scholarly journals STUDIES ON ANTIBODY PRODUCTION

1963 ◽  
Vol 117 (6) ◽  
pp. 1063-1074 ◽  
Author(s):  
Thomas F. O'Brien ◽  
Albert H. Coons

Incorporation of 5-bromodeoxyuridine (BUDR) in the culture medium for the 2nd, 3rd, or 4th day after in vitro antigen stimulation of rabbit popliteal lymph node fragments suppressed the in vitro anamnestic antibody response described previously. Before or after this 3-day period, BUDR had no measurable effect. The results suggest that the antibody response in this in vitro system depends upon cell multiplication during this period.

1963 ◽  
Vol 117 (6) ◽  
pp. 1053-1062 ◽  
Author(s):  
Thomas F. O'Brien ◽  
Maria C. Michaelides ◽  
Albert H. Coons

The in vitro anamnestic antibody response of popliteal lymph node fragments to additions of antigen closely resembles the in vivo anamnestic antibody response in its sensitivity to antigen, in the time course of antibody production, and in the sequence of appearance and the morphology of the antibody containing cells. Most of the cells responsible for antibody synthesis remain in the explant and do not migrate, although a few can be found in the outgrowing sheet of cells. The smallest concentration of bovine serum albumin which stimulates an anamnestic response in vitro is about 1 x 10–9 gm/ml.


1970 ◽  
Vol 131 (1) ◽  
pp. 93-99 ◽  
Author(s):  
Shraga Segal ◽  
Amiela Globerson ◽  
Michael Feldman ◽  
Joseph Haimovich ◽  
Michael Sela

Primary antibody response against the dinitrophenyl group has been elicited in vitro after the stimulation of normal mouse spleen explants with 2,4-dinitrophenyl (DNP)-hemocyanin or α-DNP-poly-L-lysine (PLL). Antibodies were detected in the culture medium by the inactivation of DNP-T4 phage. The specificity of the reaction was manifested by the lack of the capacity of the medium to inactivate the unmodified bacteriophage and by the inhibition of the inactivation of DNP-T4 with DNP-lysine.


1961 ◽  
Vol 114 (6) ◽  
pp. 837-856 ◽  
Author(s):  
M. Fishman

Neutralizing activity against T2 bacteriophage appeared in cultures of lymph node cells from normal rats in response to their in vitro stimulation with a cell-free filtrate derived from homogenized rat macrophages which had been incubated with T2 bacteriophage. This activity was specifically directed against T2 bacteriophage. It resided in a fraction of the culture fluid which had the salting-out properties of serum globulin. Phage neutralization was inhibited by antibody specific for rat serum gamma globulin. Antibody production against T2 bacteriophage in cultures of lymph node cells from normal animals failed to occur if (a) T2 bacteriophage alone was added, (b) if the incubation period of macrophages and T2 phage was unduly shortened, (c) if the cell-free filtrate was heated at 80–100°C for 15 minutes, (d) if more than an optimal amount of T2 bacteriophage was added to the macrophages. Additional factors which prevented the formation of antibody were the heat inactivation of the lymph node cells or the addition to the culture medium of either streptomycin or ribonuclease. Finally, it was found that macrophages and lymph node cells had to be obtained from animals of one and the same species. All essential findings on the production of antibody to T2 bacteriophage in vitro could be confirmed by substitution of the chick embryo for the tissue culture medium. The results are discussed in terms of a possible mechanism of antibody production in which an RNAse-sensitive substance resulting from the interaction of macrophages and antigen is capable of stimulating antibody synthesis in lymphocytic cells.


BIO-PROTOCOL ◽  
2019 ◽  
Vol 9 (15) ◽  
Author(s):  
Songjie Cai ◽  
Masayuki Fujino ◽  
Lina Lu ◽  
Xiao-Kang Li

2021 ◽  
Vol 13 (578) ◽  
pp. eabg5638
Author(s):  
Gerald P. Morris

Development of a human lymphoid organoid system enables in vitro modeling of immune responses and antibody formation.


1968 ◽  
Vol 127 (2) ◽  
pp. 307-325 ◽  
Author(s):  
Vera S. Byers ◽  
Eli E. Sercarz

A set of conditions has been described under which primed rabbit lymph nodes produce a secondary antibody response upon in vivo stimulation with a large dose of antigen, but are subsequently "exhausted;" that is, lymph node cultures prepared at intervals following the booster injection cannot be re-stimulated to display tertiary responses. Rabbits given 100-fold less antigen in the booster inoculum were able to give a tertiary response upon in vitro challenge. The system used permits neither induction nor continuation of a primary response to BSA in vitro. Since it could be demonstrated that no memory cells were generated by the booster injection within the intervals between in vivo injection and culture, the tertiary response in nonexhausted nodes must have been due to residual memory cells which remained untriggered by the in vivo booster injection. The unresponsive state was not caused by antibody feedback. These results are interpreted to mean that a population of memory cells can be exhausted by a supraoptimal dose of antigen, rendering the node temporarily incapable of further response. This implies that long-lived memory is not due to asymmetric division of memory cells. The source and fate of memory cells is discussed with regard to this evidence.


1970 ◽  
Vol 132 (5) ◽  
pp. 845-857 ◽  
Author(s):  
V. S. Byers ◽  
E. E. Sercarz

Induction of the immune response can only be completed after antigen is removed from the cellular environment. Primed rabbit lymph node fragments were cultured in vitro with 5 mg/ml BSA. If antigen was removed from the fragments 2 hr later, they produced a normal anti-BSA response, which was first evident 5 days later. If antigen removal was delayed for 3 days, the onset of the response was postponed for 2 to 3 days. Pulses with BUDR marked the periods of cell proliferation in both sets of cultures, and established that the postponement of antibody production was preceded by a postponement in the wave of proliferation among precursors of antibody forming cells. The similarity in avidity of antibody-containing fluids from normal and postponed cultures support the idea that the same cell population produced the response in each case. It was concluded that a reversible state of paralysis could be instituted in antigen-responsive cells, and this state did not depend upon cell-killing. The widespread incidence of temporary paralysis as an early aspect of the immune response was discussed.


Nature ◽  
1957 ◽  
Vol 179 (4565) ◽  
pp. 870-871 ◽  
Author(s):  
K. M. STEVENS ◽  
J. M. McKENNA

1991 ◽  
Vol 128 (3) ◽  
pp. 389-393 ◽  
Author(s):  
B. Houston ◽  
I. E. O'Neill

ABSTRACT Cultured chicken hepatocytes were used to investigate whether insulin and GH interact to regulate insulin-like growth factor-I (IGF-I) production in vitro. In the first set of experiments hepatocytes were preincubated for 6 h in hormone-free medium, and the effects of various combinations of insulin and GH on IGF-I production over the next 24 h were quantified by radioimmunoassay. Basal IGF-I production was 5·36 pg IGF-I/μg DNA and this was increased 1·31±0·13-fold (mean ± s.e.m.) by insulin, 1·90±0·24-fold by GH and 4·46±0·68-fold by a combination of insulin and GH. These results demonstrate that insulin and GH interact synergistically to stimulate IGF-I production in vitro. The synergism with GH occurred at physiological concentrations of insulin with half-maximal stimulation occurring at an insulin concentration of 6 ng/ml. In hepatocytes which had been exposed to insulin immediately before the start of the experiment, the presence of insulin was no longer required for maximal stimulation of IGF-I production by GH. This in-vitro system will facilitate the study of the molecular basis of the interaction between insulin and GH. Journal of Endocrinology (1991) 128, 389–393


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