scholarly journals INHIBITION OF THE SECONDARY ANTIBODY RESPONSE IN VITRO BY SALICYLATE AND GENTISATE

1966 ◽  
Vol 124 (3) ◽  
pp. 461-482 ◽  
Author(s):  
Charles Tesch Ambrose
Keyword(s):  
Antibody Response ◽  
Sodium Salicylate ◽  
Secondary Response ◽  
Secondary Antibody ◽  
Serum Free Medium ◽  
Free Medium ◽  
Antibody Synthesis ◽  
Salicylate Concentration ◽  
Salicylic Acid Concentration

Salicylate inhibition of the secondary antibody response initiated in vitro on day 0 has been studied in cultures of rabbit lymph node fragments. Levels of 1.25 to 1.5 mM (0.20 to 0.24 mg/ml) sodium salicylate present in serum-free medium throughout an 18- or 21-day culture period completely inhibit the secondary response. This inhibition is largely accomplished by the drug's action during the first 9 days, which corresponds to the inductive phase for this culture system. Relatively little inhibition is produced by adding the drug only after day 9, although over 90% of the antibody produced during a 21-day experiment is synthesized after day 9. Studies with media of different pH's show that this inhibition is more correctly a function of the nonionized salicylic acid concentration in the medium than of the total salicylate concentration. Arguments are presented against the possibility that salicylate at the levels used here inhibits antibody synthesis by uncoupling oxidative phosphorylation. Acetylsalicylic acid (aspirin) produces the same degree of inhibition in vitro as do equimolar concentrations of sodium salicylate. Gentisate (5-hydroxysalicylate) is 15-fold more effective in producing 50% inhibition than salicylate; its temporal pattern of inhibition is similar to that of salicylate.

scholarly journals THE REQUIREMENT FOR HYDROCORTISONE IN ANTIBODY-FORMING TISSUE CULTIVATED IN SERUM-FREE MEDIUM

1964 ◽  
Vol 119 (6) ◽  
pp. 1027-1049 ◽  
Author(s):  
Charles T. Ambrose
Keyword(s):  
Culture Medium ◽  
Essential Amino Acids ◽  
Secondary Response ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Secondary Antibody ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

It was previously reported from this laboratory that the secondary antibody response can regularly be elicited in vitro from fragments of rabbit lymph node node cultured in Eagle's medium supplemented with normal rabbit serum. Evidence is now presented that physiological levels of hydrocortisone (0.01 to 1.0 µM) can substitute for serum in the culture medium. However, with the omission of serum, serine (0.1 mM) must be included among Eagle's "essential" amino acids for consistent optimal antibody production. In some experiments the addition of insulin (0.5 unit/ml) and vitamin B12 (0.5 µg/ml) has further enhanced the secondary response in this serum-free medium.

scholarly journals STUDIES ON ANTIBODY PRODUCTION

1964 ◽  
Vol 120 (6) ◽  
pp. 1051-1060 ◽  
Author(s):  
William T. Butler ◽  
Albert H. Coons
Keyword(s):  
Antibody Response ◽  
Antibody Production ◽  
Experimental System ◽  
Diphtheria Toxoid ◽  
Secondary Response ◽  
Secondary Antibody ◽  
Real Difference ◽  
Antibody Synthesis ◽  
Drug Dosages

The effect of drugs upon the primary and the secondary antibody response to diphtheria toxoid in mice was studied using an experimental system previously described. Triethylenethiophosphoramide (thio-TEPA), chloramphenicol, 6-mercaptopurine, 8-azaguanine, and versenate were found to inhibit, partially or completely,"priming" for the secondary response. Thio-TEPA, chloramphenicol, and 6-mercaptopurine, in doses exceeding those effective in inhibiting priming, did not cause alteration of the secondary response when given only during the secondary response. However, when chloramphenicol and amethopterin were given for 5 days prior to and at least 5 days after the second antigen injection, slight suppression of peak secondary titers occurred. Therefore, drug dosages effective in suppressing priming had less effect on the secondary response. It thus appears that there is a real difference between "priming" and the induction of antibody synthesis.

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

1969 ◽  
Vol 130 (2) ◽  
pp. 287-297 ◽  
Author(s):  
E. B. Jacobson ◽  
G. J. Thorbecke
Keyword(s):  
Antibody Response ◽  
Lymph Nodes ◽  
Lymphoid Tissue ◽  
Slow Release ◽  
Lymphoid Cells ◽  
Secondary Response ◽  
Striking Difference ◽  
Secondary Antibody ◽  
Lymph Node Cells

Popliteal lymph nodes were obtained from rabbits 4 days to 9 months after a primary injection of diphtheria toxoid or bovine γ-globulin into the footpad. The ability of cells from these nodes to proliferate upon reexposure to antigen in vitro was compared to the height of the secondary response produced by tissue fragments. In addition, a comparison was made between the responsiveness of draining and contralateral lymph nodes. While the secondary antibody response in vitro increased markedly with the time after immunization at which the lymph nodes were taken from the animals, the degree of proliferation induced by antigen was highest with cells from lymph nodes taken early after priming (peak day 7) and was very much lower with lymph node cells taken longer than 3 wk after priming. This striking difference between these two responses has been discussed. Contralateral lymph nodes were much inferior to draining nodes in their ability to give a secondary antibody response in vitro, and never gave a detectable proliferative response. This difference became less marked with time after priming, but could still be demonstrated after 4 months. These results suggest a concentration of primed cells in the lymphoid tissue draining the site of injection, and a slow release of these cells into the circulation, to be distributed to the remaining lymphoid tissue.

Regulation of the in vitro secondary antibody response

1980 ◽  
Vol 55 (2) ◽  
pp. 302-311 ◽  
Author(s):  
Sharyn M. Walker ◽  
William O. Weigle
Keyword(s):  
Antibody Response ◽  
Secondary Antibody

scholarly journals Quantification of Internalized Silica Nanoparticles via STED Microscopy

2015 ◽  
Vol 2015 ◽  
pp. 1-16 ◽  
Author(s):  
Henrike Peuschel ◽  
Thomas Ruckelshausen ◽  
Christian Cavelius ◽  
Annette Kraegeloh
Keyword(s):  
Silica Nanoparticles ◽  
Super Resolution ◽  
Engineered Nanoparticles ◽  
Stimulated Emission ◽  
Alveolar Type ◽  
Serum Free Medium ◽  
Free Medium ◽  
Essential Information ◽  
Serum Free

The development of safe engineered nanoparticles (NPs) requires a detailed understanding of their interaction mechanisms on a cellular level. Therefore, quantification of NP internalization is crucial to predict the potential impact of intracellular NP doses, providing essential information for risk assessment as well as for drug delivery applications. In this study, the internalization of 25 nm and 85 nm silica nanoparticles (SNPs) in alveolar type II cells (A549) was quantified by application of super-resolution STED (stimulated emission depletion) microscopy. Cells were exposed to equal particle number concentrations (9.2×1010particles mL−1) of each particle size and the sedimentation of particles during exposure was taken into account. Microscopy images revealed that particles of both sizes entered the cells after 5 h incubation in serum supplemented and serum-free medium. According to thein vitrosedimentation, diffusion, and dosimetry (ISDD) model 20–27% of the particles sedimented. In comparison, 102-103NPs per cell were detected intracellularly serum-containing medium. Furthermore, in the presence of serum, no cytotoxicity was induced by the SNPs. In serum-free medium, large agglomerates of both particle sizes covered the cells whereas only high concentrations (≥ 3.8 × 1012particles mL−1) of the smaller particles induced cytotoxicity.

Hybridoma antibody production in vitro in type II serum-free medium using Nutridoma-SP supplements Comparisons with in vivo methods

1991 ◽  
Vol 145 (1-2) ◽  
pp. 213-221 ◽  
Author(s):  
Geneviève Federspiel ◽  
Kenneth C. McCullough ◽  
Ulrich Kihm
Keyword(s):  
Antibody Production ◽  
Type Ii ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

scholarly journals Enhancement of Dopaminergic Differentiation in Proliferating Midbrain Neuroblasts by Sonic Hedgehog and Ascorbic Acid

2004 ◽  
Vol 11 (1-2) ◽  
pp. 45-57 ◽  
Author(s):  
Floriana Volpicelli ◽  
Claudia Consales ◽  
Massimiliano Caiazzo ◽  
Luca Colucci-D'Amato ◽  
Carla Perrone-Capano ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Sonic Hedgehog ◽  
Molecular Mechanisms ◽  
Experimental System ◽  
Serum Free Medium ◽  
Free Medium ◽  
Ventral Mesencephalon ◽  
Mes Cells ◽  
Dopaminergic Differentiation

We analyzed the molecular mechanisms involved in the acquisition and maturation of dopaminergic (DA) neurons generated in vitro from rat ventral mesencephalon (MES) cells in the presence of mitogens or specific signaling molecules. The addition of basic fibroblast growth factor (bFGF) to MES cells in serum-free medium stimulates the proliferation of neuroblasts but delays DA differentiation. Recombinant Sonic hedgehog (SHH) protein increases up to three fold the number of tyrosine hydroxylase (TH)-positive cells and their differentiation, an effect abolished by anti-SHH antibodies. The expanded cultures are rich in nestin-positive neurons, glial cells are rare, allTH+neurons are DA, and all DA and GABAergic markers analyzed are expressed. Adding ascorbic acid to bFGF/SHH-treated cultures resulted in a further five- to seven-fold enhancement of viable DA neurons. This experimental system also provides a powerful tool to generate DA neurons from single embryos. Our strategy provides an enriched source of MES DA neurons that are useful for analyzing molecular mechanisms controlling their function and for experimental regenerative approaches in DA dysfunction.

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