Irradiated spleen cells

Spleen cells (4x10 ) were isolated to analyze the CTL response and restimulated with peptide-pulsed and irradiated spleen cells (2 X10 cells in 2 mL medium). On day 3, IL-2 (2.5 Lf/mL) was... 

OsoBA (1969 a, b) used the Marbrook system to culture varying numbers of normal mouse spleen cells (0.25-4 XIO ) mixed with irradiated spleen cells (to maintain a constant cell density) in the presence of SRC and/or ChRC. He observed that a certain fraction of cultures responded while other remained negative. 

Figure 1. Experimental Set up. Irradiated female mice were transplanted with retrovirally transduced BM cells from male donors (6 mice per vector). After 6 months of monitoring a secondary BMT took place. Spleen cells were used for the analysis of retroviral vector insertion sites (RVIS) by means of Ligation Mediated PCR (LM PCR) °. After another 6 months follow up spleen cells of the secondary recipients were again analysed for RVIS via LM-PCR.

The autoantibody production in autoimmune diseases may be attributed to the inability of Treg cells to control their synthesis. In an autoimmune model, T cells regulate the mechanisms through which B cells that were autoreactive to selfantigens do not produce autoantibodies, suggesting a role for suppressor T cells. The administration of irradiation, thymocytes, lymph nodes or spleen cells inhibits the production of autoantibodies, which is attributed to the suppressor T cells. 

Sodium fluoride did not induce reverse mutations in Salmonella typhimurium, nor did it induce gene conversion in Saccharomyces cerevisiae. A fluoride level of 0.4-1.0mgl inhibited DNA repair after irradiation of mouse spleen cells in vitro. Sodium fluoride was not mutagenic in cell cultures of human leukocytes at concentrations of 18 and 54mgl . Little or no effect was noted on chromosomes when mouse oocytes were exposed in vitro to a fluoride concentration of 200 mg 1 in media for up to 14 h. 

When a less severely immunocomprised host, that is, an adult thymec-tomized animal was used, it has been shown that suppressor cell activity can be restored with injections of thymosin or FTS and that phenotypic changes that occur in splenic T cells after thymectomy can be reversed by TP5 or FTS (Nash et al., 1981 Bach, 1977a). In an irradiated, thymectomized, and bone marrow-restored host, both bone marrow and spleen cells were induced with TF5 and Toj to provide helper cell activity for antibody production (Ahmed et al., 1979). 

In a typical experiment (Yamazaki, et al., in preparation), inbred mice of one H-2 type (e.g., H-2 ) were irradiated to destroy the entire hematopoietic system. Such mice would quickly succumb to infection if this system were not reconstituted with donor bone marrow and spleen cells. One group of irradiated animals was injected with these cells from a heterozygote (Fj ) donor (see Fig. 1). Another (control) group was injected with donor cells from individuals of the same H-2 type as itself. The success of the reconstitution was determined subsequently by cytotoxicity assay of lymph nodes. The question was, would the reconstituted animal produce odor more like his own genetic type or more like the donor type (an F heterozygote) In short, did cell replacement change the chemosensory identity of the subject in the direction of the donor type ... 

More importantly, spleen cells from GAT-primed non-responder DBA/1 mice specifically suppressed the GAT-specific PFC response of spleen cells from normal DBA/1 mice incubated with GAT-MBSA. This suppression was prevented by pretreatment of the GAT-primed spleen cells with anti-theta serum and complement or by X-irradiation (Figure 4.6). Identification of the suppressor cells as T cells was confirmed by the demonstration that the... 

Figure 4.10 DNP-ASC-primed spleen cells from A.TL, A, A.AL, A.TH and A.SW mice were depleted of T cells by in vitro treatment with anti-0 serum plus complement and then cultured with irradiated KLH-primed spleen cells from A.TL, A or (A x A.TH)Fi donors in the combinations indicated. Cells were cultured with either no antigen (not shown) or DNP-KLH.

Mice of High and Low lines were irradiated in order to suppress their immune responsiveness. This suppression is only due to the lymphoc)rtes destruction since it is reversed by the transfer of pure populations of small lymphocytes . Irradiated mice of each line received i.v. 4 x 10 spleen cells from (High x Low)Fj hybrids together with an immunizing dose of 3 x 10 SE. 

F spleen cells into irradiated High responders... 

Figure 5.12 Cell-transfer experiment production of SE agglutinin in irradiated High and Low responders after transfer of (High x Low)Fj spleen cells (4 x 10 per recipient). 3 x 10 SE are injected intravenously together with spleen cells. X rays 950 rad 24 hours before transfer. From Biozzi et al (1974). In La Genetique des Immunoglobulines et de la Reponse Immunitaire. Ann, Immunol (Inst. Pasteur) 125C, 107, by courtesy of Masson et Cie, Paris.

Mice were immunized intradermally with irradiated collagenese-dissoc-iated UV 2237 tumor cells, with or without poly(ICLC). Ten days later spleen cells were exposed to target cells, a 75Se methionine labeled clone of UV 2237, and the amount of released radioactivity released in the cultures of cells from animals that had received poly(ICLC) was measured. 3 Spleen cells from mice immunized with tumor cells show an increase in specific cytoxic activity as compared to spleen cells from mice inoculated with salt solution. Poly(ICLC) further enhanced the development of specific cytotoxic activity. 3... 

One other activity that is enhanced by the agent in mice is the mixed lymphocyte reaction (MLR). If live spleen cells from C57B1 mice are cocultivated with irradiated cells from C3H mice the live cells respond by increasing in growth rate, as measured by uptake of tritiated thymidine. If poly(ICLC) is added to the cell mixture during the cocultivation, in doses from 0.01 to 5 pg/ml., there is an increase in the MLR. At 10 pg/ml there was inhibition (6). These results are summarized in Table 6. 

The splenic foci technique was developed independently in two laboratories (Kennedy et al., 1965 Playfair et al., 1965), with the same goal to obtain an assay for the precursors of antibody-forming cells. In principle, a small inoculum of donor spleen cells and antigen (red cells) is injected into lethally irradiated syngeneic mice. After 5 or more days the host spleens are cut into many slices, each of which is further cut into separate pieces. These are placed on petri dishes containing a layer of agar with embedded red cells. Antibody is allowed to diffuse out and complement-dependent lysis is observed around some pieces. In this way, it is possible to construct a three-dimensional map of foci of antibody-producing cells. 

Fig. 5. Titration of B cells in microcultures. (From Quintans and Lefkovits, 1973.) All microcultures contained a constant number of allogeneic cells (2 X 10 per microculture). Irradiated and non-irradiated (active) nude spleen cells were mixed in different rations to give 1.8 X 10 total nude spleen cells per well. Abscissa indicates the input of active nude spleen cells per microculture. Each point is based on duplicate spot tests from 5 trays (300 microcultures). About 4.7X10 active B cells correspond to 37% negative microcultures (/ = 2.1 X 10 )...

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