Rabbit spleen cells

Rats are also suitable for the production of monoclonal antibodies on a 10-times larger scale (Galfre et al., 1979). The Y3 myeloma (K-producing) of Lou rats can be used and ascites tumors can be obtained two weeks after an i.p. injection of 0.5 ml pristane by the i.p. injection of 5 x 10 hybridoma cells. A minor disadvantage is the unreactiveness of rat IgG2a with protein A (Rousseaux et al., 1981). The use of rabbit spleen cells with mouse myelomas has not yet been successful. 

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. 

Bell, C., Dray, S. Conversion of non-immune rabbit spleen cells by ribonucleic acid of lymphoid cells from an immunized rabbit to produce IgG antibody of foreign light chain allotype. J. Immunol. 105, 541-556 (197O). 

Berberine inhibits oxidative decarboxylation of yeast pyruvic acid (310) the same dose has, however, no effect upon aerobic glycolysis, Warburg s respiratory enzymes, indophenol oxidase, etc. Berberine and tetrahydroberberine have an inhibitory effect on oxidation of (+ )-alanine in rat kidney homogenates (498). Berberine and palmatine show a specific inhibitory effect upon cholinesterase in rabbit spleen and on pseudocholinesterase in horse serum (499). Berberine inhibits cellular respiration in ascitic tumors and even in tissue cultures (500-502). The specific toxic effect of berberine on the respiration of cells of ascitic tumors in mice was described (310). The glycolysis was not found to be affected, but the uptake of oxygen was smaller. Fluorescence was used in order to demonstrate berberine in cellular granules. Hirsch (503) assumed that respiration is inhibited by the effect of berberine on the yellow respiratory enzymes. Since the tumorous tissue contains a smaller number of yellow respiratory enzymes than normal tissue it is more readily affected by berberine. Subcutaneous injections of berberine, palmatine, or tetrahydropalmatine significantly reduce the content of ascorbic acid in the suprarenals, which is not affected by hypophysectomy (504). 

In 1923 antibodies specific for a polysaccharide in the pneumococcal cell wall were detected in the serum from rabbits immunized with non-viable pneumococcal cells by Heidelberger and Avery [1], Cell wall polysaccharides from other bacteria as reported by Lancefield [2], McCarty [3], Krause [4], Pazur [5], and Karakawa [6] activated the plasma cells in the serum of animals to initiate the biosynthesis of antibodies which were specific for the carbohydrates. Such antibodies are appropriately classified as anticarbohydrate antibodies. The antibodies which are synthesized on immunization are polyclonal antibodies and consist of various numbers of isoforms. The antibodies which are synthesized by fusion of immune spleen cells and myeloma cells are monoclonal antibodies. 

Catalytic antibodies were first prepared in the mid-1980s. In one example, a rabbit or mouse was immunized with phosphonate ester A, and pure anti-A antibodies were isolated from this animal s spleen cells. Some of the anti-A antibodies were found to catalyze the hydrolysis of the ester functionality of B at rates significantly faster than background. 

Type I allergens are usually considered those macromolecules with the ability to induce specific IgE immune responses and to provoke allergic reactions in sensitized subjects. Antibody-based immunoassays are widely used for the measurement of specific major allergens in the air. Antibodies can originate from sensitized humans, animals (rabbits producing polyclonal antibodies) or fusions of myeloma cells and mouse spleen cells in culture producing monoclonal antibodies. Examples have been published using... 

A-coated Sepharose beads were usefiil for cell separation following initial incubation of the cells with IgG antibodies directed against specific cell surface markers. Surface IgG-bearing mouse spleen cells were pretreated with rabbit antibodies to mouse IgG prior to passage over the protein A-coated support. The cells of interest were then isolated by positive selection chromatography. 

Figure 4.7 Suppressive activity of column purified T cells from GAT-primed non-responder mice. Mice were primed with 10 GAT in Maalox 3 days before culture initiation. Sephadex G-200 columns to which rabbit anti-mouse Fab had been conjugated were used to fractionate T and B spleen cells. (Taken from Benacerraf, B., Kapp, J, A. and Pierce, C. W. (1974). In D. H. Katz and B. Benacerraf (eds.). Immunological Tolerance Mechanisms and Potential Therapeutic Applications, p. 507. (New York Academic Press, Inc.)...

According to more recent observations, cytallene (lid) is readily phosphorylated with ATP and 6,000-fold purified dCyd kinase from human leukemic spleen cells. In addition, cytallene (lid) but neither adenallene (11c) nor ddAdo inhibit phosphorylation of dCyd and dAdo catalyzed with dCyd kinase. The efficiency of phosphorylation of lid is lower than that of ddCyd. Results obtained with human T lymphoblastoid CEM cell line deficient in adenosine (Ado) kinase indicate that the latter enzyme may be important for phosphorylation of ddAdo. However, another study20 shows that ddAdo is not efficiently phosphorylated in a reaction catalyzed by Ado kinase from human thymus. It should also be noted that ddAdo is a substrate for dCyd kinase from the latter source whereas neither racemic adenallene (11c) nor / -enantiomer (31) inhibit phosphorylation of dAdo and dCyd catalyzed by the respective kinases from calf and rabbit thymus.21... 

This fact indicates that spleen cells possess a highly active salvage pathway similar to that found in microorganisms (Partsch 1971). But in contrast microorganisms de novo synthesis may be absent, as is the case in rabbit bone marrow cells (Thompson 1960) and human leucocytes (Scott 1962). 

Figure 1. Effect of FBI on [ H]-thymidine incorporation into mouse spleen cells in vitro. For this experiment FB1 was purchased from Dr. Merrill, A.Fl., Jr, (Emory University, Atlanta. GA). For separation T and B spleen cells, we used anti-Thyl. 2 Abs diluted 1 100 (obtained from Dr. Chervonsky, A.V., Cancer Research Center, Moscow, Russia) or anti-Ig serum (rabbit anti-mouse) diluted 1 10 (Caltag Laboratories) with Guinea pig complement (Sigma) in final concentration 1 10. Whole splenocyte suspension, T, and B cells were washed 3 times and plated at 2xl0 /w and incubated either with different doses of FBI or with known mitogens (PHA and LPS) at optimal concentrations. Control cells were untreated. After 48h incubation, 50 pCi of [ H]thymid-ine (Amersham) was added to each well for 16h. Then the cells were transferred to filter paper, each filter disk was added to scintillation vials and counted. The results (cpm) are given as S.E. from 5 experiments.

In humans the clearance rate of Hb is higher than that of HpHb (L4, Lll). Murray et al. (M6) found this also to hold for rabbits and, by studying the elimination in nephrectomized animals, they also proved that the difference was not due to urinary loss of Hb. Analysis of the organs proved that the HpHb complex and Hb were assimilated mainly in the liver and were catabolized with an early reappearance of the iron as transferrin iron within 30 minutes. The free Hb accumulated also in the tubular cells of the kidneys. No data have been published suggesting that the spleen is of any appreciable importance in this respect. No typical exponential clearance of the HpHb complex from plasma was observed (L10, Lll) in the first few experiments. Lathem and Worley (L4) found that HpHb disappeared at a simple exponential rate in 5... 

Additional information <1-7, 11, 14, 15, 17-19, 21, 30> (<7> cell-free synthesis in mRNA-dependent rabbit reticulocyte lysate system [40] <2,4,5> high activities in tissues where turnover of energy from adenine nucleotides is great, e. g. muscle [3] <1-6,11,14,15> tissue distribution [3,46] <2,5> rabbit and human carry a minimum of 2 sets of isozymes within an individual one set in muscle, erythrocytes, brain and another in liver, kidney and spleen [3]) [3, 40, 46]... 

Otani, H. and Hata, I. 1995. Inhibition of proliferative responses of mouse spleen lymphocytes and rabbit Peyer s patch cells by bovine milk caseins and their digests. J. Dairy Res. 62, 339-348. 

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