Identification of epistasis through a partial advanced intercross reveals three arthritis loci within the Cia5 QTL in mice

Johannesson, Martina; Karlsson, Jenny; Wernhoff, Patrik; Nandakumar, Kutty Selva; Lindqvist, Anna-Karin; Olsson, Lina; Cook, Andrew D.; Andersson, Åsa; Holmdahl, Rikard

doi:10.1038/sj.gene.6364155

Cited by 46 publications

(63 citation statements)

References 28 publications

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“…1 However, the macroscopic signs of arthritis in the B10.RIII strain have in our hands never been detected before day 17 after immunization. 5 Interestingly, several genes located in the congenic fragment were differently expressed and of these Chi3l3 showed the most pronounced expression difference between the two strains (FC -23.5 at day 12 in spleen). Some of the genes were differently expressed in more than one of the tissues (Chi3l3) or at more than one time point (Gstm5, Sprr2I) supporting the interpretation of a true difference.…”

Section: Resultsmentioning

confidence: 96%

“…Four of the differently expressed genes are located in the recently described Cia21 region and 14 in the Cia22 region identified in the same study. 5 The differently expressed genes in Cia22 are located in two clusters, containing six and eight genes, respectively. The expression differences for one gene with a highly significant P-value (P ¼ 0.005), Chi3l3, and one with a P-value of 0.03, Ptpn22, were tested and confirmed with real-time RT-PCR.…”

Section: Discussionmentioning

confidence: 99%

“…The production of Cia5-region congenic mice (denoted by R3) is described elsewhere. 5 Parental B10.RIII and Cia5 congenic 12-week-old mice were immunized with 100 mg bovine CII emulsified in IFA (incomplete Freund's adjuvant)(Difco, Detroit) by intradermal (i.d.) immunization at the base of the tail.…”

Section: Animal Experimentsmentioning

confidence: 99%

“…We have recently developed the partial advanced intercross (PAI) strategy to increase penetrance through genetic interactions, and at the same time accumulate valuable recombinations. 5 In addition, in order to facilitate the genetic mapping, our group and others have studied subphenotypes with higher penetrance than the originally mapped more complex phenotype, such as the production of autoantibodies and CD4/CD8 T-cell ratio. 6,7 Another approach to reduce the number of candidate genes is to investigate the expression of the genes in the QTL.…”

Section: Introductionmentioning

confidence: 99%

“…14 The Cia5 congenic strain with an RIIIS/J fragment inserted onto the B10.RIII susceptible background has a reduced severity of CIA compared to the parental strain. 5 In order to select candidate genes for further investigations, we have studied the QTL gene expression profile during the priming phase of arthritis. We initially investigated the expression differences across the genome, which gave us an indication that the Cia5 region contains several differently expressed genes.…”

Section: Introductionmentioning

confidence: 99%

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Gene expression profiling of arthritis using a QTL chip reveals a complex gene regulation of the Cia5 region in mice

et al. 2005

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One of the major quantitative trait loci (QTLs) associated with arthritis in crosses between B10.RIII and RIIIS/J mice is the Cia5 on chromosome 3. Early in the congenic mapping process it was clear that the locus was complex, consisting of several subloci with small effects. Therefore, we developed two novel strategies to dissect a QTL: the partial advanced intercross (PAI) strategy, with which we recently found the Cia5 region to consist of three loci, Cia5, Cia21 and Cia22, and now we introduce the QTL-chip strategy, where we have combined congenic mapping with a QTL-restricted expression profiling using a novel microarray design. The expression of QTL genes was compared between parental and congenic mice in lymph node, spleen and paw samples in five biological replicates and in dye-swapped experiments at three time points during the induction phase of arthritis. The QTL chip approach revealed 4 genes located in Cia21, differently expressed in lymph nodes, and 14 genes in Cia22, located within two clusters. One cluster contains six genes, differently expressed in spleen, and the second cluster contains eight genes, differently expressed in paws. We conclude the QTL-chip strategy to be valuable in the selection of candidate genes to be prioritized for further investigation.

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Section: Resultsmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Section: Animal Experimentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Gene expression profiling of arthritis using a QTL chip reveals a complex gene regulation of the Cia5 region in mice

et al. 2005

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Wnt signaling genes of murine chromosome 15 are involved in sex‐affected pathways of inflammatory arthritis

Kudryavtseva¹,

Forde²,

Pucker³

et al. 2012

Arthritis & Rheumatism

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Objective Gender disparities in rheumatoid arthritis (RA) are well documented despite the lack of any known major RA susceptibility genes mapped to sex chromosomes. Murine chromosome 15 carries the sex-affected Pgia8 locus that mediates proteoglycan-induced arthritis (PGIA); homologous human loci are associated with RA. This study uses a Pgia8 congenic strain to identify genes/mechanisms implicated in gender disparities in arthritis. Methods Gene expression analysis was performed using RNA isolated from paws of congenic males and females with collagen antibody-induced arthritis (CAIA). Data were corroborated with RT-PCR and also studied in mice prior to disease onset. Ingenuity Pathways Analysis of the expression patterns and gene functions were used to discover locus-specific and sex-affected signature transcripts. Results We found that the Pgia8 locus regulates antibody-mediated inflammatory arthritis differently in males and females. In Pgia8 congenic males, arthritis severity was 30% less (p < 0.005) than in wild-type males, but the anti-inflammatory effect was similar in wild type and congenic females. Transcriptome analysis indicated that twelve genes within the locus were significantly deregulated in arthritic joints of congenic mice; expression of these genes was also gender specific. The genes that correlated the most with arthritis severity include collagen triple helix repeat containing-1 (Cthrc1), metalloproteinase Adamts12, R-spondyn (Rspo2) and Syndecan (Sdc2) (r=0.87–0.91). The level of Cthrc1 message also correlated with that of pro-inflammatory cytokines IL-1β and IL-6. Conclusion Gender-specific disparities in RA are linked to transcriptional regulation of genes involved in cartilage degradation (Adamts12) and canonical and non-canonical Wnt signaling (Cthrc1, Rspo2, Sdc2).

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Monoclonal Antibody Production Using a New Supermacroporous Cryogel Bioreactor

Nilsang¹,

Nandakumar²,

Galaev³

et al. 2007

Biotechnol Progress

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A supermacroporous cryogel bioreactor has been developed to culture hybridoma cells for long-term continuous production of monoclonal antibodies (mAb). Hybridoma clone M2139, secreting antibodies against J1 epitope (GERGAAGIAGPK; amino acids, 551-564) of collagen type II, are immobilized in the porous bed matrix of a cryogel column (10 mL bed volume). The cells got attached to the matrix within 48 h after inoculation and grew as a confluent sheet inside the cryogel matrix. Cells were in the lag phase for 15 days and secreted mAb into the circulation medium. Glucose consumption and lactic acid production were also monitored, and during the exponential phase (approximately 20 days), the hybridoma cell line consumed 0.75 mM day-1 glucose, produced 2.48 mM day-1 lactic acid, and produced 6.5 microg mL-1 day-1 mAb during the exponential phase. The mAb concentration reached 130 microg mL-1 after continuous run of the cryogel column for 36 days. The yield of the mAb after purification was 67.5 mg L-1, which was three times greater than the mAb yield obtained from T-flask batch cultivation. Even after the exchange of medium reservoir, cells in the cryogel column were still active and had relatively stable mAb production for an extended period of time. The bioreactor was operated continuously for 55 days without any contamination. The results from ELISA as well as arthritis experiments demonstrate that the antibodies secreted by cells grown on the cryogel column did not differ from antibodies purified from the cells grown in commercial CL-1000 culture flasks. Thus, supermacroporous cryogels can be useful as a supporting material for productive hybridoma cell culture. Cells were found to be viable inside the porous matrix of the cryogel during the study period and secreted antibodies continuously. The antibodies thus obtained from the cryogel reactor were found to be functionally active in vivo, as demonstrated by their capacity to induce arthritis in mice.

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Identification of epistasis through a partial advanced intercross reveals three arthritis loci within the Cia5 QTL in mice

Cited by 46 publications

References 28 publications

Gene expression profiling of arthritis using a QTL chip reveals a complex gene regulation of the Cia5 region in mice

Gene expression profiling of arthritis using a QTL chip reveals a complex gene regulation of the Cia5 region in mice

Wnt signaling genes of murine chromosome 15 are involved in sex‐affected pathways of inflammatory arthritis

Monoclonal Antibody Production Using a New Supermacroporous Cryogel Bioreactor

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