THE ANTIGENIC LOCI OF INSULIN

1967 ◽  
Vol 45 (7) ◽  
pp. 1135-1144 ◽  
Author(s):  
S. Wilson ◽  
M. A. Aprile ◽  
L. Sasaki
Keyword(s):  
Guinea Pig ◽  
Anaphylactic Reaction ◽  
Guinea Pigs ◽  
Synthetic Peptides ◽  
Cross Reactivity ◽  
Antigenic Determinants ◽  
Diabetic Patients ◽  
Insulin Resistant ◽  
A Chain ◽  
Passive Cutaneous Anaphylaxis Test

Insulin exhibited some cross-reactivity with the isolated S-sulfo A- and B-chains in the passive cutaneous anaphylaxis test. The synthetic peptides A1–9, A10–21, and A1–21 gave a passive cutaneous anaphylactic reaction with guinea pig antiserum to natural A-chain, whereas high levels of A10–21 and A1–21 could also inhibit a subsequent insulin challenge from reacting with antiserum to insulin. The synthetic peptides B1–8, B9–30, B24–30, and B1–30 reacted with antiserum to natural B-chain but high levels of peptides (B2–8)2, B9–14, and (B17–23)2 were inactive. The symmetrical double peptide (B1–8)2 reacted with guinea pig antiserum to intact insulin as well as with antiserum to B-chain and certain sera from insulin-resistant diabetic patients. The results indicated that injection of ox insulin into guinea pigs caused production of anaphylactic antibodies mainly towards antigenic loci in the regions A10–21 and B1–8 of the insulin molecule, whereas antigenic determinants in the regions A1–9 and B24–30 were less important.

scholarly journals Immune response gene control of determinant selection. I. Intramolecular mapping of the immunogenic sites on insulin recognized by guinea pig T and B cells.

1977 ◽  
Vol 145 (3) ◽  
pp. 726-742 ◽  
Author(s):  
M A Barcinski ◽  
A S Rosenthal
Keyword(s):  
T Cell ◽  
Guinea Pig ◽  
Guinea Pigs ◽  
Cell Function ◽  
Strain Differences ◽  
Amino Acid Sequences ◽  
Immune Response Gene ◽  
Antigenic Determinants ◽  
Histocompatibility Complex ◽  
A Chain

T-cell DNA synthesis and T-helper cell function in response to isolated insulin chains and naturally occurring insulin variants was assessed in insulin immune guinea pigs. Two distinct antigenic determinants, recognized by T cells, were defined. One localized in the B chain and the other one constituted by amino acids A8, A9, and A10 of the insulin A-chain loop. Recognition of the B-chain determinant is under the control of Ir genes linked to the strain 13 major histocompatibility complex. This was shown by studying the response to isolated insulin B chain in F1(2 x 13) guinea pigs, as well as serologically defined backcrosses and outbred animals. Insulin recognition through the A-chain loop determinant is specific for strain 2 guinea pigs. These animals recognize this region of the molecule even when displaying different amino acid sequences. The strain differences observed in those antigenic sites eliciting T-cell recognition was not found at an antibody level. No differences could be detected in the ability of the different insulin variants to inhibit the binding of 125I-labeled pork insulin to strain 2 guinea pig anti-pork insulin or to strain 13 guinea pig anti-pork insulin.

Interaction of a Plasmin A-Chain/t-PA B-Chain Hybrid Enzyme with Plasma Inhibitors In Vivo and In Vitro

1990 ◽  
Vol 63 (03) ◽  
pp. 459-463 ◽  
Author(s):  
S Wilson ◽  
P Chamberlain ◽  
I Dodd ◽  
A Esmail ◽  
J H Robinson
Keyword(s):  
Guinea Pig ◽  
Plasminogen Activator ◽  
Active Site ◽  
Guinea Pigs ◽  
Fibrinolytic Activity ◽  
Pharmacokinetic Profile ◽  
Inhibitory Mechanisms ◽  
A Chain

SummaryA hybrid plasminogen activator consisting of the “A” chain of plasmin linked to the “B” chain of rt-PA was inhibited in vitro in human and guinea pig plasmas 4 to 5-fold more rapidly than its parent activator, two-chain t-PA. Using zymographic and autoradiographic techniques together with the use of immunodepleted plasma the major inhibitor was identified as aIpha-2-antiplasmin. The pharmacokinetic profile of the hybrid in guinea pigs was determined by two different methods: disappearance of fibrinolytic activity and removal of radiolabelled hybrid from the circulation. Fibrinolytic activity was cleared rapidly via inhibitory mechanisms, whilst radiolabelled material was cleared considerably more slowly due to the formation of hybrid-inhibitor complexes. When the active site of the hybrid was reversibly acylated inhibitory mechanisms were evaded and a prolonged pharmacokinetic profile of activity was observed.

scholarly journals STUDIES ON DELAYED HYPERSENSITIVITY

1965 ◽  
Vol 121 (6) ◽  
pp. 873-888 ◽  
Author(s):  
Bernard B. Levine
Keyword(s):  
Guinea Pig ◽  
Guinea Pigs ◽  
Antigenic Determinant ◽  
Cross Reactivity ◽  
Delayed Hypersensitivity ◽  
Hypersensitivity Reactions ◽  
Binding Affinities ◽  
Structural Adaptation ◽  
Skin Reactions ◽  
High Binding

Experiments carried out with several well defined antigenic systems (hapten conjugates of poly-L-lysine and guinea pig serum albumin) in guinea pigs demonstrated that: 1. Arthus reactions also manifest carrier specificity, although to a smaller extent than do delayed hypersensitivity reactions. 2. Desensitization by injection of minute doses of antigen results in moderate specific desensitization of delayed hypersensitivity without desensitization of Arthus reactivity to the same antigenic determinant. 3. Insoluble antigen-antibody complexes prepared from high affinity guinea pig antibodies can elicit specific delayed skin reactions in sensitized guinea pigs. 4. Homologous conjugates of structurally similar haptens show considerably less cross-reactivity in delayed reactions than in immediate hypersensitivity reactions to the same antigenic determinant. These experimental results are interpreted as indicating that delayed hypersensitivity reactions in the guinea pig are mediated by "antibodies" of comparatively high binding affinities. High binding affinities are achieved for these antibodies more likely by closer structural adaptation between antigen and antibody than by a larger area of specific contact.

scholarly journals Impact of feline zona pellucida glycoprotein B-derived synthetic peptides on in vitro fertilization of cat oocytes

Reproduction ◽  
2004 ◽  
Vol 127 (2) ◽  
pp. 179-186 ◽  
Author(s):  
Jennifer Ringleb ◽  
Marlies Rohleder ◽  
Katarina Jewgenow
Keyword(s):  
Zona Pellucida ◽  
Synthetic Peptides ◽  
Cross Reactivity ◽  
Antigenic Determinants ◽  
Dot Blot ◽  
Oocyte Growth ◽  
Sperm Binding ◽  
Stray Cats

Although immunocontraception based on porcine zona pellucida (ZP) proteins is widely applied in many species, it is not suitable for cat contraception due to the lack of cross-reactivity. Since the first ZP gene expressed during oocyte growth in domestic cats is ZPB, we assumed that immunization with feline ZPB (fZPB)-derived synthetic peptides may cause irreversible infertility, which would be preferable in stray cats. Thus, the present study evaluated the immunogenicity and the contraceptive potential of synthetic fZPB peptides. Antigenic epitope sequences were detected via epitope mapping using specific rabbit anti-fZP antibodies. Six peptides representing the recognized epitopes were synthesized subsequently. Two out of six peptides (ZPB amino acid residue 130–149 = P3 and 175–193 = P6) cross-reacted with anti-fZP antiserum in dot blot analysis and ELISA. Coupled to BSA, both peptides were utilized to produce specific antibodies in rats. Despite several booster injections the antibody titers monitored by ELISA did not exceed 1:5000. Both rat antisera were tested for contraceptive potential in cat in vitro maturation/in vitro fertilization (IVF). Antiserum against peptide P3 significantly inhibited sperm binding and fertilization of cat oocytes in vitro (57.3% of sperm binding; 41.5% of fertilization), whereas the inhibition by anti-P6 was not significant. Pre-incubation of sperm cells with both peptides before IVF failed to affect either sperm binding or fertilization (22.3 ± 3.7 sperm/egg vs 25.5 ± 5.8 for P3 and 20.7 ± 4.0 for P6, respectively). In conclusion, antibodies directed against one of the two identified antigenic determinants of fZPB inhibited sperm binding and IVF and therefore showed promising results as a contraceptive. However, the specific immune response and anti-fertile properties of this synthetic vaccine have to be examined in vivo to verify the suitability of its components.

scholarly journals THE RELATION BETWEEN ANTIANAPHYLAXIS AND ANTIBODY BALANCE

1936 ◽  
Vol 64 (4) ◽  
pp. 641-655 ◽  
Author(s):  
Marion C. Morris
Keyword(s):  
Guinea Pig ◽  
Intravenous Injection ◽  
Anaphylactic Reaction ◽  
Guinea Pigs ◽  
Normal Serum ◽  
Intravenous Inoculation ◽  
Lethal Doses ◽  
Circulating Antibody ◽  
Pig Serum ◽  
Increased Susceptibility

1. Sensitized guinea pigs injected with normal rabbit or guinea pig serum previous to intravenous inoculation of antigen may be protected against a few lethal doses of antigen. The protection is greater with foreign than with homologous serum and appears to be related roughly to the amount of serum introduced. 2. Sensitized guinea pigs injected with antibody-containing serum preliminary to intravenous injection of antigen, show no greater refractoriness to anaphylaxis than do those injected with normal serum. 3. Moreover, in many instances, the injection of an excess of antibody into the circulation of sensitized guinea pigs, leads to an increased susceptibility of these animals to anaphylaxis. 4. These results indicate that an excess of circulating antibody is not responsible for a state of antianaphylaxis, but on the contrary, may contribute toward the anaphylactic reaction itself.

ELISA for Guinea-pig IgG1: An Alternative to the PCA Test for Relative Allergenicity

1991 ◽  
Vol 19 (1) ◽  
pp. 8-14
Author(s):  
Annette Prentø
Keyword(s):  
Guinea Pig ◽  
Guinea Pigs ◽  
Passive Cutaneous Anaphylaxis ◽  
Relative Importance ◽  
Elisa Method ◽  
Passive Cutaneous Anaphylaxis Test

An ELISA method to replace the PCA (passive cutaneous anaphylaxis) test has been developed for use in the evaluation of relative allergenicity in guinea-pigs (Dunkin-Hartley). The method detects antigen-specific IgG1 antibodies formed in the guinea-pig during sensitisation, and correlates well with the corresponding PCA test. The relative importance of IgG1 and IgE for relative allergenicity in the Dunkin-Hartley guinea-pig strain is discussed.

scholarly journals Sensitizing Capacities and Cross-Reactivity Patterns of Some Diisocyanates and Amines Using the Guinea-Pig Maximization Test. Can p-phenylenediamine be Used as a Marker for Diisocyanate Contact Allergy?

2017 ◽  
Vol 11 (1) ◽  
pp. 87-97 ◽  
Author(s):  
Haneen Hamada ◽  
Erik Zimerson ◽  
Magnus Bruze ◽  
Marléne Isaksson ◽  
Malin Engfeldt
Keyword(s):  
Guinea Pig ◽  
Patch Test ◽  
Guinea Pigs ◽  
Contact Allergy ◽  
Cross Reactivity ◽  
The Other ◽  
Positive Patch Test ◽  
Other Hand ◽  
Test Reactions ◽  
Toxic Reactions

Background:Isocyanates are mainly considered respiratory allergens but can also cause contact allergy. Diphenylmethane-4,4′-diamine (4,4′-MDA) has been considered a marker for diphenylmethane-4,4′-diisocyanate (4,4′-MDI) contact allergy. Furthermore, overrepresentation of positive patch-test reactions top-phenylenediamine (PPD) in 4,4′-MDA positive patients have been reported.Objectives:To investigate the sensitizing capacities of toluene-2,4-diisocyanate (2,4-TDI) and PPD and the cross-reactivity of 4,4′-MDA, 2,4-TDI, dicyclohexylmethane-4,4′-diamine (4,4′-DMDA), dicyclohexylmethane-4,4′-diisocyanate (4,4′-DMDI), 4,4′-MDI and PPD.Methods:The Guinea Pig Maximization Test (GPMT) was used.Results:PPD was shown to be a strong sensitizer (p<0.001). Animals sensitized to PPD showed cross-reactivity to 4,4′-MDA (p<0.001). Animals sensitized to 4,4′-MDA did not show cross-reactivity to PPD. 8 animals sensitized to 2,4-TDI were sacrificed due to toxic reactions at the induction site and could thus not be fully evaluated.Conclusion:PPD was shown to be a strong sensitizer. However, it cannot be used as a marker for isocyanate contact allergy. On the other hand, positive reactions to 4,4′-MDA could indicate a PPD allergy. The intradermal induction concentration of 2,4-TDI (0.70% w/v) can induce strong local toxic reactions in guinea-pigs and should be lowered.

scholarly journals PROPERTIES OF GUINEA PIG 7S ANTIBODIES

1963 ◽  
Vol 117 (6) ◽  
pp. 937-949 ◽  
Author(s):  
Baruj Benacerraf ◽  
Zoltan Ovary ◽  
Kurt J. Bloch ◽  
Edward C. Franklin
Keyword(s):  
Guinea Pig ◽  
Guinea Pigs ◽  
Analytical Ultracentrifugation ◽  
Antigenic Determinants ◽  
Protein Antigens ◽  
Intraperitoneal Route ◽  
Electrophoretic Mobilities ◽  
Single Protein ◽  
Pig Serum

Guinea pigs hyperimmunized with single protein antigens or hapten conjugates emulsified in complete adjuvants produced two types of precipitating antibodies with different electrophoretic mobilities. "Slow" migrating antibody generally appeared earlier and "fast" migrating antibody later in the course of immunization. Animals initially immunized by the intraperitoneal route with hapten conjugates without adjuvants produced primarily fast migrating antibody. Purified guinea pig antibodies were also separable into slow and fast migrating components by electrophoresis in supporting media. Using suitable antisera prepared in rabbits hyperimmunized with guinea pig serum, it was demonstrated that slow and fast antibodies have both common and distinct antigenic determinants. Analytical ultracentrifugation disclosed that both antibodies have sedimentation coefficients of approximately 7S. These antibodies have been designated guinea pig 7Sγ1 and 7Sγ2.

scholarly journals INSULIN ANTIBODY VARIATIONS IN RABBITS AND GUINEA PIGS AND MULTIPLE ANTIGENIC DETERMINANTS ON INSULIN

1963 ◽  
Vol 118 (1) ◽  
pp. 55-71 ◽  
Author(s):  
Edward R. Arquilla ◽  
Jack Finn
Keyword(s):  
Guinea Pig ◽  
Antibody Production ◽  
Guinea Pigs ◽  
Genetic Factors ◽  
Insulin Antibodies ◽  
Antigenic Determinants ◽  
Antibody Molecule ◽  
Insulin Molecule ◽  
The Individual ◽  
The Relationship

1. A method is presented for measuring the degree to which insulin antibodies in one antiserum react with an insoluble insulin complex saturated with antibodies from a different antiserum. 2. Many rabbits produce antibodies which bind to portions of the insulin molecule to which antibodies from guinea pigs or other rabbits cannot bind. 3. Occasional guinea pigs produce antibodies which bind to portions of the insulin molecule to which antibodies from rabbits or other guinea pigs cannot bind. 4. Studies with labeled antisera and repeated incubations of test antisera with antibody insulin complexes demonstrate the individual antibody variations to be due to antibodies directed to different determinants and not to dissociation of antibodies from the same determinant on the insulin molecule. 5. More than one antibody molecule can simultaneously bind to an insulin molecule. 6. Insulin has a multiplicity of antigenic determinants. 7. The relationship between antigenic determinants, insulin antibodies, and neutralization of insulin by antisera is discussed. 8. The determinants to which insulin antibodies are directed appear to be characteristic for the individual rabbit or guinea pig immunized. It is postulated therefore that genetic factors direct antibody production toward specific determinants when insulin is the antigen.

The concept and operational definition of protein epitopes

1989 ◽  
Vol 323 (1217) ◽  
pp. 451-466 ◽  
Keyword(s):  
Synthetic Peptides ◽  
Foot And Mouth Disease ◽  
Operational Definition ◽  
Cross Reactivity ◽  
Antigenic Specificity ◽  
Antigenic Determinants ◽  
Immune Recognition ◽  
Segmental Mobility ◽  
X Ray Crystallography ◽  
Relative Contribution

The antigenic determinants or epitopes of a protein correspond to those parts of the molecule that are specifically recognized by the binding sites or paratopes of certain immunoglobulin molecules. Epitopes are thus relational entities that require complementary paratopes for their operational recognition. Some authors consider that the concept of epitope necessarily involves the two properties of antigenic reactivity (ability to bind to a paratope) and immunogenicity (ability to induce an immune response). Such a view creates difficulties because it makes the existence of epitopes in a protein depend on immunogenetic and regulatory mechanisms of the immunized host. The delineation of epitopes can be achieved by antigenic cross-reactivity studies or by X-ray crystallography. Both approaches require specific criteria for deciding which residues of the antigen are in contact with the paratope and are functionally part of the epitope. The relative contribution of static accessibility, segmental mobility and induced fit to immune recognition remains controversial. Each of the methods used for analysing antigenic specificity is subject to various operational constraints originating from the type of experimental probe and from the form at, sensitivity and specificity of the immunoassay used. If a protein is assumed to contain as many epitopes as the number of different monoclonal antibodies that can be raised against it, the delineation of epitopes corresponds to the summation in various hosts of the immune repertoire specific for the antigen. Neutralization epitopes are a special subclass of the epitopes of infectious agents and toxins that are specifically recognized by antibody molecules able to neutralize the biological activity of the antigen. The identification of neutralization epitopes is important for the development of synthetic vaccines because it is this type of epitope that should be mimicked by synthesis and used as a vaccine for eliciting protective immunity. The first demonstration that synthetic peptides could elicit antibodies that neutralized viral infectivity was made by Anderer and his colleagues in the 1960s in their work with tobacco mosaic virus. Nearly 20 years passed before it was shown that antibodies to synthetic peptides were also able to neutralize the infectivity of other viruses such as foot-and-mouth disease, polio and hepatitis B viruses.

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