11．The studies on the effects of gelatin and its related substances
In the present paper I review the studies on the effects of gelatin and its related substances discovered during the course of applying the Shwartzman reaction to experimental cancers and the research entailed, starting from an analysis of the Shwartzman reaction. These studies were carried out in the laboratory of Professor Motoki Yamanaka at Osaka Medical College.
It is widely accepted that antigen antibody reaction induces anaphylaxis, allergy reaction elevates permeability and that Shwartzman reaction induces thrombus formation and hemorrhagic necrosis. However, how the molecular and humoral immune responses are converted into mechanical and physical phenomena in the circulation remain unclear.
Although the basic logic of liquidity follows Hagen-Poiseuille's law, Poiseuille himself noted that blood flow does not always follow the law. In Hagen-Poiseuille's law, the inner diameter of vessels is the most significant factor. however, blood vessels with an inner diameter of 100 microns or smaller does not follow Hagen-Poiseuille's law. Assuming that blood flow follows Newton's law, then cardiac pumping would not be able to complete the circulation. The descriptions that capillary blood vessels are the second heart and that blood viscosity is low in appearance were an attempt to explain this contradiction.
It was elucidated in the laboratory of Professor Mashimo before World War II that one reason for the contradiction is derived from the fact that blood is a suspension containing solid matter, i.e. blood cells. However, in the circulation system in humans, four liters of blood are in contact with 2,000 square meters of vascular endothelium. Therefore, surface chemical forces should be considered. As blood surface tension increases, greater force is required in a centrifugal direction for the circulation.
After provocative injection for Shwartzman reaction, serum surface tension transiently elevates with simultaneous reduction in a coefficient of foaming retention at the site of the hemorrhage caused by sensitization injection. (See Table appended to reference article 3.) That is, the blood having the property of high and retained foaming, which is a surface physicochemical property of protein solution, is lowered. In addition, we found that the interface viscosity of serum and vascular endothelium is greater when in interface with sensitized endothelium than with normal endothelium.
This may explain how the injected site of sensitization can be discovered upon the provocation injection, enabling the initial step for paralyzed blood flow at the site to be taken. If it is demonstrated that immunological Shwartzman reaction1）,2）is converted to mechanical physical phenomenon through surface chemical phenomena, it would explain the mystery that necrotic hemorrhage is induced although provocation and sensitization in Shwartzman reaction is immunologically not strictly identical to each other. As serum surface tension decreases, the force of centripetal blood circulation from peripherals decreases, explaining the increase in permeability, reduction in circulating blood flow and the phenomena of oedema, allergy and anaphylaxis.
Serum surface tension tends to vary among various diseases. It is elevated in some cases of obesity and hypertension, while it is lowered in some cases of allergic disease and malignant tumor. The correlation coefficient of serum surface tension with blood pressure is 0.36 and that with age-corrected index following the Faught's formula is 0.45. It is also partially correlated with serum protein fraction. The correlation coefficient is -0.32withα1with globulin, -0.37 with α1/α2value and -0.33 with α1/Al value. I presented this topic at the meeting of the Japanese Society of Allergies and at the second meeting of the Japanese Society of Biophysics.3)
It is known that bacterial endotoxins including Shwartzman filtrate have carcinostatic activity as Coley's toxin4） and Shear's polysaccharide5）. To utilize endotoxins with their strong adverse effects in cancer therapy, I thought it necessary to develop a technique for inducing necrosis at a given point on a tumor by applying Shwartzman reaction rather than by administering endotoxins in drug form. Shwartzman reaction occurs in rabbit skin. To discover the method of inducing Shwartzman reaction at a given point in parenchymal organs, rabbit stomach was used in this study.
The addition of adjuvant to endotoxin for local injection was not successful. Hemorrhagic necrosis could be initially induced by locally injecting heat killed bacterial cells. The lesion of hemorrhagic necrosis subsequently progressed to typical gastric ulcer 6）. This technique used to prepare rabbit gastric ulcer was applied to mouse Ehrlich carcinoma. After repeated systemic administration of a minute amount of endotoxin for sensitization, cancer cells were transplanted to form a solid tumor, then heat-killed bacterial cells were injected into the tumor, which caused hemorrhagic necrosis and subsequent regression and disappearance of the tumor 7）8）. There have been many studies performed focusing on polysaccharides of bacterial endotoxin, but development for practical use was not successful in many studies, failing to separate the adverse effects and carcinostatic activity. In my experience the heat killed bacterial cells are preferable to purified endotoxin. In addition, experiments were carried out to examine if the protein contained in the heat killed bacterial cells acts as an adjuvant by changing the quantitative ratio. However, this was not the case. Since only a portion of scleroproteins is heat-resistant, I injected collagen and gelatin, and obtained a lowered transplantation rate in mouse Ehrlich carcinoma9） and lowered incidence of rat DMBA mammary cancer10）. When Ehrlich carcinoma was replanted to mice cured of Ehrlich carcinoma by applying Shwartzman-type reaction as described above, and to mice in which Ehrlich carcinoma was transplanted after injection of collagen or gelatin had been rejected, replanted Ehrlich carcinoma was not observed in these mice. However, a quantitative relationship was observed. That is, the more a graft was rejected, the stronger the anti-graft reaction became. Solid tumor was more curable than ascites carcinoma. Although anti-tumor agents tend to be more effective for ascites carcinoma than for solid tumors, in the present case this was to the contrary. Subcutaneous transplantation is rejected through coagulative necrosis, while intraperitoneal cavity transplantation induces rosette formation by macrophages, resulting in rejection of the carcinoma cells. In mice which have become unable to be grafted with Ehrlich carcinoma, sarcoma 9）180 and related skin iso-graft were also rejected (in contrast, mice in which the heart had been successfully transplanted were able to be grafted with skin in a study). This transplantation immunity is not inheritable.
Except for the tumorigenesis, cancer immunity and transplantation immunity overlap in tumor bearing, may be due to allergy. Interpreting that the presence or absence of acclimation is the difference between immunity and allergy, the immunity against transplantation induced by ingestion of heterologus gelatin is an allergic phenomenon without acclimation, showing a useful aspect of allergy.11）12）13）14）15）16）Ingestion of antigenic gelatin enhances anti-transplantation immunity through enhanced interstitial reaction.
Later, in the wake of an accident in protective vaccination, the presence of anti-gelatin lgE antibody were discovered. Oral ingestion of gelatin was effective for hemorrhage, resulting in better wound healing. Externally administered gelatin was also effective for leg ulcer, bedsore and radiation skin erosion. It has been demonstrated that the external use of insoluble gelatin powder methylene polymerized by formaldehyde at an amino group is convenient17）. When the gelatin powder, which is insoluble even in boiling water although it swells, was suspended in water and subcutaneously injected into mice using a heavy gauge needle, a globular granulation tissue was formed, incorporating the gelatin. The region in contact with the skin then disintegrated and dehydrated dry gelatin powder was discharged from the body. After the discharge of gelatin powder, Ehrlich carcinoma was transplanted but the cancer cells were unable to be grafted in the mice. That is, although the injected insoluble gelatin powder was eliminated from the body, the strong anti-transplantation immunity remained.18)
We recommend cancer patients to receive surgery, and we also recommend them to eat gelatin. We attempted to apply this treatment to cancers of the digestive organs and liver, to laryngeal cancer, hypopharyngeal cancer and lymphosarcoma, and it was effective. Gelatin in the patients' diet is easy to administer and it does not contradict other medical care or the law. Therefore, we advise gelatin ingestion as part of anti-cancer treatment. Considering the result of the Walzer's experiment19）, a portion of orally ingested macromolecules are absorbed with the macromolecular structure intact, and is therefore valuable.
- YAMANAKA et.:
ウサギ血清中のフォルスマン抗体量とシュワルツマン現象惹起注射量 との至適比関係に就いて 東京医事新誌 69（1）39,1952 (English title: Optimum ratio of rabbit serum Forssman antibody titer to amount of injection inducing the Shwartzman phenomenon. The Tokyo Medical Journal 69 (1) 39, 1952)
総 合臨臨床 4（9）1,1955(Engllsh title: Forssman antigen-antibody reaction Proposal on the significance of natural F antibody as a predisposing antibody in inflammation and independence of Shwartzman antigen - Substantial view of Shwartzman phenomenon- . Clinic all-round 4 (9) 1, 1955)
- ENOKI：Rheological Studies of the Shwartzman reaction. Bulletin of the Osaka medical school 8(2)53,1962
- Coley:A review of the influence of bacterial infection and of bacterial products (Coley's toxin) on malignant tumors in man. Acta Med Scand 45,1953
- Shear et al:Chemical treatment of tumors. Isolation of the hemorrhage-producing fraction from Serratia marcescens culture filtrate J Nat Cancer Inst 4:81,1943
医学と生物学70（4）195,1965(English title: ExperiTnental qastric ulcer in the rabbit. Medieine and Biology 70（4）195,1965
- ENOKI：Shwartzman型反応によるエールリッヒ固型癌の治療実験 医学と生物学 (English title: Experimental therapy by Shwartzman-type reaction of Ehrlich solid carcinoma in mice. Medicine and Biology 70（4）197,1965
- ENOKI：Allergic necrotizing reaction applied to mouse tumor and experimentally acquired resistance of mice to the tumor transplantation and iso-graft. JINSEN-IGAKU 11（4）197,1968
- ENOKI：ハツカネズミの移植癌に対するゼラチンならびにその重合物による 抗移植性の免疫 医学と生物学 82（2）81,1971(English title: An immunity of mice resistant to the tumor transplant by injection of gelatin and its derivatives. Medicine and Biology 82 (2) 81, 1971)
- ENOKI, NISHIZATO and NAKATA：ダイコクネズミのDMBA乳癌に対する重合ゼラチンの影響 医学と生物学 87（6）321,1973 (English title: The effect of polymerized gelatin on rat mammary carcinoma produced by the carcinogen, DMBA. Medicine and Biology 87 (6) 321, 1973)
- ENOKI：ハツカネズミの実験的獲得性の腫瘍移植抵抗性と同系皮移植の拒否現象 医学と生物学 74（3）185,1967 (English title: Experimentally acquired resistance in mice to the tumor transplantation and the rejection of the skin iso-graft. Medicine and Biology 74 (3) 185, 1967)
- ENOKI, NAKATA and NISHIZATO：Ehrlich 腹水癌における実験的免疫現象 医学と生物学 87（5）311,1973(English title: Experimentally acquired immunity of mice to Ehrlich's ascites carcinoma. Medicine and Biology 87 (5) 311, 1973)
- ENOKI, NISHIZATO and NAKATA：ハツカネズミがEhrlich癌に対して実験的獲得性に抗移植性の免疫を作る現象に与える各種マクロフアージ誘発物質の免疫誘導能力の比較 医学と生物学 87（5）311,1973(English title: A comparison of the effect of macrophage inducing substances on the production of immunity in mice to experimentally acquired resistance to Ehrlich's carcinoma transplant. Medicine and Biology 87 (5) 313, 1973)
- ENOKI：ハツカネズミの移植癌に対する同種同系、同種異系のゼラチンの抗移植性の免疫 医学と生物学 100（2）111,1980 (English title: Immunity of mice resistant to tumor transplant by injection of gelatin; mice of the same species-same strain, and the same species-different strain. Medicine and Biology 100 (2) 111, 1980)
- ENOKI：A study on allergic reaction of cancer and its application to transplantation immunity in experimental animals: The immunological approach for the teatment of cancer JINSEN-IGAKU 12（4）175,1973
- ENOKI：Immunitiy of mice resistant to tumor transplant using endotoxin and substituting endotoxin with gelatin and its related substances. Bacterial endotoxins and host response, P.203,1980 Proceedings of the 4th international congress of immunology. Statellite workshop July 1980
- ENOKI and UEDA：下腿潰瘍及び褥瘡に試みた新しい治療経験 大阪医科大学雑誌 24（1）1,1965(English title: On the curative effects of the powder or the lamina of the formogelatin to the leg ulcer and the bedsore. The Journal of Osaka Medical College 24 (1) 1, 1965)
- ENOKI：ハツカネズミの移植癌に対する不溶性のゼラチン粉末による抗移植性の免疫 医学と生物学 100（2）113,1980 (English title: Tumor transplant immunity in mice by the injection of insoluble gelatin powder. Medicine and Biology 100 (2) 113, 1980)
- Walzer：Studies in absorption of undigested proteins in human beings. J.Allrgy 6:532,1935