Limits...
B lymphocyte involvement in ankylosing spondylitis: the heavy chain variable segment gene repertoire of B lymphocytes from germinal center-like foci in the synovial membrane indicates antigen selection.

Voswinkel J, Weisgerber K, Pfreundschuh M, Gause A - Arthritis Res. (2001)

Bottom Line: In contrast to RA, neither VH6 genes nor the autoimmunity-prone VH4-34 were seen, whereas another autoimmunity-prone gene, V3-23, was predominantly used (11%).CDR3 were shorter and more variable in length than in RA.Comparable with RA and reactive arthritis, there is a biased repertoire of selected VH genes, whereas the panel of represented genes is different and less clonal expansion was observed.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Rheumatology, Universitätsklinikum Lübeck, Lübeck, Germany. Jvoswinkel@aol.com

ABSTRACT
The synovial membrane (SM) of affected joints in ankylosing spondylitis (AS) is infiltrated by germinal center-like aggregates (foci) of lymphocytes similar to rheumatoid arthritis (RA). We characterized the rearranged heavy chain variable segment (VH) genes in the SM for gene usage and the mutational pattern to elucidate the B lymphocyte involvement in AS. Cryosections from an AS-derived SM were stained for B and T lymphocytes. B cells were isolated from different areas of a focus. The rearranged VH genes were amplified by semi-nested polymerase chain reaction (PCR) using oligonucleotides specific for the six different VH families and heavy chain joining segments (JHs). PCR products were cloned and sequenced.Fifty-nine of 70 different heavy chain gene rearrangements were potentially functional. Most of the rearranged genes were mutated (range, 1-15%). Thirty of 70 products had a mutational pattern typical for antigen selection. Most of the rearranged VH genes belonged to the VH3 family (54%), consistent with data from healthy donors and patients with RA, while VH4 genes, in contrast to RA, were identified less frequently (10%) and VH5 genes were over-represented (11%). In contrast to RA, neither VH6 genes nor the autoimmunity-prone VH4-34 were seen, whereas another autoimmunity-prone gene, V3-23, was predominantly used (11%). One VH1-derived and one VH3-derived B cell clone were expanded. CDR3 were shorter and more variable in length than in RA. Comparable with RA and reactive arthritis, there is a biased repertoire of selected VH genes, whereas the panel of represented genes is different and less clonal expansion was observed.

Show MeSH

Related in: MedlinePlus

© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC30712&req=5

Mentions: DNA was subjected to a semi-nested PCR for VH genes (see supplementary material). Thirty-five cycles of PCR, using a mix of six oligonucleotides specific for the different VH families as primers at the 5' end and a mix of oligonucleotides corresponding to the known JH overlapping to a part of the intron sequence as primers at the 3' end, were followed by 35 cycles with 1 μl of the first PCR products and individual VH and JH primers (at a more 5' position) (Table 1). A similar PCR without DNA was run as a negative control.


B lymphocyte involvement in ankylosing spondylitis: the heavy chain variable segment gene repertoire of B lymphocytes from germinal center-like foci in the synovial membrane indicates antigen selection.

Voswinkel J, Weisgerber K, Pfreundschuh M, Gause A - Arthritis Res. (2001)

© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC30712&req=5

Mentions: DNA was subjected to a semi-nested PCR for VH genes (see supplementary material). Thirty-five cycles of PCR, using a mix of six oligonucleotides specific for the different VH families as primers at the 5' end and a mix of oligonucleotides corresponding to the known JH overlapping to a part of the intron sequence as primers at the 3' end, were followed by 35 cycles with 1 μl of the first PCR products and individual VH and JH primers (at a more 5' position) (Table 1). A similar PCR without DNA was run as a negative control.

Bottom Line: In contrast to RA, neither VH6 genes nor the autoimmunity-prone VH4-34 were seen, whereas another autoimmunity-prone gene, V3-23, was predominantly used (11%).CDR3 were shorter and more variable in length than in RA.Comparable with RA and reactive arthritis, there is a biased repertoire of selected VH genes, whereas the panel of represented genes is different and less clonal expansion was observed.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Rheumatology, Universitätsklinikum Lübeck, Lübeck, Germany. Jvoswinkel@aol.com

ABSTRACT
The synovial membrane (SM) of affected joints in ankylosing spondylitis (AS) is infiltrated by germinal center-like aggregates (foci) of lymphocytes similar to rheumatoid arthritis (RA). We characterized the rearranged heavy chain variable segment (VH) genes in the SM for gene usage and the mutational pattern to elucidate the B lymphocyte involvement in AS. Cryosections from an AS-derived SM were stained for B and T lymphocytes. B cells were isolated from different areas of a focus. The rearranged VH genes were amplified by semi-nested polymerase chain reaction (PCR) using oligonucleotides specific for the six different VH families and heavy chain joining segments (JHs). PCR products were cloned and sequenced.Fifty-nine of 70 different heavy chain gene rearrangements were potentially functional. Most of the rearranged genes were mutated (range, 1-15%). Thirty of 70 products had a mutational pattern typical for antigen selection. Most of the rearranged VH genes belonged to the VH3 family (54%), consistent with data from healthy donors and patients with RA, while VH4 genes, in contrast to RA, were identified less frequently (10%) and VH5 genes were over-represented (11%). In contrast to RA, neither VH6 genes nor the autoimmunity-prone VH4-34 were seen, whereas another autoimmunity-prone gene, V3-23, was predominantly used (11%). One VH1-derived and one VH3-derived B cell clone were expanded. CDR3 were shorter and more variable in length than in RA. Comparable with RA and reactive arthritis, there is a biased repertoire of selected VH genes, whereas the panel of represented genes is different and less clonal expansion was observed.

Show MeSH
Related in: MedlinePlus