Immunology 1977 33 179
Suppression and potentiation of expression of delayed-type hypersensitivity by dextran sulphate JOHANNA L'AGE-STEHR & TIBOR DIAMANTSTEIN Immunology Research Unit, Klinikunm Steglitz, Freie Universitat, Berlin, Germany
Received 14 September 1976; acceptedfor publication 30 December 1976
Summary. Dextran sulphate delays the onset, or even completely suppresses the expression in mice of DTH to SRBC when administered via a route different from that of eliciting antigen. However, DS injected together with the eliciting antigen potentiated the expression of DTH. Dextran showed no effect on DTH. Cell transfer experiments suggest that the targets for the action of DS are the accessory cells (monocytes) and not the T-effector cells. As shown, using polystyrene latex particles and lipopolysaccharide from E. coli, trapping and perhaps activation of the trapped accessory cells rather than toxic effects of DS are responsible for these phenomena. INTRODUCTION
Polyanions, e.g. dextran sulphate (DS), show a variety of immunologically relevant activities such as (a) enhancement (Diamantstein, Wagner, Beyse, Odenwald & Schultz, 1971a) as well as suppression (Diamantstein, Keppler, Blitstein-Willinger & BenEfraim, 1976) of humoral immunity; (b) mitogenicity for a subpopulation of B lymphocytes (Diamantstein, Blitstein-Willinger & Schulz, 1974); (c) changes in thymocyte reactivity to lectins (Blitstein-Willinger, Schulz & Diamantstein, 1976); (d) activation of the complement system (Hadding, Dierich, Konig, Correspondence: Professor T. Diamantstein, Hindenburgdamm 30, 1000 Berlin 45, Germany.
Limbert, Schorlemeyer & Bitter-Suermann, 1973); (e) enhancement of antigen uptake by the spleen (Diamantstein, Meinhold & Wagner, 1971b); (f) suppression of hepatic uptake of antigen, presumably due to toxicity of DS for Kupfer cells (Bradfield, Souhami & Addison, 1974) and (g) impairment of resistance of mice to Listeria monocytogenes infection (Hahn & Bierther, 1974). The effects of polyanions on T cells and on macrophages raised the question as to whether DS may influence cell-mediated immunity in vivo. To test this possibility, we examined the effect of DS (and of compounds sharing some biological activities with DS) on the delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC) in mice (Miller, MacKaness & LaGrange 1973).
MATERIALS AND METHODS Animals Specific pathogen-free female BALB/c mice (Dr Friis, Bomholdgard, Denmark) were used at 6-8 weeks of age. Test substances DS mol. wt 5 x 105 from Serva, Heidelberg; dextran (D) mol. wt 5 x 105, Pharmacia Uppsala; polystyrene latex particles 1 3-p diameter from K.W. Pfannenschmidt, Hamburg and lipopolysaccharide from E. coli 055: B5 (LPS) from Difco were used.
179
Johanna L'Age-Stehr & Tibor Diamantstein
180 Antigen
SRBC were obtained weekly from the same animal. Cells were collected and stored at 40 in Alsever's solution for 7-14 days. The cells were washed three times with saline before use. Immunization and detection of DTH Mice were sensitized either by i.v. injection of 105 SRBC suspended in 01 ml saline or by i.c. injection of 108 SRBC in 40 u1 of saline i.c. into the right hind footpad. To assess DTH, mice were challenged 4 days after immunization by injection of 108 SRBC in 40 p1 of saline i.c. into the left hind footpad. As controls, non-immunized mice were injected into the footpad with the same dose of antigen. The swelling of the footpad was measured before challenge and 24, 48 and 72 h afterwards with a dial gauge caliper (Oditest type OD 100T50, H.C. Kroeplin, Schluchtern, Germany). The results are expressed as the specific increment in units of 10-3 cm calculated as the difference in thickness between footpads receiving antigen challenge compared to those that had not.
Cell transfer Spleen cell suspensions of i.v.-sensitized mice or popliteal lymph node cells (LN) of footpad-sensitized animals were used 4 days after sensitization as a source of passive transfer cells. Cell suspensions were prepared as described earlier (L'Age-Stehr & Herzenberg 1970). One spleen equivalent of i.v.sensitized mice was transferred i.v. to normal mice which were challenged within 1 h with 108 SRBC into the footpad. Control mice received spleen cells of unsensitized donors. The draining LN-cells of footpad sensitized mice (1 -2 x 107 cells per recipient) were mixed with 108 SRBC and injected locally into the footpad of normal recipients. Control mice were injected with LN-cells of unsensitized donors. Treatment of mice with the test compounds The test compounds were given dissolved or suspended in saline by various routes and simultaneously, or 30 min before antigenic challenge, as described in results.
RESULTS
Suppression of the expression of DTH by dextran sulphate Since DS showed no marked effects on the induction
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Suppression and potentiation of expression of delayed-type hypersensitivity by dextran sulphate JOHANNA L'AGE-STEHR & TIBOR DIAMANTSTEIN Immunology Research Unit, Klinikunm Steglitz, Freie Universitat, Berlin, Germany
Received 14 September 1976; acceptedfor publication 30 December 1976
Summary. Dextran sulphate delays the onset, or even completely suppresses the expression in mice of DTH to SRBC when administered via a route different from that of eliciting antigen. However, DS injected together with the eliciting antigen potentiated the expression of DTH. Dextran showed no effect on DTH. Cell transfer experiments suggest that the targets for the action of DS are the accessory cells (monocytes) and not the T-effector cells. As shown, using polystyrene latex particles and lipopolysaccharide from E. coli, trapping and perhaps activation of the trapped accessory cells rather than toxic effects of DS are responsible for these phenomena. INTRODUCTION
Polyanions, e.g. dextran sulphate (DS), show a variety of immunologically relevant activities such as (a) enhancement (Diamantstein, Wagner, Beyse, Odenwald & Schultz, 1971a) as well as suppression (Diamantstein, Keppler, Blitstein-Willinger & BenEfraim, 1976) of humoral immunity; (b) mitogenicity for a subpopulation of B lymphocytes (Diamantstein, Blitstein-Willinger & Schulz, 1974); (c) changes in thymocyte reactivity to lectins (Blitstein-Willinger, Schulz & Diamantstein, 1976); (d) activation of the complement system (Hadding, Dierich, Konig, Correspondence: Professor T. Diamantstein, Hindenburgdamm 30, 1000 Berlin 45, Germany.
Limbert, Schorlemeyer & Bitter-Suermann, 1973); (e) enhancement of antigen uptake by the spleen (Diamantstein, Meinhold & Wagner, 1971b); (f) suppression of hepatic uptake of antigen, presumably due to toxicity of DS for Kupfer cells (Bradfield, Souhami & Addison, 1974) and (g) impairment of resistance of mice to Listeria monocytogenes infection (Hahn & Bierther, 1974). The effects of polyanions on T cells and on macrophages raised the question as to whether DS may influence cell-mediated immunity in vivo. To test this possibility, we examined the effect of DS (and of compounds sharing some biological activities with DS) on the delayed-type hypersensitivity (DTH) to sheep red blood cells (SRBC) in mice (Miller, MacKaness & LaGrange 1973).
MATERIALS AND METHODS Animals Specific pathogen-free female BALB/c mice (Dr Friis, Bomholdgard, Denmark) were used at 6-8 weeks of age. Test substances DS mol. wt 5 x 105 from Serva, Heidelberg; dextran (D) mol. wt 5 x 105, Pharmacia Uppsala; polystyrene latex particles 1 3-p diameter from K.W. Pfannenschmidt, Hamburg and lipopolysaccharide from E. coli 055: B5 (LPS) from Difco were used.
179
Johanna L'Age-Stehr & Tibor Diamantstein
180 Antigen
SRBC were obtained weekly from the same animal. Cells were collected and stored at 40 in Alsever's solution for 7-14 days. The cells were washed three times with saline before use. Immunization and detection of DTH Mice were sensitized either by i.v. injection of 105 SRBC suspended in 01 ml saline or by i.c. injection of 108 SRBC in 40 u1 of saline i.c. into the right hind footpad. To assess DTH, mice were challenged 4 days after immunization by injection of 108 SRBC in 40 p1 of saline i.c. into the left hind footpad. As controls, non-immunized mice were injected into the footpad with the same dose of antigen. The swelling of the footpad was measured before challenge and 24, 48 and 72 h afterwards with a dial gauge caliper (Oditest type OD 100T50, H.C. Kroeplin, Schluchtern, Germany). The results are expressed as the specific increment in units of 10-3 cm calculated as the difference in thickness between footpads receiving antigen challenge compared to those that had not.
Cell transfer Spleen cell suspensions of i.v.-sensitized mice or popliteal lymph node cells (LN) of footpad-sensitized animals were used 4 days after sensitization as a source of passive transfer cells. Cell suspensions were prepared as described earlier (L'Age-Stehr & Herzenberg 1970). One spleen equivalent of i.v.sensitized mice was transferred i.v. to normal mice which were challenged within 1 h with 108 SRBC into the footpad. Control mice received spleen cells of unsensitized donors. The draining LN-cells of footpad sensitized mice (1 -2 x 107 cells per recipient) were mixed with 108 SRBC and injected locally into the footpad of normal recipients. Control mice were injected with LN-cells of unsensitized donors. Treatment of mice with the test compounds The test compounds were given dissolved or suspended in saline by various routes and simultaneously, or 30 min before antigenic challenge, as described in results.
RESULTS
Suppression of the expression of DTH by dextran sulphate Since DS showed no marked effects on the induction
' - 5 -
