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The short arm is labelled ‘p’ (for petit. and. 13.XY) stained by Giemsa banding. On average there are about 52 crossovers per human male meiosis. the tip of each arm being called the telomere. Reductive cell division. Meiosis only occurs in gonadal cells and consists of two sequential divisions in which the DNA replicates only once.2). called the centromere. meiosis. The study of human variation depends upon analysis of the outcomes of matings of individuals and the scientiﬁc study of deﬁnable differences. The questions that surround human genetics are concerned with the transmission of heritable traits. Heredity represents the transmission of information required for the formation and regulation of proteins. results in cells with a half-set of 23 chromosomes (the haploid number). which are held together at the centromere (Fig. THE GENETIC BASIS OF ALLERGIC DISEASE Stephen T Holgate INTRODUCTION Medical genetics comprises the study of human variability and heredity and its application to human disease. Mitosis centromere sister chromatids prophase metaphase centromere anaphase Fig.1). few chromosomes are inherited intact from a parent and so cosegregation or linkage of genes on a chromosome will only occur if they are physically close. 21.13 Holgate 2/6/03 8:17 AM Page 189 13. there is at least one chiasma per chromosome arm. During mitosis each chromosome replicates to form a pair of sister chromatids. 189 . French for ‘small’) and the long arm ‘q’. Therefore.3). 18.2 Diagrammatic representation of mitosis (two chromosome pairs are shown). Each chromosome has a narrow waist. with the exception of the short arms of chromosomes 13–15. 13. telophase Fig. 13. Note the distinctive centromere. 13. and 22. The chromosomes which have exchanged genetic material are called recombinants. Most cells contain 46 chromosomes (the diploid number). which can be arranged into 22 pairs of autosomes and a pair of sex chromosomes – XX in the female and XY in the male (Fig. before the ﬁrst division (Fig.1 Normal male human chromosomes (46. which has a constant position within a given chromosome. 13.
6). THE GENETIC BASIS OF ALLERGIC DISEASE Meiosis sites of recombination Autosomal Dominant Inheritance I 1 II 1 2 3 4 5 6 5 6 III 1 IV 1 2 3 4 2 3 4 5 2 first meiotic division leptotene zygotene diplotene nonpenetrant individual anaphase I second meiotic division anaphase II Fig. Vertical type of pedigree with both males and females affected. 13. i. 13. the affected individual has one normal gene and one mutant gene and is heterozygous at this locus (Fig. one gene of a pair being assigned to one gamete and one to another gamete – this is Mendel’s ﬁrst law. then the other can be mapped to the same region.7).5). with pairs of hereditary elements or genes and their variants (alleles) segregating at meiosis. 13. If the disease and marker loci are on separate chromosomes. 13. The mechanisms for this are not clearly understood but seem to involve gene–gene interactions (epistasis). The parents are When two genes are close together. i. results in a characteristic pattern of inheritance (Fig. If the chromosomal location of one of the genes is known. the parents are carriers of one defective gene each. alleles at such loci have a tendency to pass together into each gamete. 13. recombinant chromosomes commonly unaffected yet each has one defective gene which will only express itself if the two defective genes are combined in the offspring. Many dominant traits exhibit variable expression. As the distance between the disease locus and a marker locus increases so the chance of recombination in the interval between them increases Autosomal Recessive Inheritance 1 2 3 4 1 2 3 4 5 6 7 1 2 3 4 5 6 7 8 9 Fig. gametes Recombination and linkage Fig. Autosomal dominant traits show a vertical type of pedigree pattern with equal numbers of males and females affected. 190 . Modes of single gene inheritance In autosomal dominant disorders the condition is produced by a mutation of one member of an autosomal gene pair. Mendel’s second law states that members of different gene pairs assort gametes independently of one another.4 Autosomal dominant inheritance. Thus. independent assortment will not occur and the disease and marker will occur together in each child unless they are separated by a crossover at meiosis. the individual is homozygous at this locus (Fig.4). In this example. independent assortment will occur and the disease and markers should be found as often together as apart in the offspring (Fig. 13.5 Autosomal recessive inheritance.e. In contrast to dominant disorders in which heterozygotes manifest the condition. which.13 Holgate 2/6/03 8:17 AM Page 190 13. they are said to be linked.e. who inherit both genes. Therefore. Expression is only seen in offspring 4 and 7. any disturbance of independent assortment deﬁned by Mendel’s second law provides an important clue that two genes are linked. INHERITANCE AND LINKAGE Mendelian inheritance Gregor Mendel (1822–84) clearly deﬁned pairs of contrasting characters and concluded that their inheritance (traits) must be particulate. on account of the uneven distribution of the sex chromosomes. If the disease and marker loci lie close together on the same chromosome.3 Diagrammatic representation of meiosis (two chromosome pairs are shown). affected individuals in the same family vary in their clinical severity. Thus. 13. Further complications are introduced in X-linked recessive and X-linked dominant disorders. in autosomal recessive disorders the affected individual has mutations in both members of a gene pair.
The relationship between q and the actual physical distance between loci depends upon several factors. (b) pedigree showing X-linked dominant inheritance. q decreases from 50% to 0% when tight linkage occurs. A q of 0. a crossover between the loci is highly likely and the disease and marker traits will occur separately in each recombinant but together in nonrecombinants. X-Linked Inheritance I II 1 III 1 IV I II 1 III 1 IV 1 2 2 3 3 4 5 6 4 7 5 8 6 9 7 2 1 2 2 3 4 3 5 4 6 5 7 6 8 2 3 4 5 a 1 2 b 1 1 3 2 4 5 6 Fig.5.13 Holgate 2/6/03 8:17 AM Page 191 INHERITANCE AND LINKAGE and the proportion of recombinant increases. Secondly. If the disease and marker loci are separated by a considerable distance on the same chromosome. (d) disease and marker loci far apart on the same chromosome mimic independent assortment and linkage cannot be detected (the recombination fraction is 50%). 13. the number of recombinants will approximate to the number of non-recombinants and the number of recombinants divided by the total number of offspring (the recombination fraction.7 Markers and linkage. Thus. (a) Independent assortment at meiosis of the disease locus (disease allele D and normal allele d) and the marker locus (alleles T and t) on different chromosomes. which obeys the rule of independent assortment. or q) will be 0. one centimorgan is equivalent to one megabase of DNA.1 (10%) corresponds to a map distance of 10 centimorgans (cM) but with increasing distance apart the apparent recombination fraction falls due to double crossovers. 13. for a distant marker trait. (c) disease and marker loci nearby on the same chromosome showing linkage (the recombination fraction is 20%). which is equivalent to 3 × 109 base pairs.6 (a) Pedigree showing X-linked recessive inheritance. (b) absence of independent assortment at meiosis for disease and marker loci very close together on the same chromosome (tight linkage). crossovers for autosomes occur more frequently in females than in males and also vary in different parts of the chromosome. Markers and Linkage parent D d T t D T parent d t D T D T d T d t D T d t a 25% b 25% 25% 25% 50% gametes parent D T d t 50% gametes parent D T d t D T d t D t d T D d D d T 10% 10% 40% 40% nonrecombinants recombinants gametes t t T c 25% 25% 25% 25% nonrecombinants recombinants gametes d 191 . As this distance decreases. Fig. or 50%. Thus. The total length of the genome is 3000 cM.
and environmental factors may Gene 1 + Gene 2 + Gene 3… + Environmental factor 1 + Environmental factor 2 +… Fig. If a disorder cannot be accounted for by alterations in known candidate genes. incomplete penetrance. Polygenic inheritance Asthma and atopy are classiﬁed as multifactorial disorders in which both environmental and genetic factors are important. based on an understanding of pathophysiology. Analysis of the segregation of a disease such as asthma within families (segregation analysis) using computer programs helps establish a possible mode of inheritance of a trait or partial phenotype. This means that in every human population. then it may be necessary to screen the entire human genome for linkage. 13. New loci that contribute to a disease such as asthma can be identiﬁed by positional cloning (Fig.8). which themselves are functionally inactive.13 Holgate 2/6/03 8:17 AM Page 192 13. bronchial hyperresponsiveness (BHR). This approach can be strengthened by information from classic genetic studies in which chromosome deletions or translocations may provide additional clues that help identify new disease-related loci. and the contribution made by environmental factors. Many genetic loci participate (Fig. it should be possible to deduce linkage to a locus that contains a gene that contributes to the clinical phenotype. GENETIC VARIATION AND SUSCEPTIBILITY Variation in DNA sequences occurs once in approximately every 200–500 base pairs. and tetranucleotide repeats. Thus. • higher NF-AT-binding afﬁnity. the risk of a polygenic disease in ﬁrst degree relatives is generally 5–15%. the thymidine-containing polymorphism is associated with the following: • a unique Nuclear Factor for Activated T cells (NF-AT)binding site separate from P0–P4 sites. a particular gene has a strong possibility of inﬂuencing a disease. This approach has been made easier by the identiﬁcation of variable nucleotide tandem repeats (VNTR) and simple di-. most genes can be expected to show variation. 192 . Inevitably this leads to disagreements between various research groups and an inability to compare results achieved using different deﬁnitions. This example explains the molecular basis of one mechanism of the IL-4-associated increase in IgE production. The term ‘candidate’ is used when. including our responses to environmental stimuli.10). and clinical history. An alteration in the rate of IL-4 secretion (produced by an appropriate polymorphism of a transcription binding site in the IL-4 gene promoter) would provide the basis for a good candidate gene (Figs 13. each exerting a small effect. As shown in the example of the cytosine to thymidine exchange. This is polygenic inheritance. 13. AND GENETICS DISEASE Deﬁning the clinical phenotype of atopy and asthma Asthma is a clinical diagnosis with no foolproof diagnostic test. THE GENETIC BASIS OF ALLERGIC DISEASE which together combine to give the end phenotype. • increased levels of IgE in vivo. GENETIC MODELLING Knowledge of the recurrence risk of a disease in relatives and curve ﬁtting of disease measures between affected and nonaffected families (commingling analysis) may point towards a mode of inheritance. Thus. Genetic heterogeneity. tri-. interleukin 4 (IL-4) represents such a candidate because it controls the switching of B cells to IgE synthesis and the maintenance of the Th2–T cell phenotype – both essential features of allergic disorders.11). In the case of allergic disorders. so surrogate markers for the disease are used including atopy.8 In complex genetic disorders there are multiple interactions between individual genes and between genes and the environment. Genetic epidemiology has provided statistical methods for measuring the effects of polymorphisms on a clinical phenotype. Polymorphisms Polymorphisms form the basis of human diversity. ASTHMA. Sequence variation (mutations) occurring in over 1% of the population are called polymorphisms and those that occur in less than 1% are rare alleles. The influence of rare alleles will be superimposed on the population distribution. the risk of developing a disease such as asthma depends upon a complex interaction among common alleles. With such highly polymorphic markers spaced at regular intervals across each chromosome (10–20 cM). Within a family the risk will be inﬂuenced by the severity of the disorder in the proband. the number of affected family members. 13. Polygenic Inheritance single gene diseases gene disease complex genetic diseases gene 1 gene 2 environmental factor 1 environmental factor 2 ALLERGY.9 and 13.
AND GENETICS Fig. is defined as a disorder of the IgE response to common allergens such as pollen. which contributes to diseases such as asthma. animal dander. NF-kB (ReIA + NFKB1) Regulation of Human IL-4 Gene Transcription Transduction of Membrane Signals Nuclear Transcription of Factors (NT-Fs) POS-I POS-II NEG-I NEG-II IRF-2 NF-IL6 (C/EBPβ) NF (P) family members NF-AT NF-Y AP-1 (fos+jun) Transcription start site −44 −25 −311−303−300−288 −239 −224 −195−172 −114−107 −88 −76 5' NRE I NRE II polymorphic region (C T) −87−80 −79−69 PRE 1 "CCAATd" ISRE "CCAATm" (Y-box) C/EBPp C/EBPm P-sequence 3' INTRONS' EXONS' OAP-40 also confound statistical analysis and make it difﬁcult to reproduce positive ﬁndings. VNTR genetic markers + linkage analysis = analyze for mutations genetic mapping physical mapping candidate genes 193 . 13.9 The transcription factor binding site of the IL-4 gene is a good allergic disease candidate because of the privotal role of IL-4 in maintaining the Th2 cytokine repertoire and B-cell switching to IgE. Problems with definition of the asthma phenotype have led researchers to study atopy.10 The association of IgE production with C→T exchange. T) – (~590. These diseases are frequently detected by a raised total serum IgE level. C) IgE (IU/dL) 146 39 ® 44 subjects. and positive skin tests to common aeroallergens. Linkage analysis using polymorphic variable nucleotide tandem repeat (VNTR) markers allows the identiﬁcation of a chromosomal region containing the disease gene.) Identification of Disease Genes by Positional Cloning families Fig. and fungi. and allergic rhinitis. 13. candidate genes are then screened for mutations. a major risk factor for the development of asthma. as characterized by a persistent IgE-mediated response to common environmental allergens. Association of IgE Production with C T Exchange Subjects 15 29 Affected + (~590. 13. international unit.0028 Fig. ASTHMA. house dust mites. Atopy.10).13 Holgate 2/6/03 8:17 AM Page 193 ALLERGY. (IU. This is followed by physical mapping of this region to identify genes within it. 13. a raised speciﬁc IgE level.11 Identiﬁcation of disease genes by positional cloning. eczema. p = 0. The promoter region is complex but at –589 a C→T exchange is associated with increased IgE production (see Fig.
A number of variables have been shown to affect both serum IgE levels and BHR. that greatly enhanced bronchial responsiveness may be present in the absence of symptoms. Airway hyperresponsiveness (methacholine PC20 FEV1 < 8 mg/mL) was found to be strongly correlated with reported asthma and wheezing and with atopy as deﬁned by a positive skin-prick test. there is documentation from longitudinal studies that BHR may not be present in some people at a time when they have unmistakable asthma symptoms and airway obstruction and. and the results available so far from genome screening projects will be brieﬂy described.13 Holgate 2/6/03 8:17 AM Page 194 13. Further evidence for the relationship between IgE levels and asthma has been provided by Sears who studied the relationship between serum total IgE levels and airway responsiveness to methacholine challenge in the presence or absence of asthma in a birth cohort of New Zealand children. An eigenvector plot (Fig. Using a physician-based diagnosis to deﬁne the presence of asthma. Linkage and strong association has been detected to speciﬁc alleles on chromosomes 5 and 11. as will a small percentage of normal subjects. asthma. Each of these regions will be discussed in more details. the separate variables derived from the clinical and laboratory data can be combined to generate quantitative asthma and atopy scores. Moreover. skin prick.12). There is clearly a link between atopy. 13. video questionnaire. and wheeze. repeatability has proven problematic. airway hyperresponsiveness. although between studies. On the basis of this. Regardless of the atopic status of the subjects or their age group. wheeze. not only in those with a reported history of asthma and wheezing but also in those without any history suggestive of asthma and rhinitis.13) illustrates the separation of individual traits into clusters associated with asthma. and migraine) were used to define an asthma and atopy score. all the children with diagnosed asthma and airway hyperresponsiveness were atopic. A signiﬁcant shift in atopy score distributions can also be seen between atopic and nonatopic individuals. although the overlap between groups is large. atopy. Mechanisms Underlying Asthma Symptoms environmental risk factors (causes) Inflammation SYMPTOMS airway hyperresponsiveness airflow limitation triggers Fig. the prevalence of asthma was closely related to the serum total IgE level standardized for age and sex. 13. particularly to house dust mite and cat.14). The prevalence of diagnosed asthma was signiﬁcantly related to the serum IgE level and airway hyperresponsiveness was still related to an allergic diathesis.12 The relationship between bronchial hyperresponsiveness and the expression of asthma symptoms. 194 . Principal component and logistic regression analyses of eight traits (IgE. conversely. Enhanced bronchial responsiveness has a strong association with clinically defined asthma and the association appears to be stronger in those with more immediate and severe symptoms and with greater treatment requirements. Furthermore. BHR. some atopic subjects with no evidence of symptomatic asthma will also demonstrate BHR according to the same criteria. Several other studies provide positive evidence for these two regions. However. Evidence for candidate genes Candidate genes (or regions) have now been investigated in a total of 13 human chromosomes. the asthma score distributions differ signiﬁcantly between asthmatics and nonasthmatics from the combined random and multiplex populations (Fig. as reﬂected by the serum total IgE level even in children who had been asymptomatic throughout their lives and had no history of atopic disease. There is a tendency to dichotomize subjects as hyperresponsive or nonresponsive on the basis of whether or not their forced expiratory volume in one second (FEV1) falls by 20% at a given dose of inhaled histamine or methacholine. atopy. 13. although the precise relationship remains a source of considerable debate (Fig. although BHR and asthma are related. and asthma Further work on the New Zealand cohort of children has looked at the relationship between airway hyperresponsiveness. different cut-off doses have been used by different workers. No asthma was present in the 177 subjects with the lowest IgE levels for their age and sex. As an alternative to studying asthma or surrogate markers (IgE or BHR) as the principal phenotype. asthma. and atopy. THE GENETIC BASIS OF ALLERGIC DISEASE The association of self-reported asthma or allergic rhinitis with serum IgE levels and skin test reactivity to allergens has been investigated in 2657 subjects in a general population study. as well as for chromosomes 6 and 14. Thus. and asthma. Bronchial hyperresponsiveness. 13. For example. one would hope to be able to discriminate clearly between asthmatics and nonasthmatics. The conclusion reached was that asthma is almost always associated with some type of IgE-related reaction and therefore has an allergic basis. or may develop after symptoms have become manifest as occurs in seasonal asthma. atopy. the two are not synonymous. It was concluded that atopy was a major determinant of airway hyperresponsiveness in children. smoking has been shown to lead to an increase in total serum IgE levels.
total serum IgE level. We did. IL-9. responses to the video and written questionnaire on wheezing in the last year. centred around the ADRB2 gene. 13.2 35. eczema. These are indicated by arrows in Figure 13. was first reported in 11 large Amish pedigrees.3 32 33.3 1 cM 1 cM 2 cM 4 cM 3 cM 3 cM 4 cM 6 cM 9 cM D5S393 D5S399 D5S500 D5S658 D5S436 D5S434 D5S470 D5S410 D5S412 D5S415 IL-4 IL-13 IL-5 IRF-1 CDC25 IL-3 CSF-2 IL-9 EGR-1 CD14 ADRB2 FGF-A GRL-1 PDGFR CSF-1R IL-12B 8 cM Chromosome 5 The 5q31–34 region of the genome contains several candidate genes implicated in the pathogenesis of asthma and in the regulation of IgE. The linkage evidence was strongest when subjects with high speciﬁc IgE were excluded from the analysis. Fig. HF. In the population of Southampton. centred around the IL-4 locus. AND GENETICS Relationship Between Asthma Partial Phenotypes RFEV IgE BHR ASI VID WZ RAST SP frequencies ASI Distribution 40 30 20 10 0 Asthmatics −9 −7 −6 −5 −4 −3 −2 −1 0 1 2 3 4 5 6 7 8 250 200 150 100 50 0 Nonasthmatics HF −9 −7 −6−5 −4 −3 −2 −1 0 1 2 3 4 5 6 7 8 EZ Fig. (FEV and FEV1. The asthma index (ASI) represents the prinicpal component derived from bronchial hyperresponsiveness (BHR) and the wheeze responses on a standardized questionnaire.13 Holgate 2/6/03 8:17 AM Page 195 ALLERGY. % predicted.15 Genetic map of chromosome 5q31–34 showing potential candidate genes including those of the IL-4 cluster and the β2-adrenergic receptor. 13. IgE and SP indicate their possible association. 13. SP.) asthma score Fig.1 31. IgE. including the IL-4 cytokine cluster (IL-3. The strongest evidence for both phenotypes in the Dutch population was more distal than the IL-4 locus. ASTHMA. The close proximity of RAST.3 34 35. 13. This makes ADRB2 a promising candidate. summated positive skin tests to common aeroallergens.1 35. IL-5. These studies on chromosome 5q do not have sufﬁcient power to produce an accurate localization of the disease gene(s) and it is highly likely that more than one important locus exists in the region including CD14 (lipopolysaccharide receptor) and α-catenin (involved in epithelial homeostasis). we were unable to identify signiﬁcant linkage to any of the seven markers typed. we have genotyped polymorphic markers within the IL-4 and IL-9 genes as well as ﬁve anonymous markers (i. UK. IL-13 and the granulocyte–macrophage colony-stimulating factor) and the β2-adrenergic receptor (ADRB2) (Fig. There is also evidence that these variants could have a functional signiﬁcance.14 Distribution of the asthma index (ASI) in a population of families with one or two affected children with asthma (top). hayfever.13 Principal component regression analysis used to examine the relationship between various asthma partial phenotypes. particularly as coding variants identified within the gene have been associated with hyperresponsiveness and total serum IgE. EZ.e. VID and WZ. IL-4. ﬁnd strong allelic association with log total 195 . summated radioallergosorbent test for speciﬁc IgE. suggesting that speciﬁc IgE responsiveness is a confounding factor in the analysis of the genetics of total serum IgE. however. The relationship between the ASI and the various partial phenotypes is displayed as an eigenvector plot.2 33.15.1 33. at loci with no known biological function) which span the 5q31–34 region. Candidate Genes on Chromosome 5q31–34 Candidate genes 5q VNTR Markers D5S421 D5S404 9 cM 31.2 31.15). Using a similar phenotype a second study also reported linkage to total IgE in 92 Dutch families. as well as to a phenotype based on BHR. Compared to a normal population (bottom). RAST. Evidence of linkage for this region to total serum IgE. Although positive linkage has been demonstrated to several of these markers in other studies.
g. and in the method of HLA typing used. e. HSP-70. Polymorphisms within the promoter regions of several of these genes have now been identiﬁed and there is evidence for association between a nucleotide substitution in the IL-4 promoter and total serum IgE (see Fig. Thus. TNF. The number of alleles indentiﬁed by 1996 is indicated. 13. and DQ) and also those that are central to the process of antigen recognition and presentation and. has now been identiﬁed in exon 7. An additional factor contributing to positive association to MHC class I and II genes through linkage disequilibrium is the presence of additional card- F G H A J C HLA gene complex B 119 36 4 E HLA class I region HLA class II region HLA class III region allelic variants 1996 62 8 25 16 128 2 4 59 TNF HSP70 C2 Bf C4A CYP21B C4B CYP21 DRA DRB Basic Structure of HLA Class I and Class II Molecules chromosome 6 class II peptide binding groove α (heavy) chain α chain β chain class I DQA DQB DPA DPB cytosol β2-microglobulin Fig. designated Leu 181 and Leu 183. polymorphisms of which are associated with asthma.and family-based studies have examined the relationship between HLA type and speciﬁc IgE responses and there is much evidence for association between certain haplotypes and individual responses to puriﬁed allergens.) 196 . Any comparisons between studies is complicated by differences in the ethnic origin of the study population. IgE at the IL-9 locus (p < 0. modulate the speciﬁcity of the immune response (Figs 13.13 Holgate 2/6/03 8:17 AM Page 196 13. which were highly predictive of atopy when inherited maternally.17 The HLA gene complex on the short arm of chromosome 6. (CYP. E237G. tumor necrosis factor. The ﬁrst demonstration was an association between HLA haplotypes and IgE responses to antigen E derived from ragweed (Ambrosia artemisifolia). This association could not be repeated in the multiplex sample but the positive linkage to other markers in the region and the biological plausibility of the candidate genes within the cytokine cluster suggest that this may not be a type I error. UK group ﬁrst reported genetic linkage of atopy to chromosome 11q13 in extended and nuclear families with the ‘atopy gene’ preferentially active in maternally derived alleles. Both population.17).003) in the random sample. signiﬁcant associations have been found with highly puriﬁed simple allergens rather than with more complex common ones.16 The basic structure of HLA class I and class II molecules. which is also associated with asthma but with no maternal effect (Figs 13. An additional substitution.19). the phenotypic deﬁnitions used. THE GENETIC BASIS OF ALLERGIC DISEASE dates. 13. This was subsequently shown to be due to a restriction of the response to a minor component of the ragweed antigen (Amb a V) by HLA-DR2. Generally. cytochrome P450 enzymes. TNFα. HSP. DP.9). heat shock protein.18 and 13. the accumulated evidence associating HLA type with speciﬁc IgE responsiveness is inconsistent and appears insufﬁcient to account for individual differences in reactivity to common allergens. Fig. Chromosome 11 The Oxford. 13. HLA Gene Complex or Chromosome 6 Chromosome 6 The major histocompatibility complex (MHC) on chromosome 6 includes genes that code for human leukocyte antigen (HLA) class II molecules (designated DR. as a consequence. The gene encoding the β subunit of the high-afﬁnity IgE receptor (FcεRI-β) was proposed as the candidate gene and determination of its coding sequence identiﬁed two amino acid substitutions within exon 6.16 and 13.
This result could not be repeated in the random Fig. ASTHMA. CA repeat Leu 181/183 E237G Rsal 1 2 3 4 Exon Number Leu 181/183 5 6 ITAM 7 D11S451 FcεR-I. the proposed maternal effect. both in the random sample. An intronic marker within the FcεRI-β gene gave weak evidence for linkage to loge IgE (p = 0. although only one marker.05) in the multiplex sample. particularly with anonymous markers. 13. between D11S527 and BHR (p < 0.007). The Leu 181.β CEN PGA D11S480 D11S97 INT2 D11S527 D11S534 TEL ATOPY LOCUS 197 . AND GENETICS Despite the biological plausibility of the FcεRI-β candidate gene. and E237G polymorphisms are shown. 13. the putative 11q linkage aroused controversy because of the broad definition of atopy used. was close to the putative atopy locus. positive results can be produced by type I errors.13 Holgate 2/6/03 8:17 AM Page 197 ALLERGY.0003) and between D11S534 and loge IgE (p = 0.19 Schematic map of chromosome 11q and the FcεRIβ gene. The association between D11S527 and BHR could not be validated in the asthma multiplex sample and in multiple association tests. Two strong allelic associations were observed. 183. Structure of the High-affinity Receptor for IgE Leu 181 Leu 183 C C human α chain N rat β chain C C human γ chains Schematic Map of Chromosome 11q and the FcεRI-β Gene Fig. Genotyping of three anonymous markers in the region produced no evidence for linkage in the Southampton random population.18 The four-chain structure of the high-afﬁnity receptor for IgE (FcεRIαβγ2). D11S480. and the lack of independent replication.
13 Holgate 2/6/03 8:17 AM Page 198 13. then it must be at a very low frequency and with limited clinical relevance to asthma. signiﬁcant linkage of IgE responses to house dust mite allergens. Single-locus. nonparametric analysis revealed linkage evidence to several markers with the strongest linkage evidence in distal 12q around the marker D12S97 (p = 0.20 Schematic representation of chromosome 12q where linkage to asthma has been found.01 to atopy score). However.5% in both our random and asthma multiplex UK populations. Multipoint locus testing has conﬁrmed linkage to these regions (Fig.05 to asthma score. while the linkage values for chromosome 12 would be classed as signiﬁcant by this convention (p < 0. they fall short of being highly signiﬁcant (p < 0. although not to BHR or log IgE. Linkage to this marker was also replicated in the random sample to the atopy score (p = 0.002) and D12S78 and loge IgE (p = 0.006) and analysis of the 12q data also identiﬁed two allelic associations in the same region – D12S366 and atopy (p = 0. Furthermore. has failed to identify a single Leu 181 or Leu 183 carrier. Using two large population samples (ascertained by separate criteria) and employing different analytical methodologies overcomes the limitations apparent in many of the other negative linkage reports but can not unequivocally conﬁrm the original linkage result.001).3% and 6% in unselected Australian and Japanese populations. However. the wild-type nucleotide sequence has been found in every case. Analysis of these data produced only weak evidence of association to BHR in the random sample (p = 0. phospholipase A2 group 1 B.0001 in the multiplex sample). chromosome 12q contains several candidate genes including Fig. together with additional 12q markers taken from published Genethon® sequences on the multiplex sample. but clearly it is important to attribute functions to FcεRI in the expression of elevated IgE levels and to asthma if the clinical relevance of the polymorphisms within the β chain is to be demonstrated. which are coded for by genes on chromosomes 14 and 7 respectively. Chromosome 12 Linkage to chromosome 12 has been identiﬁed in Southampton by genotyping a chromosome-12-speciﬁc marker set. Alterations in the wild type sequence of a gene can have biological and clinical signiﬁcance. e. signiﬁcant association between a gene polymorphism and a disease trait does not necessarily indicate a causative function but that the gene is in linkage disequilibrium with a neighboring relevant gene Chromosome 14 Together with HLA. In contrast.) Schematic Representation of Chromosome 12q D12S83 CEN IFNγ D12S92 PAH D12S82 IGF-1 D12S95 D12S366 D12S324 D12S357 TEL D12S97 D12S78 MGF NFYB NOS-1 D12S342 PLA2G1B 198 . Positive linkage has also been found in two distinct sample populations. p = 0. using direct-cycle sequencing of polymerase-chain-reactor-ampliﬁed DNA fragments encompassing exon 6 from all 120 parents of the asthma multiplex families and from both parents of 90 random families. cat. implying that a gene in the TCR-α region modiﬁes speciﬁc IgE responses. Guidelines for the interpretation of genetic linkage studies of complex traits have recently been proposed and. the T cell receptor (TCR) proteins are central to the handling and recognition of foreign antigens. uteroglobin secreted by Clara cells (CC10 or CC16). the substitution of glutamic acid for glycine at residue 237 (E237G) has been demonstrated at frequencies of 5. and total serum IgE was demonstrated in TCR-α serotypes. one each from the UK and Australia. which is a strict criterion for the conﬁrmation of a putative linkage. no reliable assay has been established for these variants and every independent investigation.05). using a variety of different methodologies.0009). 13. there was signiﬁcant linkage to both the asthma score (p = 0. There are two further reasons to think that linkage to this region is genuine. 13. mast cell growth factor.20). and analysis of the combined sample failed to reach signiﬁcance. However. positive linkage has been replicated in a second distinct population. sample. An investigation of genetic linkage between speciﬁc IgE responses to common allergens and both the TCR-α and TCR-β gene complexes identiﬁed no linkage to TCR-β serotypes. TCR complexes are usually made up of α and β chains. In both cases the presence of E237G is strongly associated with asthma (p = 0. In contrast.02). The Leu 181 substitution has been reported by the Oxford group at a frequency of 15% in an unselected UK population. (MGF. Despite publications regarding the signiﬁcance of the Leu 181 and Leu 183 substitutions.009 in both samples) and the atopy score (p = 0. The substitution occurs at a frequency of around 3. Firstly.g.005) and in the Australian population it is also associated with BHR (p = 0. the anti-inﬂammatory protein.001). If these substitutions do exist. PLA2G1B. THE GENETIC BASIS OF ALLERGIC DISEASE close by.
11. a mast cell growth factor (MGF-1). Reviewing candidate genes with known Loci-linked and Genome-wide Scan for Atopy and Asthma Phenotypes Locus D4S426 D6S276 D7S484 FCAR> D13S153 D16S289 0.22 Candidate genes with known mutations associated with asthma severity. IgE. 11.0001 p < 0. Only regions showing linkage to one or more phenotypes with p < 0. the level of signiﬁcance achieved for linkage was low. 7. there is independent evidence for linkage to chromosome 12q from a genome screen in two populations (Afro-Caribbean and Caucasian Hutterite families). although the maximum linkage determined by multipoint analysis in this study was a considerable distance proximal to the D12S97 locus.0005 G Loge IgE Skin test index Loge eosinophil count Loge slope p < 0. AND GENETICS interferon γ.05 p < 0. and 16). Due to this ratial admixture.20 0. 13. 6.10 0. 13. bronchial hyper-responsiveness. ASTHMA.05 p < 0. and 16 (Fig. Hispanics. and relatively small numbers of families in each group.00005 p < 0.0005 Atopy Fig. BHR.21). four also showed linkage in a second panel of markers (chromosomes 4. An additional whole genome screen includes the US CSGA study comprising three racial groups – Caucasians.06 p < 0. the constitutive form of the nitric oxide synthase gene (NOS-1). TNF.00 0. T-cell receptor.001) to chromosomes 4. and Afro-Caribbean. nocturnal asthma Specific IgE ?Asthma Asthma Asthma Atopy.0005 p < 0. HLA. treatment response. 13. TCR.22 shows those candidate genes with known mutations which have been proposed to be important in the development of atopy and asthma and the subphenotypes. BHR Specific IgE Atopy Fig. tumor necrosis factor. six regions. and the β subunit of the nuclear factor Y involved in transcription of HLA genes (NFYB).05 p < 0. 13. 5-LO.03 0.) 6 Low High High High Low Low Low 10 11 14 16 199 . UK) has identiﬁed six positive linkages (p < 0. 13. (BHR. Candidate Genes with Known Mutations Associated with Asthma Severity Chromosome Candidate gene Association Potential contribution to disease severity Low Low High 5 IL-4 IL-9 >2AR HLA TNF= 5-LO CC16 FcAR1-> TCR= IL-4R IgE IgE IgE.0005 p < 0.13 Holgate 2/6/03 8:17 AM Page 199 ALLERGY. Secondly.21 Loci-linked and genome-wide scan for atopy and asthma phenotypes. The recombination fraction (q) is to the subsequent locus. an insulinlike growth factor (IGF-1). The chromosome 11q13 has been implicated in the pathogenesis of asthma and atopy and a polymorphism in the esterase D protein on chromosome 13 had previously been linked to total serum IgE. Genome screens Recent advances in molecular technology and the availability of dense genetic maps has made it feasible to screen the whole genome. Of these Candidate genes in allergic disease and their potential contribution to asthma severity Figure 13.001 p < 0.001 are shown. 5lipoxygenase. A number of these projects are now underway and the ﬁrst to be completed (in Oxford. human lymphocyte antigen.
With the development of 5-LO inhibitors for the treatment of asthma. There have been many negative studies that have looked at other genes which might potentially be considered to be disease modifying but it may be premature to discard these as contributory genes given that severity was not speciﬁcally examined in many of these studies. this polymorphism confers increased levels of downregulation following agonist exposure.23 Structure of the β2-adrenoceptor showing the sites of polymorphisms at amino acids 16 and 27.13 Holgate 2/6/03 8:17 AM Page 200 13. The Boston. it will be important to determine whether polymorphisms affecting gene transcription will determine response to treatment. 13. relevant mutations do not Structure of the β2-Adrenoceptor 16 Arg or Gly NH2 27 Gln or Glu Extracellular 34 Val or Met Intracellular 164 Thr or IIe HOOC 200 . although it is possible that these ﬁndings are due to linkage disequilibrium with other genes close by on 5q31 given that both IgE and BHR have been linked to this region of 5q by other groups. Fig. although whether this effect occurs with clinically relevant doses of inhaled long-acting β-adrenoceptor agonists remains to be determined. More recently the response to long-acting β2-adrenoceptor agonists of individuals who are homozygous for the glycine 16 β2-adrenoceptor variant has been studied. As such. it is clear that the majority of these genes will be expected to be involved in the control of IgE. mutations. To date.and Th2mediated inﬂammatory responses. no speciﬁc mutations have been identiﬁed which determine steroid resistance although this work is ongoing. There is a small group of individuals who do not appear to respond to corticosteroids even at a high dose. A number of attempts have been made to deﬁne the molecular basis of corticosteroid resistance including sequencing the glucocorticoid receptor from a number of such patients. THE GENETIC BASIS OF ALLERGIC DISEASE have to be within the coding region of the glucocorticoid receptor gene itself but could be in genes for products in downstream signalling pathways.24). Subjects with this variant develop tachyphylaxis to the bronchodilator effects of longacting β-agonists following chronic dosing with oral formoterol. given that mutations in these genes would be expected to predispose to asthma per se. Such patients have been labelled ‘corticosteroid-resistant asthmatics’. The glycine 16 form of the receptor is also associated with a nocturnal fall in FEV1. Pharmacogenetic studies One obvious way in which gene factors may inﬂuence asthma severity is by determining treatment responses. it is perhaps not surprising that positive results have been obtained looking at mutations in these genes using endpoints likely to identify disease-initiating genes. In practice there appears to be a continuum of corticosteroid responsiveness and such patients may just represent one extreme end of the distribution. This possibility has been specifically examined in asthmatics with respect to responses to corticosteroids and to β2-adrenoceptor agonists to date. Other researchers have concentrated on assessing the contribution of β2-adrenoceptor polymorphism to allergic disease (Fig. which have been associated with allergic disease. In vitro.23). Initial studies demonstrated that the glutamate 27 β2-adrenoceptor polymorphism was associated with lower IgE levels and less reactive airways in asthmatic subjects (Fig. Obviously. USA group has recently described a promoter polymorphism in the 5-lipoxygenase gene which alters transcription of this leukotriene generating enzyme in in-vitro systems. 13. 13. One other study has been reported which is potentially relevant to asthma pharmacogenetics.
Daffy DL. Hill MR.1 5:389–92. A co-twin control study. Martin NG. Amelung PJ. 2 8(Suppl. It is also clear that agreement must be reached on deﬁnition of the phenotype and methods of ascertainment in order to carry out large. 1):88–9. Doull I. Bronchial hyper-responsiveness co-inherited with a major gene for atopy. 5 8:359–68. Proc Natl Acad Sci USA 1998.13 Holgate 2/6/03 8:17 AM Page 201 FURTHER READING CONCLUSION New techniques for scanning the human genome promise great advances in tracking the origins of disorders caused by multiple genes. 13. it is clear from the studies presented in this overview that we are far from understanding the genetic basis of asthma and atopy and their interactions with the environment. Mapping a disease locus by allelic association. Wilding P. Cookson WOCM. Collins A. Beasley R. Postma DS. 2 8(Suppl. Am J Respir Crit Care Med 1998. 5:959–62. Watson M. Hum Mol Genet 1996. Lawrence S. Sears MR. Genetic analysis of atopy and asthma as quantitative traits and ordered polychotamies. Clin Exp Allergy 1998. Bhattacharrga S. However. Daniels SE. Genetic susceptibility to asthma. multicenter. 5 0:815–18. HLA genetics and allergic disease. Kruglyak L. Palmer LJ. Morton NE.24 Association between reduced bronchial hyperresponsiveness (He – heterozygote) and the Glu 27 polymorphism of the β2-adrenoceptor. Liggett SB. Am J Respir Crit Care Med 1996. Association of the Glu 27 beta 2-adrenoreceptor polymorphism with lower airways reactivity in asthmatic subjects. Nat Genet 1995. A new variant of the β subunit of the high afﬁnity receptor for immunoglobulin E (FcεR1β E237G): associations with measures of atopy and bronchial hyper-responsiveness. et al. The evolution of asthma through childhood. FURTHER READING Collaborative study on the genetics of asthma. 201 . 1741–5. Bleecker ER. 3 8 3: 247–50. Allelic association of markers on chromosomes 5q and 11q with atopy and bronchial hyperresponsiveness. 1 5 3:1280–4. 5):82-9. N Engl J Med 1995. 3 3 3:894–900. Nature 1996. Howell WM. Clin Exp Allergy 1998. Doull IJM. Positive ﬁndings need to be validated in different populations selected for the presence of the disease and then conﬁrmed in a random population where the prevalence of asthma and atopy will also be expected to be signiﬁcant. Ann Hum Genet 1994. 1 1:241–7. 345:1213–14. Mitchell CA. Lancet 1995. et al. collaborative studies. 1 5 7:840–5. Lander ES. Nat Genet 1997. Investigating the asthma phenotype. Hall IP. Genetic dissection of complex traits: guidelines for interpreting and reporting linkage results. James A. Cookson WOCM. et al. Thorax 1995. Association Between Allergic Disease and the Glu 27 β2-Adrenoceptor Polymorphism 3 2 1 0 −1 −2 Gln 27 He Glu 27 Fig. A genome wide search for quantitative trait loci underlying asthma. Genetic and environmental risk factors for asthma. A genome wide search for asthma susceptibility loci in ethnically diverse populations. Lawrence S. Wheatley A.
13 Holgate 2/6/03 8:17 AM Page 202 202 .