RESUMO
OBJECTIVE: Cia3 is a locus on rat chromosome 4 that regulates severity and joint damage in collagen- and pristane-induced arthritis (CIA and PIA). This study was undertaken to refine the Cia3 gene-containing interval toward gene identification and obtain insights into its mode of action. METHODS: Five DA.F344(Cia3) subcongenic rat strains were generated and studied using the PIA and CIA models. Levels of antibodies against type II collagen (both allo- and autoantibodies) were measured. Joints and synovial tissue were collected 32 days after the induction of PIA (chronic stage) for histologic and quantitative polymerase chain reaction analysis of interleukin-1ß (IL-1ß) and matrix metalloproteinase (MMP) levels. RESULTS: Three subcongenic strains sharing the centromeric Cia3d interval were protected and 2 subcongenic strains sharing the telomeric Cia3g interval, which did not overlap with Cia3d, were also protected, developing significantly less severe CIA and PIA. Normal joint architecture was preserved in DA.F344(Cia3) and DA.F344(Cia3d) congenic rats with PIA, while DA rats had pronounced synovial hyperplasia, angiogenesis, inflammatory infiltration, and bone or cartilage erosions. The DA.F344(Cia3d) and DA.F344(Cia3g) strains had significantly lower synovial levels of IL-1ß (5-fold and nearly 2-fold, respectively [the latter not reaching statistical significance]), MMP-1 (expressed predominantly in DA rats), MMP-3 (79-fold and 8-fold, respectively), and MMP-14 (21-fold and 1.4-fold, respectively) and reduced levels of pathogenic autoantibodies against type II collagen, compared with DA rats. CONCLUSION: We have identified 2 new arthritis severity and articular damage loci within Cia3. These loci regulate pathogenic processes in 2 different models of rheumatoid arthritis, and the identification of these genes has the potential to generate new targets for therapies aimed at reducing disease severity and articular damage, and may additionally have prognostic value.
Assuntos
Artrite Experimental/imunologia , Colágeno/toxicidade , Imunossupressores/toxicidade , Terpenos/toxicidade , Animais , Animais Congênicos , Artrite Experimental/induzido quimicamente , Artrite Experimental/patologia , Autoanticorpos/imunologia , Biomarcadores/metabolismo , Colágeno/imunologia , Colágeno Tipo II/imunologia , Progressão da Doença , Expressão Gênica , Marcadores Genéticos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Articulações/efeitos dos fármacos , Articulações/patologia , Articulações/fisiopatologia , Metaloproteases/genética , Metaloproteases/metabolismo , Locos de Características Quantitativas/genética , Ratos , Ratos Endogâmicos F344 , Índice de Gravidade de Doença , Especificidade da Espécie , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/patologia , Membrana Sinovial/fisiopatologia , Terpenos/imunologiaRESUMO
BACKGROUND: Findings of small studies have suggested that short treatments with anti-CD3 monoclonal antibodies that are mutated to reduce Fc receptor binding preserve ß-cell function and decrease insulin needs in patients with recent-onset type 1 diabetes. In this phase 3 trial, we assessed the safety and efficacy of one such antibody, teplizumab. METHODS: In this 2-year trial, patients aged 8-35 years who had been diagnosed with type 1 diabetes for 12 weeks or fewer were enrolled and treated at 83 clinical centres in North America, Europe, Israel, and India. Participants were allocated (2:1:1:1 ratio) by an interactive telephone system, according to computer-generated block randomisation, to receive one of three regimens of teplizumab infusions (14-day full dose, 14-day low dose, or 6-day full dose) or placebo at baseline and at 26 weeks. The Protégé study is still underway, and patients and study staff remain masked through to study closure. The primary composite outcome was the percentage of patients with insulin use of less than 0·5 U/kg per day and glycated haemoglobin A(1c) (HbA(1C)) of less than 6·5% at 1 year. Analyses included all patients who received at least one dose of study drug. This trial is registered with ClinicalTrials.gov, number NCT00385697. FINDINGS: 763 patients were screened, of whom 516 were randomised to receive 14-day full-dose teplizumab (n=209), 14-day low-dose teplizumab (n=102), 6-day full-dose teplizumab (n=106), or placebo (n=99). Two patients in the 14-day full-dose group and one patient in the placebo group did not start treatment, so 513 patients were eligible for efficacy analyses. The primary outcome did not differ between groups at 1 year: 19·8% (41/207) in the 14-day full-dose group; 13·7% (14/102) in the 14-day low-dose group; 20·8% (22/106) in the 6-day full-dose group; and 20·4% (20/98) in the placebo group. 5% (19/415) of patients in the teplizumab groups were not taking insulin at 1 year, compared with no patients in the placebo group at 1 year (p=0·03). Across the four study groups, similar proportions of patients had adverse events (414/417 [99%] in the teplizumab groups vs 98/99 [99%] in the placebo group) and serious adverse events (42/417 [10%] vs 9/99 [9%]). The most common clinical adverse event in the teplizumab groups was rash (220/417 [53%] vs 20/99 [20%] in the placebo group). INTERPRETATION: Findings of exploratory analyses suggest that future studies of immunotherapeutic intervention with teplizumab might have increased success in prevention of a decline in ß-cell function (measured by C-peptide) and provision of glycaemic control at reduced doses of insulin if they target patients early after diagnosis of diabetes and children. FUNDING: MacroGenics, the Juvenile Diabetes Research Foundation, and Eli Lilly.
Assuntos
Complexo CD3/efeitos dos fármacos , Diabetes Mellitus Tipo 1/tratamento farmacológico , Diabetes Mellitus Tipo 1/imunologia , Hipoglicemiantes/uso terapêutico , Insulina/uso terapêutico , Muromonab-CD3/uso terapêutico , Adolescente , Adulto , Anticorpos Monoclonais Humanizados , Peptídeo C/sangue , Complexo CD3/imunologia , Canadá , Criança , Diabetes Mellitus Tipo 1/sangue , Esquema de Medicação , Toxidermias/etiologia , Europa (Continente) , Feminino , Hemoglobinas Glicadas/metabolismo , Humanos , Hipoglicemiantes/administração & dosagem , Hipoglicemiantes/efeitos adversos , Hipoglicemiantes/imunologia , Índia , Insulina/administração & dosagem , Células Secretoras de Insulina/efeitos dos fármacos , Células Secretoras de Insulina/imunologia , Israel , Masculino , México , Muromonab-CD3/administração & dosagem , Muromonab-CD3/efeitos adversos , Muromonab-CD3/imunologia , Resultado do Tratamento , Estados Unidos , Adulto JovemRESUMO
Immunogenetic mechanisms operating within the immune system are known to influence cytokine profiles and disease susceptibility. Yet the role of the individual's neurohormonal background in these processes remains undefined. Hormonal imbalances are documented in immune-related diseases, but it is unclear whether this represents a secondary phenomenon or a primary "defect" related to specific neurohormonal immune phenotype(s). We report that in a large subpopulation of healthy humans the baseline epinephrine output (but not cortisol and sex steroid hormones) correlated inversely with proinflammatory and positively with anti-inflammatory cytokine production. Thus, low vs high epinephrine excretors had a 2- to 5-fold higher TNF-alpha and IL-12 production but 2-fold lower IL-10 production induced by LPS ex vivo. In alternative settings, we found low baseline levels and profoundly blunted stress-induced epinephrine responses but high TNF-alpha levels in Lewis vs Fischer inbred rats. Additionally, isoproterenol, a beta adrenoreceptor agonist suppressed LPS-induced TNF-alpha production, with more pronounced effect in Lewis than in Fischer rats. In human monocytes, epinephrine and the beta(2) adrenoreceptor agonist fenoterol potently inhibited LPS-induced TNF-alpha and IL-12, but stimulated IL-10 production. The order of potency for hormones able to inhibit IL-12 production ex vivo was: epinephrine > norepinephrine > or = 1,25-(OH)(2) vitamin D(3) > hydrocortisone. This indicates that baseline epinephrine conditions cytokine responsiveness and through this mechanism intrinsic hypo- or hyperactive adrenal medullas in some individuals may shape opposite cytokine profiles. Since Lewis and Fischer rats have opposite susceptibility to experimental immunological diseases, this suggests that the parallel human phenotypes could be linked to differing responsiveness and susceptibility to infections and immune/inflammatory-related conditions.
Assuntos
Citocinas/biossíntese , Citocinas/imunologia , Epinefrina/farmacologia , Imunidade Inata/efeitos dos fármacos , Imunidade Inata/imunologia , Animais , Células Cultivadas , Humanos , Masculino , Fenótipo , Ratos , Ratos Endogâmicos F344 , Ratos Endogâmicos Lew , Receptores Adrenérgicos beta/metabolismo , Sistema Nervoso Simpático/efeitos dos fármacos , Sistema Nervoso Simpático/imunologia , Sistema Nervoso Simpático/metabolismoRESUMO
Osteopetrosis is caused by a heterogenous group of bone diseases that result in an increase in skeletal mass because of inadequate osteoclastic bone resorption. In the op osteopetrotic rat, the disease has been linked to a single genetic locus located at the proximal end of rat chromosome 10. In this study, we identified a 1.5-cM genetic interval that contains the mutation. We then generated an improved radiation hybrid (RH) map of this region to identify potential functional and positional candidates for the op gene. Using the rat genome radiation hybrid panel, we mapped 57 markers including 24 genes (14 that have not yet been mapped in the rat) and 10 expressed sequence tag markers. Included in the mapped genes are several candidate genes that might significantly influence the biochemical pathways involved in osteopetrosis. These include genes involved in osteoclast differentiation, apoptosis, and the functional capabilities of mature osteoclasts to resorb bone. Further analysis of the genes and expressed transcripts mapped to this region may yield important insights into the multifactorial control of osteoclast function and the mechanisms of failed bone homeostasis in diseases such as osteopetrosis, osteoporosis, and rheumatoid arthritis in which failed bone homeostasis is an instigating or exacerbating circumstance of the disease process.
Assuntos
Osteopetrose/veterinária , Mapeamento de Híbridos Radioativos , Ratos/genética , Doenças dos Roedores/genética , Animais , Reabsorção Óssea/genética , Cricetinae , Cruzamentos Genéticos , Modelos Animais de Doenças , Etiquetas de Sequências Expressas , Homeostase , Humanos , Meiose , Osteoclastos/fisiologia , Osteopetrose/genética , Ratos Endogâmicos BN , Ratos Endogâmicos Lew , Homologia de SequênciaRESUMO
BACKGROUND: The identification of genetic factors controlling stress-responsiveness should advance the understanding of susceptibility to psychiatric illness. METHODS: Rat strains, F344/NHsd and LEW/NHsd, which differ in measures of stress-responsiveness and behaviors modeling psychiatric disorders, were bred to generate F2 progeny that were used in a quantitative trait loci (QTL) analysis to identify genomic regions influencing late-afternoon corticosterone levels. RESULTS: Regions on chromosomes 4 and 10 previously identified as influencing autoimmune phenomena were the most significant QTL observed, reaching suggestive significance at the genome-wide level. Congenic animals targeting these regions with F344/NHsd deoxyribonucleic acid on a DA/Bkl genomic background demonstrated corticosterone levels approximating those of F344/NHsd rats and differing significantly from DA/Bkl rats. CONCLUSIONS: Specific genomic regions influence both corticosterone levels and stress-related disease susceptibility. These findings not only represent the first identification of QTL controlling corticosterone levels but also suggest a mechanism underlying genetic differences in stress-responsiveness.
Assuntos
Corticosterona/sangue , Genoma , Transtornos de Estresse Traumático/metabolismo , Animais , Animais Congênicos , Distribuição de Qui-Quadrado , Mapeamento Cromossômico , Corticosterona/genética , Ligação Genética , Escore Lod , Masculino , Biologia Molecular/métodos , Locos de Características Quantitativas/fisiologia , Ratos , Ratos Endogâmicos F344 , Ratos Endogâmicos Lew , Transtornos de Estresse Traumático/genéticaRESUMO
Adaptation to stressful stimuli, maintenance of homeostasis, and ultimately, survival require bidirectional feedback communication among components of the stress response and immune and endocrine systems. Substantial progress has been made in delineating molecular, cellular, and systemic physiologic mechanisms underlying this communication, particularly mechanisms that target the immune system. For example, our understanding of the immunomodulatory activities of numerous neuroendocrine mediators, such as cortisol, estrogen, testosterone, DHEA, catecholamines, corticotropin-releasing hormone, and adenosine, has advanced substantially. Substantial progress has also been made in defining how abnormalities involving these factors may contribute to the initiation, progression, and severity of autoimmune rheumatic diseases, particularly rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). For RA, the available data support the view that inflammatory and immune system inhibitory mechanisms, involving the hypothalamic-pituitary-adrenal (HPA) axis and sympathetic nervous system are deficient. Age, gender, and reproductive status acting, in part, through gonadal hormonal effects on disease susceptibility genes also appear likely to modulate the inhibitory stress response systems and immune function. Animal model data also have provided direct evidence that many autoimmune disease regulatory genes are gender influenced. For SLE, a growing body of recent data indicates that estrogens and androgens exert contrasting effects on B-lymphocytes (i.e., estrogens enhance and testosterone suppresses autoantibody production). These observations provide potential new insights into SLE pathogenesis and gender differences in prevalence. Continued investigation will refine our understanding of these observations and will uncover even more extensive interactions of the nervous, immune, and endocrine systems. Moreover, it is highly likely that improved understanding of these interactions will translate into improved therapy for the rheumatic diseases.
Assuntos
Doenças Autoimunes/fisiopatologia , Lúpus Eritematoso Sistêmico/fisiopatologia , Neuroimunomodulação , Sistemas Neurossecretores/fisiopatologia , Doenças Reumáticas/fisiopatologia , Corticosteroides/fisiologia , Animais , Doenças Autoimunes/imunologia , Retroalimentação , Feminino , Previsões , Predisposição Genética para Doença , Hormônios Esteroides Gonadais/fisiologia , Homeostase , Humanos , Lúpus Eritematoso Sistêmico/imunologia , Masculino , Neuroimunomodulação/fisiologia , Sistema Hipófise-Suprarrenal/fisiologia , Ratos , Ratos Endogâmicos F344 , Ratos Endogâmicos Lew , Ratos Nus , Doenças Reumáticas/imunologiaRESUMO
Cia5 is a locus on rat chromosome 10 which regulates the severity of collagen- and pristane-induced arthritis (CIA and PIA). To refine the region toward positional identification, Cia5 subcongenic strains were generated and studied in PIA and CIA. The protective effect of the telomeric locus Cia5a was confirmed in both models. A second arthritis severity locus (Cia5d) was identified within the most centromeric portion of Cia5. DA.F344(Cia5d) rats had a significantly lower median arthritis severity index in PIA, but not in CIA, compared with DA. On histologic analyses DA.F344(Cia5a) and DA.F344(Cia5d) congenics with PIA preserved a nearly normal joint architecture compared with DA, including significant reduction in synovial hyperplasia, pannus, angiogenesis, inflammatory infiltration, bone and cartilage erosions. Cia5 and Cia5a synovial levels of IL-1beta mRNA were reduced. Although both DA.F344(Cia5) and DA.F344(Cia5a) rats were protected in CIA, the arthritis scores of DA.F344(Cia5) were significantly higher than those of DA.F344(Cia5a), suggesting the existence of a third locus where F344-derived alleles centromeric from Cia5a contribute to increased arthritis severity. The existence of the third locus was further supported by higher levels of autoantibodies against rat type II collagen in DA.F344(Cia5) congenics compared with DA.F344(Cia5a). Our results determined that Cia5 contains three major arthritis severity regulatory loci regulating central events in the pathogenesis of arthritis, and differentially influencing CIA and PIA. These loci are syntenic to regions on human chromosomes 17q and 5q implicated in the susceptibility to rheumatoid arthritis, suggesting that the identification of these genes will be relevant to human disease.
Assuntos
Artrite Experimental/genética , Artrite Experimental/patologia , Colágeno Tipo II/administração & dosagem , Exsudatos e Transudatos , Marcadores Genéticos , Locos de Características Quantitativas/genética , Membrana Sinovial/patologia , Terpenos/administração & dosagem , Animais , Animais Congênicos , Artrite Experimental/fisiopatologia , Autoanticorpos/biossíntese , Autoanticorpos/sangue , Bovinos , Colágeno Tipo II/imunologia , Relação Dose-Resposta Imunológica , Feminino , Marcadores Genéticos/imunologia , Interleucina-1/genética , Interleucina-1/metabolismo , Masculino , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos F344 , Índice de Gravidade de Doença , Especificidade da Espécie , Membrana Sinovial/metabolismo , Membrana Sinovial/fisiopatologiaRESUMO
OBJECTIVE: To construct rats congenic for the chromosome 2 arthritis-regulatory quantitative trait locus Cia10, originally identified in a (DA x ACI)F(2) intercross rat strain that had been assessed for collagen-induced arthritis (CIA), and to determine the effect of this congenic interval on arthritis severity, joint histologic structure, and cytokine transcription in rats with pristane-induced arthritis (PIA). METHODS: A 52.6-MB interval derived from the ACI (CIA- and PIA-resistant) strain and containing the Cia10 interval was introgressed into the DA (arthritis-susceptible) background through genotype-guided congenic breeding. Homozygous male and female DA.ACI(Cia10) congenic rats were studied for their susceptibility to and severity of PIA, and were compared with same-sex DA rats. Histologic analyses were done on hind paws collected on day 32 following the pristane injection. Levels of interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNFalpha) messenger RNA (mRNA) were measured with real-time polymerase chain reaction on synovial tissues from day-32 ankles. RESULTS: Both male and female DA.ACI(Cia10) congenic rats developed a significantly milder form of arthritis, with a 95% and 92% reduction in the arthritis severity index compared with DA male and female controls, respectively (males P < or = 0.001 and females P = 0.003). DA.ACI(Cia10) congenic rat synovial tissue was more likely to preserve its normal histologic architecture, including minimal to no cartilage and bone erosions, synovial hyperplasia, and pannus formation, and reduced numbers of vessels (angiogenesis), when compared with DA synovial tissue. There was a 2.7- and 2.4-fold reduction in the amount of IL-1beta and TNFalpha mRNA, respectively, in the synovial tissue of DA.ACI(Cia10) congenic rats compared with DA rats. Sequencing analyses of complementary DNA for the Cia10-predicted candidate gene Ptpn8, the rat homolog of the rheumatoid arthritis (RA)-susceptibility gene PTPN22, revealed no polymorphisms between the DA and ACI strains. CONCLUSION: This study determined that Cia10 harbors a major autoimmune arthritis-regulatory gene. This gene regulates clinical disease severity, histologic damage, and the levels of at least two central proinflammatory cytokines. We are in the process of narrowing down the critical region for positional cloning of the Cia10 gene. The identification of this gene will provide novel targets or pathways for focused candidate-gene studies in RA.
Assuntos
Artrite Experimental/genética , Artrite Experimental/patologia , Exsudatos e Transudatos/metabolismo , Articulações/patologia , Locos de Características Quantitativas , Animais , Animais Congênicos , Artrite Experimental/induzido quimicamente , Artrite Experimental/metabolismo , Peso Corporal , Cromossomos de Mamíferos , DNA Complementar/genética , Extremidades , Feminino , Interleucina-1/genética , Complexo Principal de Histocompatibilidade/genética , Masculino , Proteína Tirosina Fosfatase não Receptora Tipo 22 , Proteínas Tirosina Fosfatases/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos , Índice de Gravidade de Doença , Membrana Sinovial/metabolismo , Terpenos , Fator de Necrose Tumoral alfa/genéticaRESUMO
Streptococcal cell wall (SCW) arthritis in rats is an experimentally-induced inflammatory model with many features that resemble rheumatoid arthritis (RA) in humans. In this unit, Lewis rats are injected with an aqueous suspension of Group A SCW streptococcal cell wall peptidoglycan-polysaccharide polymers (SCW PG-PS) and observed for the development of arthritis. The resulting arthritis is biphasic. An acute phase typically develops within 48 hr, followed 10 to 21 days later by a chronic phase which persists for months. Support protocols are included for preparing Group A PG-PS and measuring PG-PS concentration.
Assuntos
Artrite Experimental/imunologia , Parede Celular/imunologia , Streptococcus/imunologia , Animais , Parede Celular/química , Modelos Animais de Doenças , Feminino , Injeções Intraperitoneais , Polissacarídeos Bacterianos/administração & dosagem , Ratos , Ratos Endogâmicos Lew , Streptococcus/química , Streptococcus pyogenes/químicaRESUMO
Osteopetrosis results from a heterogeneous group of congenital bone diseases that display inadequate osteoclastic bone resorption. We recently mapped tl (toothless), a mutation that causes osteopetrosis in rats, to a genetic region predicted to include the rat Csf1 gene. In this study, we sequenced the coding sequence of the rat Csf1 gene to determine if a mutation in Csf1 could be responsible for the tl phenotype. Sequencing revealed a 10-base insertion in the coding sequence of mutant animals that produces a frameshift and generates a stop codon early in the mutant Csf1 coding sequence. The 41 amino acid polypeptide predicted to be produced from the Csf1 promoter would have only the first nine amino acids of the wild-type rat protein. These data suggest that osteopetrosis develops in tl/tl rats because they cannot produce functional mCsf, a growth factor required for osteoclast differentiation and activation.
Assuntos
Fator Estimulador de Colônias de Macrófagos/genética , Mutação , Osteopetrose/genética , Sequência de Aminoácidos , Animais , Animais Congênicos , Sequência de Bases , Humanos , Camundongos , Dados de Sequência Molecular , Mapeamento de Híbridos Radioativos , Ratos , Ratos Endogâmicos Lew , Ratos Mutantes , Alinhamento de SequênciaRESUMO
OBJECTIVE: To screen for potential efficacy and assess the feasibility of intravenous (IV) doxycycline as a treatment for rheumatoid arthritis (RA). METHODS: The study was a (stratified, block) randomized, double blind, 12 week, pilot trial of IV doxycycline 300 mg/day versus identical appearing IV placebo given over 2 h for 14 days. The primary comparison was to a hypothesized placebo rate of 20% as described by Paulus. If a total of 14 consecutive subjects receiving doxycycline treatment did not respond, it would be considered futile to proceed to a Phase III trial. We planned a placebo group of 14 subjects to verify the placebo response rate and estimate sample size required for a definitive Phase III trial, if such a trial was warranted based on the pilot study. American College of Rheumatology (ACR) RA response criteria were used. After 23 subjects entered, the study was closed due to recruitment difficulties. RESULTS: At baseline, mean (SD) tender joint count was 37 (11.9), swollen joint count 30 (9.6), morning stiffness 317 (319) min, and erythrocyte sedimentation rate 72 mm/h (27.5). Randomization resulted in 10 subjects receiving doxycycline and 13 receiving placebo. Treatment was stopped in 8 subjects: in 6, treatment was ineffective (one taking doxycycline, 5 placebo), and in 2, rashes occurred (one taking doxycycline, one placebo). Only one subject met ACR response criteria in the doxycycline group and none in the placebo group. Having no responders in the placebo group was consistent with placebo response rate of 20% or less. Several patients required peripherally inserted central catheters for venous access. CONCLUSION: The efficacy of IV doxycycline as a treatment for RA could not be ruled out. However, as the proportion of responders was small, it is unlikely that potential efficacy of IV doxycycline would outweigh potential disadvantages of IV administration.
Assuntos
Antibacterianos/administração & dosagem , Artrite Reumatoide/tratamento farmacológico , Doxiciclina/administração & dosagem , Adulto , Feminino , Humanos , Injeções Intravenosas , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Resultado do TratamentoRESUMO
OBJECTIVE: To evaluate the regulatory potential of genetic loci controlling Mycobacterium butyricum adjuvant-induced arthritis (Mbt-AIA) using mono- and polycongenic rats. METHODS: Of 4 quantitative trait loci (QTLs) that regulate Mbt-AIA, F344 alleles at 3 of these loci, Aia1, Aia2, and Aia3, are associated with lower arthritis severity, whereas F344 alleles at Aia4 are associated with greater arthritis severity. In this study, we constructed congenic lines by transferring 1 or more of the F344 genomic segments containing Aia1, Aia2, and Aia3 onto the DA genome. We comparatively evaluated their responses to Mbt-AIA with the responses of parental DA and F344 rats. RESULTS: Aia1, encompassing the rat major histocompatibility complex, reduced arthritis severity in monocongenic rats of both sexes. The arthritis-lowering effects of Aia2 and Aia3 were sex-influenced and were therefore observed in only males and only females, respectively. Polycongenic rats containing F344 genomic regions at Aia1, Aia2, and Aia3 developed Mbt-AIA of relatively greater severity than did F344 rats, implying that in DA and F344 rats, there could be other Mbt-AIA loci in addition to Aia1, Aia2, Aia3, and Aia4. To test the possibility that some of these Mbt-AIA-regulatory loci may colocalize with other arthritis QTLs, we evaluated Mbt-AIA in DA.F344 monocongenic rats containing collagen-induced arthritis QTLs. Cia5 (the QTL region on chromosome 10), but not Cia5a, Cia4, or Cia6, also regulated Mbt-AIA, and was named Aia5. CONCLUSION: F344 genomic regions at Aia1, Aia2, and Aia3 and the newly identified Aia5 contain genes that reduce Mbt-AIA severity in DA rats. These Mbt-AIA-regulatory loci overlap rheumatoid arthritis-susceptibility loci in humans.
Assuntos
Artrite Experimental/genética , Alelos , Animais , Animais Congênicos , Artrite Reumatoide/genética , Mapeamento Cromossômico , Cromossomos Humanos Par 17 , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Mycobacterium , Característica Quantitativa Herdável , Ratos , Ratos Endogâmicos F344 , Índice de Gravidade de Doença , Fatores Sexuais , Organismos Livres de Patógenos EspecíficosRESUMO
OBJECTIVE: To identify additional sex-specific and epistatic quantitative trait loci (QTL) regulating collagen-induced arthritis (CIA) severity overall, as well as within different stages during the disease course, in an intercross between major histocompatibility complex-identical inbred rat strains DA/Bkl (susceptible) and ACI/Hsd (resistant). METHODS: Arthritic male (DA x ACI)F2 intercross offspring (n = 143) were analyzed separately from the females (n = 184). Phenotypic extremes (maximum arthritis scores [MAS]) were genotyped and used for QTL analysis. All 327 rats were genotyped with the simple sequence-length polymorphism (SSLP) markers closest to the peak of Cia7 and Cia10, the major loci previously identified in this intercross, and with SSLPs covering chromosomes 12 and 18. Phenotypes studied were disease onset, arthritis severity scores on days 14-39, MAS, mean and cumulative arthritis scores, delayed-type hypersensitivity, and antibody responses to rat type II collagen. RESULTS: A new female-specific arthritis-severity recessive locus was identified on rat chromosome 12 (Cia25), with a maximum effect observed on day 28 (logarithm of odds [LOD] 4.7). The homozygous DA genotype at Cia25 was associated with a 45% higher median arthritis score in females. Sequencing analyses of the Cia25 candidate gene Ncf1 revealed polymorphisms between DA and ACI. The previously identified locus, Cia10, was found to be male-specific. A 2-locus interaction model analysis identified a novel recessive chromosome 18 QTL, Cia26, which was dependent on Cia7, with its maximum effect observed at later stages during the disease course (peak LOD score of 3.6 for arthritis scores on day 39). CONCLUSION: This study identified 2 novel female-specific loci, and 1 male-specific locus. Cia25 regulates MAS and disease severity during the mid-to-late stages of the disease course and may be accounted for by Ncf1 polymorphisms. Cia26 is in epistasis with Cia7 and regulates later stages of disease, suggesting an involvement in disease perpetuation and/or chronicity.
Assuntos
Artrite Experimental/genética , Epistasia Genética , Locos Secundários de Histocompatibilidade/genética , Animais , Doença Crônica , Cruzamentos Genéticos , Feminino , Genótipo , Masculino , Fenótipo , Ratos , Índice de Gravidade de Doença , Fatores SexuaisRESUMO
OBJECTIVE: Collagen-induced arthritis (CIA) is a model of inflammatory arthritis with many similarities to rheumatoid arthritis (RA). We previously mapped in F(2) offspring of CIA-susceptible DA and CIA-resistant F344 rats, 5 quantitative trait loci (QTLs) for which F344 alleles were associated with reduced CIA severity. In the present study, we sought to characterize the independent arthritis-modulating effects of these 5 QTLs. METHODS: CIA-regulatory regions were transferred from the F344 genome to the DA background or vice versa by repeated backcrossing. The arthritis-modulating effects of the transferred alleles were determined by comparing the severity of experimentally induced arthritis in congenic rats with that in DA rats. RESULTS: Congenic lines with either the F344 major histocompatibility complex (MHC) on the DA background or the DA MHC on the F344 background were resistant to CIA, confirming both MHC and non-MHC contributions to the genetic regulation of CIA. F344 alleles at the Cia3 and Cia5 regions of chromosomes 4 and 10 reduced CIA severity relative to that observed in DA rats. F344 Cia4 and Cia6 regions of chromosomes 7 and 8 failed to significantly alter CIA severity. Arthritis-modifying effects of Cia4 and Cia6 were, however, detected in pristane-induced and/or Freund's incomplete adjuvant oil-induced arthritis. The arthritis-modifying effects of the non-MHC CIA-regulatory loci differed in males and females. CONCLUSION: These congenic lines confirmed the existence and location of genes that regulate the severity of experimental arthritis in rats. Mechanisms responsible for the sex-specificity of individual arthritis-regulatory loci may explain some of the sex differences observed in RA and other autoimmune diseases in humans.
Assuntos
Animais Congênicos/genética , Modulação Antigênica/genética , Artrite Experimental/genética , Complexo Principal de Histocompatibilidade/genética , Característica Quantitativa Herdável , Animais , Animais Congênicos/imunologia , Modulação Antigênica/imunologia , Artrite Experimental/imunologia , Artrite Experimental/fisiopatologia , Colágeno/imunologia , Colágeno/farmacologia , Modelos Animais de Doenças , Feminino , Adjuvante de Freund/imunologia , Adjuvante de Freund/farmacologia , Complexo Principal de Histocompatibilidade/imunologia , Masculino , Ratos , Ratos Endogâmicos F344 , Fatores Sexuais , Terpenos/imunologia , Terpenos/farmacologia , Fatores de TempoRESUMO
OBJECTIVE: A number of non-HLA loci that have shown evidence (P < 0.05) for linkage with rheumatoid arthritis (RA) have been previously identified. The present study attempts to confirm these findings. METHODS: We performed a second genome-wide screen of 256 new multicase RA families recruited from across the United States by the North American Rheumatoid Arthritis Consortium. Affected sibling pair analysis on the new data set was performed using SIBPAL. We subsequently combined our first and second data sets in an attempt to enhance the evidence for linkages in a larger sample size. We also evaluated the impact of covariates on the support for linkage, using LODPAL. RESULTS: Evidence of linkage at 1p13 (D1S1631), 6p21.3 (the HLA complex), and 18q21 (D18S858) (P < 0.05) was replicated in this independent data set. In addition, there was new evidence for linkage at 9p22 (D9S1121 [P = 0.001]) and 10q21 (D10S1221 [P = 0.0002] and D10S1225 [P = 0.0038]) in the current data set. The combined analysis of both data sets (512 families) showed evidence for linkage at the level of P < 0.005 at 1p13 (D1S1631), 1q43 (D1S235), 6q21 (D6S2410), 10q21 (D10S1221), 12q12 (D12S398), 17p13 (D17S1298), and 18q21 (D18S858). Linkage at HLA was also confirmed (P < 5 x 10(-12)). Inclusion of DRB1*04 as a covariate significantly increased the probability of linkage on chromosome 6. In addition, some linkages on chromosome 1 showed improved significance when modeling DRB1*04 or rheumatoid factor positivity as covariates. CONCLUSION: These results provide a rational basis for pursuing high-density linkage and association studies of RA in several regions outside of the HLA region, particularly on chromosomes 1p, 1q, and 18q.