scholarly journals CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE

1968 ◽  
Vol 128 (4) ◽  
pp. 821-837 ◽  
Author(s):  
G. F. Mitchell ◽  
J. F. A. P. Miller
Keyword(s):  
Bone Marrow ◽  
Synergistic Effect ◽  
Thoracic Duct ◽  
Cell Types ◽  
Cell Interaction ◽  
Thoracic Duct Lymph ◽  
Sheep Erythrocytes ◽  
Duct Cells ◽  
Duct Lymph ◽  
Thymus Cells

The number of discrete hemolytic foci and of hemolysin-forming cells arising in the spleens of heavily irradiated mice given sheep erythrocytes and either syngeneic thymus or bone marrow was not significantly greater than that detected in controls given antigen alone. Thoracic duct cells injected with sheep erythrocytes significantly increased the number of hemolytic foci and 10 million cells gave rise to over 1000 hemolysin-forming cells per spleen. A synergistic effect was observed when syngeneic thoracic duct cells were mixed with syngeneic marrow cells: the number of hemolysin-forming cells produced in this case was far greater than could be accounted for by summating the activities of either cell population given alone. The number of hemolytic foci produced by the mixed population was not however greater than that produced by an equivalent number of thoracic duct cells given without bone marrow. Thymus cells given together with syngeneic bone marrow enabled irradiated mice to produce hemolysin-forming cells but were much less effective than the same number of thoracic duct cells. Likewise syngeneic thymus cells were not as effective as thoracic duct cells in enabling thymectomized irradiated bone marrow-protected hosts to produce hemolysin-forming cells in response to sheep erythrocytes. Irradiated recipients of semiallogeneic thoracic duct cells produced hemolysin-forming cells of donor-type as shown by the use of anti-H2 sera. The identity of the hemolysin-forming cells in the spleens of irradiated mice receiving a mixed inoculum of semiallogeneic thoracic duct cells and syngeneic marrow was not determined because no synergistic effect was obtained in these recipients in contrast to the results in the syngeneic situation. Thymectomized irradiated mice protected with bone marrow for a period of 2 wk and injected with semiallogeneic thoracic duct cells together with sheep erythrocytes did however produce a far greater number of hemolysin-forming cells than irradiated mice receiving the same number of thoracic duct cells without bone marrow. Anti-H2 sera revealed that the antibody-forming cells arising in the spleens of these thymectomized irradiated hosts were derived, not from the injected thoracic duct cells, but from bone marrow. It is concluded that thoracic duct lymph contains a mixture of cell types: some are hemolysin-forming cell precursors and others are antigen-reactive cells which can interact with antigen and initiate the differentiation of hemolysin-forming cell precursors to antibody-forming cells. Bone marrow contains only precursors of hemolysin-forming cells and thymus contains only antigen-reactive cells but in a proportion that is far less than in thoracic duct lymph.

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.

scholarly journals CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE

1968 ◽  
Vol 128 (4) ◽  
pp. 839-853 ◽  
Author(s):  
G. J. V. Nossal ◽  
A. Cunningham ◽  
G. F. Mitchell ◽  
J. F. A. P. Miller
Keyword(s):  
Bone Marrow ◽  
Immune Response ◽  
Immune Responses ◽  
Thoracic Duct ◽  
Bone Marrow Cells ◽  
Cell Interaction ◽  
Thoracic Duct Lymph ◽  
Chromosomal Marker ◽  
Sheep Erythrocytes ◽  
Marrow Cells

Two new methods are described for making chromosomal spreads of single antibody-forming cells. The first depends on the controlled rupture of cells in small microdroplets through the use of a mild detergent and application of a mechanical stress on the cell. The second is a microadaptation of the conventional Ford technique. Both methods have a success rate of over 50%, though the quality of chromosomal spreads obtained is generally not as good as with conventional methods. These techniques have been applied to an analysis of cell to cell interaction in adoptive immune responses, using the full syngeneic transfer system provided by the use of CBA and CBA/T6T6 donor-recipient combinations. When neonatally thymectomized mice were restored to adequate immune responsiveness to sheep erythrocytes by injections of either thymus cells or thoracic duct lymphocytes, it was shown that all the actual dividing antibody-forming cells were not of donor but of host origin. When lethally irradiated mice were injected with chromosomally marked but syngeneic mixtures of thymus and bone marrow cells, a rather feeble adoptive immune response ensued; all the antibody-forming cells identified were of bone marrow origin. When mixtures of bone marrow cells and thoracic duct lymphocytes were used, immune restoration was much more effective, and over three-quarters of the antibody-forming mitotic figures carried the bone marrow donor chromosomal marker. The results were deemed to be consistent with the conclusions derived in the previous paper of this series, namely that thymus contains some, but a small number only of antigen-reactive cells (ARC), bone marrow contains antibody-forming cell precursors (AFCP) but no ARC, and thoracic duct lymph contains both ARC and AFCP with a probable predominance of the former. A vigorous immune response to sheep erythrocytes probably requires a collaboration between the two cell lineages, involving proliferation first of the ARC and then of the AFCP. The results stressed that the use of large numbers of pure thoracic duct lymphocytes in adoptive transfer work could lead to good adoptive immune responses, but that such results should not be construed as evidence against cell collaboration hypotheses. Some possible further uses of single cell chromosome techniques were briefly discussed.

scholarly journals Brief Note: Marrow Repopulating Ability of Peripheral Blood Cells Compared to Thoracic Duct Cells

Blood ◽  
1968 ◽  
Vol 32 (4) ◽  
pp. 662-667 ◽  
Author(s):  
R. STORB ◽  
R. B. EPSTEIN ◽  
E. D. THOMAS
Keyword(s):  
Bone Marrow ◽  
Thoracic Duct ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Whole Body ◽  
Thoracic Duct Lymph ◽  
Peripheral Blood Cells ◽  
Hemopoietic Stem Cells ◽  
Duct Lymph ◽  
Marrow Repopulating Ability

Abstract Ten dogs were exposed to 1200 r. of whole body irradiation at a dose rate of 9.2 r./min. Five of these dogs were then given infusions of 21 to 74 x 109 autologous peripheral blood cells which had been previously stored at -80 C. 4.0 to 19.4 x 109 of these cells were lymphocytes, 0.4 to 4.9 x 109 were monocytes and 16.4 to 50.3 x 109 were granulocytes. All five dogs showed clinical or histologic evidence of bone marrow repopulation. The remaining 5 dogs were given 7 to 22 x 109 autologous thoracic duct lymphocytes. In none of these dogs was marrow repopulation observed. It was concluded that hemopoietic stem cells are not present in the thoracic duct lymph of the dog in any appreciable number.

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 THE LIFE-SPAN AND RECIRCULATION OF MARROW-DERIVED SMALL LYMPHOCYTES FROM THE RAT THORACIC DUCT

1972 ◽  
Vol 135 (2) ◽  
pp. 185-199 ◽  
Author(s):  
Jonathan C. Howard
Keyword(s):  
Bone Marrow ◽  
Life Span ◽  
B Lymphocytes ◽  
Thoracic Duct ◽  
Bone Marrow Cells ◽  
Thoracic Duct Lymph ◽  
Clear Cut ◽  
Duct Lymph ◽  
Order Of Magnitude ◽  
Marrow Cells

These experiments describe the preparation of pure marrow-derived lymphocyte suspensions from the thoracic duct of thymectomized, irradiated rats reconstituted with bone marrow cells. The majority of marrow-derived cells were small lymphocytes morphologically indistinguishable from small lymphocytes in thoracic duct lymph of normal donors. Marrow-derived small lymphocytes (B lymphocytes) were a predominantly long-lived population; the frequency of short-lived B lymphocytes in the thoracic duct was not significantly higher than the frequency of short-lived small lymphocytes in normal lymph. B lymphocytes transferred to normal recipients recirculated from blood to lymph. The first appearance of intravenously injected B lymphocytes in the thoracic duct was delayed relative to lymphocytes from normal donors and there was no clear cut modal recirculation time. Nevertheless their recirculation over a 48 hr period after transfusion was of the same order of magnitude as that of lymphocytes from normal donors.

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 Characterization of nonlymphoid cells derived from rat peripheral lymph

1983 ◽  
Vol 157 (6) ◽  
pp. 1758-1779 ◽  
Author(s):  
CW Pugh ◽  
GG MacPherson ◽  
HW Steer
Keyword(s):  
Bone Marrow ◽  
Dendritic Cells ◽  
Thoracic Duct ◽  
Tritiated Thymidine ◽  
Thoracic Duct Lymph ◽  
Nonspecific Esterase ◽  
Cytoplasmic Inclusions ◽  
Peripheral Lymph ◽  
Duct Lymph

Mesenteric lymphadenectomy in rats is followed by union of peripheral and central lymphatics, allowing the collection of intestine-derived peripheral lymph cells via the thoracic duct for several days. These cells include a proportion of nonlymphoid cells (NLC) that show irregular and heterogeneous surface morphology including long pseudopodia and veils. They stain variably for nonspecific esterase and acid phosphatase and are ATPase-positive. Their nuclei are irregular and some contain cytoplasmic inclusions, some of which show peroxidase activity and/or contain DNA. NLC have a range of densitites generally lower than that of lymphocytes. Freshly collected NLC express the leukocyte-common antigen (defined by monoclonal antibody MRC Ox 1) and Ia antigens (I-A and I-E subregion products defined by monoclonal antibodies) but they show a relative lack of other surface markers normally found on rat B or T lymphocytes (W3/13, W3/25, MRC Ox 12 (sIg), MRC Ox 19) or rat macrophages (FcR, C'R, mannose R, W3/25). In general NLC are only weakly adherent to glass or plastic. Although a subpopulation of NLC appear to have had a phagocytic past, freshly collected NLC fail to phagocytose a variety of test particles in vitro. NLC also appear incapable of pinocytosis in vitro. This heterogeneity may represent distinct subpopulations of NLC or different stages in the development of a single cell lineage. Direct cannulation of mesenteric lacteals shows that the majority of NLC are derived from the small intestine and their precursors appear to be present both in lamina propria and Peyer's patches. Kinetic studies, following irradiation or intravenous tritiated thymidine, show that the majority of NLC turn over rapidly in the intestine with a modal time of 3-5 d. Studies with bone marrow chimeras show that they are derived from a rapidly dividing precursor present in normal bone marrow. NLC occur at very low frequencies in normal thoracic duct lymph at all times following cannulation. The evidence presented suggests that NLC closely resemble mouse lymphoid dendritic cells. This conclusion is supported by evidence already obtained showing that NLC are potent stimulators of the semi-allogeneic rat primary mixed leukocyte reaction. In addition to the ceils resembling dendritic cells rare monocytoid cells are found in thoracic duct lymph of lymphadenectomized specific pathogen-free rats. The proportion of these cells increases greatly when the animals are conventionally housed. It seems probable that the physiological function of NLC is to act as accessory cells in the lymph nodes to which they normally drain. Methods for enriching NLC and thus facilitating analysis of their functions are discussed.

scholarly journals THE ORIGIN AND TURNOVER OF MONONUCLEAR CELLS IN PERITONEAL EXUDATES IN RATS

1966 ◽  
Vol 124 (2) ◽  
pp. 241-254 ◽  
Author(s):  
Alvin Volkman
Keyword(s):  
Bone Marrow ◽  
Experimental Evidence ◽  
Thoracic Duct ◽  
Mononuclear Cells ◽  
Tritiated Thymidine ◽  
Thoracic Duct Lymph ◽  
Duct Lymph ◽  
Kinetics Of

Tritiated thymidine-labeling data in individual and parabiotic rats showed that macrophages in peritoneal exudates were derived from cells in the blood which were the progeny of rapidly and continuously proliferating precursors. The characteristics of this population identify them with free macrophages studied in other sites; similarly, they can be obtained from transfused bone marrow. Cells in the exudates which were morphologically indistinguishable from small lymphocytes were also found to have the labeling features of a rapidly proliferating population in contrast with the known kinetics of the majority of small lymphocytes in blood and thoracic duct lymph. However, experimental evidence indicated that the lymphocytelike exudate cells had emigrated from the blood and that bone marrow was a source of their precursors. These findings support the concept of the heterogeneity of lymphocytes. The possible relationships among the mononuclear cells is discussed.

scholarly journals MECHANISM OF EOSINOPHILIA

1970 ◽  
Vol 131 (6) ◽  
pp. 1288-1305 ◽  
Author(s):  
Antony Basten ◽  
Paul B. Beeson
Keyword(s):  
Bone Marrow ◽  
Thoracic Duct ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Thoracic Duct Lymph ◽  
Antilymphocyte Serum ◽  
Diffusion Chambers ◽  
Duct Lymph ◽  
Diffusible Factor ◽  
Secondary Type

A possible role for the lymphocyte in the mechanism of eosinopoiesis has been examined. Procedures known to deplete or inactivate the pool of recirculating lymphocytes such as neonatal thymectomy, administration of antilymphocyte serum, and prolonged thoracic duct drainage, either singly or in combination, resulted in a highly significant reduction in the eosinophil response to trichinosis. Irradiated animals exposed to parasitic challenge did not develop eosinophilia unless reconstituted with lymphocytes as well as bone marrow cells. When "memory" cells were used instead of normal lymphocytes, a "secondary" type of eosinophil response was observed. Transfer of a primary eosinophilia was achieved adoptively with a population of living large lymphocytes from thoracic duct lymph and peripheral blood, but not with blood plasma or cell-free lymph. The potency of the active lymphocytes was not impaired by enclosing them in cell-tight diffusion chambers, indicating that they exerted an effect on bone marrow by agency of a diffusible factor. The demonstration of a role for lymphocytes in induction of the eosinophil response to this kind of stimulus supports the conclusion that eosinophilia belongs in the category of immunologic phenomena.

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