scholarly journals CELLULAR RECOGNITION BY MOUSE LYMPHOCYTES IN VITRO

1970 ◽  
Vol 131 (6) ◽  
pp. 1049-1078 ◽  
Author(s):  
W. H. Adler ◽  
T. Takiguchi ◽  
B. Marsh ◽  
R. T. Smith
Keyword(s):  
Human Serum ◽  
Cell Interaction ◽  
Mouse Cell ◽  
Culture Conditions ◽  
Lymphoid Cells ◽  
Spleen Cells ◽  
Background Stimulation ◽  
Human Sera ◽  
Stimulation Of

The media and culture conditions required for in vitro stimulation of mouse lymphoid cells are described. The medium was arginine-rich and contained heat-inactivated human serum. A component of the human sera necessary for stimulation of the cells was a natural mouse cell agglutinin, which affected both background stimulation and the degree of induced stimulation with phytohemagglutinin (PHA). Absorption of the agglutinin from the human serum rendered the medium incapable of sustaining DNA synthesis in the presence of PHA. The response to PHA of mouse spleen and thymus cells was age-dependent and, although this response was not present at birth, it rapidly rose to adult levels. Spleen cells from mice immunized with bacillus Calmette-Guérin (BCG) or sheep erythrocytes (SRBC) showed increased in vitro reactivity to added purified protein derivative (PPD) or SRBC stroma, dependent on the time of immunization. The dose response curve for the SRBC stroma stimulated, immune spleen cells is compatible with a theory of cell to cell interaction being necessary for an in vitro reaction to antigen. The possible role of the mouse cell agglutinin (AMLG) is discussed.

scholarly journals THE PROLIFERATIVE AND ANAMNESTIC ANTIBODY RESPONSE OF RABBIT LYMPHOID CELLS IN VITRO

1970 ◽  
Vol 131 (5) ◽  
pp. 970-980 ◽  
Author(s):  
G. A. Theis ◽  
G. J. Thorbecke
Keyword(s):  
Cell Interaction ◽  
Lymphoid Cells ◽  
Secondary Response ◽  
Adherent Cell ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
Absorbent Cotton ◽  
Depletion Effect ◽  
Made In

Both primary and secondary responses to sheep erythrocytes and to Brucella abortus antigen have been obtained in cultures of dispersed rabbit spleen cells. Removal of adherent cells by repeated incubation of spleen cells on absorbent cotton diminished the ability of the spleen cell suspensions to give secondary as well as primary responses in vitro. When comparing cultures made in dishes and in tubes, the loss of responsiveness after incubation on cotton was much more evident in the dish cultures. It was concluded that the cell-to-cell interaction needed for immune responses to particulate antigens in vitro was more readily interfered with when the cells were spread over a larger surface area. The proliferative response to antigen, as measured by uptake of 3H-thymidine in tube cultures of the sensitive spleen cells, appeared particularly resistant to the depletion effect of adherent cell removal. Dispersed spleen cells from sensitized mice gave a secondary response to sheep erythrocytes. This response was readily abolished by one incubation on absorbent cotton when the cells were cultured in dishes.

scholarly journals THE SECONDARY IMMUNE RESPONSE TO A HAPTEN IN VITRO

1971 ◽  
Vol 133 (5) ◽  
pp. 963-972 ◽  
Author(s):  
Norman R. Klinman
Keyword(s):  
B Lymphocyte ◽  
Cell Interaction ◽  
Antigenic Determinants ◽  
Secondary Immune Response ◽  
Spleen Cells ◽  
Marked Enhancement ◽  
Carrier Molecule ◽  
Stimulation Of

The in vitro secondary stimulation of the production of anti-hapten antibody has been analyzed with a view to elucidating the role of the carrier molecule and cell-to-cell interactions in this response. Stimulation was carried out on fragment cultures of the spleens of irradiated BALB/c mice which had been reconstituted with 3–4 x 107 spleen cells from isologous mice previously immunized with DNP-Hy. The results indicated that the response was maximized by stimulation with DNP-Hy, the homologous complex, however anti-DNP antibody production could be obtained by stimulation with DNP on several nonhomologous carriers including poly-D-lysine, a poor immunogen. The results also indicated that while DNP-Hy and DNP-nonhomologous-carrier complexes were stimulatory at equally low DNP concentrations, at DNP concentrations over 10–6 M DNP-Hy was stimulatory, while DNP on nonhomologous carriers was inhibitory. The results are interpreted as indicating that: (a) the affinity of the antigen-cell interaction is more likely determined by multivalent binding than by carrier recognition, (b) that a stimulatory interaction of a polyvalent antigen with a B-lymphocyte cannot be excluded, (c) that if cell-to-cell interaction is necessary for stimulation, then both cells may recognize the same determinant, and (d) that the marked enhancement of antigenic stimulation attributable to carrier recognition may result from stimulatory interactions of cells recognizing different antigenic determinants. A mechanism is postulated whereby stimulation is dependent on the formation of stable complexes resulting from two cells sharing in the binding of numerous antigen molecules. Cells recognizing carrier determinants would increase the probability of such interactions particularly at high antigen concentrations.

scholarly journals Autologous stimulation of human lymphocyte subpopulation.

1975 ◽  
Vol 142 (5) ◽  
pp. 1327-1333 ◽  
Author(s):  
G Opelz ◽  
M Kiuchi ◽  
M Takasugi ◽  
P I Terasaki
Keyword(s):  
T Cells ◽  
B Cells ◽  
B Lymphocytes ◽  
Human Lymphocyte ◽  
Lymphocyte Subpopulation ◽  
High Background ◽  
Background Stimulation ◽  
Most Probable Explanation ◽  
Stimulation Of

The background stimulation universally seen when lymphocytes are cultured in vitro has been shown to be markedly lowered by reducing the proportion of B lymphocytes. B-rich fractions of lymphocytes had extremely high background stimulation. It is concluded that stimulation of T cells, probably by autologous B cells, provides the most probable explanation for the findings described.

scholarly journals IMMUNE RESPONSES IN VITRO

1971 ◽  
Vol 134 (2) ◽  
pp. 395-416 ◽  
Author(s):  
Carl W. Pierce ◽  
Barbara M. Johnson ◽  
Harriet E. Gershon ◽  
Richard Asofsky
Keyword(s):  
Culture Conditions ◽  
Antibody Concentration ◽  
Spleen Cells ◽  
Immunoglobulin Class ◽  
Cell Densities ◽  
Erythrocyte Antigen ◽  
First Time ◽  
Kinetics Of

We have demonstrated for the first time that mouse spleen cells stimulated in vitro with heterologous erythrocytes developed immunoglobulin class-specific γM, γ1, γ2a+2b, and γA plaque-forming cell (PFC) responses. A modification of the hemolytic plaque technique, the addition of goat anti-mouse µ-chain antibody to the assay preparation, specifically prevented development of all γM PFC and enabled accurate and reproducible enumeration of immunoglobulin class-specific PFC after treatment with appropriate monospecific anti-globulins and complement. Culture conditions, with regard to medium, atmosphere, agitation, and spleen cell densities, were similar to those previously shown to support only γM PFC responses. Evaluation of the kinetics of appearance of PFC showed that γM PFC reached maximum numbers on days 4–5; the magnitude of this response was 3–10 times greater than γ1 γ2a+2b, or γA PFC which reached maximum numbers on days 5–6. Optimal erythrocyte antigen dose for γM PFC responses was 107/culture, whereas a dose of 106 erythrocytes/culture consistently stimulated optimal γ1 γ2a+2b, or γA PFC responses. Investigations of the effects of anti-erythrocyte antibody on γM and γG PFC responses indicated that antibody suppressed these responses by neutralizing the effective antigenic stimulus at the macrophage-dependent phase of the response. At the same antibody concentration, γG PFC responses were more effectively suppressed than γM PFC responses. Further, γG responses could be almost completely suppressed by antibody as long as 48 hr after initiation of cultures, whereas γM PFC responses could only be completely suppressed during the first 24 hr. These results were discusssed in terms of the role of antigen in the stimulation γM and γG antibody.

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.

scholarly journals Stimulation of syngeneic and allogeneic lymphoid cells by tumour cells in vitro

1975 ◽  
Vol 32 (1) ◽  
pp. 5-15
Author(s):  
M R Potter ◽  
B M Balfour
Keyword(s):  
Tumour Cells ◽  
Lymphoid Cells ◽  
Stimulation Of

scholarly journals HORMONE-LIKE ACTIVITY OF A THYMUS HUMORAL FACTOR ON THE INDUCTION OF IMMUNE COMPETENCE IN LYMPHOID CELLS

1974 ◽  
Vol 139 (1) ◽  
pp. 193-207 ◽  
Author(s):  
Abraham I. Kook ◽  
Nathan Trainin
Keyword(s):  
Adenyl Cyclase ◽  
Lymphoid Cells ◽  
Humoral Factor ◽  
Flufenamic Acid ◽  
Intracellular Camp ◽  
Immune Competence ◽  
Dibutyryl Camp ◽  
Spleen Cells ◽  
Antigenic Challenge

Experiments reported here were performed to understand the mechanism by which THF increases the immunocompetence of spleen cells from NTx mice. Dibutyryl cAMP or substances which increase intracellular levels of cAMP in lymphocytes such as Poly(A:U), theophylline, or PGE2 were shown to mimic the effect of THF and confer reactivity in an in vitro GvH response to spleen cells from NTx mice. Flufenamic acid, an antagonist to PGE2, was shown to inhibit the induction of competence by this substance. It was found that THF induces competence by activating membranal adenyl cyclase which leads to a rise in intracellular cAMP in thymus-derived cells only. These biochemical changes occur before antigenic stimulation and are unrelated to antigenic challenge. These findings indicate that THF exerts its effect via cAMP and are in agreement with the concepts which permit to classify THF as a thymus hormone.

In vitro Stimulation of Lymphoid Cells by Antilymphocytic Globulins

1969 ◽  
pp. 138-145
Author(s):  
Gert Riethmüller ◽  
Doris Riethmüller ◽  
Peter Rieber ◽  
Hans Stein
Keyword(s):  
Lymphoid Cells ◽  
Stimulation Of
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