- Email: [email protected]
Asthma Genetics
cells in nude mice. In summary, these data suggest that GPC3 is a candidate lung tumor suppressor gene, the expression of which may be regulated by tobacco exposure.
Differential Gene Expression of sFRP-1 and Apoptosis in Pulmonary Emphysema* Kazushi Imai, DDS, PhD; and Jeanine D’Armiento, MD, PhD
(CHEST 2002; 121:7S) to identify the unique molecular pathways I nthatorder are involved in emphysema, a differential display analysis was performed on lung tissue to identify the genes expressed in the lung tissue of emphysema patients but not in that of patients with healthy lungs. We detected 142 differentially expressed genes. One of these genes, secreted frizzled-related protein sFRP-1, an inhibitor of Wnt signaling, was further characterized by its role in the pathophysiology of emphysema. Although this gene was not detected in the healthy adult mouse lung, it was expressed in the distal epithelial cells of the lung during development and also in the lung in two separate mouse emphysema models. Since the Wnt pathway is involved in proliferation and apoptosis, a study was undertaken that determined that tissue from the emphysematous lung of human patients was undergoing apoptosis. The apoptotic index closely correlated with the severity of disease. In vitro tissue transfection studies have confirmed that sFRP-1 leads to apoptosis of pulmonary epithelial and endothelial cells. The identification of sFRP-1 in emphysema has provided us with novel insight into the pathophysiology of this disease. The characterization of the other 142 differentially expressed genes will allow us to further identify the changes in gene expression in this disease. These genes have been placed on array chips so as to compare the differential gene expression from patients at various clinical stages of COPD with that of smokers prior to the development of COPD.
*From the Division of Molecular Medicine, Department of Medicine, College of Physicians and Surgeons of Columbia University, New York, NY. Correspondence to: Jeanine D’Armiento, MD, PhD, Assistant Professor of Medicine, Columbia University, 630 W 168th St, P&S 9 – 449, New York, NY 10032; e-mail: [email protected]
Asthma Genetics* William Osmond Charles Cookson, MA, DPhil, MD
Asthma is the most common chronic childhood disease in developed nations and is a complex disease that has high social and economic costs. Asthma and its associated intermediate phenotypes are under a substantial degree of genetic control. Identifying the genes underlying asthma offers a means of better understanding its pathogenesis, with the promise of improving preventive strategies, diagnostic tools, and therapies. A number of chromosomal regions containing genes influencing asthma and atopy have been identified consistently by different groups, and a role for several candidate genes has been established. (CHEST 2002; 121:7S–13S) Abbreviations: HDM ⫽ house dust mite; IL ⫽ interleukin; LD ⫽ linkage disequilibrium; LPS ⫽ lipopolysaccharide; MHC ⫽ major histocompatibility complex; TCR ⫽ T-cell receptor; Th ⫽ T-helper cell; TNF ⫽ tumor necrosis factor
has become an epidemic, affecting 155 million A sthma individuals in the world. One child in seven in the
United Kingdom wheezes,1 and similar numbers suffer from the related disorder of eczema (atopic dermatitis).2,3 Asthma is due to a combination of strong genetic and environmental factors. It has risen in prevalence over the past 30 years in all Westernized societies,4 perhaps as a result of the loss of childhood infections.5–7 Many candidate gene and positional cloning studies of asthma have now been carried out. Although the number of candidate gene studies in asthma is growing rapidly, many contain small numbers of subjects and give equivocal results that do not subsequently replicate. This review will, therefore, concentrate on regions identified consistently through genetic linkage, because these by and large represent the strongest genetic effects, and candidates studied within these regions will be discussed in detail.
Genome Screens The first genome-wide screen for linkages to quantitative traits underlying asthma identified significant evidence for linkage on chromosomes 4q, 6 (near the major histocompatibility complex [MHC]), 7, 11q (containing Fc⑀RI-), 13q and 16. A replication sample of families in the same study confirmed linkage to chromosomes 4, 11, 13, and 16.8 A two-stage screen in Hutterite families from the United States found suggestive evidence for linkage and replication for loci on chromosome 5q, 12q, 19q, and 21q.9 A screen in German families identified suggestive *From the Department of Human Genetics, University of Oxford, Oxford, UK. Correspondence to: William Osmond Charles Cookson, MA, DPhil, MD, Wellcome Trust Centre for Human Genetics, Roosevelt Dr, Headington, Oxford OX3 7BN, UK; e-mail: [email protected] CHEST / 121 / 3 / MARCH, 2002 SUPPLEMENT
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evidence for linkage to asthma on chromosomes 2q (near the interleukin-1 cluster), 6p (near the MHC), 9, and 12q.10 A genome screen for responsiveness to the house dust mite (HDM) allergen found suggestive linkages to chromosomes 2q, 6p (near the MHC), and 13q, as well as chromosome 8p.11 A genome screen in American families from three racial groups found a weak linkage to broad regions that might match other studies on chromosomes 2q, 5q, 6p, 12q, 13q, and 14q.12 A two-stage genome screen in French families found replicated linkages on chromosomes 1p, 12q, and 17q.13 Thus, the loci most consistently and robustly identified by these screens are on chromosomes 5, 6, 12, and 13.
Chromosome 5 Chromosome 5q31 has been studied by many groups following an original observation of genetic linkage to total serum IgE concentrations in extended Amish pedigrees14 and the confirmation of linkage to the same region.15 The region also has been linked to eosinophil levels16 and to schistosomiasis resistance.17 The region contains several genes that modulate atopic responses, including interleukin (IL)-4, IL-13, IL-5, CD14, and granulocyte macrophage-colony-stimulating factor. Humans (and different strains of mice) seem to exhibit a constitutional preference for either cellular or humoral immune responses,5,18 which are associated with distinct cytokine profiles in T-helper cells (Ths). The profiles are classified as Th-1 or Th-2 responses, respectively. Th-2 responses are characterized by high levels of secretion of IL-4, IL-13, and IL-5, and are associated with atopy. A number of polymorphisms have been identified in IL-13 and are convincingly associated with a variation in IgE levels in large population samples.19 IL-13 enhances bronchial mucus secretion and up-regulates IgE production.20 It is in close proximity to IL-4 and is highly homologous to that gene. Polymorphisms within IL-4 have been less securely related to IgE levels or atopic disease than IL-13.21,22 Functionally important polymorphisms within the IL-4 receptor ␣ gene (on chromosome 16) are associated with atopy and asthma,23,24 although positive findings are not universal.25 CD14 is found on the surface of monocytes and macrophages, as well as in a soluble form. CD14 acts as a high-affinity ligand for bacterial lipopolysaccharide (LPS) [endotoxin], and initiates the nonspecific innate immune response to bacterial infection. A polymorphism upstream of the transcription start site for CD14 is associated with high levels of soluble CD14 and low levels of IgE.26 The prevalence of asthma correlates inversely with a rural lifestyle and high ambient levels of LPS, so it has been suggested that the CD14 interaction with LPS may be protective against allergic disease.26 The level of variation in IgE associated with any of the chromosome-5 polymorphisms is of the order of 1% or 2%, and the polymorphisms so far identified cannot be considered to have major influences on the allergic process. Genetic linkage maps of the region suggest the presence of a least two genes influencing atopy.27 Local8S
izations within the region have been imprecise, and the overall impression is that of a weak linkage to a number of adjoining loci. However, the coincidence of nonhomologous cytokine genes within a limited interval suggests that the region is involved in the coordinate regulation of cytokine responses. Sequence comparison between human and mouse of 1 megabase of the proximal cluster, including interleukins 4, 13, and 5, found 90 noncoding highly conserved sequences.28 Fifteen of these elements were found to be present in other mammals.28 The characterization of the largest element in yeast artificial chromosome transgenic mice revealed it to be a coordinate regulator of IL-4, IL-13, and IL-5.29 This work, therefore, has begun to unpick at a structural level the mechanisms for T-cell commitment to a particular cytokine profile. It also has identified a high level of conservation of controlling elements of gene expression between species. It will be of interest to see whether this level of conservation is observed in other genomic regions.
Chromosome 6 The MHC region on chromosome 6 has shown consistent linkage to asthma-associated phenotypes in several studies8 –12 and may be considered to be a major locus influencing allergic diseases. The MHC contains many molecules involved in innate and specific immunity. At the same time, the asthma phenotype is complex, containing inflammatory and allergic components. An investigation of the effects of the MHC on asthma and its related phenotypes, therefore, poses a methodological and statistical challenge. The class II genes of the MHC have recognized influences on the ability to respond to particular allergens.30 –35 Asthma and bronchial hyperresponsiveness are associated primarily with an allergy to the HDM and to a lesser degree with allergy to cat dander and molds,36,37 so that genetic control of specific IgE responses is of relevance to clinical disease. The T-cell receptor (TCR) genes, on chromosomes 7q and 14q, are also important potential genetic modifiers of the specific IgE response. Genetic linkage and allelic association have been reported between specific IgE responses and the TCR ␣/␦ locus on chromosome 14q.38,39 The class I genes of the MHC may have important effects on atopic responses, but these have not yet been adequately investigated. Similarly, the class III complement genes contain polymorphisms that may be of relevance to inflammatory or immune diseases, but which have not yet been tested in asthmatic subjects. Nonclassical MHC genes also may have an impact on asthma through nonallergic pathways, and a polymorphism in the control elements of inflammatory cytokines and their receptors is an important mechanism for flexibility in the immunoregulatory machinery.40 Tumor necrosis factor (TNF) is a potent proinflammatory cytokine that is found in excess in asthmatic airways. A polymorphism in the TNF complex is associated with variation on the expression of TNF-␣ and with the presence of asthma.41– 43 These results emphasize the inflammatory nature of the asthmatic response, as distinct from its allergic basis.
Thomas L. Petty 44th Annual Aspen Lung Conference: Pulmonary Genetics, Genomics, Gene Therapy
Chromosome 11q The linkage of atopy to a VNTR (ie, variable number of tandem repeats) polymorphism on chromosome 11q13 was first reported in 198944 and was at first disputed.45 The -chain of the high-affinity receptor for IgE (Fc⑀RI-) was subsequently localized to the region. The Fc⑀RI receptor acts as the allergic trigger on mast cells and other types of cells, and is central to the allergic response.46 The -chain is not essential for Fc⑀RI function, but it both stabilizes the surface expression of the receptor and acts as an amplifying element within it.47 Any variation in the level of the -chain expression may therefore modify receptor function. A polymorphism in Fc⑀RI- has been related to atopy,48 asthma,49 bronchial hyperresponsiveness,50,51and severe atopic dermatitis.52 A polymorphism within the gene also has been associated with levels of IgE in heavily parasitized Australian aborigines, implying a protective role for the gene in helminth infestation.53 Although coding changes have been identified within Fc⑀RI-,54,55 they are conservative and do not seem to alter gene function.56 The Ile181Leu polymorphisms identified by Shirakawa et al54 have not been found in several other studies57–59 but have been found in association with asthma in Kuwaiti Arabs60 and black South Africans.61 The difficulty in their identification nevertheless suggests that they are artifactual or in homologous sequences out of the Fc⑀RI- gene. The structural variation that causes the effect of this gene on patients with asthma, therefore, has not yet been identified. The genetic linkage and association of atopy to the locus both have been typified by a strong maternal effect,52,62,63 with preferential linkage and the transmission of maternal alleles to affected children. Maternal effects are wellrecognized in allergic disorders, and asthma, eczema, elevated serum IgE concentrations, and skin prick test positivity in children all have been accompanied by an increased prevalence of asthma or atopy in mothers.64 The preferential transmission of or linkage to alleles from either the maternal or paternal sides also has been observed for other loci influencing allergic disease, including those identified on chromosomes 13 and 16.8 Parent-of-origin effects have been noted in other immunologic disorders, including type I diabetes,65 rheumatoid arthritis,66 inflammatory bowel disease,67 and selective IgA deficiency.68 Parent-of-origin linkages also have been observed in the same diseases, so that the preferential linkage of paternal alleles is seen from the insulin locus and type I diabetes,69 and HLA alleles for selective IgA deficiency.68 These findings suggest a common underlying mechanism for parent-of-origin effects in immune disorders. The monoallelic expression of cytokines and cell-signaling molecules is recognized70 –72 and mediated by methylation,73 providing a potential mechanism for these effects.
Chromosome 12 The initial demonstration of the genetic linkage of asthma to chromosome 12q74 was followed by single-locus confirmatory studies75,76 and by several general genome screens.9,10,12,13 In addition, a genome screen in a mouse model of asthma found a linkage to bronchial hyperre-
sponsiveness on mouse chromosome 10 in the region of syntenic homology to human chromosome 12q.77 The facility with which many groups have identified a linkage to this region suggests that it is a true major atopy locus. Interferon-␥ does not seem to be responsible for the linkage. High-density mapping of the region has been begun,78 and it is likely that positional cloning of the gene will be possible.
Chromosome 13q14 A linkage of the total serum IgE to the esterase D protein polymorphism on chromosome 13q14 was reported in 1985.79 The linkage of chromosome 13q to atopy was confirmed by a genome-wide scan8 and by a singlelocus study of Japanese families.80 The same study identified potential linkage disequilibrium (LD) between disease and D13S153.80 A two-stage screen in Hutterite families from the United States found a linkage of asthma to chromosome 13q21.39 in the first-stage families but not in the second-stage families. A linkage of chromosome 13q14 to HDM allergy in children with asthma also has been observed,81 as has a linkage to children with atopic dermatitis.82 These results suggest that chromosome 13q14 also contains a major atopy locus. The same chromosomal region has been shown to be linked to total serum IgA concentrations.83 Low levels of serum IgA occur much more frequently in atopic children than in healthy children,84 and salivary IgA deficiency is more common in infants with atopic parents.85 Ig production is known to be under the control of many genes (reviewed by Corrigan86), none of which have been mapped to chromosome 13q14. The gene of interest may encode a regulatory component of the humoral immune system but, alternatively, might influence both IgA levels and atopic status by influencing the mucosal handling of allergens.
Sharing of Loci With Other Disorders Genetic studies of other disorders also may have an impact on asthma and atopy. Crohn’s disease and ulcerative colitis are inflammatory bowel diseases of unknown etiology that show familial clustering.87– 89 Genome-wide screens have implicated loci on chromosomes 3, 7, 12, and 16.90 –92 The regions on chromosomes 7 and 12 may coincide with the asthma and atopy loci on the same chromosomes. Polymorphism in the IL-1 cluster on chromosome 2 also has been shown to influence the severity of the disease.93,94 A genome-wide screen in families with rheumatoid arthritis similarly has shown linkage near the asthma locus on chromosome 2 and the TCR-␣ locus on chromosome 14.95 Linkage to type I diabetes is found near Fc⑀RI- on chromosome 11q13.96 These findings suggest that important genes or gene families may be common to several inflammatory and immune disorders.
Mendelian Disorders Asthma, eczema, and allergic diseases are associated with a number of mendelian disorders. The identification of genes causing such disorders is much easier than the CHEST / 121 / 3 / MARCH, 2002 SUPPLEMENT
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positional cloning of complex disease genes. These mendelian disorders may therefore be extremely helpful in identifying genes influencing asthma and allergy. They include Netherton’s syndrome,97,98 Job syndrome (also known as hyper-IgE syndrome or Buckley’s syndrome),99 thymic hypoplasia (DiGeorge syndrome), cellular deficiency with Igs (Nezelof syndrome),100 selective IgA deficiency, and Wiskott-Aldrich syndrome. Netherton’s syndrome is a rare recessive disease in which children are born with a severe ichthyotic dermatosis.97 Severe symptomatic atopy is a universal accompaniment of the disease.98 The gene for Netherton’s syndrome has recently been located distal to the chromosome 5 cytokine cluster,101 and the gene underlying the disorder has been identified as SPINK5.102 SPINK5 codes a multidomain serine protease inhibitor LEKTI, which is expressed in the epithelium, mucosa, and thymus.103 Common coding polymorphisms have been identified in the gene by our group, and these show associations with atopy, asthma, and eczema in children without Netherton’s syndrome. These findings therefore define a new pathway for allergic disorders. Other mendelian disorders localize to regions that may be relevant to common allergic diseases. Selective IgA deficiency has been localized to the MHC.68 Hypereosinophilia syndrome has been localized to the distal part of the chromosome 5 cytokine cluster,104 and it is of interest that acquired hypereosinophilia is associated consistently with translocations on chromosome 5q35.105,106 Hyper-IgE syndrome has been linked to the distal arm of chromosome 4q.107 A small chromosomal deletion in one child with the disease may have limited this localization to a 20-megabase interval.107 A linkage of the total serum IgE to this region has been seen in at least one genome scan (http://www.well.ox.ac.uk/asthma/public/GenomeScan/ index.html),8 suggesting that this locus also may have an affect on the normal regulation of IgE levels.
Conclusion The positional cloning of the genes underlying asthma and allergic disorders is becoming increasingly tractable. Agreed-on regions of strong genetic linkage have emerged, some of which coincide with linkages to singlegene disorders or to other immunologic diseases. The completion of the human genomic sequence and the availability of the deep public expressed sequence tag databases mean that laborious physical mapping of these loci may not be necessary. The key element in gene discovery will be the identification of robust patterns of LD between markers and disease. LD mapping of at least one atopy locus has shown that LD is irregularly distributed,108 and the challenge is to develop statistical as well as genotyping methods to handle dense local single-nucleotide polymorphism maps.
References 1 Strachan DP, Anderson HR, Limb ES, et al. A national survey of asthma prevalence, severity, and treatment in Great Britain. Arch Dis Child 1994; 70:174 –178 10S
2 Sampson HA. Pathogenesis of eczema. Clin Exp Allergy 1990; 20:459 – 467 3 Schulz-Larsen F. A genetic-epidemiologic study in a population-based twin sample. J Am Acad Dermatol 1993; 28: 719 –723 4 von Mutius E, Fritzsch C, Weiland SK, et al. Prevalence of asthma and allergic disorders among children in united Germany: a descriptive comparison. BMJ 1992; 305:1395–1399 5 Shirakawa T, Enomoto T, Shimazu S, et al. The inverse association between tuberculin responses and atopic disorder. Science 1997; 275:77–79 6 Cookson WO, Moffatt MF. Asthma: an epidemic in the absence of infection? Science 1997; 275:41– 42 7 Rook GA, Stanford JL. Give us this day our daily germs. Immunol Today 1998; 19:113–116 8 Daniels SE, Bhattacharrya S, James A, et al. A genome-wide search for quantitative trait loci underlying asthma. Nature 1996; 383:247–250 9 Ober C, Cox NJ, Abney M, et al. Genome-wide search for asthma susceptibility loci in a founder population: the Collaborative Study on the Genetics of Asthma. Hum Mol Genet 1998; 7:1393–1398 10 Wjst M, Fischer G, Immervoll T, et al. A genome-wide search for linkage to asthma: German Asthma Genetics Group. Genomics 1999; 58:1– 8 11 Hizawa N, Freidhoff L, Chiu Y, et al. Genetic regulation of Dermatophagoides pteronyssinus-specific IgE responsiveness: a genome-wide multipoint linkage analysis in families recruited through 2 asthmatic sibs; Collaborative Study on the Genetics of Asthma (CSGA). J Allergy Clin Immunol 1998; 102:436 – 442 12 The Collaborative Study on the Genetics of Asthma. A genome-wide search for asthma susceptibility loci in ethnically diverse populations. Nat Genet 1997; 15:389 –392 13 Dizier MH, Besse-Schmittler C, Guilloud-Bataille M, et al. Genome screen for asthma and related phenotypes in the French EGEA study [abstract]. Am J Respir Crit Care Med 1999; 159:A649 14 Marsh DG, Neely JD, Breazeale DR, et al. Linkage analysis of IL4 and other chromosome 5q31.1 markers and total serum immunoglobulin E concentrations. Science 1994; 264:1152–1156 15 Meyers DA, Postma DS, Panhuysen CI, et al. Evidence for a locus regulating total serum IgE levels mapping to chromosome 5. Genomics 1994; 23:464 – 470 16 Martinez F, Solomon S, Holberg C, et al. Linkage of circulating eosinophils to markers on chromosome 5q. Am J Respir Crit Care Med 1998; 158:1739 –1744 17 Marquet S, Abel L, Hillaire D, et al. Genetic localization of a locus controlling the intensity of infection by Schistosoma mansoni on chromosome 5q31– q33. Nat Genet 1996; 14: 181–184 18 Kelso A. Th1 and Th2 subsets: paradigms lost? Immunol Today 1995; 16:374 –379 19 Graves PE, Kabesch M, Halonen M, et al. A cluster of seven tightly linked polymorphisms in the IL-13 gene is associated with total serum IgE levels in three populations of white children. J Allergy Clin Immunol 2000; 105:506 –513 20 Wills-Karp M, Luyimbazi J, Xu X, et al. Interleukin-13: central mediator of allergic asthma. Science 1998; 282: 2258 –2261 21 Rosenwasser LJ, Klemm DJ, Dresback JK, et al. Promoter polymorphisms in the chromosome 5 gene cluster in asthma and atopy. Clin Exp Allergy 1995; 25:74 –78 22 Walley AJ, Cookson WO. Investigation of an interleukin-4 promoter polymorphism for associations with asthma and atopy. J Med Genet 1996; 33:689 – 692
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23 Shirakawa I, Deichmann KA, Izuhara I, et al. Atopy and asthma: genetic variants of IL-4 and IL-13 signalling. Immunol Today 2000; 21:60 – 64 24 Ober C, Leavitt SA, Tsalenko A, et al. Variation in the interleukin 4-receptor alpha gene confers susceptibility to asthma and atopy in ethnically diverse populations. Am J Hum Genet 2000; 66:517–526 25 Noguchi E, Shibasaki M, Arinami T, et al. No association between atopy/asthma and the ILe50Val polymorphism of IL-4 receptor. Am J Respir Crit Care Med 1999; 160: 342–345 26 Baldini M, Lohman I, Halonen M, et al. A polymorphism in the 5⬘ flanking region of the CD14 gene is associated with circulating soluble CD14 levels and with total serum immunoglobulin E. Am J Respir Cell Mol Biol 1999; 20:976 –983 27 Xu J, Levitt RC, Panhuysen CI, et al. Evidence for two unlinked loci regulating total serum IgE levels. Am J Hum Genet 1995; 57:425– 430 28 Loots GG, Locksley RM, Blankespoor CM, et al. Identification of a coordinate regulator of interleukins 4, 13, and 5 by cross-species sequence comparisons. Science 2000; 288: 136 –140 29 Lacy DA, Wang ZE, Symula DJ, et al. Faithful expression of the human 5q31 cytokine cluster in transgenic mice. J Immunol 2000; 164:4569 – 4574 30 Marsh DG, Meyers DA, Bias WB. The epidemiology and genetics of atopic allergy. N Engl J Med 1981; 305:1551– 1559 31 Young RP, Dekker JW, Wordsworth BP, et al. HLA-DR and HLA-DP genotypes and immunoglobulin E responses to common major allergens. Clin Exp Allergy 1994; 24: 431– 439 32 Fischer GF, Pickl WF, Fae I, et al. Association between IgE response against Bet v I, the major allergen of birch pollen, and HLA-DRB alleles. Hum Immunol 1992; 33:259 –265 33 Sparholt SH, Georgsen J, Madsen HO, et al. Association between HLA- DRB3*0101 and immunoglobulin-E responsiveness to Bet v I. Hum Immunol 1994; 39:76 –78 34 D’Amato M, Scotto d’Abusco A, Maggi E, et al. Association of responsiveness to the major pollen allergen of Parietaria officinalis with HLA-DRB1* alleles: a multicenter study. Hum Immunol 1996; 46:100 –106 35 Donfack J, Tsalenko A, Hoki DM, et al. HLA-DRB1*01 alleles are associated with sensitization to cockroach allergens. J Allergy Clin Immunol 2000; 105:960 –966 36 Sears MR, Herbison GP, Holdaway MD, et al. The relative risks of sensitivity to grass pollen, house dust mite and cat dander in the development of childhood asthma. Clin Allergy 1989; 18:419 – 424 37 Cookson WOCM, De Klerk NH, Ryan GR, et al. Relative risks of bronchial hyper-responsiveness associated with skinprick test responses to common antigens in young adults. Clin Exp Allergy 1991; 21:473– 479 38 Moffatt MF, Hill MR, Cornelis F, et al. Genetic linkage of T cell receptor ␣/␦ complex to specific IgE responses. 1994; Lancet 343:1597–1600 39 Moffatt MF, Schou C, Faux JA, et al. Germline TCR-A restriction of immunoglobulin E responses to allergen. Immunogenetics 1997; 46:226 –230 40 Daser A, Mitchison H, Mitchison A, et al. Non-classicalMHC genetics of immunological disease in man and mouse: the key role of pro-inflammatory cytokine genes. Cytokine 1996; 8:593–597 41 Moffatt MF, Cookson WO. Tumour necrosis factor haplotypes and asthma. Hum Mol Genet 1997; 6:551–554 42 Li Kam Wa TC, Mansur AH, Britton J, et al. Association between ⫺308 tumour necrosis factor promoter polymor-
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phism and bronchial hyperreactivity in asthma. Clin Exp Allergy 1999; 29:1204 –1208 Chagani T, Pare PD, Zhu S, et al. Prevalence of tumor necrosis factor-alpha and angiotensin converting enzyme polymorphisms in mild/moderate and fatal/near-fatal asthma. Am J Respir Crit Care Med 1999; 160:278 –282 Cookson WOCM, Sharp PA, Faux J, et al. Linkage between immunoglobulin E responses underlying asthma and rhinitis and chromosome 11q. Lancet 1989; 1:1292–1295 Morton NE. Major loci for atopy? [editorial]. Clin Exp Allergy 1992; 22:1041–1043 Turner H, Kinet JP. Signalling through the high-affinity IgE receptor Fc epsilonRI. Nature 1999; 402:B24 –B30 Lin S, Cicala C, Scharenberg A, et al. The Fc(epsilon)RIbeta subunit functions as an amplifier of Fc(epsilon)RIgammamediated cell activation signals. Cell 1996; 85:985–995 Hill MR, James AL, Faux JA, et al. Fc⑀RI- polymorphism and risk of atopy in a general population sample. BMJ 1995; 311:776 –779 Shirakawa T, Mao XQ, Sasaki S, et al. Association between Fc epsilon RI beta and atopic disorder in a Japanese population [letter]. Lancet 1996; 347:394 –395 van Herwerden L, Harrap SB, Wong ZY, et al. Linkage of high-affinity IgE receptor gene with bronchial hyperreactivity, even in absence of atopy. Lancet 1995; 346:1262–1265 Trabetti E, Cusin V, Malerba G, et al. Association of the FcepsilonRIbeta gene with bronchial hyper-responsiveness in an Italian population. J Med Genet 1998; 35:680 – 681 Cox HE, Moffatt MF, Faux JA, et al. Association of atopic dermatitis to the beta subunit of the high affinity immunoglobulin E receptor. Br J Dermatol 1998; 138:182–187 Palmer LJ, Pare PD, Faux JA, et al. Fc epsilon R1-beta polymorphism and total serum IgE levels in endemically parasitized Australian aborigines. Am J Hum Genet 1997; 61:182–188 Shirakawa T, Li A, Dubowitz M, et al. Association between atopy and variants of the  subunit of the high affinity immunoglobulin E receptor. Nat Genet 1994; 7:125–129 Hill MR, Cookson WOCM. A new variant of the  subunit of the high affinity receptor for immunoglobulin E (Fc⑀RIE237G): associations with measures of atopy and bronchial hyper-responsiveness. Hum Mol Genet 1996; 5:959 –962 Donnadieu E, Cookson WO, Jouvin MH, et al. Allergyassociated polymorphisms of the FcepsilonRIbeta subunit do not impact its two amplification functions. J Immunol 2000; 165:3917–3922 Duffy D, Healey S, Chenevix-Trench G, et al. Atopy in Australia [letter]. Nat Genet 1995; 10:260 Rohrbach M, Kraemer R, Liechti-Gallati S. Screening of the Fc epsilon RI-beta-gene in a Swiss population of asthmatic children: no association with E237G and identification of new sequence variations. Dis Markers 1998; 14:177–186 Dickson P, Wong Z, Harrap S, et al. Mutational analysis of the high affinity immunoglobulin E receptor beta subunit gene in asthma. Thorax 1999; 54:409 – 412 Haider M, Hijazi Z. Prevalence of high affinity IgE receptor [Fc epsilon RI beta] gene polymorphisms in Kuwaiti Arabs with asthma [letter]. Clin Genet 1998; 54:166 –167 Green S, Gaillard M, Song E, et al. Polymorphisms of the beta chain of the high-affinity immunoglobulin E receptor (Fcepsilon RI-beta) in South African black and white asthmatic and nonasthmatic individuals. Am J Respir Crit Care Med 1998; 158:1487–1492 Cookson WO, Young RP, Sandford AJ, et al. Maternal inheritance of atopic IgE responsiveness on chromosome 11q. Lancet 1992; 340:381–384 Shirakawa T, Hashimoto T, Furuyama J, et al. Linkage CHEST / 121 / 3 / MARCH, 2002 SUPPLEMENT
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between severe atopy and chromosome 11q13 in Japanese families. Clin Genet 1994; 46:228 –232 Moffatt M, Cookson W. The genetics of asthma: maternal effects in atopic disease. Clin Exp Allergy 1998; 28(suppl): 56 – 61 Warram JH, Krolewski AS, Gottlieb MS, et al. Differences in risk of insulin-dependent diabetes in offspring of diabetic mothers and diabetic fathers. N Engl J Med 1984; 311: 149 –152 Koumantaki Y, Giziaki E, Linos A, et al. Family history as a risk factor for rheumatoid arthritis: a case-control study. J Rheumatol 1997; 24:1522–1526 Akolkar PN, Gulwani-Akolkar B, Heresbach D, et al. Differences in risk of Crohn’s disease in offspring of mothers and fathers with inflammatory bowel disease. Am J Gastroenterol 1997; 92:2241–2244 Vorechovsky I, Webster AD, Plebani A, et al. Genetic linkage of IgA deficiency to the major histocompatibility complex: evidence for allele segregation distortion, parentof-origin penetrance differences, and the role of anti-IgA antibodies in disease predisposition. Am J Hum Genet 1999; 64:1096 –1109 Bennett S, Todd J. Human type 1 diabetes and the insulin gene: principles of mapping polygenes. Annu Rev Genet 1996; 30:343–370 Bix M, Locksley RM. Independent and epigenetic regulation of the interleukin-4 alleles in CD4⫹ T cells. Science 1998; 281:1352–1354 Hollander G, Zuklys S, Morel C, et al. Monoallelic expression of the interleukin-2 locus. Science 1998; 279:2118 –2121 Held W, Raulet D. Expression of the Ly49A gene in murine natural killer cell clones is predominantly but not exclusively mono-allelic. Eur J Immunol 1997; 27:2876 –2884 Young HA, Ghosh P, Ye J, et al. Differentiation of the T helper phenotypes by analysis of the methylation state of the IFN-gamma gene. J Immunol 1994; 153:3603–3610 Barnes KC, Neely JD, Duffy DL, et al. Linkage of asthma and total serum IgE concentration to markers on chromosome 12q: evidence from Afro-Caribbean and caucasian populations. Genomics 1996; 37:41–50 Nickel R, Wahn U, Hizawa N, et al. Evidence for linkage of chromosome 12q15– q24.1 markers to high total serum IgE concentrations in children of the German Multicenter Allergy Study. Genomics 1997; 46:159 –162 Wilkinson J, Thomas NS, Morton N, et al. Candidate gene and mutational analysis in asthma and atopy. Int Arch Allergy Immunol 1999; 118:265–267 Zhang Y, Lefort J, Kearsey V, et al. A genome-wide screen for asthma-associated quantitative trait loci in a mouse model of allergic asthma. Hum Mol Genet 1999; 8:601– 605 Barnes KC, Freidhoff LR, Nickel R, et al. Dense mapping of chromosome 12q13.12-q23.3 and linkage to asthma and atopy. J Allergy Clin Immunol 1999; 104:485– 491 Eiberg H, Lind P, Mohr J, et al. Linkage relationship between the human immunoglobulin E polymorphism and marker systems [abstract]. Cytogenet Cell Genet 1985; 40:622 Kimura K, Noguchi E, Shibasaki M, et al. Linkage and association of atopic asthma to markers on chromosome 13 in the Japanese population. Hum Mol Genet 1999; 8:1487– 1490 Hizawa N, Freidhoff L, Ehrlich E, et al. Genetic influences of chromosomes 5q31– q33 and 11q13 on specific IgE responsiveness to common inhaled allergens among African American families: Collaborative Study on the Genetics of Asthma (CSGA). J Allergy Clin Immunol 1998; 102:449– 453 Beyer KWU, Freidhoff L, Nickel R, et al. Evidence for linkage of chromosome 5q31– q33 and 13q12– q14 markers
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to atopic dermatitis [abstract]. J Allergy Clin Immunol 1998; 101:152 Wiltshire S, Bhattacharyya S, Faux JA, et al. A genome scan for loci influencing total serum immunoglobulin levels: possible linkage of IgA to the chromosome 13 atopy locus. Hum Mol Genet 1998; 7:27–31 Ludviksson BR, Eiriksson TH, Ardal B, et al. Correlation between serum immunoglobulin A concentrations and allergic manifestations in infants. J Pediatr 1992; 121:23–27 van Asperen PP, Gleeson M, Kemp AS, et al. The relationship between atopy and salivary IgA deficiency in infancy. Clin Exp Immunol 1985; 62:753–757 Corrigan CJ. T and B lymphocytes and the development of allergic reactions. In: Kay AB, ed. Allergy and allergic diseases. Oxford, UK: Blackwell Science Ltd, 1997; 36 –57 Rioux JD, Silverberg MS, Daly MJ, et al. Genome-wide search in Canadian families with inflammatory bowel disease reveals two novel susceptibility loci. Am J Hum Genet 2000; 66:1863–1870 Duerr RH, Barmada MM, Zhang L, et al. High-density genome scan in Crohn disease shows confirmed linkage to chromosome 14q11–12. Am J Hum Genet 2000; 66:1857–1862 Ma Y, Ohmen JD, Li Z, et al. A genome-wide search identifies potential new susceptibility loci for Crohn’s disease. Inflamm Bowel Dis 1999; 5:271–278 Satsangi J, Parkes M, Louis E, et al. Two stage genome-wide search in inflammatory bowel disease provides evidence for susceptibility loci on chromosomes 3, 7 and 12. Nat Genet 1996; 14:199 –202 Hugot JP, Thomas G. Genome-wide scanning in inflammatory bowel diseases. Dig Dis 1998; 16:364 –369 Duerr RH, Barmada MM, Zhang L, et al. Linkage and association between inflammatory bowel disease and a locus on chromosome 12. Am J Hum Genet 1998; 63:95–100 Mansfield J, Holden H, Tarlow J, et al. Novel genetic association between ulcerative colitis and the anti-inflammatory cytokine interleukin-1 receptor antagonist. Gastroenterology 1994; 106:637– 642 Parkes M, Satsangi J, Jewell D. Contribution of the IL-2 and IL-10 genes to inflammatory bowel disease (IBD) susceptibility. Clin Exp Immunol 1998; 113:28 –32 Hardwick L, Walsh S, Butcher S, et al. Genetic mapping of susceptibility loci in the genes involved in rheumatoid arthritis. J Rheumatol 1998; 24:197–198 Nakagawa Y, Kawaguchi Y, Twells R, et al. Fine mapping of the diabetes-susceptibility locus, IDDM4, on chromosome 11q13. Am J Hum Genet 1998; 63:547–556 Netherton EW. A unique case of trichorrhexis nodosa, “Bamboo hairs.” Arch Dermatol 1958; 78:483– 487 Judge MR, Morgan G, Harper JI. A clinical and immunological study of Netherton’s syndrome. Br J Dermatol 1994; 131:615– 621 Davis SD, Schaller J, Wedgwood RJ. Job’s syndrome: recurrent, cold, staphylococcal abscesses. Lancet 1966; 1:1013–1015 Knutsen AP, Wall D, Mueller KR, et al. Abnormal in vitro thymocyte differentiation in a patient with severe combined immunodeficiency-Nezelof’s syndrome. J Clin Immunol 1996; 16:151–158 Chavanas S, Garner C, Bodemer C, et al. Localization of the Netherton syndrome gene to chromosome 5q32, by linkage analysis and homozygosity mapping. Am J Hum Genet 2000; 66:914 –921 Chavanas S, Bodemer C, Rochat A, et al. Mutations in SPINK5, encoding a serine protease inhibitor, cause Netherton syndrome. Nat Genet 2000; 25:141–142 Ma¨ gert HJ, Standker L, Kreutzmann P, et al. LEKTI, a novel 15-domain type of human serine proteinase inhibitor.
Thomas L. Petty 44th Annual Aspen Lung Conference: Pulmonary Genetics, Genomics, Gene Therapy
J Biol Chem 1999; 274:21499 –21502 104 Rioux J, Stone V, Daly M, et al. Familial eosinophilia maps to the cytokine gene cluster on human chromosomal region 5q31– q33. Am J Hum Genet 1998; 63:1086 –1094 105 Jani K, Kempski H, Reeves B. A case of myelodysplasia with eosinophilia having a translocation t(5; 12) (q31; q13) restricted to myeloid cells but not involving eosinophils. Br J Haematol 1994; 87:57– 60 106 Sato H, Saito H, Ikebuchi K, et al. Biological characteristics of chronic eosinophilic leukemia cells with a t(2; 5)(p23; q35) translocation. Leuk Lymphoma 1995; 19: 499 –505 107 Grimbacher B, Schaffer AA, Holland SM, et al. Genetic linkage of hyper-IgE syndrome to chromosome 4. Am J Hum Genet 1999; 65:735–744 108 Moffatt MF, Traherne JA, Abecasis GR, et al. Single nucleotide polymorphism and linkage disequilibrium within the TCR alpha/delta locus. Hum Mol Genet 2000; 9:1011–1019
Genetic Analysis of AntigenInduced Airway Manifestations of Asthma Using Recombinant Congenic Mouse Strains* Antoon J.M. Van Oosterhout, PhD; Prescilla V. Jeurink; Peter C. Groot, PhD; Gerard A. Hofman; Frans P. Nijkamp, PhD; and Peter Demant, PhD
(CHEST 2002; 121:13S) Abbreviation: OVA ⫽ ovalbumin
asthma is a heterogeneous and genetically A llergic complex disease that is characterized by the pres-
ence of allergen-specific IgE, eosinophilic airway inflammation, and hyperresponsiveness to bronchospasmogenic stimuli. To facilitate the mapping of genes controlling complex asthma traits, we have used a powerful tool, the recombinant congenic strains of mice, which transforms a multigenic difference into a set of monogenic or oligogenic differences.1 This approach offers a higher resolution power than do the standard mouse crosses in mapping segregating quantitative trait loci and in the detection of their potential interactions. In the present study, we compared 4 strains (Ccs/DEM-5, Ccs/DEM-11, Ccs/DEM-12, and Ccs/DEM-18) from a series of 20 strains, each of which carries a random set of 12.5% of genes from the T-helper cell type 1 responder strain STS and 87.5% of
Table 1—Lung Function and Inflammation* Strain 5 11 12 18
(n ⫽ 13) (n ⫽ 4) (n ⫽ 12) (n ⫽ 8)
Penh Maximum 2.6 ⫾ 0.3 2.5 ⫾ 0.2 3.3 ⫾ 0.3 2.7 ⫾ 0.3
vs vs vs vs
⌬ED300
3.5 ⫾ 0.5† ⫺ 1.1 ⫾ 4.4 3.0 ⫾ 0.3† ⫺ 33.2 ⫾ 8.3† 3.0 ⫾ 0.4 ⫹ 6.6 ⫾ 5.1 6.0 ⫾ 1.1† ⫺ 6.3 ⫾ 3.7
Eosinophils, ⫻ 105 1.0 ⫾ 0.4 2.9 ⫾ 1.5 5.1 ⫾ 1.2 6.0 ⫾ 1.5
*Values given as mean ⫾ SEM. ED300 ⫽ dose of methacholine needed to increase baseline values of enhanced pause by 300%; Penh ⫽ enhanced pause. †p ⬍ 0.05, as determined by Student’s t test.
genes from the T-helper cell type 2 responder strain BALB/c. Mice were sensitized with ovalbumin (OVA), and later on were repeatedly challenged by the inhalation of an OVA aerosol. Before and after the OVA challenges, serum samples were obtained and the airway responsiveness to nebulized methacholine (dose range, 1.5 to 50 mg/mL) was determined. Finally, BAL was performed, and the number of leukocytes were determined (Table 1). All mouse strains showed increased serum levels of OVA-specific IgE as a result of the OVA inhalation challenge (data not shown). It appears that there is no correlation between the number of BAL eosinophils and the extent of airway hyperresponsiveness. Strain 12 displayed airway eosinophilia but a resistance to hyperresponsiveness. However, the baseline airway responsiveness of strain 12 was higher than that of the other strains. In strain 11, airway hyperresponsiveness was determined predominantly by an increased sensitivity (ie, the dose of methacholine needed to increase baseline values of enhanced pause by 300%) to methacholine, whereas in strains 5 and 18 only a significant increase in the maximal response could be observed. These data demonstrate that different asthma traits like the presence of IgE, eosinophilia, and hyperresponsiveness may be genetically dissociated. Further mapping studies will be carried out to localize the modifier genes involved in specific traits.
Reference 1 Groot PC, Moen CJ, Dietrich W, et al. The recombinant congenic strains for analysis of multigenic traits: genetic composition. FASEB J 1992; 6:2826 –2835
*From the Department of Pharmacology and Pathophysiology (Drs. Van Oosterhout, Groot, and Nijkamp, Ms. Jeurink, and Mr. Hofman), Faculty of Pharmacy, Utrecht University, Utrecht, Netherlands; Division of Molecular Genetics (Dr. Demant), Netherlands Cancer Institute, Amsterdam, Netherlands. This research was supported by a research grant (AF99.23) of the Dutch Asthma Foundation. Correspondence to: Antoon J.M. Van Oosterhout, PhD, Department of Pharmacology and Pathophysiology, Faculty of Pharmacy, Utrecht University, PO box 80.082, 3508TB Utrecht, Netherlands; e-mail: [email protected] CHEST / 121 / 3 / MARCH, 2002 SUPPLEMENT
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Differential Gene Expression of sFRP-1 and Apoptosis in Pulmonary Emphysema* Kazushi Imai, DDS, PhD; and Jeanine D’Armiento, MD, PhD
(CHEST 2002; 121:7S) to identify the unique molecular pathways I nthatorder are involved in emphysema, a differential display analysis was performed on lung tissue to identify the genes expressed in the lung tissue of emphysema patients but not in that of patients with healthy lungs. We detected 142 differentially expressed genes. One of these genes, secreted frizzled-related protein sFRP-1, an inhibitor of Wnt signaling, was further characterized by its role in the pathophysiology of emphysema. Although this gene was not detected in the healthy adult mouse lung, it was expressed in the distal epithelial cells of the lung during development and also in the lung in two separate mouse emphysema models. Since the Wnt pathway is involved in proliferation and apoptosis, a study was undertaken that determined that tissue from the emphysematous lung of human patients was undergoing apoptosis. The apoptotic index closely correlated with the severity of disease. In vitro tissue transfection studies have confirmed that sFRP-1 leads to apoptosis of pulmonary epithelial and endothelial cells. The identification of sFRP-1 in emphysema has provided us with novel insight into the pathophysiology of this disease. The characterization of the other 142 differentially expressed genes will allow us to further identify the changes in gene expression in this disease. These genes have been placed on array chips so as to compare the differential gene expression from patients at various clinical stages of COPD with that of smokers prior to the development of COPD.
*From the Division of Molecular Medicine, Department of Medicine, College of Physicians and Surgeons of Columbia University, New York, NY. Correspondence to: Jeanine D’Armiento, MD, PhD, Assistant Professor of Medicine, Columbia University, 630 W 168th St, P&S 9 – 449, New York, NY 10032; e-mail: [email protected]
Asthma Genetics* William Osmond Charles Cookson, MA, DPhil, MD
Asthma is the most common chronic childhood disease in developed nations and is a complex disease that has high social and economic costs. Asthma and its associated intermediate phenotypes are under a substantial degree of genetic control. Identifying the genes underlying asthma offers a means of better understanding its pathogenesis, with the promise of improving preventive strategies, diagnostic tools, and therapies. A number of chromosomal regions containing genes influencing asthma and atopy have been identified consistently by different groups, and a role for several candidate genes has been established. (CHEST 2002; 121:7S–13S) Abbreviations: HDM ⫽ house dust mite; IL ⫽ interleukin; LD ⫽ linkage disequilibrium; LPS ⫽ lipopolysaccharide; MHC ⫽ major histocompatibility complex; TCR ⫽ T-cell receptor; Th ⫽ T-helper cell; TNF ⫽ tumor necrosis factor
has become an epidemic, affecting 155 million A sthma individuals in the world. One child in seven in the
United Kingdom wheezes,1 and similar numbers suffer from the related disorder of eczema (atopic dermatitis).2,3 Asthma is due to a combination of strong genetic and environmental factors. It has risen in prevalence over the past 30 years in all Westernized societies,4 perhaps as a result of the loss of childhood infections.5–7 Many candidate gene and positional cloning studies of asthma have now been carried out. Although the number of candidate gene studies in asthma is growing rapidly, many contain small numbers of subjects and give equivocal results that do not subsequently replicate. This review will, therefore, concentrate on regions identified consistently through genetic linkage, because these by and large represent the strongest genetic effects, and candidates studied within these regions will be discussed in detail.
Genome Screens The first genome-wide screen for linkages to quantitative traits underlying asthma identified significant evidence for linkage on chromosomes 4q, 6 (near the major histocompatibility complex [MHC]), 7, 11q (containing Fc⑀RI-), 13q and 16. A replication sample of families in the same study confirmed linkage to chromosomes 4, 11, 13, and 16.8 A two-stage screen in Hutterite families from the United States found suggestive evidence for linkage and replication for loci on chromosome 5q, 12q, 19q, and 21q.9 A screen in German families identified suggestive *From the Department of Human Genetics, University of Oxford, Oxford, UK. Correspondence to: William Osmond Charles Cookson, MA, DPhil, MD, Wellcome Trust Centre for Human Genetics, Roosevelt Dr, Headington, Oxford OX3 7BN, UK; e-mail: [email protected] CHEST / 121 / 3 / MARCH, 2002 SUPPLEMENT
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evidence for linkage to asthma on chromosomes 2q (near the interleukin-1 cluster), 6p (near the MHC), 9, and 12q.10 A genome screen for responsiveness to the house dust mite (HDM) allergen found suggestive linkages to chromosomes 2q, 6p (near the MHC), and 13q, as well as chromosome 8p.11 A genome screen in American families from three racial groups found a weak linkage to broad regions that might match other studies on chromosomes 2q, 5q, 6p, 12q, 13q, and 14q.12 A two-stage genome screen in French families found replicated linkages on chromosomes 1p, 12q, and 17q.13 Thus, the loci most consistently and robustly identified by these screens are on chromosomes 5, 6, 12, and 13.
Chromosome 5 Chromosome 5q31 has been studied by many groups following an original observation of genetic linkage to total serum IgE concentrations in extended Amish pedigrees14 and the confirmation of linkage to the same region.15 The region also has been linked to eosinophil levels16 and to schistosomiasis resistance.17 The region contains several genes that modulate atopic responses, including interleukin (IL)-4, IL-13, IL-5, CD14, and granulocyte macrophage-colony-stimulating factor. Humans (and different strains of mice) seem to exhibit a constitutional preference for either cellular or humoral immune responses,5,18 which are associated with distinct cytokine profiles in T-helper cells (Ths). The profiles are classified as Th-1 or Th-2 responses, respectively. Th-2 responses are characterized by high levels of secretion of IL-4, IL-13, and IL-5, and are associated with atopy. A number of polymorphisms have been identified in IL-13 and are convincingly associated with a variation in IgE levels in large population samples.19 IL-13 enhances bronchial mucus secretion and up-regulates IgE production.20 It is in close proximity to IL-4 and is highly homologous to that gene. Polymorphisms within IL-4 have been less securely related to IgE levels or atopic disease than IL-13.21,22 Functionally important polymorphisms within the IL-4 receptor ␣ gene (on chromosome 16) are associated with atopy and asthma,23,24 although positive findings are not universal.25 CD14 is found on the surface of monocytes and macrophages, as well as in a soluble form. CD14 acts as a high-affinity ligand for bacterial lipopolysaccharide (LPS) [endotoxin], and initiates the nonspecific innate immune response to bacterial infection. A polymorphism upstream of the transcription start site for CD14 is associated with high levels of soluble CD14 and low levels of IgE.26 The prevalence of asthma correlates inversely with a rural lifestyle and high ambient levels of LPS, so it has been suggested that the CD14 interaction with LPS may be protective against allergic disease.26 The level of variation in IgE associated with any of the chromosome-5 polymorphisms is of the order of 1% or 2%, and the polymorphisms so far identified cannot be considered to have major influences on the allergic process. Genetic linkage maps of the region suggest the presence of a least two genes influencing atopy.27 Local8S
izations within the region have been imprecise, and the overall impression is that of a weak linkage to a number of adjoining loci. However, the coincidence of nonhomologous cytokine genes within a limited interval suggests that the region is involved in the coordinate regulation of cytokine responses. Sequence comparison between human and mouse of 1 megabase of the proximal cluster, including interleukins 4, 13, and 5, found 90 noncoding highly conserved sequences.28 Fifteen of these elements were found to be present in other mammals.28 The characterization of the largest element in yeast artificial chromosome transgenic mice revealed it to be a coordinate regulator of IL-4, IL-13, and IL-5.29 This work, therefore, has begun to unpick at a structural level the mechanisms for T-cell commitment to a particular cytokine profile. It also has identified a high level of conservation of controlling elements of gene expression between species. It will be of interest to see whether this level of conservation is observed in other genomic regions.
Chromosome 6 The MHC region on chromosome 6 has shown consistent linkage to asthma-associated phenotypes in several studies8 –12 and may be considered to be a major locus influencing allergic diseases. The MHC contains many molecules involved in innate and specific immunity. At the same time, the asthma phenotype is complex, containing inflammatory and allergic components. An investigation of the effects of the MHC on asthma and its related phenotypes, therefore, poses a methodological and statistical challenge. The class II genes of the MHC have recognized influences on the ability to respond to particular allergens.30 –35 Asthma and bronchial hyperresponsiveness are associated primarily with an allergy to the HDM and to a lesser degree with allergy to cat dander and molds,36,37 so that genetic control of specific IgE responses is of relevance to clinical disease. The T-cell receptor (TCR) genes, on chromosomes 7q and 14q, are also important potential genetic modifiers of the specific IgE response. Genetic linkage and allelic association have been reported between specific IgE responses and the TCR ␣/␦ locus on chromosome 14q.38,39 The class I genes of the MHC may have important effects on atopic responses, but these have not yet been adequately investigated. Similarly, the class III complement genes contain polymorphisms that may be of relevance to inflammatory or immune diseases, but which have not yet been tested in asthmatic subjects. Nonclassical MHC genes also may have an impact on asthma through nonallergic pathways, and a polymorphism in the control elements of inflammatory cytokines and their receptors is an important mechanism for flexibility in the immunoregulatory machinery.40 Tumor necrosis factor (TNF) is a potent proinflammatory cytokine that is found in excess in asthmatic airways. A polymorphism in the TNF complex is associated with variation on the expression of TNF-␣ and with the presence of asthma.41– 43 These results emphasize the inflammatory nature of the asthmatic response, as distinct from its allergic basis.
Thomas L. Petty 44th Annual Aspen Lung Conference: Pulmonary Genetics, Genomics, Gene Therapy
Chromosome 11q The linkage of atopy to a VNTR (ie, variable number of tandem repeats) polymorphism on chromosome 11q13 was first reported in 198944 and was at first disputed.45 The -chain of the high-affinity receptor for IgE (Fc⑀RI-) was subsequently localized to the region. The Fc⑀RI receptor acts as the allergic trigger on mast cells and other types of cells, and is central to the allergic response.46 The -chain is not essential for Fc⑀RI function, but it both stabilizes the surface expression of the receptor and acts as an amplifying element within it.47 Any variation in the level of the -chain expression may therefore modify receptor function. A polymorphism in Fc⑀RI- has been related to atopy,48 asthma,49 bronchial hyperresponsiveness,50,51and severe atopic dermatitis.52 A polymorphism within the gene also has been associated with levels of IgE in heavily parasitized Australian aborigines, implying a protective role for the gene in helminth infestation.53 Although coding changes have been identified within Fc⑀RI-,54,55 they are conservative and do not seem to alter gene function.56 The Ile181Leu polymorphisms identified by Shirakawa et al54 have not been found in several other studies57–59 but have been found in association with asthma in Kuwaiti Arabs60 and black South Africans.61 The difficulty in their identification nevertheless suggests that they are artifactual or in homologous sequences out of the Fc⑀RI- gene. The structural variation that causes the effect of this gene on patients with asthma, therefore, has not yet been identified. The genetic linkage and association of atopy to the locus both have been typified by a strong maternal effect,52,62,63 with preferential linkage and the transmission of maternal alleles to affected children. Maternal effects are wellrecognized in allergic disorders, and asthma, eczema, elevated serum IgE concentrations, and skin prick test positivity in children all have been accompanied by an increased prevalence of asthma or atopy in mothers.64 The preferential transmission of or linkage to alleles from either the maternal or paternal sides also has been observed for other loci influencing allergic disease, including those identified on chromosomes 13 and 16.8 Parent-of-origin effects have been noted in other immunologic disorders, including type I diabetes,65 rheumatoid arthritis,66 inflammatory bowel disease,67 and selective IgA deficiency.68 Parent-of-origin linkages also have been observed in the same diseases, so that the preferential linkage of paternal alleles is seen from the insulin locus and type I diabetes,69 and HLA alleles for selective IgA deficiency.68 These findings suggest a common underlying mechanism for parent-of-origin effects in immune disorders. The monoallelic expression of cytokines and cell-signaling molecules is recognized70 –72 and mediated by methylation,73 providing a potential mechanism for these effects.
Chromosome 12 The initial demonstration of the genetic linkage of asthma to chromosome 12q74 was followed by single-locus confirmatory studies75,76 and by several general genome screens.9,10,12,13 In addition, a genome screen in a mouse model of asthma found a linkage to bronchial hyperre-
sponsiveness on mouse chromosome 10 in the region of syntenic homology to human chromosome 12q.77 The facility with which many groups have identified a linkage to this region suggests that it is a true major atopy locus. Interferon-␥ does not seem to be responsible for the linkage. High-density mapping of the region has been begun,78 and it is likely that positional cloning of the gene will be possible.
Chromosome 13q14 A linkage of the total serum IgE to the esterase D protein polymorphism on chromosome 13q14 was reported in 1985.79 The linkage of chromosome 13q to atopy was confirmed by a genome-wide scan8 and by a singlelocus study of Japanese families.80 The same study identified potential linkage disequilibrium (LD) between disease and D13S153.80 A two-stage screen in Hutterite families from the United States found a linkage of asthma to chromosome 13q21.39 in the first-stage families but not in the second-stage families. A linkage of chromosome 13q14 to HDM allergy in children with asthma also has been observed,81 as has a linkage to children with atopic dermatitis.82 These results suggest that chromosome 13q14 also contains a major atopy locus. The same chromosomal region has been shown to be linked to total serum IgA concentrations.83 Low levels of serum IgA occur much more frequently in atopic children than in healthy children,84 and salivary IgA deficiency is more common in infants with atopic parents.85 Ig production is known to be under the control of many genes (reviewed by Corrigan86), none of which have been mapped to chromosome 13q14. The gene of interest may encode a regulatory component of the humoral immune system but, alternatively, might influence both IgA levels and atopic status by influencing the mucosal handling of allergens.
Sharing of Loci With Other Disorders Genetic studies of other disorders also may have an impact on asthma and atopy. Crohn’s disease and ulcerative colitis are inflammatory bowel diseases of unknown etiology that show familial clustering.87– 89 Genome-wide screens have implicated loci on chromosomes 3, 7, 12, and 16.90 –92 The regions on chromosomes 7 and 12 may coincide with the asthma and atopy loci on the same chromosomes. Polymorphism in the IL-1 cluster on chromosome 2 also has been shown to influence the severity of the disease.93,94 A genome-wide screen in families with rheumatoid arthritis similarly has shown linkage near the asthma locus on chromosome 2 and the TCR-␣ locus on chromosome 14.95 Linkage to type I diabetes is found near Fc⑀RI- on chromosome 11q13.96 These findings suggest that important genes or gene families may be common to several inflammatory and immune disorders.
Mendelian Disorders Asthma, eczema, and allergic diseases are associated with a number of mendelian disorders. The identification of genes causing such disorders is much easier than the CHEST / 121 / 3 / MARCH, 2002 SUPPLEMENT
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positional cloning of complex disease genes. These mendelian disorders may therefore be extremely helpful in identifying genes influencing asthma and allergy. They include Netherton’s syndrome,97,98 Job syndrome (also known as hyper-IgE syndrome or Buckley’s syndrome),99 thymic hypoplasia (DiGeorge syndrome), cellular deficiency with Igs (Nezelof syndrome),100 selective IgA deficiency, and Wiskott-Aldrich syndrome. Netherton’s syndrome is a rare recessive disease in which children are born with a severe ichthyotic dermatosis.97 Severe symptomatic atopy is a universal accompaniment of the disease.98 The gene for Netherton’s syndrome has recently been located distal to the chromosome 5 cytokine cluster,101 and the gene underlying the disorder has been identified as SPINK5.102 SPINK5 codes a multidomain serine protease inhibitor LEKTI, which is expressed in the epithelium, mucosa, and thymus.103 Common coding polymorphisms have been identified in the gene by our group, and these show associations with atopy, asthma, and eczema in children without Netherton’s syndrome. These findings therefore define a new pathway for allergic disorders. Other mendelian disorders localize to regions that may be relevant to common allergic diseases. Selective IgA deficiency has been localized to the MHC.68 Hypereosinophilia syndrome has been localized to the distal part of the chromosome 5 cytokine cluster,104 and it is of interest that acquired hypereosinophilia is associated consistently with translocations on chromosome 5q35.105,106 Hyper-IgE syndrome has been linked to the distal arm of chromosome 4q.107 A small chromosomal deletion in one child with the disease may have limited this localization to a 20-megabase interval.107 A linkage of the total serum IgE to this region has been seen in at least one genome scan (http://www.well.ox.ac.uk/asthma/public/GenomeScan/ index.html),8 suggesting that this locus also may have an affect on the normal regulation of IgE levels.
Conclusion The positional cloning of the genes underlying asthma and allergic disorders is becoming increasingly tractable. Agreed-on regions of strong genetic linkage have emerged, some of which coincide with linkages to singlegene disorders or to other immunologic diseases. The completion of the human genomic sequence and the availability of the deep public expressed sequence tag databases mean that laborious physical mapping of these loci may not be necessary. The key element in gene discovery will be the identification of robust patterns of LD between markers and disease. LD mapping of at least one atopy locus has shown that LD is irregularly distributed,108 and the challenge is to develop statistical as well as genotyping methods to handle dense local single-nucleotide polymorphism maps.
References 1 Strachan DP, Anderson HR, Limb ES, et al. A national survey of asthma prevalence, severity, and treatment in Great Britain. Arch Dis Child 1994; 70:174 –178 10S
2 Sampson HA. Pathogenesis of eczema. Clin Exp Allergy 1990; 20:459 – 467 3 Schulz-Larsen F. A genetic-epidemiologic study in a population-based twin sample. J Am Acad Dermatol 1993; 28: 719 –723 4 von Mutius E, Fritzsch C, Weiland SK, et al. Prevalence of asthma and allergic disorders among children in united Germany: a descriptive comparison. BMJ 1992; 305:1395–1399 5 Shirakawa T, Enomoto T, Shimazu S, et al. The inverse association between tuberculin responses and atopic disorder. Science 1997; 275:77–79 6 Cookson WO, Moffatt MF. Asthma: an epidemic in the absence of infection? Science 1997; 275:41– 42 7 Rook GA, Stanford JL. Give us this day our daily germs. Immunol Today 1998; 19:113–116 8 Daniels SE, Bhattacharrya S, James A, et al. A genome-wide search for quantitative trait loci underlying asthma. Nature 1996; 383:247–250 9 Ober C, Cox NJ, Abney M, et al. Genome-wide search for asthma susceptibility loci in a founder population: the Collaborative Study on the Genetics of Asthma. Hum Mol Genet 1998; 7:1393–1398 10 Wjst M, Fischer G, Immervoll T, et al. A genome-wide search for linkage to asthma: German Asthma Genetics Group. Genomics 1999; 58:1– 8 11 Hizawa N, Freidhoff L, Chiu Y, et al. Genetic regulation of Dermatophagoides pteronyssinus-specific IgE responsiveness: a genome-wide multipoint linkage analysis in families recruited through 2 asthmatic sibs; Collaborative Study on the Genetics of Asthma (CSGA). J Allergy Clin Immunol 1998; 102:436 – 442 12 The Collaborative Study on the Genetics of Asthma. A genome-wide search for asthma susceptibility loci in ethnically diverse populations. Nat Genet 1997; 15:389 –392 13 Dizier MH, Besse-Schmittler C, Guilloud-Bataille M, et al. Genome screen for asthma and related phenotypes in the French EGEA study [abstract]. Am J Respir Crit Care Med 1999; 159:A649 14 Marsh DG, Neely JD, Breazeale DR, et al. Linkage analysis of IL4 and other chromosome 5q31.1 markers and total serum immunoglobulin E concentrations. Science 1994; 264:1152–1156 15 Meyers DA, Postma DS, Panhuysen CI, et al. Evidence for a locus regulating total serum IgE levels mapping to chromosome 5. Genomics 1994; 23:464 – 470 16 Martinez F, Solomon S, Holberg C, et al. Linkage of circulating eosinophils to markers on chromosome 5q. Am J Respir Crit Care Med 1998; 158:1739 –1744 17 Marquet S, Abel L, Hillaire D, et al. Genetic localization of a locus controlling the intensity of infection by Schistosoma mansoni on chromosome 5q31– q33. Nat Genet 1996; 14: 181–184 18 Kelso A. Th1 and Th2 subsets: paradigms lost? Immunol Today 1995; 16:374 –379 19 Graves PE, Kabesch M, Halonen M, et al. A cluster of seven tightly linked polymorphisms in the IL-13 gene is associated with total serum IgE levels in three populations of white children. J Allergy Clin Immunol 2000; 105:506 –513 20 Wills-Karp M, Luyimbazi J, Xu X, et al. Interleukin-13: central mediator of allergic asthma. Science 1998; 282: 2258 –2261 21 Rosenwasser LJ, Klemm DJ, Dresback JK, et al. Promoter polymorphisms in the chromosome 5 gene cluster in asthma and atopy. Clin Exp Allergy 1995; 25:74 –78 22 Walley AJ, Cookson WO. Investigation of an interleukin-4 promoter polymorphism for associations with asthma and atopy. J Med Genet 1996; 33:689 – 692
Thomas L. Petty 44th Annual Aspen Lung Conference: Pulmonary Genetics, Genomics, Gene Therapy
23 Shirakawa I, Deichmann KA, Izuhara I, et al. Atopy and asthma: genetic variants of IL-4 and IL-13 signalling. Immunol Today 2000; 21:60 – 64 24 Ober C, Leavitt SA, Tsalenko A, et al. Variation in the interleukin 4-receptor alpha gene confers susceptibility to asthma and atopy in ethnically diverse populations. Am J Hum Genet 2000; 66:517–526 25 Noguchi E, Shibasaki M, Arinami T, et al. No association between atopy/asthma and the ILe50Val polymorphism of IL-4 receptor. Am J Respir Crit Care Med 1999; 160: 342–345 26 Baldini M, Lohman I, Halonen M, et al. A polymorphism in the 5⬘ flanking region of the CD14 gene is associated with circulating soluble CD14 levels and with total serum immunoglobulin E. Am J Respir Cell Mol Biol 1999; 20:976 –983 27 Xu J, Levitt RC, Panhuysen CI, et al. Evidence for two unlinked loci regulating total serum IgE levels. Am J Hum Genet 1995; 57:425– 430 28 Loots GG, Locksley RM, Blankespoor CM, et al. Identification of a coordinate regulator of interleukins 4, 13, and 5 by cross-species sequence comparisons. Science 2000; 288: 136 –140 29 Lacy DA, Wang ZE, Symula DJ, et al. Faithful expression of the human 5q31 cytokine cluster in transgenic mice. J Immunol 2000; 164:4569 – 4574 30 Marsh DG, Meyers DA, Bias WB. The epidemiology and genetics of atopic allergy. N Engl J Med 1981; 305:1551– 1559 31 Young RP, Dekker JW, Wordsworth BP, et al. HLA-DR and HLA-DP genotypes and immunoglobulin E responses to common major allergens. Clin Exp Allergy 1994; 24: 431– 439 32 Fischer GF, Pickl WF, Fae I, et al. Association between IgE response against Bet v I, the major allergen of birch pollen, and HLA-DRB alleles. Hum Immunol 1992; 33:259 –265 33 Sparholt SH, Georgsen J, Madsen HO, et al. Association between HLA- DRB3*0101 and immunoglobulin-E responsiveness to Bet v I. Hum Immunol 1994; 39:76 –78 34 D’Amato M, Scotto d’Abusco A, Maggi E, et al. Association of responsiveness to the major pollen allergen of Parietaria officinalis with HLA-DRB1* alleles: a multicenter study. Hum Immunol 1996; 46:100 –106 35 Donfack J, Tsalenko A, Hoki DM, et al. HLA-DRB1*01 alleles are associated with sensitization to cockroach allergens. J Allergy Clin Immunol 2000; 105:960 –966 36 Sears MR, Herbison GP, Holdaway MD, et al. The relative risks of sensitivity to grass pollen, house dust mite and cat dander in the development of childhood asthma. Clin Allergy 1989; 18:419 – 424 37 Cookson WOCM, De Klerk NH, Ryan GR, et al. Relative risks of bronchial hyper-responsiveness associated with skinprick test responses to common antigens in young adults. Clin Exp Allergy 1991; 21:473– 479 38 Moffatt MF, Hill MR, Cornelis F, et al. Genetic linkage of T cell receptor ␣/␦ complex to specific IgE responses. 1994; Lancet 343:1597–1600 39 Moffatt MF, Schou C, Faux JA, et al. Germline TCR-A restriction of immunoglobulin E responses to allergen. Immunogenetics 1997; 46:226 –230 40 Daser A, Mitchison H, Mitchison A, et al. Non-classicalMHC genetics of immunological disease in man and mouse: the key role of pro-inflammatory cytokine genes. Cytokine 1996; 8:593–597 41 Moffatt MF, Cookson WO. Tumour necrosis factor haplotypes and asthma. Hum Mol Genet 1997; 6:551–554 42 Li Kam Wa TC, Mansur AH, Britton J, et al. Association between ⫺308 tumour necrosis factor promoter polymor-
43
44 45 46 47 48 49 50 51 52 53
54 55
56
57 58
59 60 61
62 63
phism and bronchial hyperreactivity in asthma. Clin Exp Allergy 1999; 29:1204 –1208 Chagani T, Pare PD, Zhu S, et al. Prevalence of tumor necrosis factor-alpha and angiotensin converting enzyme polymorphisms in mild/moderate and fatal/near-fatal asthma. Am J Respir Crit Care Med 1999; 160:278 –282 Cookson WOCM, Sharp PA, Faux J, et al. Linkage between immunoglobulin E responses underlying asthma and rhinitis and chromosome 11q. Lancet 1989; 1:1292–1295 Morton NE. Major loci for atopy? [editorial]. Clin Exp Allergy 1992; 22:1041–1043 Turner H, Kinet JP. Signalling through the high-affinity IgE receptor Fc epsilonRI. Nature 1999; 402:B24 –B30 Lin S, Cicala C, Scharenberg A, et al. The Fc(epsilon)RIbeta subunit functions as an amplifier of Fc(epsilon)RIgammamediated cell activation signals. Cell 1996; 85:985–995 Hill MR, James AL, Faux JA, et al. Fc⑀RI- polymorphism and risk of atopy in a general population sample. BMJ 1995; 311:776 –779 Shirakawa T, Mao XQ, Sasaki S, et al. Association between Fc epsilon RI beta and atopic disorder in a Japanese population [letter]. Lancet 1996; 347:394 –395 van Herwerden L, Harrap SB, Wong ZY, et al. Linkage of high-affinity IgE receptor gene with bronchial hyperreactivity, even in absence of atopy. Lancet 1995; 346:1262–1265 Trabetti E, Cusin V, Malerba G, et al. Association of the FcepsilonRIbeta gene with bronchial hyper-responsiveness in an Italian population. J Med Genet 1998; 35:680 – 681 Cox HE, Moffatt MF, Faux JA, et al. Association of atopic dermatitis to the beta subunit of the high affinity immunoglobulin E receptor. Br J Dermatol 1998; 138:182–187 Palmer LJ, Pare PD, Faux JA, et al. Fc epsilon R1-beta polymorphism and total serum IgE levels in endemically parasitized Australian aborigines. Am J Hum Genet 1997; 61:182–188 Shirakawa T, Li A, Dubowitz M, et al. Association between atopy and variants of the  subunit of the high affinity immunoglobulin E receptor. Nat Genet 1994; 7:125–129 Hill MR, Cookson WOCM. A new variant of the  subunit of the high affinity receptor for immunoglobulin E (Fc⑀RIE237G): associations with measures of atopy and bronchial hyper-responsiveness. Hum Mol Genet 1996; 5:959 –962 Donnadieu E, Cookson WO, Jouvin MH, et al. Allergyassociated polymorphisms of the FcepsilonRIbeta subunit do not impact its two amplification functions. J Immunol 2000; 165:3917–3922 Duffy D, Healey S, Chenevix-Trench G, et al. Atopy in Australia [letter]. Nat Genet 1995; 10:260 Rohrbach M, Kraemer R, Liechti-Gallati S. Screening of the Fc epsilon RI-beta-gene in a Swiss population of asthmatic children: no association with E237G and identification of new sequence variations. Dis Markers 1998; 14:177–186 Dickson P, Wong Z, Harrap S, et al. Mutational analysis of the high affinity immunoglobulin E receptor beta subunit gene in asthma. Thorax 1999; 54:409 – 412 Haider M, Hijazi Z. Prevalence of high affinity IgE receptor [Fc epsilon RI beta] gene polymorphisms in Kuwaiti Arabs with asthma [letter]. Clin Genet 1998; 54:166 –167 Green S, Gaillard M, Song E, et al. Polymorphisms of the beta chain of the high-affinity immunoglobulin E receptor (Fcepsilon RI-beta) in South African black and white asthmatic and nonasthmatic individuals. Am J Respir Crit Care Med 1998; 158:1487–1492 Cookson WO, Young RP, Sandford AJ, et al. Maternal inheritance of atopic IgE responsiveness on chromosome 11q. Lancet 1992; 340:381–384 Shirakawa T, Hashimoto T, Furuyama J, et al. Linkage CHEST / 121 / 3 / MARCH, 2002 SUPPLEMENT
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between severe atopy and chromosome 11q13 in Japanese families. Clin Genet 1994; 46:228 –232 Moffatt M, Cookson W. The genetics of asthma: maternal effects in atopic disease. Clin Exp Allergy 1998; 28(suppl): 56 – 61 Warram JH, Krolewski AS, Gottlieb MS, et al. Differences in risk of insulin-dependent diabetes in offspring of diabetic mothers and diabetic fathers. N Engl J Med 1984; 311: 149 –152 Koumantaki Y, Giziaki E, Linos A, et al. Family history as a risk factor for rheumatoid arthritis: a case-control study. J Rheumatol 1997; 24:1522–1526 Akolkar PN, Gulwani-Akolkar B, Heresbach D, et al. Differences in risk of Crohn’s disease in offspring of mothers and fathers with inflammatory bowel disease. Am J Gastroenterol 1997; 92:2241–2244 Vorechovsky I, Webster AD, Plebani A, et al. Genetic linkage of IgA deficiency to the major histocompatibility complex: evidence for allele segregation distortion, parentof-origin penetrance differences, and the role of anti-IgA antibodies in disease predisposition. Am J Hum Genet 1999; 64:1096 –1109 Bennett S, Todd J. Human type 1 diabetes and the insulin gene: principles of mapping polygenes. Annu Rev Genet 1996; 30:343–370 Bix M, Locksley RM. Independent and epigenetic regulation of the interleukin-4 alleles in CD4⫹ T cells. Science 1998; 281:1352–1354 Hollander G, Zuklys S, Morel C, et al. Monoallelic expression of the interleukin-2 locus. Science 1998; 279:2118 –2121 Held W, Raulet D. Expression of the Ly49A gene in murine natural killer cell clones is predominantly but not exclusively mono-allelic. Eur J Immunol 1997; 27:2876 –2884 Young HA, Ghosh P, Ye J, et al. Differentiation of the T helper phenotypes by analysis of the methylation state of the IFN-gamma gene. J Immunol 1994; 153:3603–3610 Barnes KC, Neely JD, Duffy DL, et al. Linkage of asthma and total serum IgE concentration to markers on chromosome 12q: evidence from Afro-Caribbean and caucasian populations. Genomics 1996; 37:41–50 Nickel R, Wahn U, Hizawa N, et al. Evidence for linkage of chromosome 12q15– q24.1 markers to high total serum IgE concentrations in children of the German Multicenter Allergy Study. Genomics 1997; 46:159 –162 Wilkinson J, Thomas NS, Morton N, et al. Candidate gene and mutational analysis in asthma and atopy. Int Arch Allergy Immunol 1999; 118:265–267 Zhang Y, Lefort J, Kearsey V, et al. A genome-wide screen for asthma-associated quantitative trait loci in a mouse model of allergic asthma. Hum Mol Genet 1999; 8:601– 605 Barnes KC, Freidhoff LR, Nickel R, et al. Dense mapping of chromosome 12q13.12-q23.3 and linkage to asthma and atopy. J Allergy Clin Immunol 1999; 104:485– 491 Eiberg H, Lind P, Mohr J, et al. Linkage relationship between the human immunoglobulin E polymorphism and marker systems [abstract]. Cytogenet Cell Genet 1985; 40:622 Kimura K, Noguchi E, Shibasaki M, et al. Linkage and association of atopic asthma to markers on chromosome 13 in the Japanese population. Hum Mol Genet 1999; 8:1487– 1490 Hizawa N, Freidhoff L, Ehrlich E, et al. Genetic influences of chromosomes 5q31– q33 and 11q13 on specific IgE responsiveness to common inhaled allergens among African American families: Collaborative Study on the Genetics of Asthma (CSGA). J Allergy Clin Immunol 1998; 102:449– 453 Beyer KWU, Freidhoff L, Nickel R, et al. Evidence for linkage of chromosome 5q31– q33 and 13q12– q14 markers
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84 85 86 87
88 89 90
91 92 93
94 95 96 97 98 99 100
101
102 103
to atopic dermatitis [abstract]. J Allergy Clin Immunol 1998; 101:152 Wiltshire S, Bhattacharyya S, Faux JA, et al. A genome scan for loci influencing total serum immunoglobulin levels: possible linkage of IgA to the chromosome 13 atopy locus. Hum Mol Genet 1998; 7:27–31 Ludviksson BR, Eiriksson TH, Ardal B, et al. Correlation between serum immunoglobulin A concentrations and allergic manifestations in infants. J Pediatr 1992; 121:23–27 van Asperen PP, Gleeson M, Kemp AS, et al. The relationship between atopy and salivary IgA deficiency in infancy. Clin Exp Immunol 1985; 62:753–757 Corrigan CJ. T and B lymphocytes and the development of allergic reactions. In: Kay AB, ed. Allergy and allergic diseases. Oxford, UK: Blackwell Science Ltd, 1997; 36 –57 Rioux JD, Silverberg MS, Daly MJ, et al. Genome-wide search in Canadian families with inflammatory bowel disease reveals two novel susceptibility loci. Am J Hum Genet 2000; 66:1863–1870 Duerr RH, Barmada MM, Zhang L, et al. High-density genome scan in Crohn disease shows confirmed linkage to chromosome 14q11–12. Am J Hum Genet 2000; 66:1857–1862 Ma Y, Ohmen JD, Li Z, et al. A genome-wide search identifies potential new susceptibility loci for Crohn’s disease. Inflamm Bowel Dis 1999; 5:271–278 Satsangi J, Parkes M, Louis E, et al. Two stage genome-wide search in inflammatory bowel disease provides evidence for susceptibility loci on chromosomes 3, 7 and 12. Nat Genet 1996; 14:199 –202 Hugot JP, Thomas G. Genome-wide scanning in inflammatory bowel diseases. Dig Dis 1998; 16:364 –369 Duerr RH, Barmada MM, Zhang L, et al. Linkage and association between inflammatory bowel disease and a locus on chromosome 12. Am J Hum Genet 1998; 63:95–100 Mansfield J, Holden H, Tarlow J, et al. Novel genetic association between ulcerative colitis and the anti-inflammatory cytokine interleukin-1 receptor antagonist. Gastroenterology 1994; 106:637– 642 Parkes M, Satsangi J, Jewell D. Contribution of the IL-2 and IL-10 genes to inflammatory bowel disease (IBD) susceptibility. Clin Exp Immunol 1998; 113:28 –32 Hardwick L, Walsh S, Butcher S, et al. Genetic mapping of susceptibility loci in the genes involved in rheumatoid arthritis. J Rheumatol 1998; 24:197–198 Nakagawa Y, Kawaguchi Y, Twells R, et al. Fine mapping of the diabetes-susceptibility locus, IDDM4, on chromosome 11q13. Am J Hum Genet 1998; 63:547–556 Netherton EW. A unique case of trichorrhexis nodosa, “Bamboo hairs.” Arch Dermatol 1958; 78:483– 487 Judge MR, Morgan G, Harper JI. A clinical and immunological study of Netherton’s syndrome. Br J Dermatol 1994; 131:615– 621 Davis SD, Schaller J, Wedgwood RJ. Job’s syndrome: recurrent, cold, staphylococcal abscesses. Lancet 1966; 1:1013–1015 Knutsen AP, Wall D, Mueller KR, et al. Abnormal in vitro thymocyte differentiation in a patient with severe combined immunodeficiency-Nezelof’s syndrome. J Clin Immunol 1996; 16:151–158 Chavanas S, Garner C, Bodemer C, et al. Localization of the Netherton syndrome gene to chromosome 5q32, by linkage analysis and homozygosity mapping. Am J Hum Genet 2000; 66:914 –921 Chavanas S, Bodemer C, Rochat A, et al. Mutations in SPINK5, encoding a serine protease inhibitor, cause Netherton syndrome. Nat Genet 2000; 25:141–142 Ma¨ gert HJ, Standker L, Kreutzmann P, et al. LEKTI, a novel 15-domain type of human serine proteinase inhibitor.
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J Biol Chem 1999; 274:21499 –21502 104 Rioux J, Stone V, Daly M, et al. Familial eosinophilia maps to the cytokine gene cluster on human chromosomal region 5q31– q33. Am J Hum Genet 1998; 63:1086 –1094 105 Jani K, Kempski H, Reeves B. A case of myelodysplasia with eosinophilia having a translocation t(5; 12) (q31; q13) restricted to myeloid cells but not involving eosinophils. Br J Haematol 1994; 87:57– 60 106 Sato H, Saito H, Ikebuchi K, et al. Biological characteristics of chronic eosinophilic leukemia cells with a t(2; 5)(p23; q35) translocation. Leuk Lymphoma 1995; 19: 499 –505 107 Grimbacher B, Schaffer AA, Holland SM, et al. Genetic linkage of hyper-IgE syndrome to chromosome 4. Am J Hum Genet 1999; 65:735–744 108 Moffatt MF, Traherne JA, Abecasis GR, et al. Single nucleotide polymorphism and linkage disequilibrium within the TCR alpha/delta locus. Hum Mol Genet 2000; 9:1011–1019
Genetic Analysis of AntigenInduced Airway Manifestations of Asthma Using Recombinant Congenic Mouse Strains* Antoon J.M. Van Oosterhout, PhD; Prescilla V. Jeurink; Peter C. Groot, PhD; Gerard A. Hofman; Frans P. Nijkamp, PhD; and Peter Demant, PhD
(CHEST 2002; 121:13S) Abbreviation: OVA ⫽ ovalbumin
asthma is a heterogeneous and genetically A llergic complex disease that is characterized by the pres-
ence of allergen-specific IgE, eosinophilic airway inflammation, and hyperresponsiveness to bronchospasmogenic stimuli. To facilitate the mapping of genes controlling complex asthma traits, we have used a powerful tool, the recombinant congenic strains of mice, which transforms a multigenic difference into a set of monogenic or oligogenic differences.1 This approach offers a higher resolution power than do the standard mouse crosses in mapping segregating quantitative trait loci and in the detection of their potential interactions. In the present study, we compared 4 strains (Ccs/DEM-5, Ccs/DEM-11, Ccs/DEM-12, and Ccs/DEM-18) from a series of 20 strains, each of which carries a random set of 12.5% of genes from the T-helper cell type 1 responder strain STS and 87.5% of
Table 1—Lung Function and Inflammation* Strain 5 11 12 18
(n ⫽ 13) (n ⫽ 4) (n ⫽ 12) (n ⫽ 8)
Penh Maximum 2.6 ⫾ 0.3 2.5 ⫾ 0.2 3.3 ⫾ 0.3 2.7 ⫾ 0.3
vs vs vs vs
⌬ED300
3.5 ⫾ 0.5† ⫺ 1.1 ⫾ 4.4 3.0 ⫾ 0.3† ⫺ 33.2 ⫾ 8.3† 3.0 ⫾ 0.4 ⫹ 6.6 ⫾ 5.1 6.0 ⫾ 1.1† ⫺ 6.3 ⫾ 3.7
Eosinophils, ⫻ 105 1.0 ⫾ 0.4 2.9 ⫾ 1.5 5.1 ⫾ 1.2 6.0 ⫾ 1.5
*Values given as mean ⫾ SEM. ED300 ⫽ dose of methacholine needed to increase baseline values of enhanced pause by 300%; Penh ⫽ enhanced pause. †p ⬍ 0.05, as determined by Student’s t test.
genes from the T-helper cell type 2 responder strain BALB/c. Mice were sensitized with ovalbumin (OVA), and later on were repeatedly challenged by the inhalation of an OVA aerosol. Before and after the OVA challenges, serum samples were obtained and the airway responsiveness to nebulized methacholine (dose range, 1.5 to 50 mg/mL) was determined. Finally, BAL was performed, and the number of leukocytes were determined (Table 1). All mouse strains showed increased serum levels of OVA-specific IgE as a result of the OVA inhalation challenge (data not shown). It appears that there is no correlation between the number of BAL eosinophils and the extent of airway hyperresponsiveness. Strain 12 displayed airway eosinophilia but a resistance to hyperresponsiveness. However, the baseline airway responsiveness of strain 12 was higher than that of the other strains. In strain 11, airway hyperresponsiveness was determined predominantly by an increased sensitivity (ie, the dose of methacholine needed to increase baseline values of enhanced pause by 300%) to methacholine, whereas in strains 5 and 18 only a significant increase in the maximal response could be observed. These data demonstrate that different asthma traits like the presence of IgE, eosinophilia, and hyperresponsiveness may be genetically dissociated. Further mapping studies will be carried out to localize the modifier genes involved in specific traits.
Reference 1 Groot PC, Moen CJ, Dietrich W, et al. The recombinant congenic strains for analysis of multigenic traits: genetic composition. FASEB J 1992; 6:2826 –2835
*From the Department of Pharmacology and Pathophysiology (Drs. Van Oosterhout, Groot, and Nijkamp, Ms. Jeurink, and Mr. Hofman), Faculty of Pharmacy, Utrecht University, Utrecht, Netherlands; Division of Molecular Genetics (Dr. Demant), Netherlands Cancer Institute, Amsterdam, Netherlands. This research was supported by a research grant (AF99.23) of the Dutch Asthma Foundation. Correspondence to: Antoon J.M. Van Oosterhout, PhD, Department of Pharmacology and Pathophysiology, Faculty of Pharmacy, Utrecht University, PO box 80.082, 3508TB Utrecht, Netherlands; e-mail: [email protected] CHEST / 121 / 3 / MARCH, 2002 SUPPLEMENT
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