91
Agents and Actions vol. 9/1 (1979) Birkh~iuser Verlag, Basel
7. Histamine and Immunology Susceptibility to Passive Sensitization with Reaginic Antibodies in Four Inbred Strains of Mice by J. WYCZdLKOWSKA,T. MARCINIAKand Cz. MAgLINSKI Department of Biogenic Amines, Institute of Pharmacology, Polish Academy of Sciences, 90-136 Ldd~', Poland
Introduction It has been established that mouse IgE antibodies bind firmly to the homologous and rat mast cells, sensitizing them for anaphylactic reaction [1, 2]. Previous results suggested that not only mouse IgE antibodies but also non-specific IgE are able to fix on homologous mast cells, interfering thereby in passive sensitization with specific IgE antibodies [2, 3]. The present experiments were undertaken in order to study the possible interference of non-specific IgE in passive sensitization of mouse skin and peritoneal mast cells with specific IgE antibodies.
Materials and methods Female mice from four inbred strains (Balb/C, 129, C3H/A and C57BL/6J), 6-8 weeks old, were used for passive sensitization. Male Wistar rats weighing 200-220 g were employed for heterologous Passive Cutaneous Anaphylaxis (PCA). Reaginic serum was produced by immunization of Swiss mice with several i.p. injections of 1 /tg egg albumin, five times crystallized (Schwarz/Mann, Orangeburg), mixed with 1 mg aluminium hydroxide gel, given 2-4 weeks apart. The antisera were obtained 1 week after every booster immunization and pooled. Anti-mouse IgE antiserum was prepared as previously described [4]. Passive sensitization of mouse skin with homologous reaginic serum was performed as described [1], with a 2 h and 72 h latent period for IgG1 and IgE antibodies respectively. Heterologous PCA reaction was performed in rats [3] with a 48 h latent period. For some experiments the normal or reaginic sera were heated at 56~ for 4 h. Passive sensitization of mouse peritoneal mast cells was obtained by i.v. injection of mice with reaginic serum (20 ml/kg). Peritoneal cell suspension was taken up after 48 h, as previously described [3]. The cells were washed three times in Barron-modified Ringer medium and tested for antigeninduced histamine release by incubation with egg albumin (100 ~g/ml) at 37~ The histamine content in the samples was determined by the fluorometric method of Shore et al. omitting the extraction procedure. Histamine release was expressed as percentage of the released histamine to the total histamine after deduction of spontaneous release found in controls. In some experiments mast cells of normal mice were tested for anti-IgE-induced histamine release [4].
Results In the first experiment mice of inbred strains were tested for susceptibility to passive sensitization of the skin with mouse reaginic serum (Table 1). The results show that strains differ in susceptibility to PCA reactions induced by mouse IgE antibodies; the strains 129 and C57BL/6J were much less susceptible to long-latence (72 h) PCA reaction than were C3H/A or Balb/C mice. There was poor correlation between the susceptibility of the strains to IgEmediated reactions and their susceptibility to IgGl-mediated reactions. In the next experiment the strains were tested for susceptibility to passive sensitization of their peritoneal mast cells with IgE antibodies. Figure 1 shows the low capacity of strain 129 (as compared with other strains) for passive sensitization of peritoneal mast cells and subsequent antigeninduced histamine release. This correlated well with the low susceptibility of this strain as a recipient of IgE-induced PCA reactions (Table 1). On the contrary, Balb/C and C3H/A mice, characterized by the high susceptibility as recipients of IgE-induced PCA reactions, showed a higher
Table 1 Susceptibility to PCA reactions in four inbred strains of mice.
Mouse strain
PCA titres of reaginic seruma IgE b
Baib/C C3H/A 129 Cs7BL/6J
IgG1 c
Nonheated
Heateda
Nonheated
Heated
640 640 80 40
Agents and Actions vol. 9/1 (1979) Birkh~iuser Verlag, Basel
7. Histamine and Immunology Susceptibility to Passive Sensitization with Reaginic Antibodies in Four Inbred Strains of Mice by J. WYCZdLKOWSKA,T. MARCINIAKand Cz. MAgLINSKI Department of Biogenic Amines, Institute of Pharmacology, Polish Academy of Sciences, 90-136 Ldd~', Poland
Introduction It has been established that mouse IgE antibodies bind firmly to the homologous and rat mast cells, sensitizing them for anaphylactic reaction [1, 2]. Previous results suggested that not only mouse IgE antibodies but also non-specific IgE are able to fix on homologous mast cells, interfering thereby in passive sensitization with specific IgE antibodies [2, 3]. The present experiments were undertaken in order to study the possible interference of non-specific IgE in passive sensitization of mouse skin and peritoneal mast cells with specific IgE antibodies.
Materials and methods Female mice from four inbred strains (Balb/C, 129, C3H/A and C57BL/6J), 6-8 weeks old, were used for passive sensitization. Male Wistar rats weighing 200-220 g were employed for heterologous Passive Cutaneous Anaphylaxis (PCA). Reaginic serum was produced by immunization of Swiss mice with several i.p. injections of 1 /tg egg albumin, five times crystallized (Schwarz/Mann, Orangeburg), mixed with 1 mg aluminium hydroxide gel, given 2-4 weeks apart. The antisera were obtained 1 week after every booster immunization and pooled. Anti-mouse IgE antiserum was prepared as previously described [4]. Passive sensitization of mouse skin with homologous reaginic serum was performed as described [1], with a 2 h and 72 h latent period for IgG1 and IgE antibodies respectively. Heterologous PCA reaction was performed in rats [3] with a 48 h latent period. For some experiments the normal or reaginic sera were heated at 56~ for 4 h. Passive sensitization of mouse peritoneal mast cells was obtained by i.v. injection of mice with reaginic serum (20 ml/kg). Peritoneal cell suspension was taken up after 48 h, as previously described [3]. The cells were washed three times in Barron-modified Ringer medium and tested for antigeninduced histamine release by incubation with egg albumin (100 ~g/ml) at 37~ The histamine content in the samples was determined by the fluorometric method of Shore et al. omitting the extraction procedure. Histamine release was expressed as percentage of the released histamine to the total histamine after deduction of spontaneous release found in controls. In some experiments mast cells of normal mice were tested for anti-IgE-induced histamine release [4].
Results In the first experiment mice of inbred strains were tested for susceptibility to passive sensitization of the skin with mouse reaginic serum (Table 1). The results show that strains differ in susceptibility to PCA reactions induced by mouse IgE antibodies; the strains 129 and C57BL/6J were much less susceptible to long-latence (72 h) PCA reaction than were C3H/A or Balb/C mice. There was poor correlation between the susceptibility of the strains to IgEmediated reactions and their susceptibility to IgGl-mediated reactions. In the next experiment the strains were tested for susceptibility to passive sensitization of their peritoneal mast cells with IgE antibodies. Figure 1 shows the low capacity of strain 129 (as compared with other strains) for passive sensitization of peritoneal mast cells and subsequent antigeninduced histamine release. This correlated well with the low susceptibility of this strain as a recipient of IgE-induced PCA reactions (Table 1). On the contrary, Balb/C and C3H/A mice, characterized by the high susceptibility as recipients of IgE-induced PCA reactions, showed a higher
Table 1 Susceptibility to PCA reactions in four inbred strains of mice.
Mouse strain
PCA titres of reaginic seruma IgE b
Baib/C C3H/A 129 Cs7BL/6J
IgG1 c
Nonheated
Heateda
Nonheated
Heated
640 640 80 40
