Indirect Haemagglutination for Demonstration of Antibodies to Stratum Corneum of Skin

Abstract

A new method for preparation of soluble antigens from stratum corneum of skin is described. Homogenated callus was digested with trypsin before extraction with phenol-water. The extract sensitized sheep erythrocytes for agglutination by human sera. This agglutination was due to a true antigen-antibody reaction between stratum corneum antigen on the erythrocytes and antibodies in the human sera. The indirect haemagglutination test was as sensitive as the immune adherence test with skin sections earlier described. The antigen and antibodies involved in the two test systems are identical. All of 1,150 normal sera tested contained antibodies to stratum corneum. The titres ranged from 8 to 16,384 with mode titres of 32 and 64. Presumably, the antibodies are natural hetero- or auto-antibodies.

Heterogeneity of Antigen from Callus of Man

“…Homogenated tissue was digested with trypsin (Crystalline, Trypsin, Novo) as described earlier [10] and further processed as for PW extraction. The water phase (TPW water phase) and phenol phase (TPW phenol phase) were dialyzed and lyo philized.…”

“…Human serum from healthy blood donors was used in dividually or as a pool from at least 100 individuals. The sera were heated at 56 °C for 20 min for inactivation of complement and absorbed with sheep erythrocytes [10] unless otherwise stated. Fresh serum was absorbed similarly at 4 °C with sheep erythrocytes.…”

“…Serol ogical cross-reactivity between the antigens and bacteria isolated from callus was not demonstrated. The results of immune adherence test with sections of human skin arid unabsorbed or absorbed human serum indicated that the two water phase antigens are present in normal human skin.Recent studies have shown that normal human sera contain antibodies to stratum corneum of man [6,10]. Originally, these antibodies were demonstrated using the immune adherence test with microtome sections of human skin [7], and later by fluorescent antibody and indirect haemag glutination techniques [5,10].…”

“…Originally, these antibodies were demonstrated using the immune adherence test with microtome sections of human skin [7], and later by fluorescent antibody and indirect haemag glutination techniques [5,10]. The antigen used for sensitization of eryth rocytes was prepared by extraction of homogenates of callus with phenol water (PW) and recovered from the water phase.…”

“…The antigen used for sensitization of eryth rocytes was prepared by extraction of homogenates of callus with phenol water (PW) and recovered from the water phase. The combined use of trypsin digestion and phenol water extraction (TPW) greatly increased the M^la n d / K r o g h /T o n d e r yield of the material which contains the sensitizing component [10]. The phenol phase was not investigated.…”