scholarly journals EVIDENCE FOR IDENTITY OR CLOSE ASSOCIATION OF THE Fc RECEPTOR OF B LYMPHOCYTES AND ALLOANTIGENS DETERMINED BY THE Ir REGION OF THE H-2 COMPLEX

1974 ◽  
Vol 140 (3) ◽  
pp. 779-796 ◽  
Author(s):  
Howard B. Dickler ◽  
David H. Sachs
Keyword(s):  
B Lymphocytes ◽  
B Lymphocyte ◽  
Close Association ◽  
Surface Membrane ◽  
Fc Receptor ◽  
F1 Hybrid ◽  
Ia Antigens ◽  
Series Of Experiments ◽  
Mouse Lymphocytes ◽  
Mouse Antibody

Immunoglobulin complexes, composed of heat-aggregated human Ig, were shown to bind to mouse B lymphocytes of a variety of strains, but not to either thymocytes or thymus-derived (T) lymphocytes under a variety of conditions. It was shown that this binding was not due to either natural human antibodies against mouse nor to nonspecific binding of human Ig by mouse lymphocytes. Such complexes were shown to bind to the same sites which bind mouse antibody-antigen complexes. This site is known as the Fc receptor. The binding of Ig complexes to mouse B lymphocytes was markedly inhibited by pretreatment of the lymphocytes with anti-H-2 antisera. A series of experiments indicated the specificity of this result, including the fact that this inhibition was shown not to be due to the artifact of shedding of H-2 antibody-antigen complexes, nor to nonspecific steric inhibition. The antibodies within anti-H-2 antisera which were responsible for this inhibition were specific for alloantigens associated with the Ir region of the H-2 complex (Ia antigens). Antiserum specific for these Ia antigens produced inhibition, whereas antisera specific for antigens determined by the K or D regions of the H-2 complex did not. Evidence was obtained using F1 hybrid cells that at least some Ia antigens of both parental types are expressed on every B lymphocyte (i.e. codominant expression). These data indicate that the Fc receptor and a series of alloantigens controlled by the Ir region of the H-2 complex are identical or closely associated on the B-lymphocyte surface membrane. This observation may have implications for the mechanism of control of the immune response.

scholarly journals Detection of non-H2 antigen(s) which, like Ia antigens, are associated with the Fc receptor of B lymphocytes.

1975 ◽  
Vol 142 (3) ◽  
pp. 796-801 ◽  
Author(s):  
H B Dickler ◽  
J L Cone ◽  
M T Kubicek ◽  
D H Sachs
Keyword(s):  
T Cells ◽  
B Lymphocytes ◽  
Fc Receptors ◽  
Fc Receptor ◽  
Structure And Function ◽  
Ia Antigens ◽  
And Function

Antibodies contained in an A/J anti-B10 antiserum, when tested on lymphocytes from B10.A mice, were shown to bind to both B and T cells and to inhibit the binding of Ig complexes to the Fc receptors of B lymphocytes. These antibodies could be removed by absorption with B10.D2 lymphocytes. Similar results were obtained with lymphocytes from two to six (B6A)F1 x A/J backcross mice which had H-2a/a genotype. These data indicate that alloantigens determined by at least one non-H-2 locus are associated with or a part of Fc receptors. These antigens may be similar in structure and function to Ia antigens.

scholarly journals B lymphocyte precursors and myeloid progenitors survive in diffusion chamber cultures but B cell differentiation requires close association with stromal cells

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1418-1424 ◽  
Author(s):  
PC Kierney ◽  
K Dorshkind
Keyword(s):  
B Lymphocytes ◽  
Stromal Cells ◽  
Direct Contact ◽  
Stromal Cell ◽  
B Lymphocyte ◽  
Cell Layer ◽  
Close Association ◽  
Culture Conditions ◽  
B Lymphopoiesis ◽  
Hemopoietic Cells

Abstract The aim of this study was to investigate the relative contribution of direct contact with stromal cells v stromal cell-derived soluble mediators to the differentiation of B lymphocytes and cells from other hematopoietic lineages. This was investigated by making a comparison between hemopoietic cells grown in direct contact with stroma to those in diffusion chambers (DCs) placed over purified populations of stroma. The source of stromal cells was adherent layers from myeloid or lymphoid long-term bone marrow cultures that had been treated with mycophenolic acid, an antibiotic that depletes hemopoietic cells from the cultures but retains a functional stroma. The cells seeded into the chambers were fresh marrow cells that had been passed through two consecutive nylon wool columns to deplete cell populations capable of forming an adherent cell layer in vitro. DCs were placed in wells in which the adherent stroma, growing under myeloid or lymphoid conditions, was present. The results indicate that progenitors of granulocytes and macrophages survived and differentiated in DCs under myeloid culture conditions, as the number of cells and absolute number of CFU-GM increased over that present in the reseed population. These levels, however, were markedly less than in parallel cultures in which the cells were seeded directly onto stroma. Hematopoiesis in DCs placed over hemopoietically active stroma was not optimal, suggesting that factors were used by those hemopoietic cells closest to the stroma. A B lymphocyte precursor survived in DCs under myeloid but not lymphoid conditions, and its differentiation into B lymphocytes was dependent on close association with stromal cells; B lymphopoiesis initiated when cells from DCs grown under myeloid conditions were harvested from the chambers and seeded directly onto stroma initiated and maintained under lymphoid bone marrow culture conditions. B lymphopoiesis did not initiate if the DC from the myeloid conditions was left intact and placed directly over a lymphoid stromal cell layer in lymphoid conditions.

scholarly journals B lymphocyte precursors and myeloid progenitors survive in diffusion chamber cultures but B cell differentiation requires close association with stromal cells

Blood ◽  
1987 ◽  
Vol 70 (5) ◽  
pp. 1418-1424
Author(s):  
PC Kierney ◽  
K Dorshkind
Keyword(s):  
B Lymphocytes ◽  
Stromal Cells ◽  
Direct Contact ◽  
Stromal Cell ◽  
B Lymphocyte ◽  
Cell Layer ◽  
Close Association ◽  
Culture Conditions ◽  
B Lymphopoiesis ◽  
Hemopoietic Cells

The aim of this study was to investigate the relative contribution of direct contact with stromal cells v stromal cell-derived soluble mediators to the differentiation of B lymphocytes and cells from other hematopoietic lineages. This was investigated by making a comparison between hemopoietic cells grown in direct contact with stroma to those in diffusion chambers (DCs) placed over purified populations of stroma. The source of stromal cells was adherent layers from myeloid or lymphoid long-term bone marrow cultures that had been treated with mycophenolic acid, an antibiotic that depletes hemopoietic cells from the cultures but retains a functional stroma. The cells seeded into the chambers were fresh marrow cells that had been passed through two consecutive nylon wool columns to deplete cell populations capable of forming an adherent cell layer in vitro. DCs were placed in wells in which the adherent stroma, growing under myeloid or lymphoid conditions, was present. The results indicate that progenitors of granulocytes and macrophages survived and differentiated in DCs under myeloid culture conditions, as the number of cells and absolute number of CFU-GM increased over that present in the reseed population. These levels, however, were markedly less than in parallel cultures in which the cells were seeded directly onto stroma. Hematopoiesis in DCs placed over hemopoietically active stroma was not optimal, suggesting that factors were used by those hemopoietic cells closest to the stroma. A B lymphocyte precursor survived in DCs under myeloid but not lymphoid conditions, and its differentiation into B lymphocytes was dependent on close association with stromal cells; B lymphopoiesis initiated when cells from DCs grown under myeloid conditions were harvested from the chambers and seeded directly onto stroma initiated and maintained under lymphoid bone marrow culture conditions. B lymphopoiesis did not initiate if the DC from the myeloid conditions was left intact and placed directly over a lymphoid stromal cell layer in lymphoid conditions.

Isolation of a human B lymphocyte membrane protein with Ia-like properties

1979 ◽  
Vol 57 (1) ◽  
pp. 21-31 ◽  
Author(s):  
A. K. Sullivan ◽  
L. M. Jerry ◽  
R. L. Ikeman ◽  
R. J. Maccari ◽  
H. Li Thi ◽  
...  
Keyword(s):  
B Lymphocyte ◽  
Surface Membrane ◽  
Gel Filtration ◽  
Renal Dialysis ◽  
Peritoneal Macrophages ◽  
High Titre ◽  
Membrane Glycoprotein ◽  
Normal Peripheral Blood ◽  
Ia Antigens ◽  
Major Surface

A rapid method for isolation of a major surface membrane glycoprotein from whole, unfractionated cultured human B lymphoblasts is described. After detergent solubilization the method uses gel filtration followed by affinity chromatography on Sepharose Con A and then alkaline acrylamide gel electrophoresis. Specific, high-titre, rabbit antisera to the isolated protein reacted with cultured and normal peripheral blood B lymphocytes, as well as peritoneal macrophages from a renal dialysis patient. The antisera selectively inhibited the mixed lymphocyte reaction at high dilution. The protein reacted with a heterologous antiserum to HL-B antigens and contained subunits of MW 33 000 and 27 000. Resolution of the subunits, however, required a discontinuous SDS gel system. These properties indicate its similarity to murine Ia antigens. The protein was not associated with β2 microglobulin and showed no structural or antigenic similarity to the major erythocyte glycoprotein, glycophorin. Antisera to the protein failed to precipitate surface-radioiodinated components from similarly treated extracts of cultured human T lymphoblasts. This method now makes available a reference membrane glycoprotein from a differentiated, nucleated human cell in sufficient purity and quantity for kinetic and biosynthetic studies.

scholarly journals Inhibition of the Fc receptor of human lymphoid cells by antisera-recognizing determinants of the hla system.

1976 ◽  
Vol 143 (6) ◽  
pp. 1568-1574 ◽  
Author(s):  
B G Solheim ◽  
E Thorsby ◽  
E Möller
Keyword(s):  
B Lymphocytes ◽  
Blood Cells ◽  
Inhibitory Effect ◽  
Fc Receptor ◽  
Lymphoid Cells ◽  
Effector Cells ◽  
K Cells ◽  
Ia Antigens ◽  
D Region ◽  
Dependent Cell

The inhibitory effect of HLA antisera on Fc receptors of human lymphoid cells was investigated. The ability of lymphoid cells to form rosettes (FcRFC) with antibody-coated sheep red blood cells and to function as effector cells (K cells) in antibody-dependent cell-mediated cytotoxicity were used as assay systems. We found that antisera recognizing determinants of the HLA-A, -B, and -C series had no effect on FcRFC, while a specific inhibitory effect was observed of antisera probably reacting with determinants identical to or closely associated with those of the HLA-D series. This inhibitory effect was retained in the F(ab')2 fragments. Specific inhibition of K cells was observed with all HLA antisera, but this effect was lost in the F(ab')2 fragments. We conclude that the Fc receptor of rosette-forming lymphocytes may be closely associated with products of the HLA-D region. This is analogous to the association between the Fc receptor and the Ia antigens on murine rosette-forming B lymphocytes.

scholarly journals Individual cell-by-cell quantitation of lymphocyte surface membrane Ig in normal and CLL lymphocyte and during ontogeny of mouse B lymphocytes by immunoperoxidase assay

Blood ◽  
1980 ◽  
Vol 55 (1) ◽  
pp. 93-100 ◽  
Author(s):  
G Dighiero ◽  
E Bodega ◽  
R Mayzner ◽  
JL Binet
Keyword(s):  
B Lymphocytes ◽  
B Lymphocyte ◽  
Individual Cell ◽  
Surface Membrane ◽  
Immunoperoxidase Method ◽  
Blastic Crisis ◽  
Prolymphocytic Leukemia ◽  
Antigenic Sites ◽  
Normal Individuals ◽  
Immunoperoxidase Assay

Abstract A new quantitative immunoperoxidase method is presented for determining absolute amounts of peroxidase and, consequently, surface antigen densities of individual cells in B lymphocytes from normal individuals, from subjects with CLL and prolymphocytic leukemia, and during ontogeny of B lympocytes in the mouse. The following results were observed: (1) The density of B antigenic sites were lower on CLL than on normal B lymphocytes. (2) The B antigens density of leukemic lymphocytes varied less from cell to cell, forming a homogeneous peak on histograms. (3) In a very rare case of CLL, the antigen density was measured at the time of initial diagnosis (22,500 sites or 647 U) and during the development of a blastic crisis (135,000 sites or 2576 U). The cell by cell distribution changed from a homogeneous peak with a low number of antigenic sites per cell to a heterogeneous peak with a high number of antigenic sites per cell. (4) In prolymphocytic leukemia, the density of B antigenic sites was greater than on normal B lymphocytes and much more heterogeneous than on CLL lymphocytes. (5) During ontogeny of B lymphocytes in the mouse, maturation is associated with the appearance of a population of cells of intermediate to high Smig density. The finding of a decrease in, and altered distribution of, surface markers in CLL is compared with these ontologic findings in the mouse, and the concept that a monoclonal B lymphocyte in CLL may be arrested at a particular stage in its differentiation is discussed.

Molecular structure of the gene products of the human HLA system: isolation and characterization of HLA-A, -B, -C and Ia antigens

1978 ◽  
Vol 202 (1146) ◽  
pp. 159-175 ◽  
Keyword(s):  
Molecular Mass ◽  
Close Association ◽  
Surface Membrane ◽  
Molecular Structures ◽  
Isolation And Characterization ◽  
Ia Antigens ◽  
Mass Chain ◽  
Molecular Masses ◽  
Membrane Antigens ◽  
Covalently Linked

The major histocompatibility region of man (HLA) codes for two groups of polymorphic cell surface membrane antigens. One group comprises the products of the A, B and C loci. The second group represents the Ia (immune-associated) antigens, some of which show a close association with the D locus. The strategy employed for the isolation of these anti­gens and the establishment of their molecular structures is reviewed. The A, B and C antigens are composed of a 43000 molecular mass glycosylated polypeptide which carries the polymorphic specificities and which is non-covalently linked to a non-glycosylated polypeptide of molecular mass 12000, namely β 2 -microglobulin. Structural analyses indicate that the A and B antigens have arisen by gene duplication and that the C gene(s) probably arose from the A gene(s). The Ia antigens do not contain β 2 -microglobulin but comprise two non-covalently linked glycosylated polypeptides of molecular masses 33000 and 28000. Only the 33000 molecular mass chain is apparently a product of the HLA region.

scholarly journals B-lymphocyte Fc receptor-associated non-H-2 antigens are determined by a single polymorphic locus which is linked of the Mls locus

1977 ◽  
Vol 146 (6) ◽  
pp. 1678-1692 ◽  
Author(s):  
HB Dickler ◽  
A Ahmed ◽  
DH Sachs
Keyword(s):  
B Cells ◽  
B Cell ◽  
B Lymphocyte ◽  
Polymorphic Locus ◽  
Fc Receptors ◽  
Fc Receptor ◽  
Recessive Trait ◽  
Ia Antigens ◽  
Lymphocyte Cultures ◽  
Mixed Lymphocyte Cultures

Certain non-H-2 alloantigens are associated with murine B-lymphocyte Fc receptors in that pretreatment of spleen cells with alloantibodies against these antigens inhibited binding of Ig complexes to B-cell Fc receptors. This inhibition was specific in that: (a) as has been shown previously, the Fc portion of the alloantibody was not required to produce inhibition; and (b) antibodies against some non-H-2 antigens but not antibodies against others (including some that were expressed on B cells) caused such inhibition. Backcross experiments revealed that the B-cell Fc receptor-associated non-H-2 antigens were determined by the gene(s) of a single background locus in each of the three strains tested (A/J, B10, and CBA/J). This locus was poly-morphic in that at least four different B-cell Fc receptor:associated non-H-2 antigens were identified (one each in the A/J, B10, and CBAJJ and one antigen shared or crossreactive between the B10 and CBA/J). These antigens were primarily but not exclusively expressed on B lymphocytes as determined by immunofluorescence studies, and on the basis of capping experiments they did not appear to be identical to B-cell Fc receptors. Linkage studies revealed that the locus which determined these antigens was not linked to the albino locus nor the heavy chain allotype locus and expression was neither sex-limited nor an X-linked recessive trait. However, this locus was closely linked but not identical to the Mls locus (apparent recombination frequency 6.8 percent). Thus, two closely linked non-H-2 loci both determine the expression of antigens which have characteristics similar to Ia antigens. One locus is polymorphic and determines the expression of antigens which are primarily expressed on B cells and are specifically associated with the Fc receptors of these cells. The other (Mls locus) determines antigens which are stimulatory in mixed lymphocyte cultures. These observations suggest that there may be a second gene complex which is the analogue of the I region of the H-2 complex.

scholarly journals Regulatory Role of Fc Receptor in mIgM+ B Lymphocyte Phagocytosis in Flounder (Paralichthys olivaceus)

2021 ◽  
Vol 12 ◽  
Author(s):  
Yanbo Hao ◽  
Xiaoqian Tang ◽  
Jing Xing ◽  
Xiuzhen Sheng ◽  
Heng Chi ◽  
...  
Keyword(s):  
B Cell ◽  
B Lymphocytes ◽  
Incubation Time ◽  
B Lymphocyte ◽  
Critical Role ◽  
Fc Receptor ◽  
Control Flow ◽  
Control Group ◽  
Cell Mediated Immunity

Fc receptor (FcR) is an important opsonin receptor on the surface of immune cells, playing an important role in antibody-dependent cell-mediated immunity. Our previous work found that the FcR of flounder showed a marked expression response in phagocytizing IgM+ B cell, which suggested that FcR might participate in regulating Ig-opsonized phagocytosis. In this paper, in order to elucidate the potential role of FcR in mediating phagocytosis of IgM+ B cell, flounder anti-E. tarda serum was prepared and complement-inactivated for the use of E. tarda opsonization, and the sera of healthy flounder were used as control. Flow cytometric analysis showed that the phagocytosis rates of antiserum-opsonized E. tarda in peripheral blood mIgM+ B lymphocytes were significantly higher than the control group, and higher phagocytosis rates of mIgM+ B lymphocyte could be detected with an increasing incubation time ranging from 1 to 5 h. The phagocytosis rates of antiserum-opsonized E. tarda by mIgM+ B lymphocyte for an incubation time of 1, 3 or 5 h were 51.1, 63.0, and 77.5% respectively, which were significantly higher than the phagocytosis rates in the control groups with 40.2, 50.9, and 63.8%, respectively. While the Fc fragment of IgM on the surface of opsonized E. tarda was blocked by rabbit anti-flounder IgM polyclonal antibodies, phagocytosis rates of mIgM+ B lymphocyte decreased significantly compared with the unblocked group. Moreover, the proportion of mIgM+ B lymphocytes with higher intracellular reactive oxygen species (ROS) levels rose to 32.1% from the control level of 23.0% after phagocytosis of antiserum-opsonized E. tarda. FcγRII and Syk were found to be significantly upregulated, while FcγRIII was significantly downregulated in the mIgM+ B lymphocytes post phagocytosis. Furthermore, when FcγRII of mIgM+ B lymphocytes was blocked by the prepared antibodies, their phagocytosis rate of antiserum-opsonized E. tarda was 39.0%, which was significantly lower than the unblocked group of 54.0%. These results demonstrate that FcR plays a critical role in mediating phagocytosis and bactericidal activity of mIgM+ B lymphocytes, which would facilitate an improved understanding of the regulatory roles of FcR in phagocytosis of teleost B lymphocytes.

scholarly journals Individual cell-by-cell quantitation of lymphocyte surface membrane Ig in normal and CLL lymphocyte and during ontogeny of mouse B lymphocytes by immunoperoxidase assay

Blood ◽  
1980 ◽  
Vol 55 (1) ◽  
pp. 93-100 ◽  
Author(s):  
G Dighiero ◽  
E Bodega ◽  
R Mayzner ◽  
JL Binet
Keyword(s):  
B Lymphocytes ◽  
B Lymphocyte ◽  
Individual Cell ◽  
Surface Membrane ◽  
Immunoperoxidase Method ◽  
Blastic Crisis ◽  
Prolymphocytic Leukemia ◽  
Antigenic Sites ◽  
Normal Individuals ◽  
Immunoperoxidase Assay

A new quantitative immunoperoxidase method is presented for determining absolute amounts of peroxidase and, consequently, surface antigen densities of individual cells in B lymphocytes from normal individuals, from subjects with CLL and prolymphocytic leukemia, and during ontogeny of B lympocytes in the mouse. The following results were observed: (1) The density of B antigenic sites were lower on CLL than on normal B lymphocytes. (2) The B antigens density of leukemic lymphocytes varied less from cell to cell, forming a homogeneous peak on histograms. (3) In a very rare case of CLL, the antigen density was measured at the time of initial diagnosis (22,500 sites or 647 U) and during the development of a blastic crisis (135,000 sites or 2576 U). The cell by cell distribution changed from a homogeneous peak with a low number of antigenic sites per cell to a heterogeneous peak with a high number of antigenic sites per cell. (4) In prolymphocytic leukemia, the density of B antigenic sites was greater than on normal B lymphocytes and much more heterogeneous than on CLL lymphocytes. (5) During ontogeny of B lymphocytes in the mouse, maturation is associated with the appearance of a population of cells of intermediate to high Smig density. The finding of a decrease in, and altered distribution of, surface markers in CLL is compared with these ontologic findings in the mouse, and the concept that a monoclonal B lymphocyte in CLL may be arrested at a particular stage in its differentiation is discussed.

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