Cell interactions in concanavalin A activated cation flux and DNA synthesis of mouse lymphocytes

1980 ◽  
Vol 58 (11) ◽  
pp. 1314-1317 ◽  
Author(s):  
Trevor Owens ◽  
J. G. Kaplan
Keyword(s):  
T Cells ◽  
T Cell ◽  
Concanavalin A ◽  
Thymidine Incorporation ◽  
Spleen Cells ◽  
Con A ◽  
Activated T Cell ◽  
Constant Cell ◽  
Enriched Cultures ◽  
Cell Mitogen

Co-culture at constant cell density of nude mouse spleen cells (by themselves unresponsive to the T-cell mitogen concanavalin A (Con A)), with congenic T-enriched lymphocyte suspensions and Con A caused anomalously high activation of K+ transport (measured by 86Rb uptake) and of incorporation of thymidine into DNA; the expected dilution of these two responses by nude spleen cells did not occur. However, if the nude splenocytes were added immediately prior to assay to the enriched T cells that had been precultured in presence of Con A, the expected dilution of the activated T-cell responses occurred; both 86Rb uptake and thymidine incorporation were reduced proportionally to the degree of dilution of the T cells by the nonresponding cells. These data indicate that during co-culture in presence of Con A there is interaction between the T cells, capable of responding to the mitogens, and the nude spleen cells. Attempts to demonstrate a diffusible factor in the supernatants of stimulated T cells were unsuccessful. The measured interaction is sufficient to explain our previous paradoxical findings that enrichment of T cells as measured by membrane markers did not cause a corresponding enrichment for either cation transport or for thymidine incorporation, and that depletion of T cells in the B-enriched cultures did not cause a corresponding decrease in these two Con A induced responses.

scholarly journals Concanavalin A-inducible, interleukin-2-producing T cell hybridoma.

1980 ◽  
Vol 152 (4) ◽  
pp. 893-904 ◽  
Author(s):  
L Harwell ◽  
B Skidmore ◽  
P Marrack ◽  
J Kappler
Keyword(s):  
T Cells ◽  
T Cell ◽  
Concanavalin A ◽  
Interleukin 2 ◽  
Hybridoma Cells ◽  
Adherent Cell ◽  
Spleen Cells ◽  
Con A ◽  
Normal Spleen ◽  
Stimulation Of

The fusion of an AKR T cell tumor line to normal B6D2F1, T cells resulted in the production of a cloned T cell hybridoma (FS6-14.13) inducible with the mitogen concanavalin A (Con A). The supernate from Con A-stimulated hybridoma cells was active both in the stimulation of an anti-sheep red blood cell response by partially T cell-depleted B cells and in the stimulation of the growth of antigen-specific T cell blasts. The active principle in both assays had a molecular weight of approximately 30-40,000. These results indicated the presence of interleukin 2 (IL2) in the hybridoma supernate. The activity of the hybridoma supernate in B cell responses was dependent on the presence of adherent cells and a few contaminating T cells. On the other hand, Con A-stimulated supernates from normal spleen cells were active after either adherent cell removal or severe T cell depletion. These results suggested that IL2 was the only active helper factor in the hybridoma supernate, but that additional helper factors were present in supernates from Con A-stimulated normal spleen cells.

scholarly journals Effects of anti-Ia sera on mitogenic responses. II. Differential expression of the Ia marker on phytohemagglutinin and concanavalin A-reactive T cells.

1976 ◽  
Vol 143 (2) ◽  
pp. 372-381 ◽  
Author(s):  
J E Niederhuber ◽  
J A Frelinger ◽  
M S Dine ◽  
P Shoffner ◽  
E Dugan ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cells ◽  
Differential Expression ◽  
Concanavalin A ◽  
Cell Subpopulation ◽  
Spleen Cells ◽  
Con A ◽  
Membrane Antigens ◽  
Rabbit Complement

Genes mapping in the I region of the H-2 complex control a system of lymphocyte alloantigens (Ia) which are expressed on a subpopulation of T cells and on most B cells. Specific anti-Ia serum in the presence of rabbit complement removed the splenic T-cell subpopulation responsive to Con-A, but did not affect the response to phytohemagglutinin (PHA) or Leucoagglutinin. Antibodies specific for Ia, H-2K, or H-2D membrane antigens were used without complement to pretreat spleen cells. These antibody pretreated cells responded normally to Con-A and PHA.

scholarly journals Loss of suppressor T cells in adult NZB/NZW mice.

1976 ◽  
Vol 144 (3) ◽  
pp. 662-673 ◽  
Author(s):  
R S Krakauer ◽  
T A Waldmann ◽  
W Strober
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cell Activity ◽  
Suppressor Cell ◽  
Indicator System ◽  
Spleen Cells ◽  
Con A ◽  
Normal Cells ◽  
Suppressor T Cell ◽  
Cell Fractions

We have investigated suppressor T-cell activity in female NZB/NZW F1 mice using PWM-driven IgM biosynthesis in vitro as an indicator system. In initial we studied we observed that spleen cells from normal mice (BALB/c, C57BL/6), as well as from young (4 wk) and adult (18 wk) NZB/NZW mice, cultured in the presence of PWM synthesize 860 +/- 120 ng IgM/10(6) cells/7 days. However, when Con A (at 2 mug/ml) was added directly to the cultures (along with PWM), cells obtained from adult normal mice and young NZB/NZW mice showed a 94% suppression of IgM synthesis, whereas cells obtained from adult NZB/NZW mice were suppressed significantly less. To analyze these findings we studied the effect of Con A-induced suppressor cells (cells cultured with Con A for 24 h and washed free of Con A) on PWM-driven IgM biosynthesis. Spleen cells obtained from normal mice cultured in the presence of Con A-pulsed cells obtained from normal mice and young NZB/NZW mice showed an 83-88% suppression of PWM-driven IgM synthesis. Similarly, supernates obtained from Con A-pulsed cells of normal mice or of young NZB/NZW mice suppressed PWM-driven IgM synthesis. This suppression by Con A-pulsed cells and their supernates required T cells since T-cell fractions but not B-cell fractions eluted from anti-Fab Sephadex columns mediated suppression of co-cultured normal cells; in addition, Con A-pulsed cells treated with anti-theta and complement do not mediate suppression. These studies of Con A-induced suppressor cell activity in normal mice and young NZB/NZW mice contrast with studies of Con A-induced suppressor cell activity in adult NZB/NZW mice. We found that adult NZB/NZW Con A-pulsed cells and supernates obtained from the Con A-pulse cells had vastly decreased suppressor potential; in this case the Con A-pulse cells and supernatant fluids derived from such cells did not suppress PWM-driven IgM synthesis by normal cells. Finally, whereas spleen cells from young and adult NZB/NZW mice differ in their suppressor cell potential, cells from both sources could respond equally to suppressor signals in that Con A-pulsed normal cells or supernates derived from such cells caused equivalent suppression of PWM-driven IgM synthesis by young and adult NZB/NZW cells. These observations allow us to conclude that NZB/NZW mice lose suppressor T-cell activity as they age.

scholarly journals CELL-MEDIATED IMMUNE RESPONSES IN VITRO

1974 ◽  
Vol 140 (2) ◽  
pp. 356-369 ◽  
Author(s):  
Duane L. Peavy ◽  
Carl W. Pierce
Keyword(s):  
Immune Response ◽  
T Cells ◽  
T Cell ◽  
Immune Responses ◽  
Cell Activity ◽  
Suppressor Cell ◽  
Regulatory Control ◽  
Cytotoxic Lymphocytes ◽  
Spleen Cells ◽  
Con A

The effects of soluble concanavalin A (Con A) or Con A-activated spleen cells on the generation of cytotoxic lymphocytes (CL) in mixed leukocyte cultures (MLC) were examined. Mitogenic concentrations of soluble Con A or small numbers of Con A-activated spleen cells substantially inhibited CL responses. The suppression was partial rather than absolute and was critically dependent upon the concentration and time of addition of soluble Con A or Con A-activated spleen cells to the MLC. Suppressive effects of Con-A activated spleen cells were mediated by T cells since suppressor cell activity was abrogated by treatment of spleen cells with anti-θ serum and complement before or after Con A activation. X irradiation of spleen cells before Con A treatment also abrogated generation of suppressor cell activity. After activation by Con A, however, the function of suppressor cells was radioresistant. Although the precise mechanism(s) of suppression is, as yet, unknown, the precursors of CL must be exposed to Con A-activated cells during the early phases of the immune response for suppression to occur. Kinetic studies revealed that suppression of CL responses was not due to a failure to initiate an immune response, but represented a response which developed initially, but subsequently aborted. The relevance of these observations to the concepts of T-cell-T-cell interaction and regulatory control of immune responses by T cells is discussed.

scholarly journals Two distinct mechanisms regulate the in vivo generation of cytotoxic T cells.

1982 ◽  
Vol 156 (3) ◽  
pp. 918-923 ◽  
Author(s):  
M S Sy ◽  
S H Lee ◽  
M Tsurufuji ◽  
K L Rock ◽  
B Benacerraf ◽  
...  
Keyword(s):  
T Cells ◽  
T Lymphocytes ◽  
Concanavalin A ◽  
Cytotoxic T Cells ◽  
Cytolytic T Lymphocytes ◽  
Spleen Cells ◽  
Suppressor T Cells ◽  
Con A

Treatment of responder cells with monoclonal anti-Ly-1,2 antibodies plus complement in vitro completely eliminated their ability to generate azobenzenearsonate (ABA)-specific cytolytic T lymphocytes (CTL). However, addition of the concanavalin A-stimulated supernatants of rat spleen cells (Con A-Sup) can fully reconstitute the response. Therefore, Lyt-1,2-bearing T cells are required for the generation of ABA-specific CTL, and such requirement can be replaced by factors present in the Con A- sup. Suppressor T cells (Ts), when adoptively transferred into naive recipients, will inhibit the in vivo priming of CTL. This inhibition can also be reversed by in vitro addition of Con A-Sup. furthermore, mice serving as donors of Ts also show profound unresponsiveness when primed and restimulated in vitro. In contrast to the Ts-mediated inhibition, in vitro addition of Con A-Sup was unable to abolish the unresponsiveness observed in these cultures. Thus, we identified two unresponsive states in a hapten-specific killing system that differ in their ability to be reconstituted by Con A-Sup.

scholarly journals B cell helper factors. I. Requirement for both interleukin 2 and another 40,000 mol wt factor.

1981 ◽  
Vol 154 (5) ◽  
pp. 1681-1693 ◽  
Author(s):  
H J Leibson ◽  
P Marrack ◽  
J W Kappler
Keyword(s):  
T Cells ◽  
T Cell ◽  
B Cell ◽  
Interleukin 2 ◽  
Spleen Cells ◽  
Con A ◽  
Direct Role ◽  
Cell Responses ◽  
Helper Factor ◽  
Antibody Secreting Cells

A helper factor(s) distinct from interleukin 2 (IL-2) was shown to be present in the concanavalin A-stimulated supernatant of normal mouse spleen cells (normal Con A Sn). Spleen cells thoroughly depleted of T cells required both IL-2 and this factor to produce antibody-secreting cells in response to sheep erythrocytes, although in the presence of IL-2 and a few T cells the requirement for the factor was less apparent. The factor had an apparent approximately 40,000 mol wt. The factor was found in normal Con A Sn that had been depleted of IL-2 by absorption with IL-2-dependent T cells and was absent from Con A-stimulated supernatants of the IL-2-producing T cell hybridoma, FS6-14.13. These results indicate that multiple helper factors control the B cell response to antigen and that IL-2, in addition to its T cell growth promoting activity, plays a direct role in B cell responses.

scholarly journals T cell growth factor abrogates concanavalin A-induced suppressor cell function

1981 ◽  
Vol 153 (5) ◽  
pp. 1360-1365 ◽  
Author(s):  
R Palacios ◽  
G Moller
Keyword(s):  
T Cells ◽  
T Cell ◽  
Growth Factor ◽  
Cell Growth ◽  
Concanavalin A ◽  
Cell Function ◽  
Suppressor T Cells ◽  
Con A ◽  
Suppressor Activity ◽  
T Cell Growth Factor

Concanavalin A (Con-A)-induced suppressor T cells were found to respond to T cell growth factor (TCGF) by proliferation. TCGF abrogated the suppressor activity exerted by these cells on phytohemagglutinin (PHA)- and alloantigen- induced lymphocyte proliferation and on pokeweed mitogen (PWM)-driven immunoglobulin secretion. The Con-A-activated suppressor T cells absorbed the TCGF activity, preincubation of these active suppressor cells with TCGF abolished their suppressor activity and addition of increasing numbers of Con-A-activated T cells reverted the abrogator,/ effect of TCGF. Altogether, these findings suggest that Con-A-induced suppressor T cells exert their function by decreasing the available levels of TCGF. Cyclosporin-A (CYA), which is known to inhibit the expression of receptors for TCGF on T cells, also inhibited the suppressor activity as determined in both indicator systems, namely PHA- or alloantigen-induced DNA synthesis and PWM-induced immunoglobulin synthesis. CYA made Con-A-treated T cells unresponsive to TCGF and unable to absorb the growth factor, supporting the notion that CYA inhibits the expression of TCGF receptors on T cells, a mechanism by which this drug seems to abrogate Con-A-induced suppressor T cell function.

scholarly journals Glucocorticoids induce the expression of CD8 alpha chains on concanavalin A-activated rat CD4+ T cells: induction is inhibited by rat recombinant interleukin 4.

1992 ◽  
Vol 176 (6) ◽  
pp. 1551-1559 ◽  
Author(s):  
F Ramirez ◽  
A J McKnight ◽  
A Silva ◽  
D Mason
Keyword(s):  
T Cells ◽  
T Cell ◽  
Concanavalin A ◽  
Cd4 T Cells ◽  
Interleukin 4 ◽  
Mrna Levels ◽  
Con A ◽  
Alpha Chain ◽  
T Cell Clone ◽  
Single Positive

Rat T lymphocytes, activated in vitro with concanavalin A (Con A), were shown by flow cytofluorographic analysis to contain a population of cells that simultaneously expressed CD4 and the alpha chain of CD8. The inclusion of the glucocorticoid hormone dexamethasone in the culture medium greatly increased both the frequency of these double-positive cells and the level of CD8 alpha chain expression. The level of expression of CD4 was not affected, and the cells that expressed CD8 antigen only also remained unchanged in surface phenotype. Detailed studies demonstrated unequivocally that the CD4+ CD8 alpha + cells were not artifacts produced by the random association of single-positive cells in the flow cytofluorograph, but arose from precursors that were single-positive CD4+ cells before activation. Furthermore, Con A activation of purified CD4+ T cells, in the presence of T cell-depleted accessory cells, showed that CD8+ T cells played no role in the induction process. However, the induction of CD8 alpha chain expression on CD4+ T cells and the enhancement of this expression by dexamethasone were almost completely inhibited by rat recombinant interleukin 4 (IL-4). Detection of mRNA for rat CD8 alpha chain by Northern blot closely paralleled the cell surface expression of CD8 alpha antigen, indicating that dexamethasone and IL-4 had opposing effects on mRNA levels. In contrast, IL-4 and dexamethasone both induced CD8 alpha chain expression on a rat CD4+ T cell clone when this was activated by specific antigen, and, although the effect with IL-4 was relatively weak, it did not antagonize the effect of the glucocorticoid. The possible significance of these results is briefly discussed.

scholarly journals The capacity of mitogen-responsive T cells to home to their tissue of origin, analyzed by means of chromosome markers

1976 ◽  
Vol 143 (3) ◽  
pp. 660-671 ◽  
Author(s):  
MJ Doenhoff ◽  
AJS Davies
Keyword(s):  
T Cells ◽  
Lymph Node ◽  
Concanavalin A ◽  
Spleen Cells ◽  
Con A ◽  
Syngeneic Mouse ◽  
Lymph Node Cells ◽  
Mitogen Responsiveness ◽  
Tissue Of Origin ◽  
Syngeneic Recipient

Lance and Taub (1) showed that when radioactively labeled lymphocytes were injected into a syngeneic mouse and the lymph node cells of this animal transferred to a second syngeneic recipient, the proportion of radioactivity found in the lymph node relative to the amount present in the spleen of the secondary recipient had increased markedly. The interpretation of this result was that some lymphocytes have the capacity to home to their organ of origin. The purpose of the experiments described here was to test the homing copacity of T cells by a method that did not involve radioactive labeling. It has been shown elsewhere that some or all mouse T cells are stimulated to divide in culture by the mitogens phytohemagglutinin (PHA) and concanavalin A (Con A) (2). We therefore elected to inject karyotypically distinct lymphocytes into syngeneic recipients and to follow their subsequent distribution by culture of lymph node and spleen cells of the recipient with PHA or Con A. In this manner the homing capacities of spleen and lymph node T cells could be determined, and furthermore, the effects of labeling with chromium-51 ((51)Cr) could be assayed with respect to the persistence of mitogen responsiveness in the injected cells.

scholarly journals Inhibitory Effect of Hydroxymethylfurfural in Viability of BALB/C Mice Splenocytes

2019 ◽  
Vol 25 (1) ◽  
pp. 31-36
Author(s):  
Sevda Saleh-Ghadimi ◽  
Hamed Jafari-Vayghan ◽  
Sorayya Kheirouri ◽  
Mohammad Alizadeh
Keyword(s):  
Cell Proliferation ◽  
Dna Damage ◽  
T Cells ◽  
B Cells ◽  
Concanavalin A ◽  
Untreated Control ◽  
Inhibitory Effect ◽  
Spleen Cells ◽  
Con A ◽  
Significant Difference

Background: This study was designed to discover if hydroxymethylfurfural (HMF) exposure modifies cell proliferation and DNA damage in BALB/c mice splenocytes. Methods: Mitogenesis in T cells and B cells was induced by Concanavalin A (Con A) and lipopolysaccharide (LPS). The colorimetric tetrazolium assay was used to evaluate cell proliferation. DNA damaging consequences were evaluated via measurement of 8-hydroxy-2-deoxyguanosine (8-OHdG) level in BALB/c mice splenocytes. Results: Spleen cells proliferation elicited by ConA, was dramatically suppressed by 25, 50 and 100 mM of HMF. However, there was not any significant difference between various concentrations of HMF. The same result was observed following treatment with LPS and HMF in different concentrations. Eight-OHdG concentration was elevated significantly in HMF treated groups compared with untreated control and mitogens. Conclusion: HMF was found to have immunosuppressing and DNA damaging properties in mM concentrations in mice splenocytes.

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