IMMUNOCHEMICAL STUDY OF ANTIGENIC SPECIFICITY IN DELAYED HYPERSENSITIVITY

Delayed hypersensitivity in guinea pigs was produced by immunization. with a conjugate prepared by coupling diazotized arsanilic acid to polytyrosine. The resulting sensitivity could be demonstrated by skin test with conjugates prepared from a wide variety of tyrosine-containing proteins. Definite but smaller degrees of sensitivity could be induced with conjugates of proteins containing little or no tyrosine. The apparent absence of carrier-specificity is considered to be due to the narrowed range of immunologic response produced by immunization with polytyrosine-azobenzenearsonate. Injections of the hapten N-acetyltyrosine-azobenzenearsonate was found to suppress completely the delayed reaction attributable to the tyrosine-azobenzenearsonate group. The same hapten was only slightly effective in suppressing reactions in guinea pigs immunized with guinea pig serum albumin-azobenzenearsonate, suggesting that a broader range of specificities is involved with such antigens. Confirmation of such increased range of specificity attributable to antigenic determinants contributed by the carrier protein was obtained by desensitization studies with N-acetyltyrosine-azobenzenearsonate and guinea pig serum albumin-azobenzoate. While separately these materials produced only a slight decrease in skin reactivity to guinea pig serum albumin-azobenzenearsonate, the combination was found to give almost complete suppression.

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IMMUNOCHEMICAL STUDY OF ANTIGENIC SPECIFICITY IN DELAYED HYPERSENSITIVITY

Journal of Experimental Medicine ◽

10.1084/jem.123.2.229 ◽

1966 ◽

Vol 123 (2) ◽

pp. 229-237 ◽

Cited By ~ 78

Author(s):

Sidney Leskowitz ◽

Valerie E. Jones ◽

Soloman J. Zak

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Delayed Hypersensitivity ◽

Antigenic Specificity ◽

Antigenic Determinants ◽

Immunochemical Study ◽

Aromatic Molecules ◽

Arsanilic Acid ◽

Different Types ◽

Amino Acid Polymers

Hapten-specific delayed hypersensitivity was produced by immunization of guinea pigs with arsanilic acid conjugated to N-acetyltyrosine or other small aromatic molecules. Such hapten-specific delayed sensitivity could be passively transferred by peritoneal exudate cells. While a conjugate made from a polymer of D-amino acids was ineffective in producing sensitization, the conjugate made with D-tyrosine was effective, suggesting that the inability of D-amino acid polymers to be broken down by enzymes might be bypassed by use of the monomer. The effectiveness of such monomers in producing delayed sensitivity, but not antibody production, is consistent with a hypothesis that different types of antigenic determinants are involved in the production of each.

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IMMUNOCHEMICAL STUDY OF ANTIGENIC SPECIFICITY IN DELAYED HYPERSENSITIVITY

Journal of Experimental Medicine ◽

10.1084/jem.122.3.505 ◽

1965 ◽

Vol 122 (3) ◽

pp. 505-515 ◽

Cited By ~ 8

Author(s):

Valerie E. Jones ◽

Sidney Leskowitz

Keyword(s):

Guinea Pigs ◽

Subsequent Development ◽

Delayed Hypersensitivity ◽

Antigenic Specificity ◽

Target Cells ◽

Immunochemical Study ◽

Arsanilic Acid ◽

Specific Tolerance

Injections of various conjugates of arsanilic acid into newborn guinea pigs produced a specific tolerance in respect to subsequent development of hapten-specific delayed hypersensitivity. In general, larger polyvalent conjugates produced longer lasting and more profound suppression of delayed sensitivity than did the smaller ones. Carrier injections alone were ineffective. At lower doses of conjugate, breakthrough of tolerance occurred first with animals immunized with the heterologous carrier conjugate. The duration of tolerance produced by injection of monovalent conjugates into neonates is in contrast to the transient inhibition produced by the same conjugates in previously sensitized animals, suggesting that different target cells may be involved in these two phenomena.

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PROPERTIES OF GUINEA PIG 7S ANTIBODIES

Journal of Experimental Medicine ◽

10.1084/jem.117.6.937 ◽

1963 ◽

Vol 117 (6) ◽

pp. 937-949 ◽

Cited By ~ 133

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.

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EVIDENCE THAT THE L-ASPARAGINASE OF GUINEA PIG SERUM IS RESPONSIBLE FOR ITS ANTILYMPHOMA EFFECTS

Journal of Experimental Medicine ◽

10.1084/jem.118.1.99 ◽

1963 ◽

Vol 118 (1) ◽

pp. 99-120 ◽

Cited By ~ 177

Author(s):

J. D. Broome

Keyword(s):

Guinea Pig ◽

Guinea Pigs ◽

Serum Proteins ◽

Deae Cellulose ◽

Salt Precipitation ◽

Asparaginase Activity ◽

Pig Serum

A number of the properties of the L-asparaginase present in guinea pig serum have been examined and shown to be indistinguishable from those of the agent responsible for inhibiting cells of lymphoma 6C3HED in vivo. The patterns of instability of the enzyme to changes in temperature and pH were found to parallel closely those of the antilymphoma agent. L-Asparaginase activity was essentially absent from the serum of newborn guinea pigs and this failed to inhibit 6C3HED cells. On separating guinea pig serum proteins by salt precipitation, electrophoresis, and chromatography on DEAE cellulose, antilymphoma activity was found only in fractions which contained L-asparaginase.

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EVIDENCE FOR SPECIES' DIFFERENCES IN THE EFFECT OF SERUM γ-GLOBULIN CONCENTRATION ON γ-GLOBULIN CATABOLISM

Journal of Experimental Medicine ◽

10.1084/jem.120.5.967 ◽

1964 ◽

Vol 120 (5) ◽

pp. 967-986 ◽

Cited By ~ 13

Author(s):

Stewart Sell

Keyword(s):

Human Serum Albumin ◽

Guinea Pig ◽

Serum Albumin ◽

Human Serum ◽

Guinea Pigs ◽

Serum Proteins ◽

Elevated Serum ◽

Keyhole Limpet Hemocyanin ◽

Normal Numbers ◽

Γ Globulins

The fractional rates of catabolism of isotopically labeled mouse, human, bovine, and guinea pig γ-globulins and human serum albumin were determined in mice and in guinea pigs whose serum γ-globulin and serum albumin levels were elevated by immunization or by injections of exogenous serum proteins. These serum proteins were also followed in mice with different serum γ-globulin levels due to different bacterial environments. The fractional rates of catabolism of the labeled γ-globulins from all species tested were markedly increased in mice with elevated γ-globulins due to immunization; to injections of human, mouse, guinea pig, or rabbit γ-globulins; to exposure to supra normal numbers of bacteria in the environment. Injections of bovine γ-globulin were only partially effective, and injections of human serum albumin had no effect. The γ-globulin catabolic rates were decreased in mice with subnormal serum γ-globulin levels (germfree mice). The catabolic rate of human serum albumin was essentially the same in all mice in spite of differences in serum γ-globulin levels. In contrast, elevation of the serum γ-globulin levels by injections of exogenous γ-globulins or by hyperimmunization with keyhole limpet hemocyanin produced no change in the fractional catabolic rates of the isotopically labeled γ-globulins and labeled albumin in guinea pigs. Thus, a feedback mechanism for the control of the serum γ-globulin concentration appears to be operative in the mouse, but not in the guinea pig. Guinea pigs immunized with antigens in complete Freund's adjuvant or a saline suspension of killed E. coli had an increase in the catabolic rates of all labeled proteins tested including human serum albumin. Evidence is presented that the mechanism of this increase in catabolism is not the same as that seen in mice with elevated serum γ-globulin levels.

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DELAYED HYPERSENSITIVITY TO HAPTEN-PROTEIN CONJUGATES

Journal of Experimental Medicine ◽

10.1084/jem.115.5.1037 ◽

1962 ◽

Vol 115 (5) ◽

pp. 1037-1051 ◽

Cited By ~ 40

Author(s):

P. G. H. Cell ◽

Arthur M. Silverstein

Keyword(s):

Guinea Pig ◽

Antigenic Determinant ◽

Gamma Globulin ◽

Delayed Hypersensitivity ◽

Antigenic Determinants ◽

Carrier Protein ◽

Protein Conjugates ◽

Circulating Antibodies ◽

Human Gamma Globulin ◽

Delayed Hypersensitivity Reaction

Further data have been presented showing that the specificity of the delayed hypersensitivity reaction in the guinea pig to hapten-protein conjugates involves to a considerable degree a contribution by the protein carrier. The carrier contribution is such that sensitization to guinea pig albumin-m-azobenzenesulfonate, for example, does not result in cross-reaction with conjugates of the same hapten with unrelated proteins such as ovalbumin or human gamma globulin, nor were cross-reactions observed between conjugates prepared with the same hapten, coupled to the same protein, but by two different chemical routes, such that the point of attachment of the hapten to the protein differed. It thus appears that in this system both hapten and carrier protein are necessary, but that neither alone is in general sufficient to stimulate the delayed sensitive cell. Desensitization experiments with cross-reacting hapten-protein conjugates have suggested the presence of a multiplicity of antigenic determinants participating in the elicitation of the delayed lesion, and of a concomitant development of a heterogeneity of specificities in the population of delayed sensitive cells in the sensitized animal. The data are discussed in terms of the apparent requirement of the delayed sensitivity mechanism for a larger functional antigenic determinant than that required for interaction with circulating antibodies. Some possible explanations for this difference, and some of its consequences, are discussed.

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FURTHER STUDIES ON TYPHUS FEVER

Journal of Experimental Medicine ◽

10.1084/jem.64.5.673 ◽

1936 ◽

Vol 64 (5) ◽

pp. 673-687 ◽

Cited By ~ 3

Author(s):

Hans Zinsser ◽

Attilio Macchiavello

Keyword(s):

Tissue Culture ◽

Guinea Pig ◽

Guinea Pigs ◽

Horse Serum ◽

Tissue Cultures ◽

Practical Application ◽

Typhus Fever ◽

Culture Virus ◽

Pig Serum

1. Guinea pigs can be actively immunized against European typhus fever with homologous formalinized Rickettsia tissue cultures, provided sufficient amounts are injected. The method is suggested for practical application in man. 2. Serovaccination against European typhus fever can be successfully applied to guinea pigs by a variety of methods, the simplest of which consists of the injection of mixtures of virulent defibrinated guinea pig blood and convalescent guinea pig serum taken from 3 to 5 days after defervescence. Similar results can be obtained with mixtures in which tissue culture virus, either with convalescent guinea pig serum or with antimurine horse serum, is used. There is no indication so far that such animals become carriers. Possible application of these methods to typhus epidemics is discussed.

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STUDIES ON HYPERSENSITIVITY

Journal of Experimental Medicine ◽

10.1084/jem.113.3.571 ◽

1961 ◽

Vol 113 (3) ◽

pp. 571-585 ◽

Cited By ~ 79

Author(s):

P. G. H. Gell ◽

B. Benacerraf

Keyword(s):

Guinea Pig ◽

Delayed Hypersensitivity ◽

Carrier Protein ◽

Contact Sensitivity ◽

Delayed Reaction ◽

Immunological Mechanism ◽

Order Of Magnitude

In earlier observations with the picryl system, it was concluded that contact sensitivity was a form of delayed (cellular) hypersensitivity to conjugates of sensitizer with autologous proteins indistinguishable in its immunological mechanism from other classical forms of delayed hypersensitivity to proteins. This conclusion has been confirmed and extended with the picryl and chlorbenzoyl chloride systems. 1. It is shown that to induce a state of contact sensitivity, the minimal necessary amounts of hapten are of the same order of magnitude, whether this hapten is conjugated with protein or the free reactive chemical itself. From this, it is evident that contamination of conjugates with small amounts of unreacted sensitizer plays no part in the induction of contact reactivity by the conjugate. With the dinitrophenyl system, no contact sensitivity could be induced by the conjugates used; possible reasons for this discrepancy are discussed. 2. Animals sensitized to contact by homologous conjugate can be completely desensitized by injections of such a conjugate in large amount; a similar injection schedule has no effect on the contact sensitivity of animals sensitized with the free reactive sensitizer. 3. The capacity of heterologous (ovalbumin) conjugates to evoke anti-hapten antibodies is shown to be greater than that of homologous (guinea pig seralbumin) conjugates: the reverse is true of their capacity to induce delayed reactivity. 4. Evidence is brought forward to suggest that in animals sensitized with homologous albumin conjugates, the specificity of the delayed reaction involves more than the hapten alone, even though the carrier protein is non-antigenic on its own. The contrast with the apparent lesser specificity of the antibodies later produced is discussed.

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STUDIES ON DELAYED HYPERSENSITIVITY

Journal of Experimental Medicine ◽

10.1084/jem.121.6.873 ◽

1965 ◽

Vol 121 (6) ◽

pp. 873-888 ◽

Cited By ~ 18

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.

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