scholarly journals LYMPHOCYTE PROLIFERATION IN VITRO INDUCED BY HAPTEN AUTOLOGOUS PROTEIN CONJUGATES

1974 ◽  
Vol 139 (3) ◽  
pp. 732-753 ◽  
Author(s):  
Bent Rubin ◽  
Hans Wigzell
Keyword(s):  
Lymph Node ◽  
T Lymphocytes ◽  
Dna Synthesis ◽  
Present Article ◽  
Immune Cells ◽  
Guinea Pigs ◽  
Differential Sensitivity ◽  
Specific Response ◽  
Lymph Node Cells

Immune lymph node cells from guinea pigs respond to soluble antigen in vitro by an increase in DNA synthesis. Optimal conditions for this proliferative response were studied in the present article. Under such conditions, immune cells showed increasing responses with increasing antigen concentration in vitro, the threshold dose of activation frequently being as low as 0.02 µg per culture. In contrast, normal lymph node cells (from FCA-stimulated animals) did only respond to antigen at very high doses (20 mg/culture), and immune cell dilution studies could be performed in normal cells without changing the kinetics of the antigen specific response of immune cells. Fractionation on anti-Ig columns indicated that purified, immune T lymphocytes were quite capable of proliferating in vitro upon antigen stimulation. However, our attempts to adsorb the proliferating cells onto chemically defined immunoadsorbants failed despite the fact that immune B cells (as measured by the rosette assay) were retained almost completely by such a procedure. Purified, immune T lymphocytes from guinea pigs immunized with different antigen concentrations in vivo and/or obtained at different times after immunization were tested for a differential sensitivity toward antigen-induced DNA synthesis in vitro. However, we were not able to demonstrate any regular increase in sensitivity to antigen in vitro, and if found, it seemed to be more dependent upon the number of antigen reactive cells in the population studied rather than upon differences in the average avidity of the receptors on the cells proliferating in vitro. The results in the present article are discussed in relation to current knowledge and hypotheses on T-lymphocyte receptors.

scholarly journals The specificity of cellular immune responses in guinea pigs. I. T cells specific for 2,4-dinitrophenyl-o-tyrosyl residues.

1975 ◽  
Vol 141 (1) ◽  
pp. 42-55 ◽  
Author(s):  
C A Janeway ◽  
B E Cohen ◽  
S Z Ben-Sasson ◽  
W E Paul
Keyword(s):  
T Cells ◽  
T Lymphocytes ◽  
Immune Responses ◽  
Dna Synthesis ◽  
Guinea Pigs ◽  
Hydroxyl Group ◽  
Specific Response ◽  
Protein Carriers ◽  
Cellular Immune

Guinea pigs immunized with the hapten 2,4-dinitrophenyl (DNP) coupled directly to Mycobacterium tuberculosis of strain H37Ra (DNP-H37) show a variety of cell-mediated immune responses to DNP coupled to protein carriers. The cells responsible for this specific response are thought to be T lymphocytes for the following reasons: Guinea pigs immunized with DNP-H37 displayed delayed hypersensitivity reactions to several DNP-proteins and contact sensitivity to dinitrofluorobenzene. Peritoneal exudate lymphocytes (PELs) obtained from DNP-H37 immune animals respond to DNP-proteins with DNA systhesis and cause inhibition of macrophage migration. PELs are highly enriched in T lymphocytes and contain few immunoglobulin-bearing cells. Further depletion of immunoglobulin-bearing cells from this population does not diminish the in vitro proliferative response to antigen. Nitrophenyl conjugates of proteins lacking a paranitro group stimulated DNA synthesis poorly or not at all, indicating the importance of the paranitro group of DNP in antigen recognition by T cells in this system. In this respect, the specificity of T cells resembles that of DNP-specific antibody from the same animals. On the other hand, DNP conjugates of copolymers of glutamic acid and lysine and DNP conjugated to proteins via an interposed beta-alanyl-glycyl-glycyl spacer failed to stimulate DNA synthesis, although such compounds bind very efficiently to anti-DNP antibody. By contrast, DNP conjugates of synthetic polypeptide carriers containing as little as 7% tyrosine strongly stimulated DNA synthesis in DNP-H37 immune PELs. That the determinant responsible for this stimulation was DNP coupled to the hydroxyl group of tyrosine was shown by selective removal of DNP from tyrosine by thiolysis with 2-mercaptoethanol, which abolished their ability to stimulate T cells.

scholarly journals Lectin-driven maturation of cytotoxic effector cells: the nature of effector memory.

1977 ◽  
Vol 146 (1) ◽  
pp. 230-240 ◽  
Author(s):  
A K Chakravarty ◽  
W R Clark
Keyword(s):  
Lymph Node ◽  
Dna Synthesis ◽  
Cell Function ◽  
Cell Formation ◽  
Effector Memory ◽  
Con A ◽  
Cytotoxic Response ◽  
Lymph Node Cells ◽  
Kinetics Of

In an attempt to define further the activation of effector T-cell function with concanavalin A (Con A), we examined the ability of Con A to generate effector memory in mouse lymph node cells in vitro. In the course of these studies, it became necessary to define parameters by which memory could be defined. These parameters include length of time of exposure to signal required to generate full cytotoxic function; time of onset and kinetics of blast cell formation; requirement for DNA synthesis; sensitivity to the drug 5'-bromodeoxyuridine; and kinetics of the cytotoxic response to alloantigen. By these criteria, mouse lymph node cells exposed 12 days previously to Con A behave qualitatively differently from unprimed mouse lymphocytes. We found that the time of onset and kinetics of DNA synthesis could not be used to distinguish primary and secondary cytotoxic responses. We propose that the parameters defined in these stidues can be applied generally in determining whether a given cytotoxic response involves primed (memory) or unprimed cells.

Guinea pigs immunized with xenogeneic cells: Skin test, in vitro stimulation; cytotoxicity of lymph node cells

1973 ◽  
Vol 9 (1) ◽  
pp. 157-162 ◽  
Author(s):  
GyözöG. Petrányi ◽  
Eva Klein ◽  
Erik Svedmyr ◽  
Hans Jacobsson
Keyword(s):  
Lymph Node ◽  
Skin Test ◽  
Guinea Pigs ◽  
Lymph Node Cells

scholarly journals Passive transfer of experimental autoimmune myasthenia by lymph node cells in inbred guinea pigs.

1975 ◽  
Vol 142 (3) ◽  
pp. 785-789 ◽  
Author(s):  
R Tarrab-Hazdi ◽  
A Aharonov ◽  
O Abramsky ◽  
I Yaar ◽  
S Fuchs
Keyword(s):  
Lymph Node ◽  
Human Disease ◽  
Guinea Pigs ◽  
Cellular Response ◽  
Clinical Signs ◽  
Clinical Manifestations ◽  
Passive Transfer ◽  
Lymph Node Cells ◽  
Autoimmune Myasthenia ◽  
Experimental Autoimmune

Passive transfer of experimental autoimmune myasthenia (EAM) was performed with lymph node cells from donor guinea pigs immunized with purified acetylcholine receptor (AChR) from Torpedo californica. Recipient animals revealed the same clinical signs and electromyographic patterns as observed in actively challenged animals. These phenomena are parallel to the clinical manifestations of the human disease myasthenia gravis, in which cellular response to AChR was recently demonstrated.

scholarly journals Primary generation of cytolytic T lymphocytes in the absence of DNA synthesis.

1979 ◽  
Vol 150 (1) ◽  
pp. 196-201 ◽  
Author(s):  
H R MacDonald ◽  
R K Less
Keyword(s):  
T Lymphocytes ◽  
Dna Synthesis ◽  
Cytolytic Activity ◽  
Target Cells ◽  
Cytolytic T Lymphocytes ◽  
Spleen Cells ◽  
Con A ◽  
A Cell ◽  
Stimulation Of

The requirement for DNA synthesis during the primary differentiation of cytolytic T lymphocytes (CTL) had been investigated. CTL were induced polyclonally in vitro by stimulation of normal C57BL/6 spleen cells with concanavalin A (Con A)and their cytolytic activity was tested against 51Cr-labeled target cells in the presence of Bacto Phytohemagglutinin M. With this system, CTL activity could first be detected 48 h after exposure of spleen cells to Con A. Addition of cytosine arabinoside at concentrations sufficient to reduce DNA synthesis by 95-98% in Con A-stimulated cultures did not significantly inhibit the generation of cytolytic activity on a cell-to-cell basis. These results demonstrate that derepression of the genetic information required for the expression of CTL function can occur in the absence of detectable DNA synthesis.

Sign in / Sign up

Export Citation Format

Share Document