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Reactive Splenomegaly and Variation of Lymphoid-Plasma Cell Population Associated with a Mouse Pituitary Thyrotropic Tumor*

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Reactive Splenomegaly and Variation of Lymphoid-Plasma Cell Population Associated with a Mouse Pituitary Thyrotropic Tumor* Reactive Splenomegaly and Variation of Lymphoid-Plasma Cell Population Associated with a Mouse Pituitary Thyrotropic Tumor* EDITH E. SPROULANDRAUL GRINBERG (Rosiceli Park Memorial Institute and New York Slate Department of Health, Buffalo, New York) SUMMARY A transplantable thyrotropic pituitary tumor, subline L24a, which grew after 13 months of dormancy in an intact mouse was associated in its early passage with a variety of lymphoid tumors. On subsequent passage, slow growth over a 6- to 12- month period was regularly accompanied by striking splenomegaly without lymphoid or thymic neoplasia. The enlargement was due to a great increase in cells of the lymphoid series in the red pulp without loss of splenic architecture. In thyroidec- tomized mice plasma cells with many Russell bodies were especially numerous, and these cells were found hi hypertrophied lymphoid follicles within other organs. It is suggested that the splenomegaly and, hi earlier passage the lymphoid tumors, may represent an exaggerated antibody response to a tumor's having unusual or abnormal hormonal activity. The transplanted pituitary tumors, in turn, showed severe hemor- rhagic degeneration with peripheral small lymphocyte stasis in the lymphatics. Initial observations concerning pleomorphic reticulum through the courtesy of Dr. Jacob Furth, was studied cell- and lymphosarcomas or lymphoid hyperplasias through repeated passage for purposes of comparison. regularly accompanying a thyrotropic mouse pituitary This tumor has maintained uniform behavior with rapid neoplasm, subline L24a, have been reported (22). The autonomous growth, active secretion of TSH, and a sex original L24 pituitary tumor was developed by Dr. Jacob dependence which was lacking in the L24 line. A mam- Furth by I131thyroid ablation. At the outset of these motropic tumor, 38, from Dr. Furth's collection was experiments this tumor was an autonomous growth but carried in similar manner to provide further comparison highly responsive to suppression by thyroid hormone; of the effects on mouse lymphoid tissues of a nonthy- implants into intact mice failed to progress or remained rotropic pituitary tumor. This neoplasm grew slowly and dormant for approximately 13 months (27). Growth of irregularly during the period of this study but maintained one tumor, designated L24a, having a latent period of its hormonal effect on the mammary gland. over a year, in a thyroid-intact mouse was marked by Technical procedure.—Allexperiments were conducted pronounced lymphomogenesis or hyperplasia of cells of on (C57L X A)Fi hybrid female and male mice (hereafter the lymphoid group in all its subsequent passages. The called LAFi), obtained from the Jackson Memorial present report concerns the behavior of this tumor during Laboratory, Bar Harbor, Maine. The technical details continued passage, with detailed study of the cell types of thyroid ablation, tumor transfer, autopsy procedure, affected under altered hormonal conditions. tissue section preparation, and multiple stains were es sentially as previously described (22), with some vari MATERIALS AND METHODS ations. Tumors were implanted either intramuscularly Tumors.—The association gains interest with the find into the thigh or into the dorsal subcutaneous tissue. The ings in the third transfer of subirne L24a. In accord with fixative for all organs contained 85 per cent ethyl alcohol, the behavior of many animal tumors having spécule 5 per cent acetic acid, and 40 per cent formaldehyde, a function, continued passage has yielded a more autono modification by Lillie of Carnoy's solution which was mous growth having reduced or altered specific activity especially successful in maintaining nuclear fine structure (7). With the use of an older tumor as donor tissue it (17). Cytoplasmic RNA is best preserved if the tissues was possible to follow the effects of a less active growth remain in Lillie's fixative for not more than 2 hours and for a period of 12 months after implantation. are then transferred to 80 per cent alcohol. A second thyrotropic neoplasm, 17, also obtained Wright-Giemsa differential stain was applied to im * Supported in part by N.I.P.H. Grant CA-06855. prints of the splenic cut surface. Imprints were necessary Received for publication May 18, 1964. for more accurate identification of cell types but required 1644 Downloaded from cancerres.aacrjournals.org on October 2, 2021. © 1964 American Association for Cancer Research. SPROULAND GRINBERG—Splenomegalyand Thyrotropic Tumor 1645 correlation with sections to locate the various changes. nature. A few mice of each group died spontaneously In later studies more uniform smears of spleen cells have and were discarded. The remainder were permitted to been prepared with a fine camel's hair brush and dextran. live until moribund and were bled from the right heart or Methyl green pyronine stain1 was used to facilitate the vena cava under ether anesthesia prior to the autopsy. identification of plasma cells in imprints and in sections. All organs were weighed and sectioned. Less variegation and greater intensity of cytoplasmic The spleen weights of control mice in Group C, examined staining follows the substitution of thionine for pyronine mainly 9-12 months after the beginning of the experi in combination with methyl green.2 This preparation ment, were close to 100 nig., with the exception of one was applied to tissue sections and smears in the later instance of spontaneous myeloid leukemia (Chart 1). studies. Most of the tumor grafts grew to lethal size earlier in in Protocol of major experiment.—The subline L24a thy- tact mice (Group B), with another peak in tumor growth rotropic tumor had been carried through two passages as in the later months. Thyroidectomized mice (Group A), reported (22). The mice in the third passage were or examined largely in the 6- to 8-month period after tumor ganized into three groups: implantation, developed the largest spleens—several Group A: 30 mice radiothyroidectomized with 50 nc. above 2,000 mg. I131and given injections intramuscularly of tumor. Histologie and cytologie character of the splenopathy.—- Group B: 30 intact mice also receiving tumor. Group A: Radiothyroidectomized, tumor-bearing mice Group C : 30 control mice kept through the year of this maintained the usual splenic architecture even in the experiment. most enlarged organs (Fig. 3). The lymphoid follicles Equal numbers of male and female mice comprised were large and poorly demarcated, owing to an increase in each group. All were about 8 weeks of age and were reticulum cells, large lymphocytes, and especially plasma maintained on Rockland Farm rat diet and tap water cells in the perifollicular zone. These cells were also except prior to thyroidectomy, when Remington low prominently increased throughout the red pulp, account iodine diet3 and distilled water were substituted. ing in good part for the splenic enlargement. Mast cells were also increased 3-4 times over the number in control RESULTS4 spleens. The L24a tumor grew more slowly than previously with Plasma cells appeared in several stages of maturity, a latent period of 4-8 months, appearing first in intact recognized by their large or small, coarse, round eccentric male mice. "Reverse responsiveness" to the target organ nucleus and abundant pyrininophilic cytoplasm. The hormone has been previously noted (14). Eventually, spleens of mice bearing tumors of long duration frequently all but nine of the GOtumor grafts progressed, but those persisting 6-10 months after implantation were often mgi hemorrhagic and in good part necrotic. There were 2100 Z500 Z800 tumor cells which appeared viable in histologie sections, 00« o 1900 but the amount of active tumor in the large hemorrhagic O masses could not be evaluated. From the general pattern of behavior it was clear that 1700 the L24a thyrotropic tumor was autonomous but still influenced by thyroid and ovarian hormone secretion. 1500. LEUKEMIA TSH secretion was greatly reduced. Nevertheless, the grafted tumor was regularly accompanied by splenomegaly O of 10-30 times the normal size, with a disproportionate 52 1300 o «o o moderate response in lymph nodes and lymphoid nodules 0° ° of organs (Figs. 1, 2). Splenic enlargement occurred in 1100. all mice bearing tumors, with or without the thyroid gland, and failed to develop in the few mice given inocu 900. BL lations in which tumor did not grow. There was no cor CO relation with size of the tumor, since, in a few instances, 700. tumor nodules were found only after careful sectioning of the host's thigh muscle. Comparable observations are 500. made in regard to human endocrine tumors; minute occult nodules may have striking systemic effect of lasting 300. 1The most successful methyl green pyronine was obtained from Chroma-Gesellschaft Schmid Co., distributed by Roboz Surgical 100. CC C0„ Instrument Co., Washington, D. C. 2Contributed by personal communication from Dr. A. Roqué, -n 5 6 7 8 9 IO te Roswell Park Memorial Institute, Buffalo, New York. C CONTROL 3Obtained from the Nutritional Biochemicals Corp., Cleveland • INTACT MONTHS AFTER INOCULATION 28, Ohio. O SO JIC 1131 * Presented in part at Annual Scientific Session of American Association for Cancer Research, Toronto, Canada, May, 1963 CHART 1.—Spleen weights of LAFi mice with L24a thyrotropic (21). tumor. Downloaded from cancerres.aacrjournals.org on October 2, 2021. © 1964 American Association for Cancer Research. 1646 Cancer Research Vol. 24, October 1964 were populated by plasma cells with large protoplasmic nodes in older mice of many strains. Their presence in masses which were pyronine-positive and stained mod greater numbers in Group A mice was not related to age erately with periodic acid-Schiff reagent, indicating their and contrasted with the appearance of lymph nodes in glycoprotein nature. Similar masses were also found the control group. extracellularly and were recognized as Russell bodies, The thymus was not significantly altered in size or which have been variously interpreted as the result of histologie character in tumor-bearing mice (Fig.
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