scholarly journals The role of fetal calf serum in the primary immune response in vitro.

1977 ◽  
Vol 145 (4) ◽  
pp. 1029-1038 ◽  
Author(s):  
H G Opitz ◽  
U Opitz ◽  
H Lemke ◽  
G Hewlett ◽  
W Schreml ◽  
...  
Keyword(s):  
Immune Response ◽  
Culture Medium ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Primary Immune Response ◽  
Spleen Cells ◽  
Humoral Immune ◽  
Serum Component ◽  
Immune Response In Vitro

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.

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
Keyword(s):  
Immune Response ◽  
Cell Cultures ◽  
Primary Immune Response ◽  
Antigenic Determinants ◽  
Specific Cell ◽  
Spleen Cells ◽  
Sheep Erythrocytes ◽  
Cell Clusters ◽  
Immune Response In Vitro

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.

Effects of Actinomyces and Fusobacterium on humoral immune response in vitro

1981 ◽  
Vol 16 (5) ◽  
pp. 556-563 ◽  
Author(s):  
Hiromasa Yoshie ◽  
Takashi Mitsuma ◽  
Keniti Kozima ◽  
Kohji Hara
Keyword(s):  
Immune Response ◽  
Humoral Immune Response ◽  
Humoral Immune ◽  
Immune Response In Vitro

Forskolin effect on the cryosurvival of in vitro-produced bovine embryos in the presence or absence of fetal calf serum

Zygote ◽  
2012 ◽  
Vol 22 (2) ◽  
pp. 146-157 ◽  
Author(s):  
Daniela Martins Paschoal ◽  
Mateus José Sudano ◽  
Midyan Daroz Guastali ◽  
Rosiára Rosária Dias Maziero ◽  
Letícia Ferrari Crocomo ◽  
...  
Keyword(s):  
Culture Medium ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Similar Rate ◽  
Bovine Embryos ◽  
Embryo Survival ◽  
Vitrification Solution ◽  
Oviductal Fluid

SummaryThe objective of this study was to assess the viability and cryotolerance of zebu embryos produced in vitro with or without the addition of fetal calf serum (FCS) and forskolin (F). Embryos produced in vivo were used as a control. Presumptive zygotes were cultured in modified synthetic oviductal fluid supplemented with amino acids (SOFaa), bovine serum albumin (BSA) and with (2.5%) or without (0%) FCS. On day 6 of growth, the embryos from each group were divided into treatments with or without 10 μM F to induce embryonic lipolysis, comprising a total of four experimental groups: 2.5% FCS, 0% FCS, 2.5% + F and 0% + F. For vitrification, embryos were exposed to vitrification solution 1 (5 M EG (ethylene glycol)) for 3 min and then transferred to vitrification solution 2 (7 M EG, 0.5 M galactose solution and 18% (w/v) Ficoll 70) before being introduced to liquid nitrogen. The presence of FCS in the culture medium resulted in the production of embryos with a similar rate of damaged cells compared with in vivo-produced embryos. After vitrification, the 2.5% FCS group had a significantly higher rate of damaged cells when compared with the other groups (P < 0.05). The results of this experiment indicated that the omission of FCS and the addition of forskolin do not have deleterious effect on embryo production rates. In addition, embryos produced in the presence of FCS had greater sensitivity to cryopreservation, but this effect was reversed when forskolin was added to the medium, which improved embryo survival without affecting embryo development and quality after vitrification.

scholarly journals ANTIBODY-MEDIATED SUPPRESSION OF THE IMMUNE RESPONSE IN VITRO

1970 ◽  
Vol 131 (2) ◽  
pp. 247-274 ◽  
Author(s):  
Marc Feldmann ◽  
Erwin Diener
Keyword(s):  
Immune Response ◽  
Immune Suppression ◽  
Serum Antibody ◽  
Cellular Level ◽  
Mouse Spleen ◽  
Spleen Cells ◽  
Normal Response ◽  
In Vitro Immune Response ◽  
Immune Response In Vitro

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.

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 Feedback suppression of the immune response in vitro. I. Activity of antigen-stimulated B cells.

1980 ◽  
Vol 151 (3) ◽  
pp. 667-680 ◽  
Author(s):  
R H Zubler ◽  
H Cantor ◽  
B Benacerraf ◽  
R N Germain
Keyword(s):  
Immune Response ◽  
T Cells ◽  
B Cells ◽  
Spleen Cell ◽  
Suppressor Cells ◽  
Feedback Regulation ◽  
Spleen Cells ◽  
Humoral Immune ◽  
Feedback Suppression

Feedback regulation of the primary humoral immune response to sheep erythrocytes (SRBC) was studied in vitro. Whole spleen cells or spleen cell subpopulations were incubated with antigen for 4 d under Mishell-Dutton conditions (education) and the surviving cells tested for regulatory activity in fresh anti-SRBC spleen cell cultures assayed by measuring plaque-forming cells on day 4. The data indicate that (a) whole spleen cells educated with SRBC exert potent antigen-specific suppression in the assay culture, (b) surface Ig- (sIg-) cells (T cells) prepared by either nylon-wool separation or fractionation on rabbit anti-mouse-Ig-coated polystyrene Petri dishes failed to generate suppressive activity when educated alone, in 2-mercaptoethanol, or in the presence of additional macrophages, (c) surface Ig (sIg+) (B) cells educated alone also failed to generate suppressor cells, and (d) mixing sIg- (T) and sIg+, Lyt 123- (B) cells reconstituted the ability to induce suppressor cells under these conditions. The antigen-primed cell actually required to transfer suppression was also characterized by separating cells using anti-Ig coated dishes, by fluorescence-activated cell sorting and by anti-Lyt treatment. All these methods clearly identified sIg+ (B) and not sIg+ (T) cells as the important educated cells. It is concluded that under our conditions, T cell-dependent B cells triggered by antigen during primary in vitro cultures cause potent specific feedback suppression of humoral responses. Possible mechanisms for this suppression, including antigen blockade or anti-idiotypic responses, are discussed.

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