William L. Bayer

Autologous tumor cell vaccination combined with adoptive cellular immunotherapy in patients with Grade III/IV astrocytoma

SummaryBrain tumors are highly resistant to treatment. Their diffuse infiltrative nature and the relative inaccessibility of the brain to blood and lymph are barriers to surgical and cytotoxic treatments alike. Preclinical animal studies demonstrated that intravenously administered tumor antigen-specific T lymphocytes will reject tumors growing in the brain. Specifically activated effector T lymphocytes may be generated by in vivo immunization followed by restimulation of antigen-primed T cells with autologous tumor cells in vitro. In order to apply these findings to humans, feasibility studies of combined active immunization and specific adoptive cellular immunotherapy were performed on fifteen patients with recurrent astrocytoma. The objective was to determine whether; 1) T cells could be grown from peripheral blood of patients immunized with autologous tumor cells, and 2) whether stimulated cells could be safely readministered to patients. Patients were immunized with a combination of their own irradiated tumor cells and Bacillus of Calmette and Guerin. Two weeks later, a mononuclear cell-rich fraction of blood was obtained by leukapheresis. Mononuclear cells were cultured with irradiated autologous tumor cells and interleukin-2. Selective expansion of CD4+ and CD8+ T lymphocytes occurred. Intravenous transfer of stimulated cells to the fifteen patients on twenty-four separate occasions with or without systemic administration of interleukin-2 was tolerated with limited toxicity. The studies established the feasibility of conducting controlled studies of the anti-tumor effects of tumor antigen-specific cellular immunotherapy.

Solid‐phase ABO grouping and Rh typing

A solid‐phase adherence method (SPAM) for ABO grouping and Rh typing of red cells (RBCs) has been developed. Adherence reactions were read spectrophotometrically and interpreted by a computer. The SPAM had a 99.6 percent correlation with conventional microplate agglutination methods for ABO grouping and Rh typing. The increased sensitivity of the SPAM was demonstrated because it directly detected Du‐positive RBCs and weak subgroups of A and B.

Factors influencing monocyte recognition of human erythrocyte autoantibodies in vitro

Erythrocytes, serum, and blood monocytes from patients with erythrocyte autoantibodies were examined by in vitro serologic and cell culture procedures in order to evaluate factors modulating patient mononuclear phagocyte activity. Monocyte recognition of autoantibody‐coated erythrocytes was influenced both by antibody titer score and by the monocyte source. The results suggest that patients with low serum autoantibody titer scores and normal monocyte behavior are less likely to destroy autologous or normal donor erythrocytes than are patients with high‐serum antibody titer scores or enhanced monocyte activity. We conclude that patient, rather than normal donor monocytes, must be used in cellular assays designed to predict the fate of erythrocytes transfused into patients with erythrocyte autoantibodies.

Antigens of Human Cytomegalovirus: Electroimmunodiffusion Assay and Comparison Among Strains

The antigens of strain AD169 of human cytomegalovirus (CMV) were extracted by various methods and at different times following the appearance of cytopathic effects (c.p.e.) in infected fibroblasts. Assay with a pooled human serum in electroimmunodiffusion (EID) revealed that the most reactive preparations were obtained by shell-freeze (SF) extraction on the fourth day after 4+c.p.e. As many as 20 antigens could be detected in the original gels, most of which were stable upon storage at 4 degrees C for up to 4 weeks; of these, about 14 can be reproducibly seen on photographs. EID runs on day 4 SF preparations from high-passage CMV strains C87 and Davis and low passage recent isolates VD14, 1694 and 1723 resolved, respectively, 15, 15, 13, 11 and 11 antigens in the original gels (11, 9, 11,8 and 9 are visible in photographs). Strains 1694 and 1723 shared fewer antigens with one another and with high passage strains than were shared among the latter, whereas VD14 had relatively large numbers of antigens common to both low and high passage strains. At least six antigens were common to all strains.

Serologic Activity of Fatty Acid Dependent Antibodies in Albumin‐Free Systems

Fatty acid dependent agglutinin (FADA) refers to serum with the special ability to cause agglutination of red blood cells in the presence of certain fatty acids. The agglutinating mechanism is unclear. It has been proposed that the agglutinin reacts with albumin that has been conformationallly altered by sodium caprylate and that the immune complex is passively adsorbed onto red blood cells. This report presents data that contradicts the proposal assigning a specific role to albumin in the agglutinating mechanism. FADA were isolated by column chromatography of resolubilized euglobulin preparations. No evidence of contamination with albumin was obtained in those IgM fractions possessing FADA activity. We propose, as an alternative explanation, that the serologic activity of FADA depends upon the interaction of IgM agglutinins with haptenic fatty acids.

A Technique for von Willebrand Multimer Analysis Applicable to the Clinical Laboratory

In type I von Willebrand disease, the whole series of von Willebrand factor multimers is present in plasma. However, in type II von Willebrand disease qual itative defects of the von Willebrand factor proteins exist. Using a novel semidry horizontal gel electrophoresis ap paratus combined with a discontinuous buffer system, high-resolution analysis of von Willebrand factor multi meric composition is possible. When applied in clinical practice, this technique allows for easy discrimination be tween type I and type II defects.

Fresh-Frozen Plasma: The Lack of Need for Surgical Hemostasis in a Plasma-Exchanged Patient

Three consecutive daily plasma volume plasma exchanges were done using no coagulation factor-containing materials on a patient who then underwent thymectomy without any hemostatic problems immediately after the third exchange. Key Words: Plasma exchange-Coagulation factor-Hemostasis.