The role of lymphocytes in antibody formation I. Restoration of the haemolysin response in X -irradiated rats with lymphocytes from normal and immunologically tolerant donors

1967 ◽  
Vol 168 (1012) ◽  
pp. 229-243 ◽  
Keyword(s):  
Thoracic Duct ◽  
Immunological Tolerance ◽  
Whole Body ◽  
Sheep Erythrocytes ◽  
X Irradiation ◽  
Additional Support ◽  
Immunological Deficiency ◽  
Donor Lymphocytes

The haemolysin response of rats to an intravenous dose of 10 8 sheep erythrocytes was abolished by pretreatment with 500 rad of whole body X-irradiation. The immunological deficiency in such animals could be corrected equally well by either an injection of thoracic duct cells or by an inoculum consisting almost exclusively of small lymphocytes, obtained in each case from normal (non-immune) rats. The reversal of unresponsiveness depended upon the survival of the donor lymphocytes in the X-irradiated recipients and was not due to a non-specific restoration of the hosts’ own capacity to form antibody. Evidence for this conclusion came from experiments in which the X-irradiated recipients were themselves immunologically tolerant of sheep erythrocytes; additional support came from the inability of lymphocytes from immunologically tolerant donors to restore specific responsiveness in X-irradiated (non-tolerant) recipients. In a proportion of trials the immunological tolerance to sheep erythrocytes exhibited by thoracic duct lymphocytes from tolerant donors could be broken by incubating the cells in vitro before their injection into X-irradiated recipients. This points to the existence of individual tolerant cells in the tolerant populations of lymphocytes. Taken as a whole the experiments suggest strongly that small lymphocytes are the precursors of the cells which produce haemolysin against sheep erythrocytes in the rat.

ROLE OF NA+, K+-ATPase IN MUSCULAR ENERGY EXPENDITURE OF WARM- AND COLD-EXPOSED SHEEP

1982 ◽  
Vol 62 (1) ◽  
pp. 123-132 ◽  
Author(s):  
V. A. GREGG ◽  
L. P. MILLIGAN
Keyword(s):  
Skeletal Muscle ◽  
Energy Expenditure ◽  
Cold Exposure ◽  
Whole Body ◽  
Random Group ◽  
Key Words Skeletal Muscle ◽  
Maximum Inhibition ◽  
Functional Membrane

The role of Na+, K+-ATPase in the energy expenditure of sheep skeletal muscle and the influence of exposure to cold on this role were studied. An in vitro preparation of muscle was developed that achieved O2 availability and a functional membrane potential. A 10−6 M concentration of ouabain yielded a maximum inhibition of respiration of 38.9 ± 1.8% using muscle preparations from a random group of sheep. Whole body and muscle O2 consumptions and ouabain-sensitive muscle respiration were measured for warm- and cold-exposed sheep fed at maintenance or 1150 g of alfalfa pellets per day. Cold exposure increased whole body and muscle O2 consumption. Inhibition of respiration by ouabain was 37.6 ± 1.2% and 41.0 ± 3.6% for warm- and cold-exposed sheep fed at maintenance, and 28.5 ± 4.0% and 45.0 ± 4.0% for warm- and cold-exposed sheep fed 1150 g of alfalfa pellets per day. The increase in the ouabain-sensitive component of respiration accounted for 48–79% of the increased O2 consumption of muscle from cold-exposed sheep. It was concluded that the Na+, K+-ATPase of sheep muscle is a major means of energy expenditure and has an important role in the increased thermogenesis resulting from cold exposure. Key words: Skeletal muscle, Energy expenditure, muscle respiration, cold thermogenesis, sodium-potassium transport

THE USE OF Cr51 AS A CRITERION OF THYMOCYTE REGENERATION AFTER WHOLE BODY X-IRRADIATION

1965 ◽  
Vol 43 (1) ◽  
pp. 19-27
Author(s):  
P. V. Vittorio ◽  
P. J. Baker ◽  
S. Dziubalo-Blehm
Keyword(s):  
Age Differences ◽  
Whole Body ◽  
Quantitative Criterion ◽  
X Irradiation ◽  
Early Radiation ◽  
Population Ratio ◽  
Time Required ◽  
Different Doses ◽  
In Vitro Uptake

The uptake of Cr51 chromate by thymocytes in vitro after whole body X-irradiation can be used as a sensitive quantitative criterion not only for the evaluation of early radiation damage to these cells but also as a measure of later regeneration. The development of the radiation lesion is characterized by a reduction in the in vitro uptake of Cr51 and the later regeneration of new cells by an increase in Cr51 uptake which is probably due to increased uptake of Cr51 by the young newly formed cells in the damaged tissue which is proliferating in an attempt to repair the damage. The return of the Cr51 uptake to normal is an indication of the time required for the cell population (ratio of young to older cells) to return to normal. By this technique the effect of different doses of X-irradiation on the regeneration of thymocytes has been demonstrated. Treatment with AET before whole body X-irradiation (400 r) indicated that less regeneration was necessary but the recovery time remained unchanged. Age differences produced a change in the extent of repair or regeneration but no change in recovery time.Spermatozoa showed evidence of early damage after whole body X-irradiation. This damage increased with time with no evidence of increased regeneration or repair.

The role of lymphocytes in antibody formation II. the influence of lymphocyte migration on the initiation of antibody formation in the isolated, perfused spleen

1967 ◽  
Vol 168 (1012) ◽  
pp. 244-262 ◽  
Keyword(s):  
Antibody Response ◽  
Lymphoid Tissue ◽  
Antibody Formation ◽  
Central Area ◽  
Lymphocyte Migration ◽  
Sheep Erythrocytes ◽  
Paradoxical Finding ◽  
X Irradiation

The aim of these studies was to examine the possible immunological significance of the process of lymphocyte recirculation. For this purpose a technique for perfusing the isolated spleen of the rat was developed. Antigen (sheep erythrocytes) was added to the perfusate and, after 3 to 6 h periods of perfusion, the spleens were cut into fragments and implanted into the peritoneal cavities of X-irradiated syngeneic recipients to determine the magnitude of the haemolysin response. The lymphocyte concentration in the blood perfusing the spleen was varied within wide limits to determine whether a migration of lymphocytes through the spleen influenced its ability to respond to antigenic stimulation. Normal spleens perfused for 3 h with blood containing a normal concentration of lymphocytes gave substantial haemolysin titres in the recipients when sheep erythrocytes had been added initially to the perfusate. Spleens depleted of lymphocytes in vivo by either X-irradiation or chronic drainage from a thoracic duct fistula produced very low titres after perfusion with lymphocyte-free blood and antigen, but their responses were restored by adding lymphocytes to the perfusate. The antibody response of normal spleens was significantly depressed following perfusion for 3 h with lymphocyte-free blood to which antigen had been added initially. However, normal responses were obtained in this experiment if the addition of antigen to the perfusate was delayed for 5 h and perfusion then continued for one further hour. The delay allowed lymphocytes to migrate from the normal spleens into the perfusate and to build up the concentration in the initially lymphocyte-free blood to normal levels. The conclusion drawn from this paradoxical finding was that the magnitude of the haemolysin response depended upon the concentration of lymphocytes in the perfusate at the time of antigen addition and not upon the total number of lymphocytes in the spleen. The rate of migration of small lymphocytes from the blood into the spleen was found to be directly proportional to the concentration of lymphocytes in the perfusate. The most acceptable explanation of all the data is that the magnitude of the haemolysin response is proportional to the concentration of lymphocytes in a limited compartment of the splenic lymphoid tissue into which lymphocytes have recently migrated from the blood. This compartment is probably located in the central area of the periarteriolar lymphocyte sheath.

Cytoarchitecture of the Xenopus thymus following gamma-irradiation

Development ◽  
1987 ◽  
Vol 100 (1) ◽  
pp. 95-105
Author(s):  
JH Russ ◽  
JD Horton
Keyword(s):  
Gamma Irradiation ◽  
Tolerance Induction ◽  
Cell Types ◽  
Whole Body ◽  
Thymic Stromal Cells ◽  
Normal Architecture

This paper describes in vitro and in vivo attempts to deplete the 4- to 8-month-old Xenopus laevis (J strain) thymus of its lymphocyte compartment. Gamma irradiation (2-3000 rad) of the excised thymus, followed by two weeks in organ culture, is effective in removing lymphocytes, but causes drastic reduction in size and loss of normal architecture. In contrast, in vivo whole-body irradiation (3000 rad) and subsequent in situ residence for 8-14 days proves successful in providing a lymphocyte-depleted froglet thymus without loss of cortical and medullary zones. In vivo-irradiated thymuses are about half normal size, lack cortical lymphocytes, but still retain some medullary thymocytes; they show no signs of lymphocyte regeneration when subsequently organ cultured for 2 weeks. Light microscopy of 1 micron, plastic-embedded sections and electron microscopy reveal that a range of thymic stromal cell types are retained and that increased numbers of cysts, mucous and myoid cells are found in the thymus following whole-body irradiation. In vivo-irradiated thymuses are therefore suitable for implantation studies exploring the role of thymic stromal cells in tolerance induction of differentiating T lymphocytes.

scholarly journals THE ROLE OF LYMPHOCYTES IN THE SENSITIZATION OF RATS TO RENAL HOMOGRAFTS

1965 ◽  
Vol 122 (2) ◽  
pp. 347-360 ◽  
Author(s):  
S. Strober ◽  
J. L. Gowans
Keyword(s):  
Lymph Nodes ◽  
Thoracic Duct ◽  
Femoral Vein ◽  
F1 Hybrid ◽  
Duct Cells ◽  
Degree Of Sensitization ◽  
Spleen And Lymph Nodes

In order to study the role of blood-borne small lymphocytes in the sensitization of rats to renal homografts 2 techniques for the perfusion of isolated rat kidneys were employed: (a) the in vitro perfusion of kidneys with thoracic duct cells suspended in either an artificial medium or in blood; the perfusates were then injected into rats syngeneic with the lymphocyte donors; (b) the in vivo perfusion of kidneys with blood issuing from the femoral artery and returning to the femoral vein of living rats. The degree of sensitization conferred on the recipients by the perfusates was assessed by applying a skin homograft from the kidney donor and scoring the epithelial necrosis at 6 days. The in vitro experiments indicated that parental strain thoracic duct cells, which had passed through an F1 hybrid kidney could confer upon a parental rat sensitivity to an F1 skin graft. Several perfusions with radioactively labelled lymphocytes showed that the injected cells migrated to the lymph nodes and spleen of the recipients Labelled large pyroninophilic cells were occasionally seen in the spleen and lymph nodes of recipients, and it was suggested that these had arisen from the injected cells. Although the in vitro perfusions with blood indicated that renal homografts might sensitize their hosts within 1 hour, the in vivo perfusions suggested that about 5 to 12 hours were required. The more rapid sensitization in vitro was possibly due to the more frequent opportunity for contact between lymphocytes and kidney vascular endothelium which was afforded by the conditions in vitro.

scholarly journals In vitro tolerance induction of primed, IgD-negative murine spleen cells.

1981 ◽  
Vol 153 (3) ◽  
pp. 653-664 ◽  
Author(s):  
S M Walker ◽  
W O Weigle
Keyword(s):  
B Cells ◽  
Tolerance Induction ◽  
Keyhole Limpet Hemocyanin ◽  
Immunological Tolerance ◽  
Target Cells ◽  
Spleen Cells ◽  
Murine Spleen ◽  
In Vitro Response

The above observations demonstrated induction of immunological tolerance in vitro in primed IgD-, IgG+ B cells. In these studies, addition of trinitrophenylated (TNP) turkey gammaglobulin (TGG) or TNP ovalbumin conjugates suppressed the secondary in vitro response in mice primed with TNP keyhole limpet hemocyanin (TNP-KLH). Suppression was not a reflection of a shift in kinetics of the antibody response, was not dependent on suppressor T cells, and could only be eliciate when conjugate was added within 4 h of addition of TNP-KLH moreover, preincubation of the primed spleen cells with TNP-TGG for 20 h at 37 degrees C, followed by extensive washing, was as effective in inhibiting the response to TNP-KLH as when TNP-TGG was present throughout the 5 d of culture, reflecting induction of a tolerant state. Amounts of conjugate in the concentration range that have been shown by others to tolerize immature or neonatal B cells or mature B cells that have been stripped of surface IgD were sufficient to induce tolerance. The target cells being tolerized did not bear IgD, as determined by B cell depletion and blocking procedures with anti IgD. Whether the lack of surface IgD on the primed cells contributed to the relative ease of tolerance induction was not established by these studies, but the advantages of using primed B cells to examine further the role of surface IgD in tolerance susceptibility was discussed.

scholarly journals The effect of whole-body X-irradiation of guinea pigs on liver ribosomes

1968 ◽  
Vol 109 (4) ◽  
pp. 495-505 ◽  
Author(s):  
E. J. Hidvégi ◽  
J. Holland ◽  
Elisabeth Bölöni ◽  
P. Lónai ◽  
F. Antoni ◽  
...  
Keyword(s):  
Amino Acid ◽  
Guinea Pigs ◽  
Sucrose Density Gradient ◽  
Whole Body ◽  
Radiation Doses ◽  
Amino Acid Incorporation ◽  
Acid Incorporation ◽  
X Irradiation ◽  
Liver Ribosomes

1. The size distribution of aggregates of liver ribosomes and their protein-synthesizing ability in vitro were studied shortly after X-irradiation of guinea pigs. 2. Sucrose-density-gradient analysis of the mitochondrial supernatant after treatment with deoxycholate revealed a gradual increase in the number of polysomes, reaching a maximum between 9 and 15 hr. after irradiation. At that period the amount of ribonucleoprotein particles reached a level 25–30% above the control. This finding was confirmed by analytical-ultracentrifugal analysis and electron microscopy. Experiments were conducted to exclude the possibility that the enrichment of polysomes in the irradiated animals had occurred during the isolation procedure. 3. The protein-synthesizing ability of total ribosomal particles was measured in vitro. This showed an increase in amino acid incorporation parallel to the progressive enrichment of polysomes. At radiation doses of up to 1000r. the protein-synthesizing capacity was dependent on the radiation dose: the higher the dose the higher the amino acid incorporation, reaching 40–60% above the control at the period of maximal polysome enrichment. Amino acid incorporation remained at this level after radiation doses of between 1000 and 3000r. The enhanced protein-synthesizing activity was due solely to the increase in the proportion of polysomes, since irradiation was without effect on the activity of single ribosomes. 4. The results of the experiments are discussed in the light of our knowledge of the effect of radiation on protein synthesis.

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