Forssman Antiserum Research Articles

Ｓｕｐｅｒｏｘｉｄｅ ｄｉｓｍｕｔａｓｅ（ＳＯＤ）のＤＤＳ

Superoxide anion (O2-) acts as an exacerbating factor in various disease conditions. Superoxide dismutase(SOD), a scavenger of O2-, has been expected to become a therapeutic tool for treating many diseases. However, to attain an adequate clinical effect, it is necessary to prolong the biological half-life of SOD and to increase its affinity for cell membrane by using a drug delivery system. Several chemically modified forms of SOD, including polyethylene glycol binding SOD and pyran-SOD, have not yet been approved for clinical use. In the present study, we lecithinized recombinant human SOD by covalently binding four molecules of a lecithin derivative to each SOD dimer. The blood half-life after the intravenous(i.v.)injection of lecithinized-SOD into rats was doubled compared to that of unmodified SOD. Lecithinized-SOD also had a 10-fold higher affinity for several cells. The i. v. injection of Forssman antiserum is known to induce a biphasic airway resistance and it is suggested that O2- is involved in the second phase of this response. We examined the effects of lecithionized-SOD on this phase of respiratory resistance using guinea pigs. Unmodified SOD was ineffective at i. v. doses of 1, 000∼30, 000 U/kg, whereas over a dose range of 10∼1, 000 U/kg, lecithinized-SOD demonstrated a dose-dependent inhibitory effect. Therefore, we expect PC-SOD to be safe and pharmacologically very potent in humans, with the potential for use in various clinical applications.

Effects of lecithinized superoxide dismutase (PC-SOD) on respiratory resistance induced by Forssman antiserum in guinea pigs

Effects of lecithinized superoxide dismutase (PC-SOD) on respiratory resistance induced by Forssman antiserum in guinea pigs

モルモット肺傷害に及ぼすＬｉｐｏｓｏｍｅ化 Ｒｅｃｏｍｂｉｎａｎｔ ｈｕｍａｎ ｓｕｐｅｒｏｘｉｄｅ ｄｉｓｍｕｔａｓｅの影響

We investigated the effect of liposome-r-h-SOD on the lung injury induced by Forssman antiserum in guinea pigs. Intravenous injection of Forssman antiserum produced a biphasic increase in pulmonary resistance in guinea pigs. Liposome-r-h-SOD blocked this biphasic increase in a dose-dependent manner. Exudate and hemorrhage in the alveolar spaces and connective tissues were blocked by liposome-r-h-SOD. Liposome-r-h-SOD was more effective when it was injected 30 min prior to challenge than 5 min prior to challenge. On the other hand, free r-h-SOD or a mixture of free r-h-SOD and empty liposome did not block the lung injury. SOD activity in porcine endothelial cells cultured with liposome-r-h-SOD increased in a dose-dependent and time-dependent manner, while it did not increase with free r-h-SOD. Exogenous SOD was detected by immunoperoxidase staining in endothelial cells of arterioles and capillaries and in alveolar epithelial cells of the lung of guinea pigs injected with liposome-r-h-SOD. These findings suggest that superoxide radical may take part in the lung injury induced by Forssman antiserum. Liposome-r-h-SOD, which adheres to and/or is endocytosed by endothelial cells, may protect the lung from oxygen radicals.

Interference of levamisole with forssman shock

The anti-helmintic drug levamisole has been used as an adjunct in the treatment of some immunologic defects including cancer. Recently, it has been shown that this drug inhibits thromboxane synthetase as well. Since Forssman shock in guinea pig is used as a model for pulmonary thromboembolism involving thromboxane Az, we studied the interference of levamisole with broncho-constriction, thrombocytopenia, endothelial cells and pulmonary damage induced by Forssman antiserum. Levamisole inhibited dose-dependently the pathological changes produced by Forssman antiserum raising the possibility that levamisole may be effective for the treatment of pulmonary thromboembolism in man.

Role of thromboxane receptors in Forssman shock in guinea pigs

Forssman shock is a bronchospastic reaction mounted in guinea pigs on intravenous administration of an antiserum obtained from rabbits immunized against sheep erythrocytes. The involvement of thromboxane receptors in Forssman shock was determined with SQ 30,741, which was characterized as a selective antagonist of these receptors in guinea pig airways in vitro and in vivo. A volume of antiserum producing consistent, sublethal bronchoconstriction was given either alone (control) or 3 min after SQ 30,741 (0.03, 0.3, or 1.0 mg/kg iv) to urethan-anesthetized guinea pigs. In controls, maximum reductions in dynamic compliance (-59 +/- 6%, P less than 0.01) and increases in airways resistance (383 +/- 97%, P less than 0.01) were detected 1 min after antiserum. Both responses were significantly inhibited by SQ 30,741, either partially at 0.03 mg/kg or completely at 0.3 mg/kg. An accompanying thrombocytopenia was not abated by SQ 30,741. In separate experiments, bronchospasm was reduced by aerosol administration of 0.1% SQ 30,741 and completely prevented by aspirin (10 mg/kg iv). When Forssman antiserum was injected in lethal quantities to other guinea pigs, SQ 30,741 (1 mg/kg iv) attentuated only the resistance component of bronchospasm and did not prevent death. These data demonstrate that thromboxane receptor stimulation is a pivotal step in the pulmonary manifestations of sublethal Forssman shock but is less crucial in more severe forms of the reaction.

Forssman antigen in BK virus-induced tumor cell lines.

A complement-mediated microcytotoxicity assay was used to detect Forssman antigen in BK virus (BKV)-induced tumor cells. BKT-1B, a transplantable hamster tumor cell line, contained Forssman antigen as did some other hamster cell lines tested for comparison. A rat embryo cell line transformed by BKV in vitro was negative for Forssman antigen. The presence of forssman antigen in hamster cell lines was confirmed by absorption experiments of rabbit forssman antiserum with the tumor cells, although in quantitative terms the results obtained by the two techniques were somewhat different. Immunization of rabbits with tumor cells also induced production of Forssman antibody; a much weaker response was observed in hamsters and rats.

MECHANISMS OF FORSSMAN‐INDUCED BRONCHOSPASM AND THEIR INHIBITION

1 The bronchospasm induced in the guinea-pig by the injection of Forssman antiserum was biphasic in nature in both the sublethal and the lethal reaction. 2 The development of both phases of the bronchospasm in the sublethal reaction was dependent upon the presence of the intact complement system and circulating platelets. In the lethal reaction the phase II bronchospasm did not appear to depend on these factors. 3 The compounds used in this study inhibited phase I bronchospasm of the sublethal reaction in the order, methysergide greater than indomethacin greater than aspirin = sulphinpyrazone and phase II in the order, indomethacin greater than sulphinpyrazone greater than aspirin. Methysergide was inactive. 4 Aspirin, indomethacin and sodium salicylate all prevented the inhibitory action of sulphinpyrazone in reducing the phase II bronchospasm of the sublethal reaction in the order, indomethacin greater than sodium salicylate greater than aspirin, when the drugs were administered prior to sulphinpyrazone. 5 The inhibitory action of aspirin on the sulphinpyrazone effect could be prevented by administering sulphinpyrazone before aspirin. All drug-induced inhibitions of sulphinpyrazone by aspirin, indomethacin and sodium salicylate were dose-dependent.

Inhibition of anaphylactic histamine release by Forssman antiserum. II. Mechanism of inhibition.

The mechanism was studied of the inhibitory effect of IgG Forssman antibody on anaphylactic sensitization in the guinea pig lung. Two possibilities were considered to explain the inhibitory effect: either the Forssman antibody-antigen interaction occurred on the mast cell, allowing the F(ab′)<sub>2</sub> portion to interfere with the Fc receptor(s) for anaphylactic antibody; or the interaction took place adjacent to the mast cell allowing the Fc portion to occupy the receptors for anaphylactic antibody. These studies were made possible in part by developing a method for preparing mast cell suspensions from guinea pig lung. The mast cells were viable but released histamine only on treatment with compound 48/80 and not with anaphylatoxin, soluble immune complexes or with active or passive anaphylactic sensitization. Mast cells incubated with Forssman antibody and complement showed no evidence of injury as judged by electron microscopy, suggesting that they did not contain Forssman antigen. The absence of Forssman antigen from mast cells was substantiated further by studies of mixed agglutination and fluorescence microscopy. Also, pepsin digestion of Forssman antibody resulted in a loss of the ability to inhibit the histamine-releasing activity of soluble immune complexes, indicating that the F(ab′)<sub>2</sub> portion alone was not sufficient to sustain the inhibitory process. It was concluded that the inhibitory effect of IgG Forssman antibody was due to union with interstitial Forssman antigen adjacent to mast cells, allowing the Fc portion of the Forssman antibody to irreversibly occupy the mast cell Fc receptor(s) for anaphylactic antibody.

The use of 3H serotonin release from mast cells of the mouse as an assay for mediator liberation.

The release of 3H 5-HT from murine mast cells is shown to be a simple reproducible method for studying the activation of such cells by various agents. 3H-serotonin was taken up by peritoneal cell suspensions in vitro and was released by antigen, Concanavalin A, Forssman antiserum, anti-mouse immunoglobulin, or a polypeptide antibiotic, Cinnamycin.

Complement-Mediated Tissue Damage: Contribution of the Classical and Alternate Complement Pathways in the Forssman Reaction

Abstract The contribution of the classical and alternate complement pathways to complement-mediated tissue damage in the Forssman reaction was studied in a strain of C4D guinea pigs with a nonoperative classical pathway, but an intact alternate pathway. It was found that the C4D guinea pigs were resistant to the Forssman reaction, but that reconstitution of the classical pathway by the administration of normal guinea pig serum as a source of C4 restored the deficient animals' susceptibility to the toxic effects of injected Forssman antiserum. C4D animals with a restored C4 level only developed the Forssman reaction if the Forssman antiserum was injected many hours after the injection of C4. Thus, a period of time was required for C4 to distribute within the extravascular compartment. These results indicate that interaction of anti-Forssman antibody with extravascular Forssman antigen and complement is necessary for lethality, and that vascular endothelial damage cannot alone account for the pathogenesis of the Forssman reaction, as previously believed. The results also indicate that intravascular C4 is not freely available to tissue sites.

Heterophile Antibodies and Tissue Injury

Forssman antiserum was inoculated into the skin of normal guinea pigs. Within moments endothelial and perivascular cells exhibited a spectrum of ultrastructural lesions, culminating in overt necrosis of these elements. At the same time platelets and PMN-leukocytes adhered to damaged vascular surfaces and formed thrombotic obstructions in the microcirculation. Platelets and PMN-leukocytes also appeared to be degranulating, liberating lysosomal and other biologically active constituents from the cells. These data support the concept that vascular structures are primary target tissues and suggest that PMN-leukocytes and platelets may be pathogenetic determinants in the response to Forssman antiserum.

Lethality of a Forssman antiserum for mice.

SummaryMice can be killed by an intravenous injection of a rabbit antiserum to boiled guinea pig kidney. Evidence is presented that these deaths are due to Forssman antibodies.

PRIMARY SERUM TOXICITY AS DEMONSTRATED BY THE CHICKEN EMBRYO

1. The 3 day old chicken embryo removed from its shell is a suitable test object for the demonstration of primary serum toxicity. Addition of normal rabbit type sera as well as Forssman antiserum causes the vascular network to contract and the embryo sinks in the yolk and dies. 2. Only sera of animals of the so called rabbit type produce this phenomenon. Sera of the guinea pig type are ineffective. 3. Heating to 51 degrees C. destroys the complement content of normal human serum as also its effectiveness to produce the vascular phenomenon. 4. Up to the present it has not been possible to reactivate heat-inactivated normal serum by the addition of complement, while inactivated Forssman antiserum can be easily reactivated. 5. The vascular phenomenon of the chicken embryo is produced not only by the addition of a mixture of Forssman antiserum and complement but also by separate addition of both components. 6. Guinea pig type sera, containing dissolved Forssman antigen, are not only ineffective but actually exert an inhibitory influence on effective rabbit type sera as well as on Forssman antiserum.