Acta Radiologica: Therapy, Physics, Biology
ISSN: 0567-8064 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/ionc17
Thyroid Hormones and Lymphoid Cells G. Lundell & H. Blomgren To cite this article: G. Lundell & H. Blomgren (1975) Thyroid Hormones and Lymphoid Cells, Acta Radiologica: Therapy, Physics, Biology, 14:6, 552-560, DOI: 10.3109/02841867509132697 To link to this article: https://doi.org/10.3109/02841867509132697
Published online: 08 Jul 2009.
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(DIRECTOR: STOCKHOLM, SWEDEN.
FROM RADIUMHEMMET, GENERAL DEPARTMENT
PROF. J. EINHORN), S-104 01
THYROID HORMONES AND LYMPHOID CELLS G . LUNDELL and H. BLOMGREN
Hyperplasia of the peripheral lymphatic tissue, thymic enlargement and lymphocytosis of the bone marrow have been observed in patients with hyperthyroidism & PETERSON 1927, MARGOLIS 1931, HAMand thyroiditis (CROTTI1922, BALDRIDGE MAR 1936, AXELROD & BERMAN 1951, DOUGHERTY 1952, IRVINE & SUMERLING 1965, MICHIEet coll. 1967). Similar changes were, however, not observed in the lymphatic system of patients with non-toxic goitres (MICHIEet coll.). Since the amount of thymic tissue decreased following antithyroid drug therapy, it is conceivable that hyperplasia of the thymus was a direct consequence of raised levels of circulating thyroid hormones (MICHIEet coll.). Recently it has been shown that the incorporation of 14C-thymidine of lymphoid cells in vitro is increased in patients with hyperthyroidism as compared with euthyroid individuals, and that these increased values were normalized after 1311 therapy (LUNDELLet coll. 1976). The increased levels of 14C-thymidine incorporation by lymphoid cells in vitro were not caused by a shift in the proportion of thymus dependent lymphocytes (T-cells) and non-T-lymphocytes (mainly B-cells) in hyperthyroid as compared with euthyroid individuals (LUNDELL et coll. 1975). Moreover, there is documentation demonstrating that the peripheral lymphocytes in some patients with hyperthyroidism have increased size (HERNBERG 1954) and increased RNA-content (SMITHet coll. 197I). Thymolymphatic hyperplasia may also ~~
This investigation was supported by the Cancer Society in Sweden. Submitted for publication 23 July 1975.
552
THYROID HORMONES AND LYMPHOID CELLS
553
be induced in experimental animals by the administration of thyrotrophic or thyroid hormones (GR~~GOIRE 1942, REINHARDT & WAINMAN 1942, GYLLENSTEN 1953, LUNDIN 1958, ERNSTROM & GYLLENSTEN 1959, ERNSTROM 1963). These findings strongly indicate that thyroid hormones induce mitosis of lymphocytes in vivo. The present investigation was performed to find out whether thyroid hormones induced DNA-synthesis of human lymphoid cells in vitro.
Material and Methods Separation of lymphoid cells. Venous blood was obtained from healthy euthyroid individuals. It was defibrinated by gentle agitation in a beaker containing glass pearls. Separation of nucleated cells was performed according to the method described by COULSON & CHALMERS (1964). The number of lymphoid cells was determined in a Biirker chamber after crystal violet staining. The cell-suspensions used for the experiments contained a minimum of 60 per cent of lymphocytes. Thyroid hormones. The sodium salts of levo-triiodothyronine (L-TJ and levothyroxine (L-T,) were obtained from Nyegaard and Co. AB, Stockholm, Sweden, sodium dextro-thyroxine (D-T,) from Travenol Laboratories Inc., Ill., USA, and sodium dextro-triiodothyronine (D-T,) from E. Merck Svenska AB, Stockholm, Sweden. The thyroid hormones were suspended in sterile water, the pH was adjusted to 11.9 with sodium hydroxide, and after millipore filtration were immediately used. Thyroid hormones in twofold dilution starting with 0.1 mg (dilution 1/1) were added to the cell cultures. In a few experiments, higher concentrations of the hormones were used. In each case, however, the added volume was 0.1 ml. In some experiments (Tables 1, 2) sodium hydroxide was added to control cultures at a dose equivalent to that present at a hormone dilution of 1/1. Sodium iodide. In order to test the eventual influence upon the lymphoid cell responsiveness to iodide liberated through a possible decomposition of the thyroid hormones, sodium iodide (pro analysi, E. Merck AG., Darmstadt, Germany) was added to the cell cultures as described for the thyroid hormones. Cell cultures. 2.0 x los lymphoid cells were cultured in screw cap glass tubes with an inner diameter of 15.5 mm and with rounded bottoms (Johnson & Jorgensen Ltd., London, England). Each contained 1.0 ml of Eagle’s Minimal Essential Medium supplemented with Earle’s salts (MEM) supplemented with 100 units of penicillin and 150 pg of streptomycin per ml and 10 per cent human serum from AB-positive blood donors. The serum was made complement deficient by incubation at 56°C for 30 minutes before use, and stored at -20°C. Varying amounts of thyroid hormone served as controls. After 4 days incubation at 37°C in a humidified atmosphere containing 5 per cent CO,, each culture received 0.1 ml of MEM containing 0.4
554
G. LUNDELL AND H. BLOMGREN
Table 1 Influence of thyroid hormones on the 14C-thymidine incorporation of lymphoid cells. Mean values standard errors (S. E.) were calculated from I0 cultures. Dilution 111 corresponds to 0.1 mg of L-T4 or D-T, per tube, The number of cpm for the control tubes without added NaOH, was 18 173 kS.E. 1308
Hormone
Dilution
Incorporation of thymidine, No. of counts/min Mean
S.E. 1741 1298 1144
L-T, L-TI L-T4
111 112
114
27 498 20 060 17 292
D-T4 D-T, D-T.,
1/1 1 /2 114
26 950 22 276 17 995
1128 1035 1281
Control, no hormone, NaOH added
-
17 072
I354
Level of significance for the difference to control, p
< 0.00 I -
ISSN: 0567-8064 (Print) (Online) Journal homepage: http://www.tandfonline.com/loi/ionc17
Thyroid Hormones and Lymphoid Cells G. Lundell & H. Blomgren To cite this article: G. Lundell & H. Blomgren (1975) Thyroid Hormones and Lymphoid Cells, Acta Radiologica: Therapy, Physics, Biology, 14:6, 552-560, DOI: 10.3109/02841867509132697 To link to this article: https://doi.org/10.3109/02841867509132697
Published online: 08 Jul 2009.
Submit your article to this journal
Article views: 45
View related articles
Citing articles: 3 View citing articles
Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=ionc17
(DIRECTOR: STOCKHOLM, SWEDEN.
FROM RADIUMHEMMET, GENERAL DEPARTMENT
PROF. J. EINHORN), S-104 01
THYROID HORMONES AND LYMPHOID CELLS G . LUNDELL and H. BLOMGREN
Hyperplasia of the peripheral lymphatic tissue, thymic enlargement and lymphocytosis of the bone marrow have been observed in patients with hyperthyroidism & PETERSON 1927, MARGOLIS 1931, HAMand thyroiditis (CROTTI1922, BALDRIDGE MAR 1936, AXELROD & BERMAN 1951, DOUGHERTY 1952, IRVINE & SUMERLING 1965, MICHIEet coll. 1967). Similar changes were, however, not observed in the lymphatic system of patients with non-toxic goitres (MICHIEet coll.). Since the amount of thymic tissue decreased following antithyroid drug therapy, it is conceivable that hyperplasia of the thymus was a direct consequence of raised levels of circulating thyroid hormones (MICHIEet coll.). Recently it has been shown that the incorporation of 14C-thymidine of lymphoid cells in vitro is increased in patients with hyperthyroidism as compared with euthyroid individuals, and that these increased values were normalized after 1311 therapy (LUNDELLet coll. 1976). The increased levels of 14C-thymidine incorporation by lymphoid cells in vitro were not caused by a shift in the proportion of thymus dependent lymphocytes (T-cells) and non-T-lymphocytes (mainly B-cells) in hyperthyroid as compared with euthyroid individuals (LUNDELL et coll. 1975). Moreover, there is documentation demonstrating that the peripheral lymphocytes in some patients with hyperthyroidism have increased size (HERNBERG 1954) and increased RNA-content (SMITHet coll. 197I). Thymolymphatic hyperplasia may also ~~
This investigation was supported by the Cancer Society in Sweden. Submitted for publication 23 July 1975.
552
THYROID HORMONES AND LYMPHOID CELLS
553
be induced in experimental animals by the administration of thyrotrophic or thyroid hormones (GR~~GOIRE 1942, REINHARDT & WAINMAN 1942, GYLLENSTEN 1953, LUNDIN 1958, ERNSTROM & GYLLENSTEN 1959, ERNSTROM 1963). These findings strongly indicate that thyroid hormones induce mitosis of lymphocytes in vivo. The present investigation was performed to find out whether thyroid hormones induced DNA-synthesis of human lymphoid cells in vitro.
Material and Methods Separation of lymphoid cells. Venous blood was obtained from healthy euthyroid individuals. It was defibrinated by gentle agitation in a beaker containing glass pearls. Separation of nucleated cells was performed according to the method described by COULSON & CHALMERS (1964). The number of lymphoid cells was determined in a Biirker chamber after crystal violet staining. The cell-suspensions used for the experiments contained a minimum of 60 per cent of lymphocytes. Thyroid hormones. The sodium salts of levo-triiodothyronine (L-TJ and levothyroxine (L-T,) were obtained from Nyegaard and Co. AB, Stockholm, Sweden, sodium dextro-thyroxine (D-T,) from Travenol Laboratories Inc., Ill., USA, and sodium dextro-triiodothyronine (D-T,) from E. Merck Svenska AB, Stockholm, Sweden. The thyroid hormones were suspended in sterile water, the pH was adjusted to 11.9 with sodium hydroxide, and after millipore filtration were immediately used. Thyroid hormones in twofold dilution starting with 0.1 mg (dilution 1/1) were added to the cell cultures. In a few experiments, higher concentrations of the hormones were used. In each case, however, the added volume was 0.1 ml. In some experiments (Tables 1, 2) sodium hydroxide was added to control cultures at a dose equivalent to that present at a hormone dilution of 1/1. Sodium iodide. In order to test the eventual influence upon the lymphoid cell responsiveness to iodide liberated through a possible decomposition of the thyroid hormones, sodium iodide (pro analysi, E. Merck AG., Darmstadt, Germany) was added to the cell cultures as described for the thyroid hormones. Cell cultures. 2.0 x los lymphoid cells were cultured in screw cap glass tubes with an inner diameter of 15.5 mm and with rounded bottoms (Johnson & Jorgensen Ltd., London, England). Each contained 1.0 ml of Eagle’s Minimal Essential Medium supplemented with Earle’s salts (MEM) supplemented with 100 units of penicillin and 150 pg of streptomycin per ml and 10 per cent human serum from AB-positive blood donors. The serum was made complement deficient by incubation at 56°C for 30 minutes before use, and stored at -20°C. Varying amounts of thyroid hormone served as controls. After 4 days incubation at 37°C in a humidified atmosphere containing 5 per cent CO,, each culture received 0.1 ml of MEM containing 0.4
554
G. LUNDELL AND H. BLOMGREN
Table 1 Influence of thyroid hormones on the 14C-thymidine incorporation of lymphoid cells. Mean values standard errors (S. E.) were calculated from I0 cultures. Dilution 111 corresponds to 0.1 mg of L-T4 or D-T, per tube, The number of cpm for the control tubes without added NaOH, was 18 173 kS.E. 1308
Hormone
Dilution
Incorporation of thymidine, No. of counts/min Mean
S.E. 1741 1298 1144
L-T, L-TI L-T4
111 112
114
27 498 20 060 17 292
D-T4 D-T, D-T.,
1/1 1 /2 114
26 950 22 276 17 995
1128 1035 1281
Control, no hormone, NaOH added
-
17 072
I354
Level of significance for the difference to control, p
< 0.00 I -
