Heterogeneity of Antigen from Callus of Man

Abstract

The question of whether one or more antigens are involved in the reaction between antibodies in human serum and callus of man was investigated. Callus was collected from an individual with congenital hyperkeratosis. Preparations were obtained by extraction of homogenated callus with (1) phenol water (PW), and (2) the combined use of trypsin digestion and phenol water extraction (TPW). The phenol and water phases were studied using indirect haemagglutination technique. All preparations sensitized erythrocytes for agglutination by human serum except for the phenol phase of the PW preparation. Absorption and inhibition experiments showed that one antigen present in the TPW water phase was also present in the PW water phase. In addition, the PW water phase contained another antigen. In the phenol phase of the TPW preparation a third antigen was demonstrated. Serological cross-reactivity between the antigens and bacteria isolated from callus was not demonstrated. The results of immune adherence test with sections of human skin and unabsorbed or absorbed human serum indicated that the two water phase antigens are present in normal human skin.

Species Specificity of Skin Antigens Prepared by Phenol Water Extraction

“…Similar reactivity and cross-reactivity of sera was previously reported [11] and verified in this study using the immune ad herence test with skin sections. The agreement between the results ob tained in the two tests again indicates that the antibodies active in indi rect haemagglutination are also involved in the immune adherence test [16]. The cross-reactivity of sera could be mediated either by antibodies to common antigenic determinants, antibodies to species-specific deter minants or to both.…”

“…We reported previously [16] that in addition to the TPW water phase, the PW water and TPW phenol phases from callus contain ery throcyte-sensitizing antigens. These findings were verified in this study.…”

“…The failure to detect activity in the PW water or TPW phenol phase of skin from the animals may be explained quantita tively. The results of earlier studies have indicated that the antigens in volved originate from stratum corneum [11,16]. This constitutes a very small part of the guinea pig and rabbit whole skin used for extraction but forms the major part of callus of man.…”

“…Recently, we reported that at least three different antigens can be demonstrated in extracts prepared from callus by phenol water and that two of these are present in normal skin [16]. So far, only extracts from human skin tissue have been studied.…”

Studies in Immunodermatology

“…More aggressive methods are needed to disrupt the horny layer structure and to ex pose its normally hidden components capable of reacting with SC antibodies in indirect hemagglutina tion, i.e., the method described by Krogh et al [17], combining proteolytic digestion with trypsin and sub sequent extraction with phenol and water (the socalled trypsin-phenol water method). Absorption and hemagglutination inhibition studies of Maelandet al [21] and Krogh and Tender [20] give evidence that at least three different SC antigens reacting in indirect hemagglutination are present in callus. Several au thors reported on possible differences in the chemical composition existing between normal horny layer and callus [8,9], By comparison of water and phenol phases obtained from phenol-water and trypsin-phe nol water extractions, serologic differences between SC, callus and psoriatic scales were also found by Krogh and Tonder [20], None of the antigenic prepa rations obtained by trypsin-phenol water methods is active against SC antibodies detectable by indirect IF on normal human skin, normal monkey skin, or the cut edge of callus, unless the latter is degraded by treatment with a crude trypsin preparation (contain ing both trypsin and chymotrypsin) as shown by the present studies.…”