An immunofluorescent technique for the detection of lymphocyte alloantigens

Galton, Jane; Iványi, Juraj

doi:10.1016/0022-1759(77)90076-x

Cited by 16 publications

(6 citation statements)

References 11 publications

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“…The cell recipients, which were given 850 rad of Xirradiation (57 rad min"i) 1 day before the cell transfer, were of genotypes B^B^ and fiisgis Demonstration of la-like antigens. One week after hatching-that is, 2 weeks after the cell transferthe cell recipients were killed, and the cells in the bursa of Fabricius were studied for reaction with anti-la^ and anti-Ia^^, using triple-layer immunofluorescence [6], In the chicken, la-like antigens are detected on the bursa cells from day 10 of incubation onwards; chicken la-like alloantigens (products of the B-L locus of chicken major histocompatibility or B complex) are by cellular expression and molecular weight analogous to la antigens of mammalian species [5,16], In brief, 20 ^1 of bursa cell suspension (10' cells/ml) was incubated for 30 min at 4°C with either antl-Ia-or anti-la'" 0300-9475/82/0900-0265 $02.00 © 1982 Blackwell Scientific Publications 265 diluted 1:20. After two washes with Dulbecco's phosphate-buffered saline (PBS) at 4°C, 20 /xl of rabbit antiserum to chicken IgG Fc, diluted 1:400, was added, and the cells were incubated for 30 min at 4°C.…”

Section: Methodsmentioning

confidence: 99%

Migration of Prebursal Stem Cells from the Early Chicken Embryo to the Yolk Sac

et al. 1982

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Allogeneic yolk sac-embryo chimaeras were constructed by association of B15B15 yolk sac and B2B2 embryo on day 2 of incubation. Five days later yolk sac cells from the chimaeras were injected intravenously into 14-day-old irradiated embryos, using recipients of B2B2 and B15B15 genotypes. One week after hatching, cells in the bursa of Fabricius and peripheral blood erythrocytes were studied for Ia-like antigens and B alloantigens, respectively, to determine whether they were derived from the embryo or yolk sac part of the chimaera. The results obtained demonstrate that prebursal and erythropoietic stem cells migrate from the early embryo to the yolk sac during the 2nd to the 7th day of incubation. They also exclude the de novo generation of prebursal stem cells in the yolk sac.

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Section: Methodsmentioning

confidence: 99%

Migration of Prebursal Stem Cells from the Early Chicken Embryo to the Yolk Sac

et al. 1982

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“….5 (8) Days after cell 12 1,3±1,6 (17) 1,4±1, 9 (15) O,5±O,5 (11) 0,1 ±0,5 (14) 0,4 ±0,9 (12) 0 (10) 14,1 ±8,5 (12) injection 16 6,0±8,7 4,4±5,2 2,O±3,l 0,9 ±1,1 l,7±3,l 0 I3,O±8,O (17) (21) (11) (16) (15) (14) (16) 25 4,0±4,8(ll) 2,8±3,5 (12) 5,7±3,4(7) 4,4 ±4,0 (8) 2,2±3,4 (14) 0 (11) 9,0±4,7 (14) * For experimental details see Table III, t Thymus cells from 3-week-old normal chickens, X Figure in parentheses refers to the number ofehickens, § Untreated normal control birds age-matched with the test birds.…”

Section: No Sign Of Graft-versus-host Reaction In the Test Birds Was mentioning

confidence: 99%

Maturation of Bursal Stem Cells within Allogeneic or Syngeneic Bursal Microenvironment: Surface Determinants and Capacity for Germinal Centre Formation

Vainio

1979

Scand J Immunol

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Maturation of bursal stem cells in an allogeneic environment was studied. The occurrence of surface IgM, surface IgG, and Ia-like antigens on B cells after differentiatoin within an allogeneic bursa was studied. Furthermore, the immediate capacity of B cells to form germinal centres after differentiation in an allogeneic bursa was studied by injecting them with histocompatible T cells into "test-tube" birds. The results indicate that the switch to surface-IgG-bearing cells occurs in an allogeneic bursa in a similar manner as in a syngeneic bursa. Studies of the occurrence of Ia-like antigens on bursa cells demonstrate that they retain their original Ia-like antigens, even during maturation in an allogeneic host. Third, the results demonstrate that after differentiation within an allogeneic environment bursa cells have acquired the ijmediate capacity to cooperate with histocompatible T cells as measured by germinal centre formation in "test-tube" birds.

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“…A triple-layer immunofluorescence technique as described by Galton and Ivanyi [20] was used with some modifications. First, lo6 cells were incubated with 20 y1 alloantiserum diluted 1 : 50 for 30 min at 4"C, washed twice, incubated with the predetermined dilution of goat anti-chicken Fcy antiserum (1 : 160) (Pel-Freez Biologicals, Rogers, AR), washed twice and incubated with the 1 : 10 diluted FITC-conjugated rabbit anti-goat IgG (Miles-Yeda).…”

Section: B-l+ Cellsmentioning

confidence: 99%

“…Single-cell suspensions from the spleen and BM were prepared as described earlier [17,191 and lymphocytes were isolated by the Ficoll-Isopaque (Pharmacia Fine Chemicals, Uppsala, Sweden) centrifugation. PB lymphocytes were isolated from heparinized whole blood as described by Galton and Ivanyi [20].…”

Section: Preparation and Transplantation Of Cellsmentioning

confidence: 99%

Immune capacity of the chicken bursectomized at 60 hours of incubation: transplantation of bone marrow cells of the bursectomized chickens into cyclophosphamide‐treated newly hatched recipients

Jalkanen

1983

Eur J Immunol

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Chickens bursectomized at 60 h of incubation are known to be able to produce immunoglobulins (Ig) but not specific antibodies. In the present work bone marrow (BM) cells of 2- and 10-week-old embryonically bursectomized (Bx) chickens were transferred to newly hatched cyclophosphamide-treated chickens for the purpose of studying whether the transplanted cells home into the bursa and gain a capacity to produce specific antibodies. BM cells of normal 2- and 10-week-old chickens were transferred as controls. Cells of 2-week-old control (Co) and of 2- and 10-week-old Bx chickens could to some extent reconstitute serum Ig of the recipients, but were totally incapable of homing into the bursa and of restoring the specific antibody production. Only BM cells of 10-week-old Co chickens could restore the production of specific antibodies without homing into the bursa, indicating their postbursal nature. These findings indicate (a) that 2- and 10-week Bx BM cells have irreversibly bypassed the bursal phase of education, (b) that also the normal BM at the age of 2 weeks contains cells that are capable of Ig but not of specific antibody synthesis and (c) that bursal microenvironment is not necessary for isotype switch, but is essential for creation and expansion of the antibody repertoire.

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An immunofluorescent technique for the detection of lymphocyte alloantigens

Cited by 16 publications

References 11 publications

Migration of Prebursal Stem Cells from the Early Chicken Embryo to the Yolk Sac

Migration of Prebursal Stem Cells from the Early Chicken Embryo to the Yolk Sac

Maturation of Bursal Stem Cells within Allogeneic or Syngeneic Bursal Microenvironment: Surface Determinants and Capacity for Germinal Centre Formation

Immune capacity of the chicken bursectomized at 60 hours of incubation: transplantation of bone marrow cells of the bursectomized chickens into cyclophosphamide‐treated newly hatched recipients

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