Immunotolerance to Insulin and Freund's Adjuvant

1971 ◽  
Vol 49 (8) ◽  
pp. 865-866 ◽  
Author(s):  
P. J. Moloney ◽  
M. A. Evans

Prior injections of incomplete Freund's Adjuvant with saline did not suppress antibody formation by ox insulin. This finding is in contrast to that of Jankovic (Colloq. Int. Cent. Nat. Rech. Sci. 116, 187 (1963)), namely that prior injection of complete Freund's Adjuvant suppressed antibody formation to bovine serum albumin. The essential factor in the induction of tolerance to insulin by injection of maleyl insulin in incomplete Freund's Adjuvant is maleyl insulin.

1974 ◽  
Vol 52 (3) ◽  
pp. 196-202 ◽  
Author(s):  
Lee J. Grota ◽  
Gregory M. Brown

Serotonin, N-acetyl serotonin, and 5-methoxy-N-acetyl serotonin (melatonin) were conjugated to bovine serum albumin (BSA) using formaldehyde. The molar ratio of hapten to protein was determined spectrophotometrically. Spectrophotometric data indicated that serotonin and N-acetyl serotonin but not melatonin were conjugated to bovine serum albumin. Selected hapten–protein conjugates were suspended in Freund's adjuvant and injected into rabbits. Antisera were harvested monthly and screened by double immunodiffusion. Immunodiffusion and inhibition tests indicated that antibodies raised to serotonin–BSA reacted with serotonin and 5-methoxytryptamine but failed to cross react with N-acetyl serotonin or melatonin. Inhibition tests indicated that antibodies to N-acetyl serotonin – BSA reacted with N-acetyl serotonin and cross reacted with melatonin but not with serotonin or 5-methoxytryptamine.


1963 ◽  
Vol 117 (6) ◽  
pp. 1035-1051 ◽  
Author(s):  
Maria C. Michaelides ◽  
Albert H. Coons

Rabbits were injected into the hind foot with diphtheria toxoid and bovine serum albumin. Fragments of popliteal lymph node taken from them several months later were placed in plasma-clot cultures with Eagle's medium. When antigen was added to the culture fluid, anamnestic antibody responses occurred regularly. When the antigen was diphtheria, responsiveness remained for 4 days after the beginning of the culture. When it was bovine serum albumin, responsiveness lasted for about 8 days. Once an anamnestic response had begun, antibody formation continued for 4 weeks or more. High concentrations of bovine serum albumin (0.5 mg/ml) did not inhibit the response. When both antigens were used to stimulate the same culture, it was found that the two responses were independent.


1970 ◽  
Vol 132 (1) ◽  
pp. 66-76 ◽  
Author(s):  
D. C. Benjamin ◽  
W. O. Weigle

Rabbits made unresponsive to BSA at birth were given two courses of immunization with various cross-reacting albumins at 3 months of age. Normal control rabbits, of equivalent age and weight, were similarly immunized. Sera obtained 7 days after the last injection were assayed for binding and precipitating antibody to six albumins and for their avidity for BSA. No significant differences were found between unresponsive and normal rabbits in the amount of antibody reacting with any of the six albumins used. This was the case regardless which albumin was used to terminate the unresponsive state. Avidity differences were seen and seemed to depend on the antigen used and not on the immunological status of the animal. The simultaneous injection of small amounts of BSA inhibited the termination of unresponsiveness. These results were discussed in the light of the more recent theories of the termination of unresponsiveness and of antibody formation.


1968 ◽  
Vol 128 (4) ◽  
pp. 715-728 ◽  
Author(s):  
Vera S. Byers ◽  
Eli E. Sercarz

A concentration of 5 mg/ml bovine serum albumin (BSA) prevents the in vitro elicitation of a secondary response in primed rabbit popliteal lymph nodes, if it is left in contact with the node fragments for the first 6 days of culture. No antibody formation can be detected at any time during the culture period in most cases, although occasional fragments are resistant to inhibition. Reducing the exposure time to the first 3 days of culture delays the peak of the antibody response. The inhibition is antigen specific. Reconstruction experiments demonstrate that the inhibition is not due to antigen masking of the antibody. Even shortly after optimal stimulation, the addition of 5 mg/ml BSA to the fragments was not able to prevent a normal antibody response. The implications of these findings are that (a) a high antigen concentration suspends the memory cell in a reversibly paralyzed state, (b) memory cells have a heterogeneous susceptibility to inhibition, (c) once induced, the antibody response cannot be inhibited by antigen overloading, (d) unresponsiveness in a primed animal can be due to either exhaustion of the memory cell population or paralysis of the memory cell.


2020 ◽  
Author(s):  
Ningchao Du ◽  
Yang Li ◽  
Min Xu ◽  
Changlin Wu ◽  
Xiaolan Chen ◽  
...  

Collagen-induced arthritis (CIA) mouse model is currently the most widely used and reliable autoimmune model to study rheumatoid arthritis. In this model, we used bovine type II collagen (CII) and complete Freund’s adjuvant (CFA) or incomplete Freund’s adjuvant (IFA) to form emulsifier, and mice were injected intradermal to induce autoimmune arthritis (CIA). In this model, we ground bovine collagen type II (CII) with complete Freund’s adjuvant (CFA) or incomplete Freund’s adjuvant (IFA) to form emulsifiers, and intradermal injection in mice induced autoimmune arthritis (CIA). This article describes the mouse, CFA strains, key emulsification, anesthesia, and injection immune techniques, as well as the incidence, date of onset, score, pathological results of arthritis. The total time for preparation of reagents and immunization of 20 mice was about 2-2.5 hours. In this protocol, we induced a high incidence of CIA with DBA/1J in genetically susceptible mice and assessed the severity and pathology of the disease, at the same time we found that CII also can induced enteritis, including ileitis and colitis. The initial symptoms of arthritis appear in the 24-26 days of the experiment, that is, 3-5 days after the second immunization, the peak period of inflammation was 30-36 days, the arthritis incidence about 90-100%, at the same time, the incidence of enteritis and arthritis were the same, small intestinal inflammation was more severe, but the duration was short; while the colonic inflammation was mild, and the duration was longer than enteritis, we named it collagen induced inflammations (CIIs).


1971 ◽  
Vol 134 (4) ◽  
pp. 1046-1061 ◽  
Author(s):  
Dov Theo Golan ◽  
Yves Borel

Treatment of adult mice with dinitrophenyl (DNP) bound to isogeneic serum resulted in a specific inability to respond to DNP after challenge with DNP-keyhole limpet hemocyanin (KLH) in complete Freund's adjuvant. The unresponsiveness to the hapten had all the characteristics of immunologic tolerance: it had a definite induction time; it was transient but could be maintained by additional injections of the tolerogen; it was antigen specific and dose dependent. In addition, the induction of tolerance to DNP is dependent on the nature of the carrier. Two main conclusions can be drawn from these data: DNP conjugates of three homopolymers of lysine were found to be nonimmunogenic in mice, yet tolerogenic. Thus, antigenicity is not necessary to induce tolerance. Among the various carriers tested, isogeneic 7S immunoglobulin (IgG) was found to be the most effective to induce and maintain tolerance to the hapten. This suggests that IgG may have a function other than its usual role as an immunoglobulin.


Author(s):  
G. D. Gagne ◽  
M. F. Miller

We recently described an artificial substrate system which could be used to optimize labeling parameters in EM immunocytochemistry (ICC). The system utilizes blocks of glutaraldehyde polymerized bovine serum albumin (BSA) into which an antigen is incorporated by a soaking procedure. The resulting antigen impregnated blocks can then be fixed and embedded as if they are pieces of tissue and the effects of fixation, embedding and other parameters on the ability of incorporated antigen to be immunocyto-chemically labeled can then be assessed. In developing this system further, we discovered that the BSA substrate can also be dried and then sectioned for immunolabeling with or without prior chemical fixation and without exposing the antigen to embedding reagents. The effects of fixation and embedding protocols can thus be evaluated separately.


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