CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE

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.

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CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE

Journal of Experimental Medicine ◽

10.1084/jem.128.4.821 ◽

1968 ◽

Vol 128 (4) ◽

pp. 821-837 ◽

Cited By ~ 406

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.

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CELL TO CELL INTERACTION IN THE IMMUNE RESPONSE

Journal of Experimental Medicine ◽

10.1084/jem.128.4.839 ◽

1968 ◽

Vol 128 (4) ◽

pp. 839-853 ◽

Cited By ~ 131

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.

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

Blood ◽

10.1182/blood.v16.2.1133.1133 ◽

1960 ◽

Vol 16 (2) ◽

pp. 1133-1144 ◽

Cited By ~ 25

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.

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

Journal of Experimental Medicine ◽

10.1084/jem.135.2.185 ◽

1972 ◽

Vol 135 (2) ◽

pp. 185-199 ◽

Cited By ~ 119

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.

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THE CARRIAGE OF IMMUNOLOGICAL MEMORY BY SMALL LYMPHOCYTES IN THE RAT

Journal of Experimental Medicine ◽

10.1084/jem.124.5.1017 ◽

1966 ◽

Vol 124 (5) ◽

pp. 1017-1030 ◽

Cited By ~ 115

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.

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BIOLOGICAL CHARACTERISTICS OF T AND B MEMORY LYMPHOCYTES IN THE RAT

Journal of Experimental Medicine ◽

10.1084/jem.137.5.1275 ◽

1973 ◽

Vol 137 (5) ◽

pp. 1275-1292 ◽

Cited By ~ 41

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.

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MATURATION OF B LYMPHOCYTES IN THE RAT

Journal of Experimental Medicine ◽

10.1084/jem.138.6.1331 ◽

1973 ◽

Vol 138 (6) ◽

pp. 1331-1344 ◽

Cited By ~ 48

Author(s):

Samuel Strober ◽

Jeanette Dilley

Keyword(s):

B Cells ◽

B Lymphocytes ◽

Thoracic Duct ◽

Time Course ◽

Turnover Rate ◽

Bone Marrow Cells ◽

Thoracic Duct Lymph ◽

Duct Cells ◽

Turn Over ◽

Marrow Cells

The migration pattern, tissue distribution, and turnover rate of unprimed and primed B lymphocytes involved in the adoptive anti-DNP response was studied. The adoptive primary response restored by unprimed spleen or thoracic duct cells passaged through an intermediate host (intravenous injection and subsequent collection in the thoracic duct lymph) was markedly diminished as compared with that restored by unpassaged cells. On the other hand, the adoptive response restored by passaged spleen or thoracic duct cells from DNP-primed donors was greater than or the same as that restored with unpassaged cells, respectively. This suggests that unprimed B cells change from nonrecirculating to recirculating lymphocytes after exposure to antigen. Studies of the adoptive anti-DNP response restored by unprimed or primed bone marrow cells showed little change in the time-course or amplitude of the response restored by either population of cells. The relative inability of marrow cells to carry immunological memory was related to the inability of recirculating memory cells to penetrate the marrow. The turnover rate of unprimed and primed B cells was investigated by treating the cell donors with [3H]thymidine for 48 h before removal of thoracic duct or spleen cells. The adoptive anti-DNP response restored by unprimed or primed cells was not affected by [3H]thymidine treatment. This indicates that both populations of cells turn over slowly. However, our previous studies show that unprimed B cells involved in the adoptive antibody response to ferritin turn over rapidly. The different findings are discussed in the context of antigen-dependent B-cell maturation.

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MECHANISM OF EOSINOPHILIA

Journal of Experimental Medicine ◽

10.1084/jem.131.6.1288 ◽

1970 ◽

Vol 131 (6) ◽

pp. 1288-1305 ◽

Cited By ~ 235

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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