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Schultz-Dale reaction in mouse trachea
Schultz-Dale
Reaction in Mouse Trachea
GARY A. KOPPEL, KLAUS D. HAISCH, STEPHENM. SPAETHE,JON R. SCHMIDTKE, AND JEROMEH. FLEISCH
A method was developed to induce contraction of immunologically sensitized mouse trachea by antigen (Schultz-Dale reaction). The response was mediated by immunoglobulin (Ig) E antibody directed against either the hapten DNP, the hapten carrier conjugate DNP-keyhole limpet hemocyanin (KLH), or the unmodified carrier KLH. Tracheal contractions were elicited by DNP-KLH, KLH, or DNPbovine serum albumin (BSA) but not by DNP or BSA alone. This procedure represents a useful index of in vitro anaphylaxis in mouse airway smooth muscle. Key Words:
Schultz-Dale;
Mouse; Trachea
The Schultz-Dale reaction describes the contractile response of an immunologically sensitized segment of smooth muscle to antigen challenge (Schultz, 1910; Dale, 1913; Chand and Eyre, 1978). The contraction results from the liberation of mediators of anaphylaxis. Histamine, serotonin, various prostaglandins, and slow reacting substance of anaphylaxis (SRS-A) are among the agents postulated to be released from lung as a result of such an antigen-antibody interaction (Austen, 1977). Although previous investigations have studied antigen-induced contraction of airway smooth muscle from various laboratory animals, none appears to have involved immunoglobulin (lg) E-mediated responses of murine bronchial smooth muscle. This has been due, in part, to the inability of helically cut strips of mouse trachea or bronchi to contract in vitro. A few years ago, Hooker et al. (1977) developed a simple technique whereby drug-induced responses of mouse tracheal rings could be measured in vitro. A similar prdcedure was used ‘suticessfully to obtain Schultz-Dale reactions in ,guinea pig bronchi (Fleisch et al., 1976). With this as a basis, we set forth to document an antigen-induced IgE-mediated contraction of isolated mouse trachea. The present work demonstrates an antigen specific contraction of trachea from mice actively sensitized to DNP-KLH. The response was DNP ahd keyhole limpet hemocyanin (KLH) specific, in that tissues contracted when challenged with DNPbovine serum albumin (BSA), KLH, and DNP-KLH but not to BSA. The s,en&tizing antibody produced a positive passive cutaneous anaphylactic response (PCA) in the rat. Antiserum subjected to heat inactivation at 56°C for 45 min did not cause a PCA response, suggesting the involvement of an IgE antibody. Studies designed to asFrom
the Lilly
Address
and Company, Received
Research
requests
for
Indianapolis,
December
Laboratories,
reprints
to:
Indiana
1, 1980;
Eli Lilly and Company,
Dr.
Jerome
H. Fleisch,
Lilly
Indianapolis, Research
Indiana. Laboratories,
MC9&,’
Eli Liliy
46285.
revised
and accepted February
10,1981.
39 Journal of Pharmacological 0 1981 Elsevier
North
Methods
Holland,
6, 3943
(1981)
Inc., 52 Vanderbilt
Avenue, New York, NY 10017
MM)-5402/81/050039$02.50
40
G. A. Koppel et al.
certain the primary mediator sive.
responsible
for tracheal contractions
were inconclu-
MATERIALS AND METHODS Female Balb/c mice, 8 to 12 weeks old, (Charles River Laboratories) were carrier preimmunized by intraperitoneal (ip) administration of 2 ~g keyhole limpet hemocyanin (KLH) precipitated with 4 mg aluminum hydroxide gel (alum) on day -7. The carrier primed mice were subsequently hapten-carrier immunized with 2 t.rg DNP-KLH in 2 mg alum administered ip on day 0. On day 14, mice were killed by CO2 asphyxiation, bled by cardiac puncture, and trachea excised for pharmacologic analysis using the method of Hooker et al. (1977). Serum was stored at -20°C for later use. Trachea were freed of excess fat and connective tissue, and transferred to tissue supports constructed from two 1 in. 30 gauge disposable stainless steel hypodermic needles. The tissues were then suspended in 10 ml isolated tissue baths containing Krebs’-bicarbonate solution of the following composition in millimoles per liter: KCI, 4.6; CaCI,*2H,O, 2.5; KH2P04, 1.2; MgS04*7H20, 1.2; NaCI, 118.2; NaHCO,, 24.8; and dextrose 10.0. As with our experiments on guinea pig bronchi (Fleisch et al., 1976), indomethacin, IOF M, was incorporated into the solution to enhance antigen-induced mediator release (Walker, 1973). Temperature was maintained at 37°C with a constant temperature circulating unit and the bathing solution was aerated with 95% O2 and 5% COZ. Isometric measurements were made with a Grass FT 03C force-displacement transducer recorded on a Grass polygraph as changes in grams of force. Contractions were maximized with respect to applied force. Optimal initial force for mouse trachea was 1 gram. Tissues were allowed to equilibrate for at least 1 hr before drug effects were elicited. Anti-(DNP-KLH) and anti-DNP antibodies were assayed by PCA reactions in the rat (Mota and Wong, 1969). Fifty microliter serial dilutions of the antibodies were injected intradermally into the backs of shaved male Sprague-Dawley rats (Harlan Industries). IgE reaginic activity of anti-(DNP-KLH) or anti-DNP was elicited 4 hr later by intravenous (iv) injection of either 1 mg DNP-KLH or 1 mg DNP-BSA, respectively, in 1 ml of 1% Evans blue dye in saline. PCA titers represent the reciprocal of the highest dilution, yielding a 5 mm bluing reaction 30 minutes after antigen challenge. The following drugs were used: keyhole limpet hemocyanin (KLH) (Calbiochem, San Diego, CA); crystallized bovine serum albumin, carbamylcholine, indomethacin (Sigma Chemical Co., St. Louis, MO). DNP,-KLH and DNP,,-BSA were prepared according to a slight modification of published procedures (Benacerraf and Levine, 1962; Katz et al., 1970). Subscripts refer to the number of moles of DNP per mole of subunit of KLH (100,000 daltons) and per mole of BSA. RESULTS Schultz-Dale
Reactions
DNP-KLH produced a concentration-dependent contraction of mouse trachea (Fig. 1). The response usually started within 1 min and was maximal by 2 to 3 min.
Schultz-Dale
DNPKLH,
Reaction in Mouse Trachea
@ml
FIGURE 1. Contraction of sensitized mouse trachea to DNP-KLH. Responses are plotted as percent of maximal carbachol contraction. Each value is mean of the number of determinations in parentheses 2 SE.
A similar response pattern was seen with KLH. Responses to these antigens were tachyphylactic, making it difficult to obtain a full dose-response curve on a single tissue. We circumvented this by testing one concentration of antigen on a single tissue and comparing it to a maximal contraction to carbamylcholine (carbachol). Thus, a large number of animals were needed to construct a dose-response curve. The EDs0 for DNP-KLH was 5 x lo-’ g/ml, with the maximal response at lop6 g/ml representing 45% of the highest contraction to carbachol. The antibodies responsible for the Schultz-Dale phenomenon recognized not only KLH and DNP-KLH specificities, but also the DNP specific determinant, as evidenced by the ability of DNP-BSA to contract the trachea (Fig. 2). DNP unconjugated to carrier protein and BSA failed to elicit a response. Tracheal responses to DNP-BSA were similar to those elicited by DNP-KLH. The maximal response was 44% of that to carbachol and the ED5,, was 2 x IO-’ g/ml. Characteristics
of Schultz-Dale
Antibody
Sera from mice used in the above experiments were pooled in groups of five animals and characterized using the rat PCA. The anti-(DNP-KLH) titers of mice immunized for 14 days were 120 and titers for anti-DNP were 60. Sera subjected to 56°C for 45 min did not mediate PCA responses, indicating that the antibody was of the IgE class. Sera from mice immunized with DNP-KLH and bled 7 days later had anti-DNP and anti-(DNP-KLH) IgE PCA titers less than 30. Antigen did not contract trachea from these animals.
41
42
G. A. Koppel et al.
FIGURE 2. Contraction of sensitized mouse trachea to DNP-BSA. Responses are plotted as percent of maximal carbachol contraction. Each value is mean of the number of determinations in parentheses + SE.
Mediators
Responsible
for Tracheal Contractions
Pharmacologic studies with various receptor blocking agents (pyrilamine, atropine, FPL55712) were carried out in an attempt to identify the mediators responsible for the tracheal contractions to antigen. Our results were inconsistent, mainly due to technical problems. Among them, the inability to use a single tissue as its own control, the variability of the Schultz-Dale reaction, and the difficulty inherent in blocking an antigen-induced contraction with pharmacologic antagonists. DISCUSSION Many smooth muscles from immunologically sensitized animals contract to specific antigen in vitro (Schultz, 1910; Dale, 1913; Chand and Eyre, 1978). Despite the central role of the mouse in immunological research, few studies have adequately examined the Schultz-Dale phenomenon in this species. Fink and Rothlauf (1955) and Fink (1956) evaluated this form of in vitro anaphylaxis in isolated mouse uterus. They postulated release of endogenous serotonin as mediating the uterine contraction to antigen. Our study’s main objective was the development of a pharmacological technique to produce an antigen-induced IgE-mediated contraction of mouse trachea. This, in turn, might eventually lead to an understanding of how mediator release occurs from mouse lung. The immunization procedure used was shown in the past (Katz et al., 1970) and confirmed in the present study to produce an IgE antibody against DNP and DNP-KLH. The antibody titers correlated with the Schultz-Dale. response. Titers of 30 did not elicit an antigen-induced smooth muscle contraction, whereas
Schultz-Dale
Reaction in Mouse Trachea
contractions were obtained in tissues from animals with titers of 60 and 120. Full dose-response curves to the antigens were elicited, but only one active concentration of antigen was tested on a single tissue because of desensitization caused by repeated administration of antigen. Preliminary experiments did not define the substances responsible for contraction of mouse trachea. However, our previous study (Hooker et al., 1977) showed mouse trachea devoid of histamine receptors thereby ruling out this amine from consideration. Despite the limitations cited above, this technique represents a successful attempt to elicit an antigen-induced contraction of mouse airway smooth muscle. Further refinements will be necessary to enable exact quantitation of the data. However, even at this stage of development much can be gained with this new procedure. For example, it is now possible to investigate bronchial Schultz-Dale reactions in mice that have different genetic characteristics. Such experiments will help determine some of the immunologic and genetic factors responsible for contraction of airway smooth muscle by antigen. In summary, we have developed an IgE-mediated DNP, DNP-KLH, and KLH specific Schultz-Dale reaction in mouse trachea. Even though the mediators released in response to antigen are unknown, we now have a useful qualitative and possibly even quantitative index of in vitro anaphylaxis of mouse trachea. Since the mouse can be manipulated by various immunological procedures, future studies should provide information as to the nature of and mechanisms involved in release of the mediators of anaphylaxis from mouse bronchial tissue.
REFERENCES Austen KF (1977) Structure and function of chemical mediators derived after activation of mast cells. In Asthma: Physiology, Immunopharmacology, andTreatment. Eds. LM Lichtenstein, and KFAusten. New York: Academic Press, pp. 111-130. Benacerraf B, Levine BB (1962) Immunological specificity of delayed and immediate hypersensitivity reactions. / Exp Med 115:1023. Chand N, Eyre P (1978) The Schultz-Dale a review. Agents Actions 8:171-184.
reaction:
Dale HH (1913) The anaphylactic reaction of plain muscle in the guinea pig. / Pharmacol fxp Ther 4:167-223. Fink MA (1956) Anaphylaxis in the mouse: possible relation of the Schultz-Dale reaction to serotonin release. Proc Sot Exp Biol Med 92:673-675. Fink MA, Rothlauf MV (1955) In vitro anaphylaxis in the sensitized mouse uterus. Proc Sot Exp Biol Med 90:477-480. Fleisch JH, Calkins PJ, Troxell TC, Hooker CS (1976) Inhibition of antigen-induced mediator release
from guinea pig lung by alcohols. Am Rev Resp Dis 114:1107-1112. Hooker CS, Calkins PJ, Fleisch JH (1977) On the measurement of vascular and respiratory smooth muscle responses in vitro. Blood Vessels 14:1-11. Katz DH, Paul WE, Coidl EA, Benacerraf B (1970) Carrier function in anti-hapten immune responses. I. Enhancement of primary and secondary anti-hapten antibody responses by carrier preimmunization. / Exp Med 132:261. Mota I, Wong D (1969) Homologous and heterologous passive cutaneous anaphylactic activity of mouse antisera during the course of immunization. Life Sci 8:813. Schultz WH (1910) Physiological studies in anaphylaxis. I. The reaction of smooth muscle of the guinea pig sensitized with horse serum. / Pharmacol Exp Ther 1:549-567. Walker JL (1973) The regulatory function of prostaglandins in the release of histamine and SRSA from passively sensitized human lung tissue. Adv Biosci 9:235-240.
43
Reaction in Mouse Trachea
GARY A. KOPPEL, KLAUS D. HAISCH, STEPHENM. SPAETHE,JON R. SCHMIDTKE, AND JEROMEH. FLEISCH
A method was developed to induce contraction of immunologically sensitized mouse trachea by antigen (Schultz-Dale reaction). The response was mediated by immunoglobulin (Ig) E antibody directed against either the hapten DNP, the hapten carrier conjugate DNP-keyhole limpet hemocyanin (KLH), or the unmodified carrier KLH. Tracheal contractions were elicited by DNP-KLH, KLH, or DNPbovine serum albumin (BSA) but not by DNP or BSA alone. This procedure represents a useful index of in vitro anaphylaxis in mouse airway smooth muscle. Key Words:
Schultz-Dale;
Mouse; Trachea
The Schultz-Dale reaction describes the contractile response of an immunologically sensitized segment of smooth muscle to antigen challenge (Schultz, 1910; Dale, 1913; Chand and Eyre, 1978). The contraction results from the liberation of mediators of anaphylaxis. Histamine, serotonin, various prostaglandins, and slow reacting substance of anaphylaxis (SRS-A) are among the agents postulated to be released from lung as a result of such an antigen-antibody interaction (Austen, 1977). Although previous investigations have studied antigen-induced contraction of airway smooth muscle from various laboratory animals, none appears to have involved immunoglobulin (lg) E-mediated responses of murine bronchial smooth muscle. This has been due, in part, to the inability of helically cut strips of mouse trachea or bronchi to contract in vitro. A few years ago, Hooker et al. (1977) developed a simple technique whereby drug-induced responses of mouse tracheal rings could be measured in vitro. A similar prdcedure was used ‘suticessfully to obtain Schultz-Dale reactions in ,guinea pig bronchi (Fleisch et al., 1976). With this as a basis, we set forth to document an antigen-induced IgE-mediated contraction of isolated mouse trachea. The present work demonstrates an antigen specific contraction of trachea from mice actively sensitized to DNP-KLH. The response was DNP ahd keyhole limpet hemocyanin (KLH) specific, in that tissues contracted when challenged with DNPbovine serum albumin (BSA), KLH, and DNP-KLH but not to BSA. The s,en&tizing antibody produced a positive passive cutaneous anaphylactic response (PCA) in the rat. Antiserum subjected to heat inactivation at 56°C for 45 min did not cause a PCA response, suggesting the involvement of an IgE antibody. Studies designed to asFrom
the Lilly
Address
and Company, Received
Research
requests
for
Indianapolis,
December
Laboratories,
reprints
to:
Indiana
1, 1980;
Eli Lilly and Company,
Dr.
Jerome
H. Fleisch,
Lilly
Indianapolis, Research
Indiana. Laboratories,
MC9&,’
Eli Liliy
46285.
revised
and accepted February
10,1981.
39 Journal of Pharmacological 0 1981 Elsevier
North
Methods
Holland,
6, 3943
(1981)
Inc., 52 Vanderbilt
Avenue, New York, NY 10017
MM)-5402/81/050039$02.50
40
G. A. Koppel et al.
certain the primary mediator sive.
responsible
for tracheal contractions
were inconclu-
MATERIALS AND METHODS Female Balb/c mice, 8 to 12 weeks old, (Charles River Laboratories) were carrier preimmunized by intraperitoneal (ip) administration of 2 ~g keyhole limpet hemocyanin (KLH) precipitated with 4 mg aluminum hydroxide gel (alum) on day -7. The carrier primed mice were subsequently hapten-carrier immunized with 2 t.rg DNP-KLH in 2 mg alum administered ip on day 0. On day 14, mice were killed by CO2 asphyxiation, bled by cardiac puncture, and trachea excised for pharmacologic analysis using the method of Hooker et al. (1977). Serum was stored at -20°C for later use. Trachea were freed of excess fat and connective tissue, and transferred to tissue supports constructed from two 1 in. 30 gauge disposable stainless steel hypodermic needles. The tissues were then suspended in 10 ml isolated tissue baths containing Krebs’-bicarbonate solution of the following composition in millimoles per liter: KCI, 4.6; CaCI,*2H,O, 2.5; KH2P04, 1.2; MgS04*7H20, 1.2; NaCI, 118.2; NaHCO,, 24.8; and dextrose 10.0. As with our experiments on guinea pig bronchi (Fleisch et al., 1976), indomethacin, IOF M, was incorporated into the solution to enhance antigen-induced mediator release (Walker, 1973). Temperature was maintained at 37°C with a constant temperature circulating unit and the bathing solution was aerated with 95% O2 and 5% COZ. Isometric measurements were made with a Grass FT 03C force-displacement transducer recorded on a Grass polygraph as changes in grams of force. Contractions were maximized with respect to applied force. Optimal initial force for mouse trachea was 1 gram. Tissues were allowed to equilibrate for at least 1 hr before drug effects were elicited. Anti-(DNP-KLH) and anti-DNP antibodies were assayed by PCA reactions in the rat (Mota and Wong, 1969). Fifty microliter serial dilutions of the antibodies were injected intradermally into the backs of shaved male Sprague-Dawley rats (Harlan Industries). IgE reaginic activity of anti-(DNP-KLH) or anti-DNP was elicited 4 hr later by intravenous (iv) injection of either 1 mg DNP-KLH or 1 mg DNP-BSA, respectively, in 1 ml of 1% Evans blue dye in saline. PCA titers represent the reciprocal of the highest dilution, yielding a 5 mm bluing reaction 30 minutes after antigen challenge. The following drugs were used: keyhole limpet hemocyanin (KLH) (Calbiochem, San Diego, CA); crystallized bovine serum albumin, carbamylcholine, indomethacin (Sigma Chemical Co., St. Louis, MO). DNP,-KLH and DNP,,-BSA were prepared according to a slight modification of published procedures (Benacerraf and Levine, 1962; Katz et al., 1970). Subscripts refer to the number of moles of DNP per mole of subunit of KLH (100,000 daltons) and per mole of BSA. RESULTS Schultz-Dale
Reactions
DNP-KLH produced a concentration-dependent contraction of mouse trachea (Fig. 1). The response usually started within 1 min and was maximal by 2 to 3 min.
Schultz-Dale
DNPKLH,
Reaction in Mouse Trachea
@ml
FIGURE 1. Contraction of sensitized mouse trachea to DNP-KLH. Responses are plotted as percent of maximal carbachol contraction. Each value is mean of the number of determinations in parentheses 2 SE.
A similar response pattern was seen with KLH. Responses to these antigens were tachyphylactic, making it difficult to obtain a full dose-response curve on a single tissue. We circumvented this by testing one concentration of antigen on a single tissue and comparing it to a maximal contraction to carbamylcholine (carbachol). Thus, a large number of animals were needed to construct a dose-response curve. The EDs0 for DNP-KLH was 5 x lo-’ g/ml, with the maximal response at lop6 g/ml representing 45% of the highest contraction to carbachol. The antibodies responsible for the Schultz-Dale phenomenon recognized not only KLH and DNP-KLH specificities, but also the DNP specific determinant, as evidenced by the ability of DNP-BSA to contract the trachea (Fig. 2). DNP unconjugated to carrier protein and BSA failed to elicit a response. Tracheal responses to DNP-BSA were similar to those elicited by DNP-KLH. The maximal response was 44% of that to carbachol and the ED5,, was 2 x IO-’ g/ml. Characteristics
of Schultz-Dale
Antibody
Sera from mice used in the above experiments were pooled in groups of five animals and characterized using the rat PCA. The anti-(DNP-KLH) titers of mice immunized for 14 days were 120 and titers for anti-DNP were 60. Sera subjected to 56°C for 45 min did not mediate PCA responses, indicating that the antibody was of the IgE class. Sera from mice immunized with DNP-KLH and bled 7 days later had anti-DNP and anti-(DNP-KLH) IgE PCA titers less than 30. Antigen did not contract trachea from these animals.
41
42
G. A. Koppel et al.
FIGURE 2. Contraction of sensitized mouse trachea to DNP-BSA. Responses are plotted as percent of maximal carbachol contraction. Each value is mean of the number of determinations in parentheses + SE.
Mediators
Responsible
for Tracheal Contractions
Pharmacologic studies with various receptor blocking agents (pyrilamine, atropine, FPL55712) were carried out in an attempt to identify the mediators responsible for the tracheal contractions to antigen. Our results were inconsistent, mainly due to technical problems. Among them, the inability to use a single tissue as its own control, the variability of the Schultz-Dale reaction, and the difficulty inherent in blocking an antigen-induced contraction with pharmacologic antagonists. DISCUSSION Many smooth muscles from immunologically sensitized animals contract to specific antigen in vitro (Schultz, 1910; Dale, 1913; Chand and Eyre, 1978). Despite the central role of the mouse in immunological research, few studies have adequately examined the Schultz-Dale phenomenon in this species. Fink and Rothlauf (1955) and Fink (1956) evaluated this form of in vitro anaphylaxis in isolated mouse uterus. They postulated release of endogenous serotonin as mediating the uterine contraction to antigen. Our study’s main objective was the development of a pharmacological technique to produce an antigen-induced IgE-mediated contraction of mouse trachea. This, in turn, might eventually lead to an understanding of how mediator release occurs from mouse lung. The immunization procedure used was shown in the past (Katz et al., 1970) and confirmed in the present study to produce an IgE antibody against DNP and DNP-KLH. The antibody titers correlated with the Schultz-Dale. response. Titers of 30 did not elicit an antigen-induced smooth muscle contraction, whereas
Schultz-Dale
Reaction in Mouse Trachea
contractions were obtained in tissues from animals with titers of 60 and 120. Full dose-response curves to the antigens were elicited, but only one active concentration of antigen was tested on a single tissue because of desensitization caused by repeated administration of antigen. Preliminary experiments did not define the substances responsible for contraction of mouse trachea. However, our previous study (Hooker et al., 1977) showed mouse trachea devoid of histamine receptors thereby ruling out this amine from consideration. Despite the limitations cited above, this technique represents a successful attempt to elicit an antigen-induced contraction of mouse airway smooth muscle. Further refinements will be necessary to enable exact quantitation of the data. However, even at this stage of development much can be gained with this new procedure. For example, it is now possible to investigate bronchial Schultz-Dale reactions in mice that have different genetic characteristics. Such experiments will help determine some of the immunologic and genetic factors responsible for contraction of airway smooth muscle by antigen. In summary, we have developed an IgE-mediated DNP, DNP-KLH, and KLH specific Schultz-Dale reaction in mouse trachea. Even though the mediators released in response to antigen are unknown, we now have a useful qualitative and possibly even quantitative index of in vitro anaphylaxis of mouse trachea. Since the mouse can be manipulated by various immunological procedures, future studies should provide information as to the nature of and mechanisms involved in release of the mediators of anaphylaxis from mouse bronchial tissue.
REFERENCES Austen KF (1977) Structure and function of chemical mediators derived after activation of mast cells. In Asthma: Physiology, Immunopharmacology, andTreatment. Eds. LM Lichtenstein, and KFAusten. New York: Academic Press, pp. 111-130. Benacerraf B, Levine BB (1962) Immunological specificity of delayed and immediate hypersensitivity reactions. / Exp Med 115:1023. Chand N, Eyre P (1978) The Schultz-Dale a review. Agents Actions 8:171-184.
reaction:
Dale HH (1913) The anaphylactic reaction of plain muscle in the guinea pig. / Pharmacol fxp Ther 4:167-223. Fink MA (1956) Anaphylaxis in the mouse: possible relation of the Schultz-Dale reaction to serotonin release. Proc Sot Exp Biol Med 92:673-675. Fink MA, Rothlauf MV (1955) In vitro anaphylaxis in the sensitized mouse uterus. Proc Sot Exp Biol Med 90:477-480. Fleisch JH, Calkins PJ, Troxell TC, Hooker CS (1976) Inhibition of antigen-induced mediator release
from guinea pig lung by alcohols. Am Rev Resp Dis 114:1107-1112. Hooker CS, Calkins PJ, Fleisch JH (1977) On the measurement of vascular and respiratory smooth muscle responses in vitro. Blood Vessels 14:1-11. Katz DH, Paul WE, Coidl EA, Benacerraf B (1970) Carrier function in anti-hapten immune responses. I. Enhancement of primary and secondary anti-hapten antibody responses by carrier preimmunization. / Exp Med 132:261. Mota I, Wong D (1969) Homologous and heterologous passive cutaneous anaphylactic activity of mouse antisera during the course of immunization. Life Sci 8:813. Schultz WH (1910) Physiological studies in anaphylaxis. I. The reaction of smooth muscle of the guinea pig sensitized with horse serum. / Pharmacol Exp Ther 1:549-567. Walker JL (1973) The regulatory function of prostaglandins in the release of histamine and SRSA from passively sensitized human lung tissue. Adv Biosci 9:235-240.
43