiac fibroblasts, and cardiomyocytes in the heart. We isolated Tomato-positive nonmyocytes from these mice and found that these cells can differentiate into cardiomyocytes and other cell types in vitro. Furthermore, cardiac-specific deletion of both Twist1 and Twist2 resulted in an age-dependent lethal cardiomyopathy. These findings reveal an essential contribution of Twist to long-term maintenance of cardiac function and support the concept of slow, lifelong renewal of cardiomyocytes from a Twist-dependent cell lineage in the adult heart.

scholarly journals Morphologic and Autoradiographic Observations of H3-Thymidine-Labeled Thoracic Duct Lymphocytes Cultured in Vivo

Blood ◽  
1960 ◽  
Vol 16 (2) ◽  
pp. 1133-1144 ◽  
Author(s):  
JOHN C. SCHOOLEY ◽  
IRWIN BERMAN
Keyword(s):  
Thoracic Duct ◽  
Temporal Pattern ◽  
Generation Time ◽  
Plasma Cells ◽  
Cell Types ◽  
Thoracic Duct Lymph ◽  
Duct Lymph ◽  
The Mean ◽  
Apparent Evidence

Abstract 1. The behavior of mouse and rat thoracic duct lymphocytes cultivated in diffusion chambers implanted into the peritoneal cavity of recipient mice and rats has been described. 2. The temporal pattern of labeling of cultured thoracic duct lymphocytes labeled with H3-thymidine has been described. From an analysis of this pattern and the changes in the mean grain count of the different classes of lymphocytes a maximum generation time for large and medium lymphocytes of 15 and 24 hours has been calculated. The results of these experiments favor an origin of small lymphocytes from the division of large and medium lymphocytes. 3. Some evidence for the transformation of thoracic duct lymph cells into monocytoid cells was found. In homologous cultures of labeled thoracic duct lymph cells and unlabeled bone marrow apparent evidence for transformation of labeled cells into plasma cells was found. The data suggest that neither the monocytoid cells nor the plasma cells arose necessarily from small lymphocytes. It was concluded that some unidentified cells, presumably the largest cells which are normally present in thoracic duct lymph, can be transformed into these other cell types when appropriately stimulated.

scholarly journals Differential regulation of spleen cell-mediated eosinophil and neutrophil-macrophage production

Blood ◽  
1980 ◽  
Vol 55 (3) ◽  
pp. 489-493 ◽  
Author(s):  
SH Bartelmez ◽  
WH Dodge ◽  
DA Bass
Keyword(s):  
Growth Factor ◽  
Contrast Media ◽  
Spleen Cell ◽  
Trichinella Spiralis ◽  
Cell Types ◽  
Differential Regulation ◽  
Spleen Cells ◽  
Secretory Products ◽  
Kinetics Of

Abstract Nonadherent spleen cells of mice infected with Trichinella spiralis released growth stimulatory factors (GSFs) in vitro when challenged with excretory/secretory products of muscle stage larvae. The assay of GSF was based on proliferation of normal, nonadherent syngeneic marrow cells in liquid tube cultures. Media conditioned for 1 day by challenged spleen cells stimulated eosinophil production but failed to stimulate production of other cell types. In contrast, media conditioned for 5 days supported eosinophil, neutrophil, and macrophage production. The kinetics of cell production were also different. Eosinophil production started within 1 day, reached a peak at day 2, and was down to control levels by day 4. In contrast, neutrophil/macrophage production began between 2 and 4 days and reached a peak at 6--8 days. The short duration of eosinophil production was evidently due to depletion of growth-factor-responsive cells.

scholarly journals CELL INTERACTIONS IN THE IMMUNE RESPONSE IN VITRO

1972 ◽  
Vol 135 (5) ◽  
pp. 1049-1058 ◽  
Author(s):  
Marc Feldmann
Keyword(s):  
Thoracic Duct ◽  
Glass Bead ◽  
Gamma Globulin ◽  
Antibody Responses ◽  
Cell Populations ◽  
Spleen Cells ◽  
Cooperative Response ◽  
Exact Function ◽  
Immune Response In Vitro

The requirement for macrophages in thymus-dependent antibody responses was studied in vitro. Three different macrophage-deficient cell populations were studied: spleen cells passed through a glass bead column at 37°C, spleen cells cultured with specific antimacrophage serum, and thoracic duct lymphocytes. These cell populations from mice primed to dinitrophenylated (DNP) fowl gamma globulin were unable to respond to the homologous conjugate in vitro. DNP-reactive B cells were present in normal proportions, since all three macrophage-depleted populations responded normally to macrophage-independent and thymus-independent DNP flagella. Carrier-reactive T cells were present, as the helper capacity of carrier-primed spleen cells was the same as carrier-primed lymphocytes, and thoracic duct lymphocytes are a well-established source of helper cells. The inhibition of the cooperative response was thus due to removal of macrophages, and this was proven by restoration of thymus-dependent anti-DNP responses by small numbers of anti-θ-treated peritoneal exudate cells. These results suggest that macrophages are essential in cell collaboration, While their exact function in cell collaboration is not yet known, the above observation suggests that the mechanism of T-B collaboration involves the surface of macrophages.

scholarly journals THE RATE OF DIVISION OF ANTIBODY-FORMING CELLS DURING THE EARLY PRIMARY IMMUNE RESPONSE

1968 ◽  
Vol 127 (5) ◽  
pp. 983-1002 ◽  
Author(s):  
Donald A. Rowley ◽  
Frank W. Fitch ◽  
Donald E. Mosier ◽  
Susan Solliday ◽  
Lionel W. Coppleson ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Drug Treatment ◽  
Cell Cultures ◽  
Cell Types ◽  
Primary Immune Response ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
Cycle Times ◽  
Blocking Agents

Mitotic blocking agents, colchicine or Velban, were used to estimate cycle times of spleen cells which release hemolysin for sheep erythrocytes (plaque-forming cells). The cells were obtained either from rats immunized with sheep erythrocytes or from cultures of mouse spleen cells immunized in vitro with the same antigen. 2, 3, or 4 days after immunization, animals or cell cultures were treated with mitotic blocking agents for periods of time ranging from 2.5 to 7 hr; plaque-forming cells were then enumerated. Decreased numbers of plaque-forming cells were found after such treatment. The extent of reduction was a function of duration of the drug treatment and the method of immunization, but was independent of the time after immunization. The evidence presented is consistent with premises that: (a) plaque-forming cells in mitosis do not release sufficient antibody to be detected, (b) mitotic blocking agents, by arresting plaque-forming cells in metaphase, prevent not only detection of these cells but also the increase in number of plaque-forming cells which would have resulted from cell division, (c) mitotic blocking agents do not affect release of antibody by cells in interphase. Cell cycle times, based on the extent of reduction of plaque-forming cells per unit time of drug treatment, were estimated using a mathematical model appropriate for an exponentially increasing population of cells. Cell cycle times estimated using the mitotic blocking agents agreed well with cell doubling times calculated from the increase in plaque-forming cells occurring 1–4 days after immunization. Increased responses produced by higher antigen doses or treatment of immunized animals with an adjuvant resulted from an increased rate of division of responding cells and their progeny. The results are consistent with a cell selection theory of antibody formation. Antigenic stimulation causes relatively few cells to proliferate and to synthesize antibody; apparently the magnitude of the response is dependent primarily on the rate of division of responding cells. It is suggested on the basis of observations of in vitro-immunized cell cultures that the rate of division of responding cells may be dependent on the rate of interaction between two cell types, both of which are essential for the in vitro plaque-forming cell response.

scholarly journals HEMOPOIETIC SPLEEN COLONY STUDIES

1967 ◽  
Vol 126 (5) ◽  
pp. 819-832 ◽  
Author(s):  
J. L. Curry ◽  
J. J. Trentin
Keyword(s):  
Bone Marrow ◽  
Cell Types ◽  
Slight Reduction ◽  
Normal Bone Marrow ◽  
Mixed Cell ◽  
Colony Forming Unit ◽  
Normal Bone ◽  
Endogenous Spleen Colonies

The effects of phytohemagglutinin (PHA) were studied in irradiated mice to see if a definite myeloproliferative effect could be demonstrated in vivo. The data obtained suggested the following conclusions. PHA treatment of the bone marrow donor only, causes a consistent but slight reduction in transplantable spleen colony-forming unit (CFU) content of the bone marrow 24 hr after the last PHA injection, but no change was found in the proportion of the various colony types. PHA treatment of the irradiated recipient of normal bone marrow causes no change in the number of spleen colonies. However, 8-day colonies are only about half normal size, are much more likely to be of mixed cell types, contain many large undifferentiated blastoid cells, but fewer transplantable CFU. The spleen sinusoids are packed with hemopoietic cells. Spleen colonies developing in hosts receiving daily injections of PHA show, in addition to the usual spectrum of cell types, a high proportion of unusual blastoid cells resembling the PHA transformed peripheral lymphocytes seen in vitro. The function of these cells is not known, but they may represent augmented proliferation and/or transformation of stem cells. PHA administered after irradiation significantly increased the number of endogenous spleen colonies, and, at certain doses of irradiation, improved postirradiation survival. PHA administered before irradiation had no effect on the number of endogenous spleen colonies formed, or on postirradiation survival. On the basis of these and other data, possible modes of action of PHA are discussed.

scholarly journals THE CARRIAGE OF IMMUNOLOGICAL MEMORY BY SMALL LYMPHOCYTES IN THE RAT

1966 ◽  
Vol 124 (5) ◽  
pp. 1017-1030 ◽  
Author(s):  
James L. Gowans ◽  
Jonathan W. Uhr
Keyword(s):  
Thoracic Duct ◽  
Immunological Memory ◽  
Antibody Responses ◽  
Thoracic Duct Lymph ◽  
Primary Immunization ◽  
Duct Cells ◽  
Duct Lymph ◽  
Secondary Type ◽  
Rapid Antibody

Lymphocytes were obtained from the thoracic duct of rats 1½ to 15 months after primary immunization with a single dose of bacteriophage ϕX 174. An intravenous injection of these lymphocytes conferred on heavily X-irradiated rats the ability to form antibody in a secondary-type manner after a first injection of ϕX. Negligible responses were obtained after cell transfer if the recipients were not challenged with antigen. Thoracic duct cells from some immunized donors were incubated in vitro for 24 hr before transfer in order to destroy selectively the large, dividing lymphocytes. The responsiveness conferred on X-irradiated recipients by such "incubated" inocula was then compared with that given by equal numbers of "fresh" thoracic duct cells. In all such comparisons the recipients of the "incubated" cells gave higher and more rapid antibody responses. It was concluded that the cells in thoracic duct lymph which carried immunological memory were small lymphocytes.

scholarly journals Generation of continuous large granular lymphocyte lines by interleukin 2 from the spleen cells of mice infected with Moloney leukemia virus. Involvement of interleukin 3.

1987 ◽  
Vol 166 (4) ◽  
pp. 833-849 ◽  
Author(s):  
M Hattori ◽  
T Sudo ◽  
M Iizuka ◽  
S Kobayashi ◽  
S Nishio ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Lines ◽  
Cdna Probe ◽  
Interleukin 2 ◽  
Cell Lineage ◽  
Constitutive Expression ◽  
Morphological Characteristics ◽  
Spleen Cells

Continuous cell lines could be reproducibly established by culturing spleen cells from adult mice injected with MLV-producer cells or directly infected with Mo-MLV with rIL-2, whereas the culture of normal splenic cells with rIL-2 induced only transient and limited proliferation resulting in no such lines. All of the lines showed morphological characteristics as LGL with Thy-1+,Lyt-1-,L3T4-,Lyt-2-,AsGM1+,FcR gamma+ phenotype without exception, and most of them exhibited typical NK-patterned cytotoxicity. Analysis of reverse transcriptase activity of the culture supernatants as well as Southern hybridization of the DNA from the lines using an Mo-MLV-specific cDNA probe indicated no evidence of retroviral replication or proviral integration, suggesting that the generation of cell lines reflected a reactive process and viral infection was not directly responsible. It was subsequently revealed that Thy-1+,Lyt-1+,Lyt-2- spleen cells from mice infected with Mo-MLV in vivo spontaneously produced surprising amounts of IL-3 in vitro, leading to the possibility that IL-3 was responsible for the generation of lines. The possibility was directly supported by the observation that continuous lines with identical characteristics could be generated completely in vitro by sequential stimulation with rIL-3 and rIL-2 from normal spleen cells without any involvement of Mo-MLV. The C beta gene of TCR was shown to be rearranged in all the lines examined, indicating the LGL lines were all genetically committed to T cell lineage. Unlike the situation in normal splenic populations expanded by rIL-2, where the expression of IL-2-R was progressively lost, constitutive expression of high-affinity-IL-2-R was observed in all the lines and thus, this was considered to explain the unlimited proliferation of them in response to rIL-2 alone. These results suggested the probable role of IL-3 in the regulation of growth and differentiation of a set of LGL committed to T cell lineage. The possible implications of the phenomenon in the regulation of hematopoiesis as well as in the control of Mo-MLV-induced leukemogenesis were discussed.

scholarly journals IDENTIFICATION OF MARROW-DERIVED AND THYMUS-DERIVED SMALL LYMPHOCYTES IN THE LYMPHOID TISSUE AND THORACIC DUCT LYMPH OF NORMAL RATS

1972 ◽  
Vol 135 (2) ◽  
pp. 200-219 ◽  
Author(s):  
Jonathan C. Howard ◽  
S. V. Hunt ◽  
J. L. Gowans
Keyword(s):  
Thoracic Duct ◽  
Lymphoid Tissue ◽  
Germinal Centers ◽  
Thoracic Duct Lymph ◽  
Lymphoid Tissues ◽  
Uridine Uptake ◽  
Duct Lymph ◽  
Lymphocyte Populations ◽  
Two Populations

These experiments show that small lymphocytes from the thoracic duct of rats are normally a mixture of thymus-derived and marrow-derived cells, and define the traffic areas in lymphoid tissues through which the two populations recirculate. Thoracic duct lymphocytes were labeled in vitro with uridine-3H and their histological distribution in the lymphoid tissues of normal recipients was demonstrated by radioautography. Labeled lymphocytes occupied two adjacent areas distinguished by a marked difference in the intensity of labeling; heavily labeled cells were found in thymus-dependent traffic areas of lymphocyte recirculation, while lightly labeled cells localized in the thymus-independent follicular areas around germinal centers. A corresponding heterogeneity of uridine uptake among small lymphocytes from normal donors was demonstrated by sedimentation at 1 g; slowly sedimenting cells incorporated little uridine and localized in follicular areas after transfusion while rapidly sedimenting cells incorporated more uridine and localized in thymus-dependent areas after transfusion. Experimentally prepared marrow-derived small lymphocytes behaved in sedimentation studies and after transfusion like a pure population of the lightly labeled small lymphocytes in normal lymph. Artificially reconstituted mixtures of marrow-derived and thymus-derived lymphocytes were qualitatively indistinguishable from normal lymphocyte populations.

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