Small Molecular Immunomodifiers of Microbial Origin

Preface

Small Molecular Weight Immunomodifiers Produced by Microorganisms: Their Screening and Discoveries, and the Genetics of Microbial Secondary Metabolites

Genetics of Microbial Secondary Metabolites

A Biochemical Screening Method for Small Molecular Weight Immunomodifiers

Structures and Properties of Bestatin, Amastatin, Forphenicine, and Esterastin


1. Bestatin


2. Amastatin


3. Forphenicine


4. Esterastin

Conclusion with Future Prospect

Summary

Activity of Bestatin: Enhancement of Immune Responses and Antitumor Effect

Effect of Bestatin on Immune Responses


1. Effect of Bestatin on Delayed-Type Hypersensitivity (DTH) to Sheep Red Blood Cells (SRBC)


2. Effect of Bestatin on Delayed-Cutaneous Hypersensitivity (DCH) to Oxazolone

Mitogenicity of Bestatin on Murine Splenic Lymphocytes

Antitumor Effect of Bestatin on Murine Transplantable Tumors


1. Effect of Bestatin on Syngeneic Tumors, IMC Carcinoma, and Gardner's Lymphosarcoma


2. Antitumor Effect of Bestatin in Combination with Antitumor Antibiotics Against Fast-Growing Tumors


3. Effect of Bestatin on Induction of Skin Cancer by Painting 20-Methylcholanthrene (20-MC) on Mouse Skin

The Effect of Bestatin on Colony Formation of Murine Bone Marrow Progenitor Cells in Vitro

Discussion and Conclusion

Summary

Effect of Bestatin on DNA and RNA Metabolism in T-Cells

Studies in the Animal System


1. Effect of Bestatin Pretreatment on the Incorporation Rate of [3H]-Thymidine (dThd) in Lymphoid and Non-Lymphoid Tissues


2. Influence of Bestatin Pretreatment on the Incorporation Rate of [3H]-dThd into Fractionated Lymphocytes


3. in Vivo Activation of DNA Polymerase α in Lymphocytes by Bestatin


4. Distribution of Terminal Deoxynucleotidyl Transferase Activity in Lymphoid Tissue


5. Conclusion

Studies with Cells in Culture


1. Activation of DNA Polymerase α


2. Effect on Polysome Assembly


3. Conclusion

Remarks

Summary

Levamisole and Bestatin in Immunoefficient Aged Mice

Materials and Methods


1. Modalities of Drug Administration


2. Assay for Macrophage-Mediated Cytostasis


3. Spleen Cell Responses to Mitogens


4. Antibody Response


5. Delayed-Type Hypersensitivity Reaction


6. Antibody-Dependent Cell-Mediated Cytotoxicity

Results


1. Effect on Macrophage Activation


2. Effect on Lymphocyte Responsiveness to Mitogens


3. Effect on Suppressor Cell Activity


4. Effect on Antibody Response to Various Antigens


5. Effect on Delayed-Type Hypersensitivity


6. Effect on Antibody-Dependent Cell Cytotoxicity


7. Effect of the Repeated Administration of Levamisole and Bestatin on the Lifespan and the Spontaneous Tumor Incidence

Discussion

Summary

The Influence of Bestatin on the Lymphoid System in the Human

Materials and Methods


1. Patients


2. Preparation of Cell Suspensions


3. Rosette Tests


4. Natural Killer (NK)-Cell Activity of Lymphocytes


5. Antibody-Dependent Cellular Cytotoxicity (ADCC) of Lymphocytes


6. Lectin-Dependent Cellular Cytotoxicity (LDCC) of Lymphocytes


7. Mitogenic Substances


8. Lymphocyte Cultures


9. Blood Sampling and Experimental Design


10. Statistical Methods

Results


1. in Vitro Incubation of Lymphoid Cells from Healthy Donors with Bestatin


2. Bestatin Therapy in Advanced Cancer


3. Immunological Follow-Up in Individual Patients

Discussion

Summary

Effect of Bestatin on Natural Killer Activity

Materials and Methods

Results

Discussion

Summary

Effects of An Aminopeptidase Inhibitor (Bestatin) on Human Lymphocytes I. Effect on Pokeweed Mitogen-Induced in Vitro Immunoglobulin Production

Materials and Methods


1. Bestatin


2. Lymphocytes


3. Separation of T- and B-Lymphocytes


4. Sera for Lymphocyte Treatment


5. Serum Treatment of Lymphocytes


6. Incubation of Lymphocytes


7. Quantitation of Ig in the Supernatant


8. Statistical Analysis

Results


1. Effects of Bestatin on Ig Production by the Lymphocytes from Healthy Individuals


2. Effects of Bestatin on Ig Production by Lymphocytes Stimulated by Various Concentrations of PWM


3. Effects of Bestatin on Ig Production by Lymphocytes Treated with Sera from Stomach Cancer Patients


4. Effects of Bestatin on the Helper Activity of T-Cells Treated with Cancer Sera


5. Ability of Bestatin to Block the Suppressive Effect of Cancer Sera on Ig Production by Lymphocytes

Dicussion

Summary

Effect of Bestatin on Bone Marrow

Materials and Methods

Results

Discussion

Summary

Preliminary Phase I Study of Bestatin in Lung Cancer Cases

Clinical Findings

Changes in Immunological Parameters


1. Rosette Tests in Microtest Plate (2)


2. Responses to Mitogens

Summary

Phase I Study of Bestatin: (I) A Clinical Study for Determination of the Optimal Dose of Bestatin

Methods


1. Subjects


2. Medication Schedule and Collection of Specimens


3. Parameters Examined and Examination Schedule

Results


1. Changes in Immunological Parameters in the Low Dosage, Low Pretreatment Value Group


2. Changes in Immunological Parameters in the Low Dosage, High Pretreatment Value Group


3. Changes in Immunological Parameters in the High Dosage, Low Pretreatment Value Group


4. Changes in Immunological Parameters in the High Dosage, High Pretreatment Value Group


5. Changes in Immunological Parameters in the Group at 90 mg Only Once


6. Adverse Effects of Bestatin

Discussion

Summary

Phase I Study of Bestatin: (II) Clinical Trial of Bestatin in Malignant Skin Tumors

Materials

Dosage Schedule

Results


1. Malignant Melanoma


2. Squamous Cell Carcinoma


3. Basal Cell Carcinoma


4. Adenocarcinoma


5. Malignant Lymphoma and Allied Diseases


6. Soft Tissue Sarcoma

Discussion

Summary

Phase I and Preliminary Phase II Clinical Trials of Bestain

Materials and Methods

Results

Discussion

Summary

Clinical Experiences in the Use of Bestatin in the Treatment of Locally Advanced Cancers: Case Report

Summary

Clinical Experience with Bestatin in Head and Neck Cancer

The Mode of Administration, Cases and the Results

Summary

Clinical and Experimental Study of Bestatin for Esophageal Cancer

Clinical Investigation


1. Materials and Methods


2. Results

Animal Experiment


1. Materials and Methods


2. Results

Discussion

Summary

Clinical Effect of Bestatin on Malignant Tumors of Female Genital Organs

Methods

Results


1. Carcinoma of the Cervix


2. Malignant Tumors of the Ovary

Conclusion

Summary

Pharmacokinetics and Metabolism of Bestatin in Humans and Rats by Gas Chromatography-Mass Spectrometry

Materials and Methods


1. Samples


2. Extraction and Clean-Up Procedure from Human Serum


3. Isolation of Metabolites in Rat Urine


4. Preparation of Derivatives

Results and Discussion


1. Identification of Rat Urinary Metabolites by Radiochromatography


2. Identification of Rat Urinary Metabolites by Use of GC-MS


3. Quantitation of Metabolites after Bestatin Administration

Summary

Appendix Methods of Testing Human Immune Responses

Ontogeny of the Lymphatic System

Determination of the Cellular Composition of Lymphocyte Populations

Determinations of the Proliferative Responses of Lymphocytes to Specific and Nonspecific Mitogenic Stimuli in Vitro

Studies on the Functional Activity of B-Lymphocytes

Migration Inhibition Assay for the Detection of Specifically-Sensitized Lymphocytes

Cytotoxic Functions of Lymphocytes

Detection of Antitumor Immunity

Summary