A strain difference in the daily rhythm of glyceraldehyde-3-phosphate dehydrogenase activity in the mouse

Journal of Comparative Physiology A - A/J mice differ from C57BL/6J mice in the time of the daily peak of activity of glyceraldehyde-3-phosphate dehydrogenase (GAPD) in thymus and in thyroid....     

Strain dependent response of circadian rhythms during exposure to continuous illumination

Enzymes activities were measured, at three hours intervals, during 30 hours, in various tissues of C57BL/6J and A/J male mice. The measurements, were carried out on mice which were exposed for two, five and twenty one days to continuous illumination. Identical measurements were performed also on mice which were kept in alternating 14 hours light: 10 hours dark. Activity patterns of each group were analysed to test the presence, or absence, of rhythm characteristics. The results of the experiments with C57BL/6J have been previously reported. The comparison of the results, which were obtained from the two strains revealed that under exposure to alternating light: dark conditions all activity patterns exhibited a significant circadian rhythm. Except for one enzyme (thymus GAPD), the times of peak activity (acrophase) were identical for all other examined enzymes, in both strains. On the other hand when the two strains were exposed to continuous illumination they differed in their response to the effect of continuous light. The activity of the same enzyme exhibited different periodicity and/or different acrophase in each of the two strains. This variability reflects the existence of genetic differences, between the strains in the free running behavior of these enzymes' activity rhythms.     

Deletion of self-reactive T cells in nude mice grafted with neonatal allogeneic thymus

The cellular basis of tolerance induction has been investigated in BALB/c(H-2d, thy 1.2, M1s1b2a) nude mice grafted with thymus of neonatal AKR/J mice(H-2k,Thy1.1,M1s1a2b). The spleen cells from nude mice grafted with AKR/J thymus showed a significantly decreased level of primary cytotoxic T cell response when stimulated with AKR/J cells, although these cells lysed well target cells of a third party C57BL/6 when stimulated with C57BL/6 cells. Consistent with CTL responses, T cells bearing V beta 6, that is important for recognizing M1s1a-encoded products of the thymic phenotype, were virtually abolished in the spleen and lymph node cells of nude mice 8 wk after grafting with AKR/J thymus. However, a substantial number of V beta 6-bearing T cells were detected in the peripheral organs of nude mice 23 wk after grafting with AKR/J thymus and in those of nude mice grafted with AKR/J fetal thymus depleted of macrophages/dendritic cells by incubating with 2'-deoxyguanosine in vitro before grafting. On the other hand, T cells bearing V beta 3, which are selectively related to M1s2a-encoded products of the host phenotype, were expressed neither on the peripheral T cells of nude mice grafted with AKR/J thymus at any stage after grafting nor on those of nude mice grafted with 2'-deoxyguanosine-treated AKR/J thymus. These data suggested that both V beta 6 and V beta 3 T cells were eliminated in the thymus of nude mice grafted with AKR/J thymus, presumably on the basis of interaction with both of graft-derived persisting and host-derived hemopoietic cells in the thymus and that thymic epithelium appears to have little capacity to eliminate T cells reactive to minor lymphocyte stimulating-encoded products.     

B lymphocyte colony-forming cells in the SJL/J mouse.

Mercatoethanol-induced B lymphocyte cloning in semi-solid agar has been used to study lymphocyte colony formation by cells from the SJL/J mouse thymus. From the 3rd month of life, the SJL/J mouse thymus. From the 3rd month of life, the SJL thymus develops an increasing frequency of cells forming B lymphocyte colonies in agar. The peak frequency in 6- to 12-month-old mice was one colony per 1000 to 2000 cultured thymus cells. In contrast, 10 to 100 times lower frequencies were found in the thymus of five other inbred mouse strains. The rise in B lymphocyte colony-forming cells correlated well with the age-related rise in Ig-positive cells and approximately 50% of the colony cells reacted with anti-micron-serum indicating the B lymphocyte nature of the colony cells. Colony-forming cells from the thymus showed higher sensitivity than colony-forming spleen cells to cortisol and irradiation. Cell transfer experiments and thymus grafting suggested that the increased frequency of colony-forming cells in the thymus is caused by development of special thymus-seeking B lymphocytes in ageing SJL/J mice. Finally, B lymphocyte colony-forming cells were found to be more frequent in the thymus, spleen, and lymph nodes from healthy aged mice than in lymphoid organs from mice with spontaneous reticulum cell tumors.     

Thyroid hormone levels in the developing obese-hyperglycemic syndrome.

Examination of the thyroid hormone levels of C57BL/6J ob/ob mice and their lean littermates between the ages of 6 days and 22 weeks revealed that obese mice have significantly reduced hormone concentrations between 10 and 21 days of age. Thereafter, the values remained equal to, or above those or their lean littermates. The present data indicate that the hypometabolism and abnormal thermal regulation of weanling mice could be a result of diminished thyroid hormone levels, but that other mechanisms must be responsible for the persistence of these abnormalities in adult ob/ob mice.     

Role of neuroendocrine hormones in murine T cell development. Growth hormone exerts thymopoietic effects in vivo.

Growth hormone (GH) and other neuroendocrine mediators have been implicated previously in T cell development. We therefore examined thymic development in DW/J dwarf mice. DW/J mice lack acidophilic anterior pituitary cells and consequently are totally deficient in the production of GH, as well as other neuroendocrine hormones. DW/J dwarf mice had greatly hypoplastic thymi that continued to decrease in size as the mice aged. Characterization of the different T cell subpopulations within the thymi of dwarf mice indicated a deficiency in CD4+/CD8+ double-positive thymocytes. This deficiency of progenitor cells became more complete as the mice aged culminating in the total disappearance of this subpopulation in some dwarf mice > 3 mo of age. Analysis of the lymph nodes indicated that a population of double-positive (CD4/CD8) T cells appeared in some mice concurrent with the disappearance of double-positive cells in the thymus suggesting that these thymocytes may have migrated to the periphery. However, peripheral T cells from dwarf mice did exhibit Ag-specific responses indicating that these mice have functional T cells. Treatment of the mice with recombinant human GH, which binds both murine growth hormone receptors and murine prolactin receptors, or ovine GH, which binds murine growth hormone receptors but not murine prolactin receptors, resulted in an increase in thymic size and the reappearance of the CD4+/CD8+ double-positive cells within the thymus. Additionally, after GH treatment, the double-positive cells disappeared from the lymph nodes. The thymi of mice treated with GH failed to attain normal size but did develop a normal distribution of T cell progenitors. Thus, GH exerts significant thymopoietic effects in vivo. Neuroendocrine hormones may be important for normal T cell differentiation to occur within the murine thymus.     

电磁脉冲对C57BL/6J 胸腺的作用

目的:观察电磁脉冲Electromagnetic Pulse(EMP)对C57BL/6J小鼠胸腺的影响。方法:50只C57BL/6J小鼠按体重区组随机化分为对照组和辐照组,每组25只。EMP每天照射400次,连续照射7天,照后1d(天)、3d、7d、14d、28d共5个时间点杀取胸腺。2只辐照组和2只对照组杀取的胸腺做HE染色,观察其病理改变;3只辐照与3只对照组的小鼠,杀后称取小鼠的体重和胸腺的重量,计算胸腺指数;然后提取T淋巴细胞进行计数;同时取小鼠外周血检测其中的IL-4的水平。结果:照后1d胸腺的切片没有明显改变。7天后,胸腺开始有出血,结构不清;胸腺指数呈现递减的趋势,但辐照组与对照组没有的差异没有统计学意义;T淋巴细胞数的变化也呈现先减后增的趋势,在第1d、14d、28d辐照组与对照组的差异没有统计学意义,在第3d和7d辐照组的细胞数小于对照组的细胞数(P<0.05);辐照组与对照组的外周血IL-4水平的差异也没有统计学意义。结论:电磁脉冲对雄性Balb/c小鼠胸腺结构造成一定的损伤,但胸腺指数改变不显著,T淋巴细胞数量增加。表明EMP对胸腺有一定的作用,但是胸腺不是EMP作用的敏感器官。     

Effect of cortisone in delineating thymus cell subsets in advanced age.

The mixed lymphocyte reactivity of thymus cells from normal and cortisone-treated, young and old C57BL/6J mice was evaluated. The results indicated that while in the young mice cortisone treatment caused a 12-fold increase in the reactivity of thymus cells, it had no effect on the response of thymus cells from old mice. The significance of the disappearance of cortisone-resistant mature thymus cells in old mice is discussed regarding the general decline in immunocompetence and the increase of autoimmune manifestations in old age.     

Suppressive effects of Lactobacillus casei cells,a bacterial immunostimulant,on the incidence of spontaneous thymic lymphoma in AKR mice

 The mean survival age of female AKR/J mice was significantly prolonged, the enlargement of thymus was markedly suppressed, and the proliferation of ecotropic and recombinant murine leukemia viruses (MuLV) was markedly inhibited when 8-week-old female AKR/J mice were injected intraperitoneally (i. p.) with heat-killed Lactobacillus casei cells twice weekly for 8 weeks. In contrast, such actions of heat-killed L. casei cells were not seen in 20-week-old female AKR/J mice. The leukemogenic activity of the cell-free extract of thymus from adult female AKR/J mice in newborn female AKR/J mice was drastically reduced by i. p. treatment with heat-killed L. casei cells. The difference in adjuvant effectiveness of heat-killed L. casei cells on 8- and 20-week-old animals may be dependent on the difference in the enhancing activity of the cell-mediated immune systems between the groups induced by heat-killed L. casei cells, and, as a result, on the difference in the degree of proliferation of ecotropic and recombinant MuLV in thymus, which consequently causes thymic lymphoma. Received: 13 February 1996 / Accepted: 18 April 1996     

A positive difference in nature of envelopes of thymus- and uterus-derived leukemia viruses of AKR mice.

Thymus-derived leukemia virus of AKR/J mice was inactivated by anti-theta antiserum. But it was not inactiviated by the antiserum which had been absorbed with intact thymus cells of AKR/J or RF/J mice, and by anti-uterus-derived leukemia virus antiserum. In contrast, uterus-derived leukemia virus of the strain was not inactivated by anti-theta antiserum, but was neutralized by anti-uterus-derived leukemia virus antiserum. The results suggest the possibility that some constitutents of the envelope of thymus-derived leukemia virus are derived from the plasma membrane of thymus cells of AKR mice at the time of budding and that such constituents are not associated with the enveloped of uterus-derived leukemia virus.     

Association of recombinant murine leukemia viruses of the class II genotype with spontaneous lymphomas in CWD mice

We determined the phenotype and genotype of murine leukemia viruses associated with the development of spontaneous nonthymic lymphomas in the high-leukemia mouse strain CWD/J. By T1 oligonucleotide fingerprint analysis of the viral RNA, the ecotropic viruses recovered from the spleen or thymus of preleukemic CWD/J mice were found to represent the progeny of the two endogenous ecotropic proviruses present in this strain. Polytropic murine leukemia viruses were produced by tissues from one-half of the leukemic mice, and fresh tumor cells from one of the two animals tested expressed recombinant envelope glycoproteins. The genomic structure of the recombinant viruses resembled those of class II polytropic viruses of NFS X Akv mice and differed from those of class I recombinant viruses that are commonly isolated from other high-leukemia strains such as AKR and HRS. Acquired retroviral sequences with the structural features of class II recombinant proviruses were detected in the DNA from each CWD/J tumor by the Southern blot technique. Finally, the injection of a mixture of CWD/J ecotropic and class II recombinant polytropic viruses into neonatal CWD/J mice accelerated the onset of lymphoma, whereas the endogenous ecotropic virus was inactive in these assays.     

Relationship between genetic differentiation of the thymus in mice of different strains and malignant growth. IV. Genetic analysis of the thymic index in mice]

Relative thymus weight was estimated in C3H/He, C57BL/6 mice, their F1 and backcross hybrids, as well as in the progeny of complete diallel crosses between the BALB/c, C3H/He, C57BL/6 and AKR/J strains. On the basis of the analysis of these measurements, a conclusion is drawn that this character is inherited with incomplete dominance of smaller relative weight. The genes determining greater thymus weight are concentrated in the genetic pool of AKR/J and C57BL/6 strains. These genes are characterized by some degree of recessivity with respect to the genes determining smaller thymus weight which are concentrated in the genetic pool of C3H/He and BALB/c strains. The highest concentration of the "plus" and "minus" genes is found in the genetic pools of AKR/J and C3H/He strains respectively.     

The expression and chromatin structure of the chicken glyceraldehyde-3-phosphate dehydrogenase gene in mouse cells

Chicken glyceraldehyde-3-phosphate dehydrogenase gene (GAPD) and thymidine kinase gene (TK) were co-transfected into mouse LMTK- cells by the calcium phosphate precipitation technique. Four of the eight hypoxanthine/aminopterin/thymidine-containing medium-resistant, TK+ transfectants were shown to produce different amounts of chicken glyceraldehyde-3-phosphate dehydrogenase by zymogram analysis. Subcloning and further analysis revealed that the chicken GAPD was stably inherited and that its enzyme subunits randomly combined with mouse subunits in heterotetramers. Although the contribution of chicken enzyme varied from approximately 30 to approximately 90% of the total glyceraldehyde-3-phosphate dehydrogenase activity with a proportional increase in total activity in the different subclones, it did not appear to affect the expression of mouse endogenous glycolytic enzymes since there was no distinct change in the levels of either mouse glyceraldehyde-3-phosphate dehydrogenase mRNA nor mouse phosphoglycerate kinase enzyme activity. The levels of chicken GAPD copy number, mRNA, and enzyme apparently were generally correlated in the different subclones, suggesting that the chicken GAPD in the mouse cells were expressed constitutively. In situ hybridization revealed that the transfected genes were integrated into mouse chromosomes in one cluster, and the locations of these clusters were different in different clones. Chromatin structure analyses of the chicken GAPD in four different transfectants revealed three DNase I-hypersensitive sites located around 0.2, 2.0, and 3.4 kilobases upstream from the 5' side of the gene. These sites are also present in the same locations in chicken lymphoblastoid cells (Kuo, M. T., Iyer, B., and Schwartz, R. J. (1982) Nucleic Acids Res 10, 4565-4579), indicating the dominant transmission of DNase I-hypersensitive cleavage sites in the transfected gene.     

Evidence for the existence of distinct heterogeneity among the peripheral CD4-CD8- T cells from MRL-lpr/lpr mice based on the expression of the J11d marker, activation requirements, and functional properties

Autoimmune-susceptible, MRL-lpr/lpr (lpr) mice develop a profound lymphadenopathy resulting from the accumulation of CD4-CD8- (double-negative, DN) cells in peripheral lymphoid organs. The source and the mechanism of this abnormal accumulation of cells is still unknown. Recently, we reported that a significant number (approximately 35%) of the CD4-CD8- cells expressed J11d, a marker expressed by immature thymocytes but not by mature functional peripheral T cells. In the present study, we investigated the phenotype, growth requirements, and functional properties of purified J11d+ and J11d- subpopulations. Using the mAb, F23.1, which recognizes a TCR determinant encoded by the V beta 8 gene family, it was observed that approximately 30% of the J11d+ and J11d- DN cells expressed this determinant. Further studies on the thymus revealed that J11d+ DN cells from lpr thymus also contained F23.1+ cells (approximately 25%), whereas, similar cells from normal MRL(-)+/+mice were all F23.1-, consistent with earlier reports in other normal strains. Further phenotypic studies revealed that the peripheral J11d+ and J11d- cells from lpr mice were similar in expressing CD3, Ly-5 (B220), and Ly-24 (Pgp-1) determinants. When stimulated with phorbol myristic acetate (PMA) and recombinant IL-2 (rIL-2), only J11d- cells but not J11d+ cells responded by proliferation. However, in the presence of calcium ionophore (A23187) and PMA, both J11d+ and J11d- subpopulations proliferated by producing and responding to endogenous IL-2 but not IL-4. The lymph node T cells from 1-month-old MRL-lpr/lpr mice responded strongly when stimulated with PMA + rIL-4 or PMA + rIL-6. In contrast both J11d+ and J11d- subpopulations failed to respond when similarly stimulated. The J11d+ but not J11d- cells demonstrated spontaneous cytotoxic activity against the NK-sensitive YAC-1 tumor targets. The J11d- cells did not exhibit cytotoxic potential in spite of culture with PMA + rIL-2. Even after repeated culture in vitro with PMA + A23187 or PMA + rIL-2, both J11d+ and J11d- subpopulations failed to express the mature phenotype bearing CD4 and/or CD8 antigens. The present study demonstrates the expansion of unique J11d+, alpha beta-TCR+, DN T cells with cytotoxic potential in lpr mice and further suggests the existence of phenotypic and functional heterogeneity among the abnormal lpr DN cells.     

The effect of several lymphocytotoxic agents on murine leukemogenesis

5-(3,3-dimethyl-l-triazeno) imidazole-4-carboximide (DTIC) induced lymphomas with a 70 day lag period and with an incidence greater than 90% in both AKR/J and outbred Swiss Albino mice. On the other hand, treatment with cortisone acetate and 5'-azathioprine prolonged survival of AKR/J male mice. Treatment with all three agents reduced the population of medium-sized lymphocytes in the thymus within two days and additionally, cortisone and DTIC led to a reduction i in spleen weight.     

Evolutionary comparison of murine and human delta T-cell receptor deleting elements

The gene for the T-cell antigen receptor (TCR) delta chain is a gene within a gene, being located in the TCR alpha chain gene in both mice and humans. The human delta locus is flanked by delta deleting elements that undergo preferential rearrangement in the thymus, resulting in deletion of internal delta coding segments. The mouse has conserved analogous elements, m delta Rec and m phi J alpha, which separate delta from alpha and undergo a m delta Rec/m phi J alpha rearrangement in polyclonal thymus. The 5' element, m delta Rec, which is an isolated heptamer-spacer-nonamer (h-s-n), lies within 200 kb of D delta 1, and displays two areas of nearly 80% homology to human delta Rec. The downstream element, m phi J alpha, lies 12.5 kb 3' to C delta, lacks the consensus amino acids for J alpha, and retains 80% homology to human phi J alpha. Cells from murine neonatal thymus show three prominent m delta Rec rearrangements consisting of the m delta Rec/m phi J alpha recombination, a delta Rec/D delta 1/D delta 2/J delta 1 recombination, and two hybrid recombinations. A consequence of the m delta Rec/M phi J alpha rearrangement is a deletion of internal D delta and J delta coding segments that would prevent their incorporation into alpha TCR products. The conservation of noncoding deleting elements flanking the delta TCR in mice and humans is similar to the evolutionarily preserved kappa deleting element of the B-cell lineage and argues for an important role in receptor utilization.     

Anomalous occurrence of immunoreactive calcitonin cells in the thymus of the rat

Summary In a study of the effect of pinealectomy on thyroid C-cell number, 8 animals out of 66 were found to have thymic tissue in close association with the thyroid. Cells containing immunoreactive calcitonin were found in all of the thyroids but in only one of the 8 pieces of thymus. These cells found in a piece of thymic tissue associated with the right thyroid lobe were located immediately under the capsule and did not form or associate with follicles. Unlike the other animals the rat with thymic calcitonin cells had an unequal distribution of C-cells between the left and right thyroid lobes, but the total number of thyroidal C-cells was the same as that of the other rats. Since the thymus proper was not examined in these 66 animals, ten additional rats were taken for such a study. Thyroid-associated thymic tissue was found in three of these, but none of these thymi showed any immunoreactive cells.Financial support by the Deutsche Forschungsgemeinschaft (grant Vol 35/7) is gratefully acknowledged     

Age-related changes in the cellular immune response of lymph node and thymus cells in long-lived mice.

Relative capabilities of lymph node and thymus cells from (C57BL/6J) × Balb/cJ) F₁-hybrid mice and from (C57BL/6J × 129J)F₁—hybrid mice 6, 20, and 30 mo of age to respond in the mixed lymphocyte reaction were assessed. Both the direct response and the ability to synergize with partner lymph node or thymus cells were studied. Lymph node cells demonstrated an age-related decline both in responding directly to stimulation by allogeneic cells, and in ability to synergize with syngeneic thymocytes. Thymus cells showed an age-related increase in response to direct stimulation, and no clear-cut age-related change in synergizing capability. In these long-lived mouse strains at least one cause of the age-related decline in cellular immunity relates to developing deficiencies in the recirculating lymphoid pool, probably T₂ cells.     

The efficiency and direction of thymus changes after whole-body exposure of mice to the weak electromagnetic field are determined by the initial status of the thymus

The work presents results of the experimental study on thymus changes developing after whole-body exposure of mice to ultralow power pulse-modulated electromagnetic field (carrying frequency 2.39 GHz, modulating pulses with frequency 4 Hz, duration of impulses 0.025 sec, average power density 60 mW/cm2, absorbed dose 0.086 J/g or 0.172 J/g). It was shown that a percent of the microwave induced increase or decrease of thymus mass and the number of cells in the organ (y) are determined by the initial mass or number of cells in thymus accordingly to equation of linear regression: (yx = 215-2.25x, where x is the thymus mass of control animals (in a range 31-63 mg) and (yx = 178.6-41x, where x is the initial number of cells in thymus (in a range 0.6 x 10(8)-2.6 x 10(8)) reduced by a factor of 10(8).     

H2 control of natural T regulatory cell frequency in the lymph node correlates with susceptibility to day 3 thymectomy-induced autoimmune disease

Day 3 thymectomy (D3Tx) results in a loss of peripheral tolerance mediated by natural regulatory T cells (nTregs) and development of autoimmune ovarian dysgenesis (AOD) and autoimmune dacryoadenitis (ADA) in A/J and (C57BL/6J × A/J) F(1) hybrids (B6A), but not in C57BL/6J (B6) mice. Previously, using quantitative trait locus (QTL) linkage analysis, we showed that D3Tx-AOD is controlled by five unlinked QTL (Aod1-Aod5) and H2. In this study, using D3Tx B6-Chr(A/J)/NaJ chromosome (Chr) substitution strains, we confirm that QTL on Chr16 (Aod1a/Aod1b), Chr3 (Aod2), Chr1 (Aod3), Chr2 (Aod4), Chr7 (Aod5), and Chr17 (H2) control D3Tx-AOD susceptibility. In addition, we also present data mapping QTL controlling D3Tx-ADA to Chr17 (Ada1/H2), Chr1 (Ada2), and Chr3 (Ada3). Importantly, B6-ChrX(A/J) mice were as resistant to D3Tx-AOD and D3Tx-ADA as B6 mice, thereby excluding Foxp3 as a susceptibility gene in these models. Moreover, we report quantitative differences in the frequency of nTregs in the lymph nodes (LNs), but not spleen or thymus, of AOD/ADA-resistant B6 and AOD/ADA-susceptible A/J, B6A, and B6-Chr17(A/J) mice. Similar results correlating with experimental allergic encephalomyelitis and orchitis susceptibility were seen with B10.S and SJL/J mice. Using H2-congenic mice, we show that the observed difference in frequency of LN nTregs is controlled by Ada1/H2. These data support the existence of an LN-specific, H2-controlled mechanism regulating the prevalence of nTregs in autoimmune disease susceptibility.