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Biomedical subjects

H N EISEN

Publications and source records attributed to H N EISEN.

At least 37 records · Page 2Linked to original sources

Specific inhibition of wheal-and-erythema responses with univalent haptens and univalent antibody fragments.

Wheal-and-erythema responses were studied in normal human volunteers and in a single human subject who is sensitive to the 2,4-dinitrophenyl group. In the normal subjects, reactive skin sites were established by intradermal injection of purified rabbit antibody specific for the 2,4-dinitrophenyl group. In both the active and passively sensitized subjects, wheal-and-erythema was elicited by intradermal injection of a 2,4-dinitrophenyl protein, but not by injection of the same conjugate mixed with certain low molecular weight 2,4-dinitrophenyl haptens or with univalent fragments split by papain from anti-2,4-dinitrophenyl antibody. The latter fragments, unlike intact, bivalent, antibody, do not sensitize normal human skin sites. From these and other observations it is concluded that the wheal-and-erythema response in human skin requires mutually multivalent antigen and antibody. This requirement suggests that multimolecular complexes, containing at least 2 antigen and 2 antibody molecules, are essential in the pathogenesis of this allergic response.

Animals↗

Some immunochemical properties of penicillenic acid. An antigenic determinant derived from penicillin.

Under mild conditions, benzylpenicillin in aqueous solution rearranges to benzylpenicillenic acid, which reacts, under suitable conditions, to form stable disulfides and mercaptides. Through reaction with native proteins or especially with proteins that are enriched in SH groups ("thiolated"), penicillenic acid forms, ma mixed disulfide linkages, stable highly substituted conjugates. These conjugates are potent antigens, inducing the formation of substantial amounts of antibodies specific for the penicillenic acid structure. The implications of these findings for penicillin hypersensitivity in man are discussed.

Antibodies↗

A study of the distribution of 2,4-dinitrobenzene sensitizers between isolated lymph node cells and extracellular medium in relation to induction of contact skin sensitivity.

Although induction of contact skin sensitivity by low molecular weight 2,4-dinitrobenzenes requires the formation in vivo of 2,4-dinitrophenyl-proteins, analogous protein conjugates prepared in vitro are unable to induce this hypersensitive state. Low molecular weight 2,4-dinitrobenzenes are concentrated by isolated lymph node cells, but a representative 2,4-dinitrophenyl-protein conjugate (2,4-dinitrophenyl-bovine serum albumin) was not taken up to a detectable extent by these cells. It is inferred that there exist large quantitative differences in the extent to which dinitrophenyl-proteins are localized within cells following the administration to an intact animal of (a) those simple dinitrobenzenes which are both concentrated by lymph node cells and have the capacity to form protein conjugates in vivo, and (b) 2,4-dinitrophenyl-protein conjugates prepared in vitro. It is suggested that this difference could account for the fact that a varietyof 2,4-dinitrophenyl-proteins prepared in vitro are unable to induce contact skin sensitivity.

Animals↗

The effect of antigenic stimulation on incorporation of phosphate and methionine into proteins of isolated lymph node cells.

Isolated lymph node cells incorporate inorganic orthophosphate into a protein fraction. The phosphorylated product is a phosphoprotein. The rate of phosphate incorporation into phosphoprotein was determined in cells isolated from regional lymph nodes at varying times after antigen injection. The rate was unaltered on the 3rd day, but was enhanced on the 4th day after injection. Parallel results were obtained with L-methionine incorporation into the same gross protein fraction. Possible relationships between antibody formation and the observed enhancement in phosphate incorporation into phosphoprotein are discussed.

Antibody Formation↗

Elicitation of allergic contact dermatitis in the guinea pig; the distribution of bound dinitrobenzene groups within the skin and quantitative determination of the extent of combination of 2,4-dinitro-chlorobenzene with epidermal protein in vivo.

When one or two drops of a dilute, non-irritating solution of 2,4-dinitrochlorobenzene (DNCB) is applied to a small area of skin of the intact guinea pig, about 20 per cent of the applied material, or some derivative of it, is soon excreted in urine. In normal, as well as in specifically sensitized guinea pigs, DNCB at the site of local application becomes rapidly bound to skin protein through primary chemical bonds. Twenty-four hours after application roughly half of the material present at the local skin site is still extractable with organic solvents. Of the non-extractable dinitrophenyl groups, about 99 per cent are in epidermis, and about 85 per cent are substituted in epsilon-NH(2) groups of lysine residues. Only traces of bound dinitrophenyl groups were observed in the corium. It is uncertain whether these are formed in situ, or are experimental contaminants, or are migratory epidermally formed conjugates. Even when DNCB is injected intradermally it combines predominantly with overlying epidermis and with epidermal components of hair follicles, but only slightly with corium. The 2,4-dinitrophenyl conjugates which are localized in the deeper, viable half of the epidermis, close to the epidermal-dermal junction, are inferred to be the agents responsible for specifically evoking the allergic response in sensitized animals. Conjugates which are situated in the outer, cornified half of the epidermis are shown to be incapable of eliciting the allergic response. The results furnish a basis for interpreting a common pattern of lesions in allergic contact dermatitis as it occurs spontaneously in man.

Animals↗

Studies of hypersensitivity to low molecular weight substances. II. Reactions of some allergenic substituted dinitrobenzenes with cysteine or cystine of skin proteins.

2,4-dinitrophenylsulfenyl chloride (DSCl) and 2,4-dinitrophenylthiocyanate (DSCN) elicited allergic reactions of the delayed type when applied to the skin of guinea pigs and of human beings who had been sensitized by prior exposure to 2,4-dinitrofluorobenzene (DF). DSCl and DSCN, together with 2,4-dinitrobenzene sulfonate (DSO(3)), constitute a clearly defined group of allergenic dinitrophenyl compounds in that they all combined with skin protein in vivo through reaction with cysteine or cystine. In vitro, these compounds combine with free SH groups, and with -S-S- groups of hair and epidermis, but not with -S-S- groups of oxidized glutathione or of bovine gamma globulins. DSO(3), DSCl, and DSCN did not react with amino groups in vivo, but did react with protein amino groups in vitro at pH values of about 10. Another group of dinitrophenyl compounds (DF, DCl, and DBr) previously had been shown to combine with lysine epsilon-NH(2) groups of epidermal proteins. In the present work it was found that these compounds do not react with the disulfide groups of these proteins, either in vivo or in vitro. Moreover, they did not seem to react with SH groups of viable skin, although they are highly reactive with sulfhydryl in vitro. This apparent discrepancy between reactivity with SH groups in vitro and in vivo may be due to the fact that the chromatographic technique employed was relatively insensitive for the sulfhydryl derivative. When a compound of either group was applied to the skin surface, dinitrophenyl-amino acids were recovered from the epidermis but not from the dermis. The results are discussed from the viewpoint of the epidermal localization of dinitrophenyl-protein conjugates.

Allergens↗

Elicitation of delayed allergic skin reactions with haptens; the dependence of elicitation on hapten combination with protein.

Eight 2,4-dinitrophenyl compounds with a high degree of configurational uniformity were examined in regard to their ability to elicit delayed allergic skin reactions in guinea pigs and men who had been sensitized by previous exposure to 2,4-dinitrofluorobenzene. Four of the eight compounds produced reactions (elicitors) and four others did not (non-elicitors). It was found that the elicitors combined with protein in vitro and in vivo. The non-elicitors, however, failed to combine in either situation. It is concluded that a necessary condition for the elicitation of delayed allergic skin reactions by haptens is the combination of the latter with skin protein through the formation of bonds of the covalent type. No choice is yet possible amongst the several possible explanations which are advanced to account for the obligatory character of protein combination.

Animals↗