scholarly journals Differential effects of polyadenylic: polyuridylic acid and lipopolysaccharide on the generation of cytotoxic T lymphocytes.

1978 ◽  
Vol 147 (5) ◽  
pp. 1355-1362 ◽  
Author(s):  
P R Narayanan ◽  
G Sundharadas
Keyword(s):  
Cytotoxic T Lymphocytes ◽  
Cytotoxic Lymphocytes ◽  
Adherent Cells ◽  
Spleen Cells ◽  
Cytotoxic Cells ◽  
Cytotoxic Response ◽  
Two Agents ◽  
Polyadenylic Acid ◽  
Polyuridylic Acid ◽  
Lines Of Evidence

In a mixed leukocyte culture (MLC) reaction of allogenic mouse spleen cells differing for H-2K or H-2D, only a weak cytotoxic response is generated. This cytotoxic response is augmented significantly if bacterial lipopolysaccharide (LPS), 5 microgram/ml, or polyadenylic acid (poly A):polyuridylic acid (poly U), 20 microgram/ml, is present in the culture. The cytotoxic cells generated in the presence of these two agents are specific for sensitizing H-2K or H-2D antigen. Two lines of evidence suggest that these two agents exert their effect at different steps in the development of cytotoxic lymphocytes: (a) the effect of poly A:U depends on the presence of adherent cells, whereas the effect of LPS is independent of the presence of adherent cells and (b) LPS promotes the development of cytotoxic cells when ultraviolet light-treated stimulating cells are used in the MLC whereas poly A:U does not.

scholarly journals THE EFFECT OF 2-MERCAPTOETHANOL ON MURINE MIXED LYMPHOCYTE CULTURES

1974 ◽  
Vol 139 (4) ◽  
pp. 1025-1030 ◽  
Author(s):  
Michael J. Bevan ◽  
Ruth Epstein ◽  
Melvin Cohn
Keyword(s):  
Fetal Calf Serum ◽  
Calf Serum ◽  
Mouse Spleen ◽  
Adherent Cells ◽  
Spleen Cells ◽  
Cytotoxic Cells ◽  
Cytotoxic Response ◽  
Lymphocyte Cultures ◽  
Mixed Lymphocyte Cultures

Mouse spleen cells which have been depleted of adherent cells do not respond to allogeneic lymphocytes in vitro. Their cytotoxic response can be restored by inclusion of mercaptoethanol in the medium. Mercaptoethanol is shown to have a stimulatory effect also on the response of normal (unseparated) spleen cells to alloantigens. The enhancement of the DNA-synthetic and cytotoxic response is similar, varying from 3.5–15-fold. Cytotoxic cells also appear in unmixed lymphocyte cultures in the presence of mercaptoethanol and fetal calf serum. The specificity of these background cytotoxic cells is not known.

scholarly journals Requirement for non-T cells in the generation of cytotoxic T lymphocytes in vitro. I. Use of nude mice as source of non-T cells.

1976 ◽  
Vol 144 (1) ◽  
pp. 241-258 ◽  
Author(s):  
R M Schilling ◽  
R A Phillips ◽  
R G Miller
Keyword(s):  
T Cells ◽  
Cytotoxic T Lymphocytes ◽  
Nude Mice ◽  
Culture Conditions ◽  
Cytotoxic Lymphocytes ◽  
Spleen Cells ◽  
Cell Component ◽  
Athymic Nude Mice ◽  
Cytotoxic Cells

The ability of small numbers of LN cells to produce cytotoxic lymphocytes on in vitro culture with allogeneic stimulator cells is greatly augmented by the addition of spleen cells from athymic nude mice. The possibility that the synergism is a result of improved culture conditions or a "feeder effect" is excluded. All cytotoxic cells found in these cultures are shown to be T cells and to arise from precursors contained in the LN-cell component. The nude spleen cell component appears to be providing a required non-T cell which has been lost from the LN component through dilution. Synergism between the two components can occur whether they are syngeneic or allogeneic provided that both can recognize the same alloantigens in the stimulator population.

scholarly journals An estimation of the frequency of precursor cells which generate cytotoxic lymphocytes.

1976 ◽  
Vol 143 (6) ◽  
pp. 1562-1567 ◽  
Author(s):  
M A Skinner ◽  
J Marbrook
Keyword(s):  
Immune Response ◽  
Culture System ◽  
Precursor Cells ◽  
Cytotoxic Lymphocytes ◽  
Spleen Cells ◽  
Cytotoxic Response ◽  
Cell Mediated Immune Response

The cell-mediated immune response has been generated in vitro with a polyacrylamide culture system which allows the segregation of foci (clones?) of cytotoxic lymphocytes. Using the method of limiting dilutions, the frequency of precursor cells in CBA spleen cells able to generate a cytotoxic response against DBA mastocytoma is estimated at 1 per 1,700 cells.

scholarly journals Quantitative studies on the precursors of cytotoxic lymphocytes. VI. Second signal requirements of specifically activated precursors isolated 12 h after stimulation.

1980 ◽  
Vol 151 (1) ◽  
pp. 12-19 ◽  
Author(s):  
M E Lalande ◽  
M J McCutcheon ◽  
R G Miller
Keyword(s):  
Cytotoxic T Lymphocytes ◽  
T Lymphocyte ◽  
Mixed Lymphocyte Reaction ◽  
Cytotoxic T Lymphocyte ◽  
Cytotoxic Lymphocytes ◽  
Spleen Cells ◽  
Quantitative Studies ◽  
Partial Activation ◽  
Lymphocyte Cultures ◽  
Mixed Lymphocyte Cultures

It is shown that, in a mixed lymphocyte reaction, the production of cytotoxic T lymphocytes (CL) from cytotoxic T lymphocyte precursors (CLP) requires two signals which are separated time. Using a flow cytometer-cell sorter and a vital, fluorescent DNA stain, Hoechst 3342, CLP specific for the stimulator cells can be separated from other CLP and from stimulator cells 12 h after initiation of mixed lymphocyte cultures. These CLP are in a state of partial activation and can produce CL in the absence of stimulator cells if a second signal in the form of a concanavalin A-induced spleen cell supernate factor is added. Specific CL are also generated when the partially activated CLP are cultured with both nude spleen cells and stimulator cells. In this case it appears that an interaction between the stimulator cells and the nude spleen cells leads to production of the second signal.

scholarly journals GENERATION OF CYTOTOXIC T LYMPHOCYTES IN VITRO

1974 ◽  
Vol 140 (3) ◽  
pp. 703-717 ◽  
Author(s):  
Jean-Charles Cerottini ◽  
Howard D. Engers ◽  
H. Robson MacDonald ◽  
K. Theodor Brunner
Keyword(s):  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Relative Number ◽  
Fold Increase ◽  
Culture Conditions ◽  
Target Cells ◽  
Spleen Cells ◽  
Response Curves ◽  
Cytotoxic Response

Mouse cytotoxic T lymphocytes (CTL) were generated in mixed leukocyte cultures (MLC) using spleen cells as responding cells and irradiated allogeneic spleen cells as stimulating cells. Cytotoxicity was assessed by a quantitative 51Cr assay system and the relative frequency of CTL in individual cell populations was estimated from dose-response curves. Inclusion of 2-mercaptoethanol in the MLC medium resulted in a 20–40-fold increase in the relative number of CTL generated at the peak of the response. Under these culture conditions, cell-mediated cytotoxic activity was detectable in MLC populations as early as 48 h after the onset of the cultures. When spleen cells from mice immunized with allogeneic tumor cells 2–4 mo previously were cultured with irradiated spleen cells of the same alloantigenic specificity (MLC-Imm), it was found that the cell-mediated cytotoxic response was detectable earlier and reached higher levels than that observed in a primary MLC. At the peak of the response, MLC-Imm populations were observed to lyse up to 50% of the target cells within 3 h at a lymphocyte: target cell ratio of 0.3:1. Immunological and physical characterization of the effector cells generated in MLC-Imm indicated that they were medium to large-sized T lymphocytes. Altogether, these studies suggested the existence of an anamnestic cell-mediated cytotoxic response in MLC-Imm.

scholarly journals Generation of primary cytotoxic lymphocytes against non-major histocompatibility complex antigens by anti-Ia serum plus complement-treated lymphocytes.

1980 ◽  
Vol 151 (5) ◽  
pp. 1305-1310 ◽  
Author(s):  
C E Hayes ◽  
S Macphail ◽  
F H Bach
Keyword(s):  
Major Histocompatibility Complex ◽  
Suppressor Cell ◽  
Target Cells ◽  
Cytotoxic Lymphocytes ◽  
Major Histocompatibility ◽  
Cytotoxic Cells ◽  
Major Histocompatibility Complex Antigens ◽  
Cytotoxic Response ◽  
Histocompatibility Complex

The results presented in this paper demonstrate that responding cells that remain after anti-Ia serum plus complement (C) treatment generate a highly significant in vitro cytotoxic response against minor histocompatibility complex antigens. The cytotoxic response appears to be antigen specific in that target cells of strains other than the sensitizing strain are not lysed, or lysed to a lesser extent. The cytotoxic cells are susceptible to anti-Thy-1 plus C lysis. Anti-Ia serum may function by removing an unprimed suppressor cell, although other mechanisms cannot be ruled out.

scholarly journals Polyribonucleotide synthesis by subfractions of microsomes from rat liver

1968 ◽  
Vol 109 (2) ◽  
pp. 229-238 ◽  
Author(s):  
N. M. Wilkie ◽  
R. M. S. Smellie
Keyword(s):  
Acetic Acid ◽  
Rat Liver ◽  
Transfer Rna ◽  
Light Fraction ◽  
Free Ribosome ◽  
Polyadenylic Acid ◽  
Insoluble Material ◽  
Polyuridylic Acid ◽  
Microsome Fraction

1. The microsome fraction of rat liver has been fractionated and the ability of the fractions to incorporate ribonucleotides into polyribonucleotides has been studied. Activity was found in the rough-surfaced vesicle (light) fraction and in the free-ribosome fraction and this latter activity has been examined. 2. The free-ribosome fraction contains ribosome monomers, dimers and trimers together with some higher oligomers and ferritin. In addition to catalysing the incorporation of ribonucleotides into acid-insoluble material it contains diesterase activity. It catalyses the incorporation of UMP from UTP, but not UDP, AMP from ATP and CMP from CTP into polyribonucleotide material, and for UTP the product appears to be a homopolymer not more than eight units long attached to the ends of primer polyribonucleotide strands. 3. The activity could not be removed from the free-ribosome fraction by washing or by isolation in the presence of ethylenediaminetetra-acetic acid. 4. Partially hydrolysed polyuridylic acid but not polyadenylic acid could serve as a primer for the incorporation of UMP, but some activity was always associated with an endogenous primer. 5. Analysis of RNA extracted from the free-ribosome fraction after incubation with [3H]UTP showed the presence of 28s, 18s, 5s and transfer RNA types, but no radioactivity was associated with any of these RNA fractions.

Peritoneal adherent cells inhibit the generation of cytotoxic T lymphocytes with prostaglandin-mediated system

1982 ◽  
Vol 66 (1) ◽  
pp. 195-201 ◽  
Author(s):  
Yasuhiro Koga ◽  
Kazuto Taniguchi ◽  
Chiharu Kubo ◽  
Kikuo Nomoto
Keyword(s):  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Adherent Cells

Dual effect of meningococcal antigens on a T cell dependent immune response

1983 ◽  
Vol 29 (12) ◽  
pp. 1619-1625 ◽  
Author(s):  
Brian G. Sparkes
Keyword(s):  
Immune Response ◽  
Inhibitory Activity ◽  
Sheep Erythrocyte ◽  
Subsequent Development ◽  
Plaque Formation ◽  
Adherent Cells ◽  
Spleen Cells ◽  
Dual Effect ◽  
Adjuvant Activity ◽  
Nonadherent Cells

Meningococcal antigens (MA) showed adjuvant activity when administered to mice at the same time as antigen (sheep erythrocyte (SE)), by increasing the splenocyte plaque-forming response in a dose-related manner. However, when SE were given 1 day after MA administration, the subsequent plaque formation was diminished from normal in proportion to the dose of MA injected. Splenocytes taken from mice up to 5 days after MA injection actively inhibited plaque formation when mixed with splenocytes immunized with SE 4 days earlier. Two days after MA injection the nonspecific inhibition of plaque formation was mainly due to adherent spleen cells, while at 5 days nonadherent cells had acquired the inhibitory activity. It appears that it is the degree of activation of adherent cells resulting from the timing and dosage of MA which modulates the subsequent development and secretion of antibody-forming cells.

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