Donor spleen cells

Interferon, given to mice, inhibited the graft-versus reaction. Contrary results were seen with interferon inducers. In a graft-versus-host test. Cantor, et al., showed that the interferon inducer poly(I)-poly(C) could enhance the activity of the donor spleen cells used in the test by as much as 3-fold. Poly (A)-poly (U) exerted a similar but lesser effect. In this study interferon gave equivocal results. Comparably, Turner, et al., showed that poly(I)-poly(C) increased rather than decreased allograft rejection. 

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. 

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.

Monoclonal antibodies are produced as a result of immortalizing and expanding the individual antibody secreting cells artificially in tissue culture (2). Cells grown in this way all have identical epitope specificity and because they are derived from single clones, their product is known as monoclonal antibody. Cells that secrete monoclonal antibodies are known as hybridomas and are typically derived by fusion of two cell types. B-lymphocytes, which have the capacity to make antibody, are obtained from a donor spleen and are physically fused to a tumor cell line, which is immortal. The resulting hybridom as are immortal and produce antibody into the synthetic medium in which they are growing. 

In 1973, Cotton et al. successfully fused cells of two plasmacytoma lines to produce hybrid cells capable of synthesizing both myeloma proteins. Subsequently, hybrid cells derived by fusion of a murine myeloma with spleen cells from appropriately immunized donors were shown to se-... 

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 ... 

Fig, 1, Constitution of radiation chimeras. Inbred homozygous mice (H-2 type bb) were given 940 rads of gamma radiation, destroying the hematopoietic (blood-forming) system. Shortly thereafter, the mice were intravenously injected with 45-75 x 10 bone marrow and spleen cells, from either identical donors (left) or heterozygous donors (H-2 type bk). Injected cells re-established the hematopoietic system as shown by cytotoxicity tests conducted 5-11 weeks later. 

Briles and Krause carried out adoptive transfers using 10 spleen cells from SWR/J mice that had produced antibody of restricted heterogeneity to group A streptococcal carbohydrate (118). Recipient mice produced antibodies that were identical to those of the donor by the criteria of electrophoretic mobility and idiotype. 

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.

Figure 5.8 Adoptive immunization experiment production of SE agglutinins after transfer of pre-immunized spleen cells of High or Low responder into normal recipients of the opposite line. Donor immunization 5 x 10 SE intravenously 14 and 4 days before transfer. Transfer one spleen equivalent per recipient intraperitoneally. From Biozzi et al (1971). In Progress in Immunology, p. 529, by courtesy of Academic Press, New York.

A number of experiments have been reported by Bell and Dray (1969, 1970, 1971a, b, 1972, 1973) in which the authors describe conversion of non-immune rabbit spleen cells by RNA from immunized rabbit to produce IgM and IgG of donor light chain allotype. Furthermore, spleen cells from non-immunized rabbits were converted to antibody-forming cells by incubation with RNA extracts of lymph node cells obtained from rabbits immunized with SRC. If the RNA was extracted from lymph nodes of rabbits immunized 5 days previously, IgM anti-SRC antibody-forming cells were produced, while if the donor of RNA was immunized 18 to 24 days previously, IgG anti-SRC antibody-forming cells were detected in the converted spleen cells. 

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