scholarly journals CELL INTERACTIONS IN THE PRIMARY IMMUNE RESPONSE IN VITRO: A REQUIREMENT FOR SPECIFIC CELL CLUSTERS

1969 ◽  
Vol 129 (2) ◽  
pp. 351-362 ◽  
Author(s):  
Donald E. Mosier

Mouse spleen cells were found to associate in cell clusters during the primary immune response to sheep erythrocytes in vitro. About 10% of the cell clusters had the following unique properties; (a) they contained most, if not all, antibody-forming cells, (b) they contained only cells forming antibody to one antigen when cell cultures were immunized with two antigens, (c) the cells in clusters reaggregated specifically after dispersion, and (d) the specific reaggregation of clusters appeared to be blocked by antibody to the antigen. The integrity of cell clusters was required for the proliferation of antibody-forming cells, and prevention of clustering by mechanical means or by excess antibody blocked the immune response. Antibody and antigenic determinants on the surfaces of cells probably provide the basis for interaction. The unique microenvironment of cell clusters was essential for the primary immune response in vitro.

1968 ◽  
Vol 127 (5) ◽  
pp. 983-1002 ◽  
Author(s):  
Donald A. Rowley ◽  
Frank W. Fitch ◽  
Donald E. Mosier ◽  
Susan Solliday ◽  
Lionel W. Coppleson ◽  
...  

Mitotic blocking agents, colchicine or Velban, were used to estimate cycle times of spleen cells which release hemolysin for sheep erythrocytes (plaque-forming cells). The cells were obtained either from rats immunized with sheep erythrocytes or from cultures of mouse spleen cells immunized in vitro with the same antigen. 2, 3, or 4 days after immunization, animals or cell cultures were treated with mitotic blocking agents for periods of time ranging from 2.5 to 7 hr; plaque-forming cells were then enumerated. Decreased numbers of plaque-forming cells were found after such treatment. The extent of reduction was a function of duration of the drug treatment and the method of immunization, but was independent of the time after immunization. The evidence presented is consistent with premises that: (a) plaque-forming cells in mitosis do not release sufficient antibody to be detected, (b) mitotic blocking agents, by arresting plaque-forming cells in metaphase, prevent not only detection of these cells but also the increase in number of plaque-forming cells which would have resulted from cell division, (c) mitotic blocking agents do not affect release of antibody by cells in interphase. Cell cycle times, based on the extent of reduction of plaque-forming cells per unit time of drug treatment, were estimated using a mathematical model appropriate for an exponentially increasing population of cells. Cell cycle times estimated using the mitotic blocking agents agreed well with cell doubling times calculated from the increase in plaque-forming cells occurring 1–4 days after immunization. Increased responses produced by higher antigen doses or treatment of immunized animals with an adjuvant resulted from an increased rate of division of responding cells and their progeny. The results are consistent with a cell selection theory of antibody formation. Antigenic stimulation causes relatively few cells to proliferate and to synthesize antibody; apparently the magnitude of the response is dependent primarily on the rate of division of responding cells. It is suggested on the basis of observations of in vitro-immunized cell cultures that the rate of division of responding cells may be dependent on the rate of interaction between two cell types, both of which are essential for the in vitro plaque-forming cell response.


1972 ◽  
Vol 25 (5) ◽  
pp. 345-353 ◽  
Author(s):  
MASANOBU SUGIMOTO ◽  
SHIN-ICHI TAMURA ◽  
TAKESHI KURATA ◽  
YASUYUKI EGASHIRA

1977 ◽  
Vol 145 (4) ◽  
pp. 1029-1038 ◽  
Author(s):  
H G Opitz ◽  
U Opitz ◽  
H Lemke ◽  
G Hewlett ◽  
W Schreml ◽  
...  

The mode of action of 2-mercaptoethanol (2-ME) on the primary immune response in vitro was investigated. Fetal calf serum (FCS) was preincubated with 2-ME and lyophilized to remove free 2-ME. This 2-ME-treated FCS was able to substitute the function of adherent cells in the primary immune response against sheep red blood cells (SRBC) in vitro; Fractionation of 2-tme-treated FCS on a Sephadex G-100 column showed that 2-ME acted on a high molecular serum component which after activation, could substitute for macrophages. In order to obtain a humoral immune response against SRBC in vitro, spleen cells require selected FCS. These "good" sera could be distinguished from "deficient" sera by their higher content of this 2-ME-activated factor. The height of the in vitro immune response to SRBC was dependent on the amount of activated factor added to the culture medium. FCS normally required in the culture medium could be completely replaced by the factor-containing fraction without deleterious effect on the culture medium. The factor should be added to the spleen cells during the first 24 h of culture and remain there for 72 h in order to obtain an optimal immune response. The factor could be partially absorbed by spleen cells but not by SRBC. The relationship between macrophage, 2-ME, and FCS in eliciting an in vitro primary immune response is discussed.


1970 ◽  
Vol 131 (2) ◽  
pp. 247-274 ◽  
Author(s):  
Marc Feldmann ◽  
Erwin Diener

Antibody-mediated suppression of the in vitro immune response to polymerized flagellin of Salmonella adelaide and to sheep erythrocytes was studied at the cellular level. Normal mouse spleen cells, preincubated in vitro with mixtures of antigen and antibody for short periods of time before being washed, did not respond to an optimal antigenic challenge in vitro, whereas similar cells treated with antibody alone gave a normal response. The degree of immune suppression was found to depend on the time of preincubation. Significant immune suppression could be induced in as short a time as 15 min, whereas profound suppression (90%) required the incubation of cells with mixtures of antigen and antibody for 4–6 hr. Mouse spleen cells treated similarly were also unable to respond subsequently to the antigen upon transfer to lethally irradiated hosts, as measured at both the level of the antigen-reactive cell and that of serum antibody production. These results were taken as evidence that in vitro an effect of antibody-mediated suppression occurred at the level of the immunocompetent cell. Similarities between immune tolerance and antibody-mediated suppression in vitro were described, and the significance of the findings discussed in the light of current concepts of the mechanism of antibody-mediated suppression.


1967 ◽  
Vol 126 (2) ◽  
pp. 395-405 ◽  
Author(s):  
Erna Möller

Nonimmune lymphoid cells were capable of causing cytotoxicity of H-2 incompatible mouse tumor cells in vitro in the presence of PHA, whereas syngeneic cells were not. Semisyngeneic and X-irradiated (1500–3000 R) F1 hybrid lymphoid cells were cytotoxic for target cells derived from one of the parental strains. In addition, parental nonimmune and X-irradiated lymphoid cells damaged hybrid target cells. It was concluded that one component of cytotoxicity was not related to an induction of a primary immune response in vitro, since F1 hybrid cells are not capable of reacting immunologically against parental type target cells. It seemed probable that cytotoxicity was caused by target cell confrontation with antigenically and/or structurally incompatible lymphoid cells. This conclusion was strengthened by the demonstration that isoantibodies produced in the target strain and directed against the allogeneic lymphoid cells specifically suppressed cytotoxicity. Isoantibodies reacting against some but not all of the antigenic determinants of the lymphoid cells differentiating them from the target cells did not suppress cytotoxicity. The specific suppression of cytotoxicity by specific isoantibodies against the lymphoid cells support the allogeneic inhibition concept.


1978 ◽  
Vol 20 ◽  
pp. 131-141 ◽  
Author(s):  
J. Kettman ◽  
S. Ben-Sasson ◽  
J.U. Rudin

1970 ◽  
Vol 16 (7) ◽  
pp. 623-627 ◽  
Author(s):  
H. Finger ◽  
P. Emmerling ◽  
M. Büsse

In this study we determined at both the cellular and humoral level whether or not the primary immune response of mice can be significantly enhanced by administration of a bacterial adjuvant after the primary immunization with sheep erythrocytes. As compared to the immunization of mice with 8 × 106 sheep erythrocytes alone, the simultaneous injection of 3 × 109Bordetella pertussis cells and 8 × 106 sheep erythrocytes resulted in an accelerated and prolonged multiplication of hemolysin-forming spleen cells. The adjuvant effect was also documented by increased production of serum hemolysins. When the bacterial adjuvant was given 6, 12, or 24 h after the primary antigenic stimulus, however, neither increased plaque counts nor enhanced serum hemolysin titers were detectable. These findings agree with the concept that B. pertussis cells cause multiplication of antigen-sensitive target cells or affect the initial stages of differentiation of these cells.


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