Wayne State University School of Medicine - Yearbook (Detroit, MI)

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INTRA-ERYTHROCYTIC LOCUS OF HEMOGLOBIN USING FLUORESCENT ANTIBODY TECHNIQUES As part of a continuing project in the Department of Biochem- istry to identify the intra-erythrocytic locus of hemoglobin and the subcellular site of hemoglobin synthesis, new techniques were developed replacing those previously reported CNelson and Orten, 1966 Yearbookj. Realizing that acetone extraction of the heme of hemoglobin, countercurrent separation of the alpha and beta chains and lyophillyzation led to a denatured product unsuitable for use as an antigen. The following procedure was adopted. Human erythrocytes were washed, lysed, and the hemoglobin purified by fractional precipitation with ammonium sulfate. The alpha and beta chains of hemoglobin were separated using pH gradient elution over a carboxymethylcellulose column. The beta chains were then concentrated and injected into the foot pads and subcutaneously into the backs of Dutch rabbits. After a suit- 'able period, the rabbits were sacrificed and the gamma globulin fraction containing the anti-human-beta-chain antibodies was iso- lated. The purity of the preparation was tested by immunoelectro- phoretic techniques and the titer of antibodies established. Finally, human erythrocytes were incubated with the rabbit anti-beta chain antisera, washed, then re-incubated with sheep anti-rabbit globulin-fluorescein conjugated antisera. These cells were then examined under the fluorescent microscope where pre- liminary results indicate that the beta chains Cand therefore the hemoglobinl are located uniformly throughout the mature erythro- cytes. It will be the purpose of a subsequent project to determine the sites of hemoglobin synthesis in the erythrocyte precursors using this method. Roger F. Suchyta james M. Orten, Ph.D.

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DELAYED HYPERSENSITIVITY REACTIONS IN PATIENTS WITH MALIGNANT DISEASE It is well known that in severely debilitated patients from whatever cause, for example, old age, infections and malignan- cies, there is a marked decline in the delayed hypersensitivity reaction. However, little work has been done to determine whether this decreased response Chypoergy or anergyj is non- specific. In other words, it has not been clearly shown if the immunological competence of patients is compromised by the specific disease process or the non-specific effects of debility. This question has been of more interest because there is evidence to suggest that cancer patients may suffer from a poor delayed hypersensitivity reaction due to a primary action ofthe malignancy on the immunological system. Hughes and MacKay have shown that depressed tuberculin response in cancer pa- tients have reversed themselves after curative treatment from surgery. They also correlated a depression in the tuberculin response with the early stages of cancer, when there was no detectable systemic spread of the tumor. It was thought worthwhile, therefore, to investigate the delayed hypersensitivity reactions to tuberculin, mumps, C. albicans, and dinitrobenzene in cancer patients contrasted with those in matched patients with no malignancy. Peter P. Wong V. Vaitkevicius, MD. 156