VH gene family utilization in colonies derived from B and pre-B cells detected by the RNA colony blot assay.

Wu, Gillian E.; Paige, Christopher J.

doi:10.1002/j.1460-2075.1986.tb04672.x

Cited by 124 publications

(78 citation statements)

References 30 publications

(52 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Based upon analysis of 2,343 colonies from the F, mice we found that the percent utilization detected was intermediate between the parental patterns in every VH family for which parental differences had been noted (9) . This point is best illustrated by comparing F, and parental h respect to the VHJ558/V H7183 ratio ( Fig.…”

Section: Results and Discussmentioning

confidence: 89%

“…We have used the RNA colony blot assay to characterize the VH family usage of B cell colonies derived from surface Ig* (sIg+) spleen cells (CFU-B) and sIgfetal liver cells (CFU-pre-B) with eight VH family-specific DNA probes (9) . We found that VH family usage was independent of whether the cell that initiated colony formation was a CFU-B or CFU-pre-B.…”

Section: Results and Discussmentioning

confidence: 99%

“…Approximately 5 ug of genomic DNA was digested with Eco RI fractionated on a 0 .8% agarose gel and transferred to GeneScreenPlus filters. Triplicate blots were assayed for hybridization to "1P-labeled VHQ52, VH7183, and VHJ606 gene family probes (9). After hybridization and washing the blots were exposed for 7 d with intensifying screens.…”

Section: Results and Discussmentioning

confidence: 99%

“…Colony-forming B cells were grown as previously described (8)(9)(10) from the spleen cells of adult mice obtained from the breeding facilities of the Basel Institute for Immunology . The RNA blots were hybridized with CA and eight VH family-specific DNA fragments as previously described (8), and the resulting filters were autoradiographed for 3-14 d. The VH family-specific probes were isolated from appropriately dig using standard procedures (9).…”

Section: Methodsmentioning

confidence: 99%

“…The RNA blots were hybridized with CA and eight VH family-specific DNA fragments as previously described (8), and the resulting filters were autoradiographed for 3-14 d. The VH family-specific probes were isolated from appropriately dig using standard procedures (9). The plasmids were kindly provided by F. Alt, G. YancoPoulos, E. Dzierzak, R. Riblet, P. Brodeur, R. Perlmutter, J. Adams, and G. Lennox.…”

Section: Methodsmentioning

confidence: 99%

See 4 more Smart Citations

VH gene family utilization is regulated by a locus outside of the VH region.

Paige

1988

The Journal of Experimental Medicine

View full text Add to dashboard Cite

The Ig heavy chain is composed of four regions, VH, DH, JH, and CH. Each region is encoded by discrete gene segments which, in the germline state, are widely separated on the chromosome (1). During differentiation along the B cell pathway, three of these gene segments, namely, V, D, and J, become juxtaposed, forming a complete heavy chain gene with the ensuing deletion of the intervening DNA sequences. In the mouse, there are >100 germline-encoded heavy chain VH gene segments categorized into at least nine families based upon sequence homology to representative DNA probes (2-4). Whereas the genetic mechanism involved in the joining process has received much attention, little is known concerning the basis for the selection of a particular VH gene segment Materials and MethodsColony-forming B cells were grown as previously described (8-10) from the spleen cells of adult mice obtained from the breeding facilities of the Basel Institute for Immunology . The RNA blots were hybridized with CA and eight VH family-specific DNA fragments as previously described (8), and the resulting filters were autoradiographed for 3-14 d. The VH family-specific probes were isolated from appropriately dig using standard procedures (9). The plasmids were kindly provided by

show abstract

Section: Results and Discussmentioning

confidence: 89%

Section: Results and Discussmentioning

confidence: 99%

Section: Results and Discussmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

VH gene family utilization is regulated by a locus outside of the VH region.

Paige

1988

The Journal of Experimental Medicine

View full text Add to dashboard Cite

show abstract

Development of the early B cell population inXenopus

Mußmann¹,

Courtet²,

Pasquier³

1998

Eur. J. Immunol.

View full text Add to dashboard Cite

Like mammals, the amphibian Xenopus uses combinatorial joining of the immunoglobulin V, D and J elements and multiple rearrangements to generate its B cell repertoire. Xenopus larvae hatch 2 days after fertilization and individuals are under pressure to develop an immune repertoire when the number of available cells is small (approximately 5 and 200 IgM-positive cells on days 5 and 11 after fertilization, respectively). In the liver, in a first phase of differentiation spanning days 5-12 after fertilization before immunological competence, the heavy (H) chain locus starts rearranging followed by the light (L) chain locus 3 days later. By immunohistology the first B cells expressing H and L chain are detectable on day 10. Despite the small number of cells available and the lack of external antigen selection at these early stages, the repertoire is heterogeneous. The V H families are used stepwise, although their genes are interspersed in the genome. The earliest family used (V H 1) is homologous to the V H 3 family of human and to the V H 7183 of the mouse which are also overrepresented in early mammalian development. In the second phase, from day 12-13 onwards, the spleen differentiates and the animal becomes immunologically competent. The V, D and J usage is similar to that of adults although VDJ junctions lack N nucleotides until metamorphosis. A preferential reading frame for D and one specific DJ junction are overrepresented during this second phase. The visible bias toward homology-based junction results in fact from selection after rearrangement.

show abstract

Autoantibodies, ly‐1, and immunoglobulin v gene expression in hybridomas obtained from young and from old new zealand black mice

Fidanza

Mayer

Zaghouani

et al. 1990

Arthritis & Rheumatism

View full text Add to dashboard Cite

We obtained a large number of hybridomas from l-month-old and 16-month-old New Zealand black mice to study the fine specificities of the autoantibodies produced, the expression of Ly-1, and the expression of the immunoglobulin V gene families in this autoimmune strain. Analysis of the autoantibody specificities yielded 2 major classifications: those specific for a single autoantigen and those that exhibited multispecific binding. Among the multispecific antibodies, 2 categories were found: an antigen-inhibitable group and an antigennoninhibitable group. A large proportion of V,J558 and V,7183 gene families was observed in hybridomas obtained from l-month-old mice, and in hybridomas obtained from 16-month-old mice, there was a large proportion of V,J558 and V,36-60 gene families. Among the autoantibody K chains secreted by the hybridomas, there was a higher frequency of the VJ, VJ3, and V Q gene families. Autoantibodies were produced by both the Ly-l+ and the Ly-l-B cell subsets.The New Zealand black (NZB) strain of mice is prone to the development of a lupus-like syndrome and autoimmune hemolytic anemia. This strain is therefore an excellent animal model for studying the immuno- chemical and molecular characteristics of both organspecific and non-organ-specific autoantibodies. Whereas Coombs anti-erythrocyte autoantibodies represent typical organ-specific pathogenic autoantibodies (1 ), anti-DNA, anti-Sm, antihistone, and anti-rheumatoid factor (RF) autoantibodies represent non-organ-specific autoantibodies that are associated with various systemic autoimmune diseases (2). There is evidence that a discrete subset of B cells bearing the CDS antigen (Ly-1 in mice and Leu-1 in humans) is the major contributor to the production of certain autoantibodies (3,4). It has been shown that the Ly-I B cell subset is larger in the NZB strain of mice (3.5) and that aged NZB mice show an unusual clonal expansion of Ly-1 B cells exhibiting premalignant properties, such as autonomous proliferation resistant to immunoregulatory signals (6,7). NZB mice, then, are a good model for studying the cellular origin of autoantibodies as well.In the past. the autoreactivity of antibodies was defined by the ability of a heterogenous population of immunoglobulin molecules to bind autoantigens. These Ig were isolated from the sera of patients with autoimmune diseases or from animals prone to developing autoimmune diseases. The development of hybridoma technology and sensitive cellular immunologic techniques has permitted a detailed analysis of the self-reactivity of homogenous populations of antibodies. Using these techniques, investigators have shown that an important proportion of antibodies is multispecific (8.9) and that these antibodies have a moderate affinity to g d i t e r ) for self antigens and for foreign antigens (10).Studies of the immunochemical and molecular properties of monoclonal antibodies can provide important information on the origin of autoantibodies. In particular, we can now examine the properties of the

show abstract

VH gene family utilization in colonies derived from B and pre-B cells detected by the RNA colony blot assay.

Cited by 124 publications

References 30 publications

VH gene family utilization is regulated by a locus outside of the VH region.

VH gene family utilization is regulated by a locus outside of the VH region.

Development of the early B cell population inXenopus

Autoantibodies, ly‐1, and immunoglobulin v gene expression in hybridomas obtained from young and from old new zealand black mice

Contact Info

Product

Resources

About