In Vitro Studies on Radiation Lymphoid Recovery of Mouse Spleen

Irradiated rabbits given allogeneic bone marrow cells from normal adult donors responded to an injection of sheep red blood cells by forming circulating antibodies. Their spleen cells were also capable of forming many plaques using the hemolysis in gel technique, and were also capable of undergoing blastogenesis and mitosis and of incorporating tritiated thymidine upon exposure to the specific antigen in vitro. However, irradiated rabbits injected with allogeneic bone marrow obtained from rabbits injected with sheep red blood cells 24 hr prior to sacrifice (primed donors) were incapable of mounting an immune response after stimulation with sheep red cells. This loss of reactivity by the bone marrow from primed donors is specific for the antigen injected, since the immune response of the irradiated recipients to a non-cross-reacting antigen, the horse red blood cell, is unimpaired. Treatment of the bone marrow donors with high-titered specific antiserum to sheep red cells for 24 hr prior to sacrifice did not result in any diminished ability of their bone marrow cells to transfer antibody-forming capacity to sheep red blood cells. The significance of these results, with respect to the origin of the antigen-reactive and antibody-forming cells in the rabbit, is discussed.

Section: Discussionsupporting

confidence: 84%

Cells Involved in the Immune Response

Abdou

Richter

1969

“…(17,18). Globerson has reported that nonrespondent cells could be made to respond in vitro, provided that they were first stinmlated with the mitogen PHA (24,25). It may be that the process described here as priming is in reality not an immune phenomenon, but rather a tissue culture requirement for cycling cells.…”

Section: Discussionmentioning

confidence: 95%

In Vitro and in Vivo Studies of the Immune Response to Sheep Erythrocytes Using Partially Purified Cell Preparations

Haskill

Byrt

Marbrook

1970

The BSA density-gradient technique for separating mouse spleen cells into partially purified populations has been used to compare the responsiveness of such populations to SRBC using in vivo and in vitro techniques. Two major populations were distinguished, one of which responded very well in vivo with an exponential dose response and poorly in vitro (fraction 3), and another which responded in vivo and in vitro with a linear dose response (fraction 2). A light density, radiation-resistant component was identified which markedly stimulated the response of fraction 3 in vitro, and a density gradient profile was obtained for this cell which did not correspond with a macrophage profile. A high density, radiation-sensitive cell was identified which stimulated the response of PFC precursors in lighter regions of the gradient. The activity of this cell could be replaced using thymus cells. A density profile for the PFC precursor cell was obtained by assaying small numbers of spleen cell fractions in the presence of an excess of the two auxiliary cell types.

“…Preparation of Tissue Explants and Culture Technique.--Methods were identical with those described in preceding reports (16)(17)(18)(19). Transfilter combinations of tissues were prepared according to Grobstein's technique ( 19,20,21) using a chicken plasma clot to fasten tissue'~ in the desired position.…”

Section: Methodsmentioning

confidence: 99%

“…It was therefore decided to test whether splenomegaly can be induced by irradiated spleens following interaction with bone marrow, since such tissue combinations of irradiated spleen and bone marrow had been previously shown to become lymphoid (18). BALB/C-irradiated spleen fragments, combined with bone marrow, derived from syngeneic origin, were cultivated for 3 days.…”

Section: Methodsmentioning

confidence: 99%

“…A survey of lymphoid radiation recovery of mouse spleen (18) indicated that REACTIVATION IN VITRO OF IMMUNOCONIPETENCE lymphoid regeneration depends upon interaction of the irradiated spleen and bone marrow, and that this is enhanced by the thymus. The present study extended these experiments to analysis of immunological reactivity, attempting to test (a) what is the tissue origin of immunocompetent cells, (b) what is the role of the thymus in the process of regeneration of immune reactivity, and (e) whether immune reactivity and lymphoid development are separable systems.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Reactivation in Vitro of Immunocompetence in Irradiated Mouse Spleen

Globerson

Auerbach

1967

Self Cite

The restoration of immunological competence to irradiated adult mouse spleen was studied using an in vitro assay system. Competence of the spleen to evoke a graft versus host reaction in vitro was lost upon sublethal irradiation (550 R). Regeneration of immune competence occurred when such spleens were grown for 2–3 days in the presence of thymus tissue, but not when grown in the presence of a variety of other tissues. This thymic activity was demonstrated to occur across an intervening Millipore filter barrier. When lethal doses of irradiation were used, reactivation of immune competence did not occur unless both thymus and bone marrow were present. In this experimental situation the bone marrow appeared to supply the immunocompetent cells.