Paul L. Wolf

A NEW METHOD OF ACTIVE IMMUNISATION TO AUTOLOGOUS HUMAN TUMOUR TISSUE

Abstract Fourteen volunteers with far-advanced malignant tumours were treated by a method in which an attempt was made to immunise them against their own tumours. The method consisted of chemically coupling the tumour cell to a highly antigenic foreign protein (rabbit gamma-globulin) with bisdiazobenzidine and reinjecting this complex back into the tumour donor. Of the fourteen patients treated, two are alive and have been tumour-free for about 4 years. Biopsy of their tumours revealed necrotic changes. Three patients have shown stabilisation or slow progression of tumour; nine patients have died. Antibodies in high titres directed toward the tumours, were demonstrated by various means in thirteen of these patients.

Production of active immunity to malignant neoplastic tissue. Chemical coupling to an antigenic protein carrier

C3H mice bearing spontaneous mammary adenocarcinomas and methylcholanthrene induced squamous cell carcinomas were immunized actively to their own tumors when their tumor cells were altered, thereby rendering them antigenic. The tumor cells were altered by coupling of human gamma globulin to a tumor cell suspension utilizing bis‐diazobenzidine as a coupling agent. The altered cells then were re‐injected into the animal from which they were taken with complete Freunds adjuvant. An altered host‐tumor relationship was evidenced in the study animals by retardation of tumor growth grossly and increased amounts of necrosis, lymphocytic infiltration, hydropic degeneration and fibrosis on histologic section. Antibodies to tumor cells were detected in the study group following immunization by microprecipitin, Ouchterlony and immunofluorescent techniques. Antibodies were not detectable in sera of control animals immunized with unaltered tumor cells alone in complete Freunds adjuvant.

The indigogenic reaction for histochemical demonstration of sulfatase.

Summary The indigogenic principle has been extended to the histochemical demonstration of sulfatase. The advantage of this substrate is that it affords a precise enzyme localization with no or very slight diffusion and also a simple and direct method for demonstration of this enzyme without the need for a coupling reaction. We are indebted to Mrs. Georgette M. Dunlap for stenographic assistance.

Application of the indigogenic principle for the histochemical demonstration of ribonuclease

Es wurde ein neues Indolsubstrat für die histochemische Demonstration von alkalischer RNase II und saurer RNase II synthetisiert. Dieses Verfahren ist besser, schneller und präziser als die bisherigen Kupplungstechniken.

Substrates for cytochemical demonstration of enzyme activity. IV. Kinetics of the hydrolysis of thymidine-5'(5-bromo-4-chloroindol-3-YL) phosphate by snake venom phosphodiesterase.

Abstract 1. 1. A comparison of the action of purified spleen phosphodiesterase II (orthophosphoric diester phosphohydrolase, EC 3.1.4.1) on two synthetic substrates thymidine 3′-(p-nitrophenyl) phosphate (III) and thymidine 3′-(5-bromo-4-chloro-3-indol-3-yl) phosphate (V) is described. The utility of the latter (V) derives from a chromogenic reaction sequence that leads to the highly colored 5,5′-dibromo-4,4′-dichloro-indigo (VII) via aerobic oxidation of the enzymically released intermediate 5-bromo-4-chloroindoxyl (VI). It is demonstrated that the rate of oxidation of VI to VII is significantly greater at pH 7.0 than enzymic release of VI. Therefore the oxidation step is not rate determining. 2. 2. The rates of enzymic hydrolysis of III and V, effected at a substrate concentration of 150 μg/ml in 4.1 M acetate (pH 5.5) in accord with established procedures, are of the same order of magnitude (370 and 235 μmoles/h per mg protein, respectively). The Km with III is about 0.25·10−3 and differs by a factor of approx. 10 from that (3·10−3) recorded by Razzell and Khorana. The difference is ascribed to the improved method of purification which affords a stable enzyme free of contaminants. 3. 3. A comparison of the kinetic parameters Km and V for the two substrates indicates that V is an acceptable substrate for assay of phosphodiesterase II.

The Selective Irradiation Of Canine Lymph Nodes By Means Of Intralymphatic Injection Of 32p

Intralymphatic injection of radioactive chromic phosphate produced a severe destruction of a majority of lymph nodes, and subsequent marked lymphopenia in dogs. Selectivity of radiation was evidenced by histological study of other major organs and blood counts. Antibody production of these animals against human serum albumin was significantly inhibited; however, the reactivity to the allografted heart was not altered.

Erythroleukemic phase of mouse myeloproliferative syndrome

Smears were made of blood and cell suspensions of thymus, spleen, bone marrow, left axillary lymph node, liver, kidney and brain of mice with spontaneous leukemia. They were stained either with Leishmans stain or specifically for hemoglobin with Ralphs stain. The blood and hematopoietic organs from mice with virus‐induced leukemia (Stansly, 1963) were also examined. The presence of large numbers of immature megaloblastoid cells and maturing normoblasts in the hematopoietic organs and the extensive infiltration of these cells into the other organs emphasize the erythroleukemic nature of the disease. The presence of immature myeloid elements in many of the smears suggests that the erythroleukemia is part of a myeloproliferative syndrome.

Specific anti-antibody in transplantation

Durch ein immunologisches Dreieck-System (Hund-Kaninchen-Hund) wurde ein Anti-Antikörper bei Hunden erzeugt und seine Wirkung in vitro und in vivo demonstriert. Die Anti-Antikörper-Methode bietet neue Möglichkeiten, dem Problem der Transplantat-Abstossung und der Therapie der Krankheiten immunologischen Ursprungs näherzukommen.

A histochemical study of immune destruction of the lymphopoietic system

Eine auffallende Zunahme von β-Galactosidase kommt im lymphoetischen Säuger-Gewebe im Zusammenhang mit der immunologischen Auflösung dieses Systems vor. Die hernach auftretende Proliferation retikulärer Zellen steht im Zusammenhang mit der Regeneration dieses Organs. Diese Zellen enthalten eine reichliche Anzahl lysosomatischer Enzyme (z.B. β-Galactosidase).

Studies on the Transplanted Heart. Role of Gamma-Globulin in Transplantation Immunity.

Summary Sensitization of the recipient animal with spleen homogenate resulted in a significant increase in the serum gamma globulin fraction and subsequent accelerated rejection of the graft. In exceptional cases in which serum gamma-globulin level did not increase, the accelerated rejection was absent. Treatment with 6-MP reduced the gammaglobulin level and the grafts in these animals showed prolonged survival. Reduction of gamma-globulin by exchange transfusion failed to prolong survival time of the graft. These results are in accord with the view that humoral factors present in the gammaglobulin fraction play an important role in homograft rejection.