Studies in Steroid Metabolism. XXVI. Steroid Isolation Studies in Human Leukemia

Abstract

The normal and abnormal physiology of white blood cells can be characteristically influenced by the hormonal environment. Adrenocortical hormones administered as such, or secreted under stimulation by ACTH have been shown to influence the distribution, mass, and functional activity of lymphoid tissue (1-6). The course and incidence of leukemia in test animals are intimately related to alterations in adrenal or gonadal function (7-12) and the striking, albeit temporary, amelioration achieved by adrenocortical or adrenocorticotropic hormones in human leukemia, is well known (13-16). Although decreased urinary ketosteroids have been demonstrated in leukemia (17, 18), excretion of urinary corticoids has been generally unaltered (18). However, few data are available relative to specific gonadal or adrenal hormone metabolites in human leukemia.We have applied, to the study of this problem, the precise methods of isolation, identification, and quantitation of individual urinary steroids developed within the past decade (19-23). The detailed ketosteroid patterns of six subjects with chronic lymphatic leukemia during control periods and of one subject with acute myelogenous leukemia during a control interval and while on ACTH therapy are reported. The results give evidence of decreased secretory activity of both gonads and adrenal glands in these subjects. The simultaneous development of refractoriness to ACTH therapy in one patient, with evidence of progressive adrenal dysfunction as measured by the urinary steroids, raises, in addition, an interesting question about the relation between clinical relapse in this disease, and the hormonal environ- METHODSThe urinary ketosteroid patterns of six subjects with chronic lymphatic leukemia were studied. Five of these subjects were male (M45, B58, S59, H62, B68), and one was female (H55). In addition, the urinary ketosteroids of one subject with acute myelogenous leukemia (R29) were studied during the course of two successive therapeutic trials with ACTH. Metabolic balance studies were conducted during these periods and the results have already been reported (Subject R. R. [24]). All urine collections were short term (5 to 16 days) except for that of subject M45 (195 days). The urinary steroid conjugates were hydrolyzed immediately after collection, by a combination of methods employing enzymatic (beef liver fi-glucuronidase), cold and/or hot acid hydrolysis (Methods A, B, C, or D [22]).Neutral steroid extracts were separated into "ketonic" and "non-ketonic," and "a-ketonic" and "fi-ketonic" fractions by the usual techniques (19). The "cz-ketosteroid" fractions were further analyzed by adsorption and partition chromatography employing silica gel columns. Alumina and/or magnesium silicate columns were employed in certain fractionations. Individual steroid fractions were identified by infrared spectrometry and quantitated by the modified Zimmermann reaction.The further details of the analytical procedures employed in these studies have already been described elsewhere (19,22). V...

Urinary Excretion of 17-Hydroxy-20-Oxosteroids in Normal Health and in Rheumatoid Arthritis

“…Moreover, the detection of 17-ahydroxypregnanolone (I) in normal urine (Dobriner and others, 1954), together with the evidence here presented, clearly show that, contrary to earlier indications (Dobriner, 1951;Dobriner and others, 1951), an abnormal excretion of (I) has not been demonstrated in rheumatoid arthritis. We wish to emphasize that present knowledge of the steroid excretion pattern in rheumatoid arthritis, or for that matter in any other condition, is only fragmentary, and hence that the conclusions here reached cannot be construed as arguments against a hypothetical causal relation between a derangement of steroid metabolism, or, in particular, between a defect in the adrenocortical function and rheumatoid arthritis.…”

“…In the course of their outstanding investigations into the urinary excretion of steroids, Dobriner (1951) and Dobriner and others (1951) reported the isolation of 17-a-hydroxypregnanolone (I) from the urine of four out of five patients with rheumatoid arthritis. The daily excretion of this compound (I) recorded for one patient only amounted to 1 5 mg.…”

A clinical and pathological study of resistance to infection in chronic lymphatic leukemia

“…Dobriner et al 4 have found a decreased excretion of ACS breakdown products in patients with chronic lymphatic leukemia. This implies an abnormality in chronic lymphatic leukemia to some extent corrected by ACS.…”

“…This abnormality may be either a deficiency of a specific adrenal hoimone concerned with lymphocyte metabolism, in a manner comparable to hy~)oaItlosteronisni without Addison's disease, or an end organ abnormality in the utilization of ACS by lymphoid tissue. Dobriner et al 4 have found a decreased excretion of ACS breakdown products in patients with chronic lymphatic leukemia. Doughertyh has shown that lymph node fibroblasts and some lymphocytes are capable of converting cortisone (compound E) to hydrocortisone (compound 17).…”

Study of the Genetic and Extra-Genetic Determinants of Alpha-Ketosteroid Production in Man

“…These steroid patterns differ quantitatively from person to person (1, 2) but are reproducible and constant (7) for a given individual over periods of time. Although subject to alteration by such influences as endocrine (8)(9)(10) and nonendocrine disease (11,12) or by exogenous hormone administration (2, 8, 9, 13, 14) the principal determinants of these patterns under physiological conditions apparently remain well defined. The results of the present study indicate that both genetic and extra-genetic factors play a significant role in determining the characteristics of the steroid pattern in man.…”