S.P. Lerman

Properties of reticulum cell sarcomas in sjl/j mice. Iii. Promotion of tumor growth in irradiated mice by normal lymphoid cells.

Abstract Previous irradiation (660–730 R) or cyclophosphamide treatment (400 mg/kg) of SJL/J mice markedly inhibited their ability to support growth of syngeneic intravenously (iv) injected reticulum cell sarcoma (RCS). Normal syngeneic lymphoid cells (5 × 10 7 ) injected iv after irradiation and 1 day prior to RCS cells partially restored the tumor growth in spleen and lymph nodes of irradiated mice as determined 5 to 7 days after RCS injection, but normal SJL/J serum injections had no effect. The cell type crucial for this effect appeared to be a macrophage, but some of the observations suggested the need for cell cooperation between T cells and a macrophage-like, bone marrow derived cell in this phenomenon. Pretreatment of tumor recipients with macrophage inhibiting agents, such as silica, trypan blue and carrageenan, also tended to inhibit tumor growth, particularly when combined with a dose of irradiation (460 R) which by itself had only a limited effect. However, macrophage stimulating agents did not protect against the irradiation effect.

Properties of reticulum cell sarcomas in SJL/J mice: IV. Minimal development of cytotoxic cells despite marked proliferation to syngeneic RCS in vivo and in vitro☆

Abstract Lymphoid cells from normal SJL/J mice gave high proliferative responses but failed to develop cytotoxic activity to γ-irradiated cells from syngeneic transplantable reticulum cell sarcomas (X-RCS). In spite of a vigorous in vivo proliferative response to X-RCS, cytotoxic activity was never generated to detectable levels in vivo. After repeated injections of X-RCS, spleen and, to a lesser degree, lymph node cells acquired the ability to give moderate secondary cytotoxic responses in vitro upon co-culture with X-RCS. This immunity was T-cell mediated and specific for RCS although it did not distinguish between different transplantable RCS lines. SJL/J mice also developed resistance to RCS growth after injection of X-RCS, which showed a transient RCS-line-specific component. (SJL/J × C57B1/6)F1 mice showed 60% less RCS growth than did SJL/J mice, and their lymphoid cells gave slightly lower proliferative responses than did cells from SJL/J mice, whereas (SJL/J × BALB/c)F1 mice showed little tumor growth, and their spleen cells proliferated only minimally to X-RCS. B10.S mice allowed moderate RCS growth. Cytotoxic activity was generated in co-cultures with X-RCS of immunized F1 spleen cells even after a single immunization in vivo but not in cultures of normal F1 cells with X-RCS.

Properties of reticulum cell sarcomas in sjl/j mice. II. Fate of labeled tumor cells in normal and irradiated syngeneic mice.

Abstract Transplantable reticulum cell sarcoma (RCS) cells were labeled with 3 H-uridine or 3 H-thymidine in vitro and injected intravenously into normal and irradiated syngeneic SJL/J mice. RCS cells exhibited typical B cell migration characteristics in peripheral lymphoid organs in both normal and irradiated recipients, localizing in follicles in a pattern resembling that of labeled normal bone marrow cells. However, over the first 72 hr after transfer, RCS cells diluted their label much less in irradiated than in normal recipients, reflecting their inability to proliferate in the irradiated hosts. The presence of unlabeled tumor cells did not significantly affect the distribution of labeled normal bone marrow or lymph node cells in the recipients. Thus, RCS fails to grow in irradiated recipients in spite of undisturbed homing characteristics and in the absence of any evidence of cytotoxic influences from the host.

Induction of primary and inhibition of secondary antibody response to hapten by hapten conjugates of type III pneumococcal polysaccharide

Abstract Tri- or dinitrophenylated pneumococcal polysaccharide type III (TNP- or DNP SIII)) induced a primary 19S anti-TNP response without generating immunological memory to the hapten in LAF 1 mice. Hapten-hemocyanin (TNP-KLH) or hapten conjugates of B. abortus organisms (DNP-BA) induced both 19S and 7S primary responses and memory to the hapten. Spleen cells from mice immunized with TNP-KLH or DNP-BA did not give adoptive memory responses upon challenge with hapten-SIII and, in fact, were inhibited from responding to their homologous hapten conjugates by simultaneous injection of hapten-SIII. Incubation of TNP-KLH-primed spleen cells for as short as 5 min at 0 °C with 10 μg of TNP-SIII per milliliter virtually abolished their ability to give 19S and 7S memory responses to TNP-KLH upon transfer into irradiated recipients. It is suggested that a difference in avidity and/or number of anti-TNP receptors per cell between virgin and primed B cells may be an important factor in determining whether the cells will be stimulated or inhibited by exposure to hapten-SIII. Another factor may be a difference between virgin and memory cells in their requirement for T-cell help.

Properties of reticulum cell sarcomas in sjl/j mice. Vii. Nature of normal lymphoid cells proliferating in response to tumor cells.

Abstract Treatment of SJL lymph node, spleen, and thymus cells with anti-Ly 1.2 serum and C caused a marked reduction in the usually observed T-cell proliferation in response to syngeneic X-irradiated transplantable reticulum cell sarcoma cells. By contrast, treatment of lymphoid cells with anti-Ly 2.2 serum and C either failed to affect or increased the proliferative response. It is concluded that the Ly 1 cell is the major T-cell subpopulation which proliferates in response to RCS.

Transfer of agammaglobulinemia in the chicken. I. Generation of suppressor activity by injection of bursa cells.

Abstract Spleen cells from adult agammaglobulinemic (bursectomized) chickens taken 1 to 3 weeks after an injection of histocompatible bursa cells can inhibit the adoptive antibody response to B. abortus of normal spleen or bursa cells in irradiated recipients. Spleen cells from Aγ chickens not injected with bursa cells generally do not. Moreover, bursectomized chickens which have been reconstituted with spleen cells within the first week after hatching do not respond with suppressor cell formation upon bursa cell injection. This apparent “autoimmunization” with bursa cells induces suppressor T cells which are only minimally sensitive to treatment with mitomycin C or to 5000 R γ irradiation. The suppressor activity is neither induced nor potentiated by concanavalin A in vivo . It is much stronger in spleen than in thymus cells and appears to be macrophage independent and to require intact cells. The cell component which stimulates the suppressor activity is more pronounced on bursa than on spleen cells, and is at most present to a very limited extent on bone marrow, thymus, or peritoneal exudate cells. It is better represented in comparable cell numbers of Day 17 than of Day 14 embryonic bursa. The inducing cell component is present in the membrane fraction of disrupted bursa cells. Immunization with bursa cells from B locus histoincompatible chickens leads to suppressor activity against histocompatible bursa cells. Although the removal of Ig-bearing cells from bursa greatly diminishes its immunizing capacity, injection of serum IgM and IgG does not induce suppressor cells. It is suggested that tolerance to a B-cell antigen is lacking in adult Aγ chickens, resulting in an autoimmune response upon exposure to B cells. The B-cell antigen may be a cell surface-specific form of Ig, a complex of Ig and a membrane component, or a differentiation antigen which appears simultaneously with Ig during ontogeny.

Growth of SJL/J-derived RCS as related to its ability to induce T cell proliferation in the host. II. Negative influence of H-2d1.

Backcross SJL x (SJL x BALB/c)F1 and (SJL x BSVS)F1 mice were examined for their ability to support growth of transplantable SJL lymphoma (reticulum cell sarcoma (RCS). A marked linkage to H-2 was noted in that H-2s/d backcross mice failed to support tumor growth, while H-2s/s backcross mice showed approximately 70% of the growth seen in SJL mice, as judged by lymph node and spleen weights. Spleen cells obtained from backcross mice by splenectomy were examined for their ability to give proliferative responses to gamma-RCS cells, whereafter individual splenectomized mice were also examined for their ability to support lymphoma growth. Both properties showed a similar degree of linkage to H-2 and to each other, although there seemed to be a segregating non-H-2 BALB gene which also exerted an additional, less marked negative influence on the proliferative responses. It is suggested that the proliferative response in vivo may contribute to the lymphoma growth and that the presence of H-2d is inhibitory. (SJL x BSVS)F1 mice gave excellent proliferative responses and supported growth of RCS to approximately 80% of those of controls. These results confirm previous conclusions on the negative effect of H-21d in F1 hybrids on both phenomena.

Transfer of agammaglobulinemia in the chicken. II. Characterization of the target of suppression.

Abstract The T-independent adoptive primary and secondary responses of FP chicken spleen cells to B. abortus (BA) were suppressed by simultaneously transferred histocompatible spleen cells from agammaglobulinemic (Aγ) chickens previously injected with bursa cells. The response of spleen cells transferred with BA 2 days prior to suppressor cells was partially inhibited. Highly significant suppression of both 19 S and 7 S antibody formation was seen during the second week after memory cell transfer. When suppressor cells and primed spleen cells were transferred together and challenged with BA 2 weeks later the secondary responses were more inhibited in recipients that showed persistently decreased serum IgM than in those showing recovery toward normal IgM levels. Transfer of histocompatible suppressor cells into surgically bursectomized (SBX). irradiated, 4-week-old SC and FP strain recipients caused reduction or complete disappearance of serum IgM and lowered IgG levels, which correlated well with a decrease in plasma cell numbers in spleen and trident. Many recipients of both strains, examined 1 to 2 weeks after transfer, completely lacked plasma cells from gut-associated lymphoid tissues and from spleen. When such animals had been sensitized to BA prior to transfer and were challenged after recovery from suppression, they showed good secondary responses. Experiments with serially transferred memory cells using suppressed intermediate hosts also indicated survival of such cells in the face of marked suppression of antibody synthesis. Bursa follicles that were not destroyed by irradiation did not show any effect of the suppressor cells, although plasma cells often disappeared from the interfollicular areas. Intact irradiated recipients showed a greater tendency toward recovery from suppression than did SBX recipients. Although Ig-bearing cells in spleen and peripheral blood of suppressed animals were significantly lower than in irradiated SBX controls, they never disappeared as completely as did plasma cells and serum IgM. The results suggest a direct effect of suppressor cells on antibody-forming cells with a less marked effect on their precursors.

Accessory cell requirements for lymphoma growth in vitro and in irradiated mice

Abstract Growth of the transplantable B-cell lymphoma, PU-5, is markedly diminished in γ-irradiated as compared to normal BALB/c mice. Transfer of bone marrow, but not of lymph node or peritoneal exudate cells, partially restored the ability of irradiated mice to support lymphoma growth. In vitro growth of PU-5 cells is promoted by silica-sensitive, adherent cells, bearing surface Ia antigen and present in peritoneal exudates, spleen and lymph node, but not in bone marrow. Their action on PU-5 growth can be shown only in rocking cultures; the cells do not have to be histocompatible, they act synergistically with 2-mercaptoethanol (2-ME) in the medium. The growth-promoting action in vitro is decreased 24 hr after γ-irradiation of the adherent cells in vitro . Growth of transplantable reticulum cell sarcoma in SJL/J mice has previously also been shown to be inhibited by prior irradiation of the host and to be restored by transfer of lymphoid cells including a phagocytic component, but in the present studies no consistent growth-promoting effect of accessory cells on reticulum cell sarcomas has been shown in vitro . Both lymphomas are stimulated by the presence of 2-ME in stationary cultures. The relationship between the in vivo and in vitro lymphoma growth-promoting activities of macrophage-like cells is discussed.

Lymphoid-Cell Cooperation in Immune Responses of the Chicken

Our knowledge of cellular interactions in the immune response of chickens is in a primitive stage as compared with the degree of sophistication with which such interactions are being interpreted for mouse cells. Relatively little use has been made of the unique position that the bursa occupies in the ontogeny of B cells, as initially shown by the experiments on bursectomy by Glick et al. (1956), Mueller et al. (1960), and Warner et al. (1962). Although there is still some difference of opinion about this (Jankovic et al., 1975), it seems likely that the chicken has no other source of virgin B cells (Kincade et al., 1973) and that the bursa is, from approximately day 13 until day 19 of ontogeny, the only site of B-cell formation and Ig synthesis (Thorbecke et al., 1968; Kincade and Cooper, 1971). The lymphoid follicles in the bursa arise from the influx of hematopoietic cells into the epithelial site near the cloaca (Moore and Owen, 1965, 1966). This influx starts during the 8th day and continues until at least the 15th day of incubation (Houssaint et al., 1976). Once cells have differentiated in the bursa, they have different surface antigens from those of cells in the thymus (Albini and Wick, 1974; Galton and Ivanyi, 1977; Donnelly et al., 1975; McArthur et al., 1971; Forget et al., 1970), are slightly larger (Sherman and Auerbach, 1966), and have a somewhat different chromatin distribution (Olson et al., 1973).