Tissue Culture Studies of Cytoplasmic Inclusion Bodies in Lymph Nodes of Hodgkin's Disease. ∗

In the literature, the etiology of Hodgkin's disease has been ascribed to numerous agencies. Among these, bacterial and virus infections have been included.

This report is based on nodes from 35 cases of Hodgkin's disease, obtained surgically from Memorial Hospital, New York. The lymph nodes were from early and late stages of the disease. As controls, normal lymph nodes from 12 radical mastectomy cases were used, also nodes from various lymphomas (20 lymphosarcomas, 15 leukemias) and from 23 metastatic carcinomas and 10 lymphadenitis cases.

The nodes were cut into small fragments (explants, about 1 cu mm in size) from which tissue cultures were prepared in the usual way. The medium consisted of a mixture of fowl plasma, human serum and chick embryo extract. In some cases the serum was from patients with advanced stages of Hodgkin's disease.

The cultures were maintained for a period varying from a few days to several weeks. After 24 hours′ incubation, granulocytes, eosinophils, and lymphocytes had migrated from the explant. After 48 hours the outgrowth contained also macrophages, reticulocytes, and fibrocytes. After 48 to 72 hours of incubation there also appeared, on the periphery of the explants, large multinucleated giant cells with oval nuclei. The nuclei tended to surround a relatively large, grayish and granular legion generally occupying the central portion of the cell. These cells, identified as Reed-Sternberg cells, were found in every case of true Hodgkin's disease and were absent in the other lymphomas and in the normal lymph nudes. The longer the tissue cultures were maintained, the more numerous and larger were these cells.

Brilliant cresyl blue (1:50,000), used as a vital dye for virus cell inclusions, stained the granules of the central body of the Reed-Sternberg cell within 15 minutes after exposure of the culture to the dye.