The Induction of Delayed Sensitivity to 2.4-Dinitrophenyl Conjugates in Guinea Pigs Sensitized with DNCB

Abstract

Delayed reaction to DNP conjugate prepared in vitro from the extract of homologous epidermis could be clearly detected on intradermal injection in guinea pigs sensitized by epicutaneous application of DNCB and intradermal injection of Freund’s complete adjuvant to the previously applied area, but not to DNP guinea pig serum. The animals developed the most intense delayed reaction to DNP conjugate prepared from the subcellular small granule fraction of homologous epidermis. There was significant difference in the reactivity of this conjugate and DNP-soluble fraction of the epidermis. The animals also showed positive reactions to DNP conjugates prepared from the extracts of homologous heart, liver, kidney and heterologous (human) epidermis. This finding is explainable by cross-reactivity which exists between these conjugates and DNCB conjugated in vivo to autologous epidermis.

Isolation of Dinitrophenyl-Skin Protein Conjugates by Immunoadsorbent Chromatography

“…The data presented in this paper support and extend previous observations that solu ble DNP-epidermal protein conjugates play an essential role in the development and eli citation of contact sensitivity [16][17][18][19][20][21]. Such conjugates are capable of eliciting specific lymphocyte blastogenesis and lymphokine production in vitro.…”

Delayed Hypersensitivity to Hapten-Skin Protein Conjugates in Guinea Pigs Sensitized to Benzo(a)pyrene

“…The im portance of carrier proteins in the skin is well known [11][12][13][14][15][16] and was demonstrated in the present study by the inability of guinea pig albumin to satisfy the carrier re quirement in vivo (table III). The failure of several conjugates to elicit cell-mediated hy persensitivity reactions suggests that the position or type of linkage between the car cinogen and carrier protein are important factors.…”

Direct Macrophage Migration Inhibition Test in DNCB Contact Sensitized Guinea Pigs

“…As these discrepancies might result from differences in the preparation of the conjugates, we varied the source of the microsomal fraction (mixed organs in figure 2c; lymphoid cells in figures 3a, b; epidermis in figures 3c, d, e), the DNP level of the conjugate (1-10 mg DNFB/g of carrier material) and the way of conjugation (presence or absence of living cells). The conjugates derived from the epidermis proved to be the most inhibitive in the MMIT of contact-sensitized guinea pigs, but we ob tained better results with much lower levels of conjugation than those used by Nakagawa and T anioku [14] for skin testing ( fig. 3d).…”

“…16 h later the guinea pigs were depilated and killed. The epidermis was collected from the isolat ed skin by scraping, then homogenized and fractionated in PBS according to N akagawa and T anioku [14]. The 1-hour and 5,000 g supernatant was dialyzed and lyophilized resulting in a so-called 'live' conjugate.…”

“…For a long time it has been accepted that the anti gen used for testing should optimally resemble the antigen used for sen sitization [5,6], As many hapten-carrier conjugates are thought to play a role in contact sensitization, much effort has been made to search for conjugates optimally active in the induction and elicitation of contact hypersensitivity in vivo [1,11,14,[16][17][18]. Conjugates containing mi crosomal fractions have proved to be particularly effective both in skin testing and MMIT of contact-sensitized guinea pigs [3,14,15]. Also, whole cell conjugates prepared from lymphocytes or erythrocytes have been succesfully used by Levis et al [10], and Geczy and Baumgarten [8], in the lymphocyte transformation test.…”