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 Suppression of in vitro cytotoxic response by macrophages due to induced arginase.

1977 ◽  
Vol 146 (3) ◽  
pp. 665-672 ◽  
Author(s):  
J T Kung ◽  
S B Brooks ◽  
J P Jakway ◽  
L L Leonard ◽  
D W Talmage
Keyword(s):  
Fetal Calf Serum ◽  
Calf Serum ◽  
Host Reaction ◽  
Minimum Essential Medium ◽  
Peritoneal Cells ◽  
Cytotoxic Response ◽  
Complete Reversal ◽  
Nonadherent Cells ◽  
Partial Reversal

Arginine was found to be completely depleted from cell-free supernates of mixed leukocyte cultures suppressed by the addition of excess macrophages. Partial reversal of macrophage-mediated suppression was accomplished by daily addition of a cocktail containing arginine and nonamino acid nutrients. Complete reversal of the suppression was accomplished by the addition of arginine and glucose to the medium and the nonadherent cells after their separation from the adherent macrophages. A marked increase in the enzyme arginase was found in macrophages that had been cultured 24 h in vitro in Eagle's minimum essential medium plus 10% fetal calf serum, in peritoneal cells activated by prior injection of thioglycollate, and in one spleen activated by a graft vs. host reaction.

scholarly journals Antigen recognition. IV. Discrimination by antigen-binding immunocompetent B cells between immunity and tolerance is determined by adherent cells.

1976 ◽  
Vol 143 (4) ◽  
pp. 805-821 ◽  
Author(s):  
E Diener ◽  
N Kraft ◽  
K C Lee ◽  
C Shiozawa
Keyword(s):  
B Cell ◽  
Short Pulse ◽  
Tolerance Induction ◽  
Mouse Spleen ◽  
Adherent Cells ◽  
Antigen Receptors ◽  
Spleen Cells ◽  
A Cells ◽  
Tolerant State

Mouse spleen cells capable of specifically binding intrinsically tritium-labeled polymerized flagellin (POL) (labeling by biosynthesis of flagellar protein) via IgM receptors were found to comprise a distinct population of about 20-50 cells per 10(6) lymphocytes. Evidence is presented that the majority of mouse spleen cells binding tritium-labeled POL undergoes blastogenesis after antigen capping, antigen shedding, and receptor reformation. Under conditions of tolerance induction in vitro, however, loss of antigen from the cell surface was inhibited. Such inhibition of antigen redistribution and shedding was reversed by a short pulse of colchicine and new antigen receptors were formed. In spite of this, colchicine had no effect on the tolerant state. However, tolerance could be broken, regardless of presence or absence of the alkaloid, with radioresistant theta-negative accessory (A) cells (adherent cells) from normal but not from tolerant spleen cell populations. "Tolerant" A cells, although they were incapable of cooperating in a response to POL, were capable of participating in a response to a second unrelated antigen. It is concluded that tolerance to POL in vitro is induced by mechanisms other than the physical blocking of bone marrow-derived (B) cell receptors by antigen. Most likely, the discrimination by the B cell between a tolerogenic and immunogenic signal is mediated by A cells.

scholarly journals THYMUS-INDEPENDENT (B) CELL PROLIFERATION IN SPLEEN CELL CULTURES OF MOUSE RADIATION CHIMERAS STIMULATED BY PHYTOHEMAGGLUTININ OR ALLOGENEIC CELLS

1972 ◽  
Vol 136 (4) ◽  
pp. 962-967 ◽  
Author(s):  
P.-F. Piguet ◽  
P. Vassalli
Keyword(s):  
T Cell ◽  
Spleen Cell ◽  
Cell Cultures ◽  
Bone Marrow Cells ◽  
Spleen Cells ◽  
Radiation Chimeras ◽  
Lymphocyte Cultures ◽  
Mixed Lymphocyte Cultures

Spleen cell cultures of radiation chimeras (thymectomized, lethally irradiated mice repopulated with bone marrow cells and thymocytes bearing different chromosomal markers) were stimulated by phytohemagglutinin (PHA) and F1 allogeneic spleen cells. Karyotypic analyses showed a marked predominance of T mitoses on the 2nd and 3rd days of culture followed by a strong predominance of B mitoses on the 4th and 5th days. Analysis of cells undergoing their first mitoses showed that the majority of T mitoses on day 3 resulted from continuous T cell division, and that most cells entering their first mitoses at that time were of B type. Mixed lymphocyte cultures (MLC) of chimeras immunized against allogeneic spleen cells showed sometimes, but not always, a response different from "primary" MLC, with an earlier and stronger predominance of BM mitoses. The role of stimulated T cells in the induction of B mitoses was shown by (a) the incapacity of T-depleted spleen cells to be stimulated by PHA or in primary or secondary MLC, and (b) the restoration of the mitotic response of B cells to PHA by adding to the T cell-depleted culture either a very small number of T cell (identified by their different karyotype: "in vitro chimeras") or the cell-free supernatant of a 24 hr MLC.

scholarly journals Differential effects of polyadenylic: polyuridylic acid and lipopolysaccharide on the generation of cytotoxic T lymphocytes.

1978 ◽  
Vol 147 (5) ◽  
pp. 1355-1362 ◽  
Author(s):  
P R Narayanan ◽  
G Sundharadas
Keyword(s):  
Cytotoxic T Lymphocytes ◽  
Cytotoxic Lymphocytes ◽  
Adherent Cells ◽  
Spleen Cells ◽  
Cytotoxic Cells ◽  
Cytotoxic Response ◽  
Two Agents ◽  
Polyadenylic Acid ◽  
Polyuridylic Acid ◽  
Lines Of Evidence

In a mixed leukocyte culture (MLC) reaction of allogenic mouse spleen cells differing for H-2K or H-2D, only a weak cytotoxic response is generated. This cytotoxic response is augmented significantly if bacterial lipopolysaccharide (LPS), 5 microgram/ml, or polyadenylic acid (poly A):polyuridylic acid (poly U), 20 microgram/ml, is present in the culture. The cytotoxic cells generated in the presence of these two agents are specific for sensitizing H-2K or H-2D antigen. Two lines of evidence suggest that these two agents exert their effect at different steps in the development of cytotoxic lymphocytes: (a) the effect of poly A:U depends on the presence of adherent cells, whereas the effect of LPS is independent of the presence of adherent cells and (b) LPS promotes the development of cytotoxic cells when ultraviolet light-treated stimulating cells are used in the MLC whereas poly A:U does not.

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 Fetal calf serum enhances in vitro production of Bos taurus indicus embryos

2011 ◽  
Vol 75 (3) ◽  
pp. 429-433 ◽  
Author(s):  
F.G. Leivas ◽  
D.S. Brum ◽  
S.S. Fialho ◽  
W.P. Saliba ◽  
M.T.T. Alvim ◽  
...  
Keyword(s):  
Fetal Calf Serum ◽  
Bos Taurus ◽  
Calf Serum ◽  
In Vitro Production ◽  
Bos Taurus Indicus ◽  
Vitro Production

scholarly journals Specific elimination of cytotoxic effector cells. I. Adsorptive behavior of effectors and their precursors and spleen cell monolayers.

1976 ◽  
Vol 144 (4) ◽  
pp. 996-1008 ◽  
Author(s):  
J R Neefe ◽  
D H Sachs
Keyword(s):  
Mouse Spleen ◽  
Spleen Cells ◽  
Effector Cells ◽  
Cell Monolayers ◽  
Specific Suppression ◽  
Normal Spleen ◽  
Adsorptive Behavior ◽  
Cytotoxic Effector ◽  
Primed Mouse

Monolayers formed of normal mouse spleen cells attached to polystyrene coated with poly-L-lysine were tested for their ability to bind specifically antigen-reactive cells in normal or primed mouse spleen. 88 to greater than 98% of the activity of cytotoxic populations was removed by a single adsorption. However, normal spleen cells or spleen cells previously primed in vitro could not be depleted of their capacity to be sensitized, even when adsorption effectively removed all residual cytotoxic activity from the same previously primed population. In fact, exposure to an immunoadsorbent augmented the ultimate cytotoxicity generated in a nonspecific fashion. This augmentation was especially dramatic in the case of a previously primed population and may have reflected the removal of a nonspecific suppressor. If antigen-reactive precursors cannot be removed efficiently by adsorption, other approaches to the generation of tolerant lymphoid populations, such as specific suppression of precursor differentiation must be sought.

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