Multiple labelling technique used for kinetic studies of activated human B lymphocytes

A cell transfer assay system was developed to study the precursors of bone marrow-associated (B) lymphocytes in the adult mouse. The rationale of the assay is to inject into irradiated mice a cell suspension depleted of B lymphocytes, to wait a period of time to let precursor cells differentiate to B lymphocytes, then to correlate the number of B cells present in the recipient mice with the number of precursor cells injected. The assay as described was shown to be linear in the range of 105–3 x 106 fractionated bone marrow cells. Kinetic studies indicated that precursor cells start producing detectable numbers of B cells within 3 days after transplantation; B cell activity then increases with a doubling time of 24 hr. Physical characterization of that precursor cell has shown that it is lighter and sediments faster than small lymphocytes. Precursor cells were found in bone marrow and spleen but could not be detected in peripheral lymph nodes. Results of physical analysis also indicate that the precursors of B lymphocytes described here may not be pluripotent stem cells for the immune system.

Download Full-text

51Cr Labelling of Normal Human T and B Lymphocytes for Kinetic Studies in Vivo

Scandinavian Journal of Haematology ◽

10.1111/j.1600-0609.1978.tb02463.x ◽

2009 ◽

Vol 20 (4) ◽

pp. 319-329 ◽

Cited By ~ 3

Author(s):

Viggo Jønsson ◽

Bjarne Egelund Christensen

Keyword(s):

B Lymphocytes ◽

Kinetic Studies ◽

T And B Lymphocytes ◽

Normal Human

Download Full-text

Altered Fine Structure of B Lymphocytes Correlated with Defective Terminal Differentiation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010007237x ◽

1973 ◽

Vol 31 ◽

pp. 386-387

Author(s):

Dale E. Bockman ◽

L. Y. Frank Wu ◽

Alexander R. Lawton ◽

Max D. Cooper

Keyword(s):

Immune Deficiency ◽

B Lymphocytes ◽

Plasma Cells ◽

Present Report ◽

Specific Antigen ◽

Terminal Differentiation ◽

Second Stage ◽

Two Stages ◽

Deficiency Diseases ◽

Cell Line Generation

B-lymphocytes normally synthesize small amounts of immunoglobulin, some of which is incorporated into the cell membrane where it serves as receptor of antigen. These cells, on contact with specific antigen, proliferate and differentiate to plasma cells which synthesize and secrete large quantities of immunoglobulin. The two stages of differentiation of this cell line (generation of B-lymphocytes and antigen-driven maturation to plasma cells) are clearly separable during ontogeny and in some immune deficiency diseases. The present report describes morphologic aberrations of B-lymphocytes in two diseases in which second stage differentiation is defective.

Download Full-text

Ferritin-Conjugated Antibody for the Demonstration of Isoantigens on the Surface of Murine Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100099982 ◽

1968 ◽

Vol 26 ◽

pp. 48-49

Author(s):

T. Aoki ◽

J. Izard ◽

U. Hämmerling ◽

E. de Harven ◽

L. J. Old

Keyword(s):

Cell Surface ◽

Gamma Globulin ◽

Leukemia Cells ◽

Labelling Technique ◽

Direct Labelling ◽

Labelling Method

Although a variety of viral and cellular antigens have been demonstrated by ferritin-labeled antibody, this technique has not been used to locate isoantigens on the surface of nucleated cells. The recognition of several systems of isoantigens on the surface of thymocytes, lymphocytes and leukemia cells of the mouse and the ease with which these cells can be obtained in free suspension led us to consider the ferritin-labelling method to determine the amount and location of these isoantigens on the cell surface. Because of the problems involved in the direct labelling of mouse gamma globulin by ferritin, we have chosen an indirect labelling technique (i.e. ferritin-conjugated rabbit anti mouse γG)to detect localization of mouse isoantibody.

Download Full-text

Flavin radicals, conformational sampling and robust design principles in interprotein electron transfer: the trimethylamine dehydrogenase-electron-transferring flavoprotein complex

Biochemical Society Symposium ◽

10.1042/bss0710001 ◽

2004 ◽

Vol 71 ◽

pp. 1-14

Author(s):

David Leys ◽

Jaswir Basran ◽

François Talfournier ◽

Kamaldeep K. Chohan ◽

Andrew W. Munro ◽

...

Keyword(s):

Electron Transfer ◽

Dimensional Space ◽

Three Dimensional ◽

Kinetic Studies ◽

Molecular Assembly ◽

Conformational Sampling ◽

Trimethylamine Dehydrogenase ◽

Key Residues ◽

Electron Transferring Flavoprotein ◽

Electron Transfer Complex

TMADH (trimethylamine dehydrogenase) is a complex iron-sulphur flavoprotein that forms a soluble electron-transfer complex with ETF (electron-transferring flavoprotein). The mechanism of electron transfer between TMADH and ETF has been studied using stopped-flow kinetic and mutagenesis methods, and more recently by X-ray crystallography. Potentiometric methods have also been used to identify key residues involved in the stabilization of the flavin radical semiquinone species in ETF. These studies have demonstrated a key role for 'conformational sampling' in the electron-transfer complex, facilitated by two-site contact of ETF with TMADH. Exploration of three-dimensional space in the complex allows the FAD of ETF to find conformations compatible with enhanced electronic coupling with the 4Fe-4S centre of TMADH. This mechanism of electron transfer provides for a more robust and accessible design principle for interprotein electron transfer compared with simpler models that invoke the collision of redox partners followed by electron transfer. The structure of the TMADH-ETF complex confirms the role of key residues in electron transfer and molecular assembly, originally suggested from detailed kinetic studies in wild-type and mutant complexes, and from molecular modelling.

Download Full-text