Potential viral pathogenic mechanism for new variant
inflammatory bowel disease
V Uhlmann*, C M Martin*, O Sheils, L Pilkington, I Silva, A Killalea, S B Murch,
J Walker-Smith, M Thomson, A J Wakefield, J J O’Leary
J Clin Pathol: Mol Pathol 2002;55:84–90
Aims: A new form of inflammatory bowel disease (ileocolonic lymphonodular hyperplasia) has been
described in a cohort of children with developmental disorder. This study investigates the presence of
persistent measles virus in the intestinal tissue of these patients (new variant inflammatory bowel
disease) and a series of controls by molecular analysis.
Methods: Formalin fixed, paraffin wax embedded and fresh frozen biopsies from the terminal ileum
were examined from affected children and histological normal controls. The measles virus Fusion (F)
and Haemagglutinin (H) genes were detected by TaqMan reverse transcription polymerase chain reac-
tion (RT-PCR) and the Nucleocapsid (N) gene by RT in situ PCR. Localisation of the mRNA signal was
performed using a specific follicular dendritic cell antibody.
Results: Seventy five of 91 patients with a histologically confirmed diagnosis of ileal lymphonodular
hyperplasia and enterocolitis were positive for measles virus in their intestinal tissue compared with five
of 70 control patients. Measles virus was identified within the follicular dendritic cells and some lym-
phocytes in foci of reactive follicular hyperplasia. The copy number of measles virus ranged from one
to 300 000 copies/ng total RNA.
Conclusions: The data confirm an association between the presence of measles virus and gut patho-
logy in children with developmental disorder.
pathology includes ileocolonic lymphonodular hyperplasia
and non-specific colitis, which manifests as neither Crohn’s
disease nor ulcerative colitis. The histological and clinical
aspects of this new disorder have been reported previously.2It
has been postulated that reactive follicular hyperplasia in ileal
tissue biopsies of affected children may reflect the persistence
of viral antigen at this site.2Preliminary immunohisto-
chemical data suggested the presence of measles virus (MV)
antigen in the extracellular matrix of the midgut mucosal
lymphoid tissue in affected children.2
MV belongs to the family of single stranded paramyxovi-
ruses. It is the causative agent for several diseases including
subacute sclerosing panencephalitis (SSPE) and measles
inclusion body encephalitis.3Measles ranked as one of the
leading causes of childhood mortality in the late 20th
century.4In developing countries, 1 million deaths each year
are related to MV infections.
n apparently new form of immune mediated inflamma-
tory bowel disease in a cohort of children with develop-
mental disorder has been described.1The intestinal
“Measles virus is the causative agent for several diseases
including subacute sclerosing panencephalitis and mea-
sles inclusion body encephalitis”
Our study examines a possible association between MV and
the above condition. To achieve this aim, several molecular
biological techniques have been used to identify, localise, and
measure MV from terminal ileum biopsies in children with
ileocolonic lymphonodular hyperplasia and developmental
MATERIALS AND METHODS
Patients and RNA extraction
All patient samples were provided by the department of
gastroenterology, Royal Free Hospital, London, UK. Ileal lym-
phoid tissues from 91 affected children were examined
(median age, 7 years; range, 3–14; 77 boys). Developmentally
normal paediatric controls (n = 70; range, 0–17 years; 47
boys) included:19 children with normal ileal biopsies,13 chil-
dren with mild non-specific chronic inflammatory changes,
three children with ileal lymphonodular hyperplasia (LNH)
investigated for abdominal pain, eight children with Crohn’s
disease, one child with ulcerative colitis, and 26 children who
had undergone appendicectomy for abdominal pain including
MV positive control material included two cases of SSPE
and MV infected Vero cells.Negative control material included
uninfected Vero cells, and human tissues, control RNA
extracted from Raji cells (Applied Biosystems, Foster City,
California, USA) and normal peripheral blood mononuclear
Total RNA was extracted from fresh frozen biopsies,periph-
eral blood mononuclear cells,and MV infected and uninfected
Vero cell lines using the Ultraspec-11 RNA isolation system
(Biotecx Laboratories, Houston, Texas, USA). Total RNA was
extracted from formalin fixed, paraffin wax embedded tissues
using the Purescript® RNA isolation kit (Gentra Systems,
Minneapolis, Minnesota, USA).
Abbreviations: AP, alkaline phosphatase; BCIP,
bromochloroindoylphosphate; DIG, digoxigenin; F, fusion;
H, haemagglutinin; IL, interleukin; LNH, lymphonodular hyperplasia;
MV, measles virus; N, nucleocapsid; NBT, nitrobluetetrazolium;
PCR, polymerase chain reaction; RT, reverse transcription; SSC, saline sodium
citrate; SSPE, subacute sclerosing panencephalitis; Th1, T helper cell type 1
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
*These authors share first authorship.
See end of article for
Professor J J O’Leary,
Department of Pathology,
Hospital, Dublin 8, Ireland;
Accepted for publication
8 November 2001
Solution phase RT-PCR
Polymerase chain reaction (PCR) primers and probes to
conserved regions of the MV Nucleocapsid (N), Haemaggluti-
nin (H), and Fusion (F) genes were designed using Primer
Express Software Version 1.5 (ABI Prism; Applied Biosys-
tems).The specificity of selected sequences was checked using
the NCBI Blast program (www.ncbi.nml.nih.gov/blast). Table
1 shows the MV primer and probe sequences, amplicon sizes,
and GenBank accession numbers used for designing PCR
primers and oligonucleotide probes. In some instances primer
sets overlap with each other (for example, the sequence of
amplicon N1 overlaps partially with the N2 PCR amplicon).
For in situ PCR,oligonucleotide probes were modified at the
5′ end by the addition of a biotin moiety and for Southern blot
analysis probes were labelled at the 3′ end with digoxygenin.
For TaqMan quantitative reverse transcription PCR (RT-PCR),
probes were dual labelled with the fluorescent molecule FAM
at the 5′ end and the quencher TAMRA at the 3′ end.
Purified MV RNA (HU2) was used as a positive control to
optimise PCR assays. The following optimal reaction condi-
tions were used for each 25 µl reaction: 0.4mM dNTPs, 0.4µM
forward and reverse primers, 2.5mM magnesium acetate, 5 U
rTth DNA polymerase,0.01 U AmpErase,and 1× EZ buffer.The
EZ buffer consisted of 50mM bicine, 125mM potassium
acetate, 40% (wt/vol) glycerol (pH 8.2) (Applied Biosystems);
50 ng of extracted RNA was used for each reaction. The
following RT-PCR thermal cycling conditions were used on a
9700 PCR thermocycler (Applied Biosystems): 50°C for two
minutes, 58°C for 30 minutes, 95°C for five minutes, then 40
cycles of 94°C for 20 seconds,59°C for 20 seconds,and 72°C for
20 seconds, followed by an extension step at 72°C for 10 min-
Southern blot analysis
To confirm reaction specificity, solution phase RT-PCR was
for MV by TaqMan RT-PCR (see below). MV F and H gene
amplicons from MV infected Vero cells, SSPE brain, and ileal
template control, were examined by Southern blotting5using
sequence specific probes (table 1). MV specific oligonucleotide
probes were labelled at the 3′ end with digoxigenin using a DIG
cals, Mannheim, Germany), hybridised to the Southern blot at
50°C in 5× saline sodium citrate (SSC) with 0.2% sodium
dodecyl sulfate for one hour,washed once in 5× SSC at 50°C for
and once in 0.2× SCC at room temperature for 10 minutes.
Hybrids were detected using the DIG luminescent detection kit
(Roche Molecular Biochemicals).
Real time quantitative RT-PCR based on the 5′ nuclease assay
was performed on an ABI 7700 Sequence detector (Applied
Biosystems).6 7Sequence specific PCR primers and TaqMan
probes were designed using Primer Express software as
described above. All quantitative PCRs were prepared in a
dedicated facility in a class 2 laminar flow bench hood using
dedicated pipettors and aerosol resistant pipette tips.Template
Measles virus primer and probe sequences
5′ TCA GTA GAG CGG TTG GAC CC 3′
5′ GGC CCG GTT TCT CTG TAG CT 3′
5′ GAG TCG AGG AGA AGC CAG GG 3′
5′ GCT GGA CTC CGA TGC AGT GT 3′
5′ TTC ATC GGG CAG CCA TCT AC 3′
5′ CTC TGA GGT GTC CTC AGG CC 3′
5′ TGG GCA CCA TTG AAG GAT AA 3′
5′ AAC CGT GTG TGA TCA ATG GC 3′
5′ TGA CTC GTT CCA GCC ATC AA 3′
5′ TGG GTC ATT GCA TTA AGT GCA 3′
5′ CCC ACC GGT CAA ATC CAT T 3′
5′ CCC TCG TGC AGT TAT TGA GGA 3′
5′ GAA GGT GAA GGT CGG AGT 3′
5′ GAA GAT GGT GAT GGG ATT TC 3′
5′ CAA ACA GAG TCG AGG AGA AGC CAG GGA 3′
5′ CCG CAG AGA TCC ATA AAA GCC TCA GCA C 3′
5′ CTG CAC GAG GGT AGA GAT CGC AGA ATA CAG 3′
5′ CCG ACT CTT GCC CTT CGA AC 3′
Measles virus N gene primers and probes were designed based on the following Genbank sequence entries
(accession numbers): X16565, S58435, NC_002494, NC_002496, X01999, U03661, U03658, and
U03656. Measles virus H gene primers and probes were designed based on the following GenBank
sequence entries: X16565, U03649, U03654, U03669, U03660, U03671, U03667, Z80793,
AF045204, Z66517, and Z80816. Measles virus F-gene primers and probes were designed based on the
following Genbank sequence entries: X16565, U03655, U03666, U03648, U03662, U08146, U03657,
U03651, U03659, AJ133108, X16567.
(HU2 control measles virus RNA) amplified under optimised reaction
conditions. Lanes 1 and 10, 100 base pair DNA ladder (Gibco-BRL,
Paisley, Scotland, UK). Lanes 2 and 3, Fusion (F) gene cDNA
amplicons for two different F gene primer sets. Lanes 4 and 5,
Nucleocapsid (N) gene cDNA amplicons for two different N gene
primer sets. Lanes 6 and 7, Haemagglutinin (H) gene cDNA
amplicons for two different H gene primer sets. Lane 8, positive
control human pyruvate dehydrogenase PCR product. Lane 9, no
template control for lane 2.
Gel electrophoresis of measles virus cDNA amplicons
Pathogenesis of new variant inflammatory bowel disease 85
RNA was prepared and added to the PCR mastermix in a
TaqMan RT-PCR was performed using EZ TaqMan RT PCR
reagents according to the manufacturer’s instructions (Ap-
plied Biosystems). RT-PCR reactions were performed in dupli-
cate under the following conditions for each 25 µl reaction:1×
EZ buffer, 3mM MnOAc2, 200 nmol of each primer, 100 nmol
of TaqMan probe, 0.01 U of AmpErase, 0.1 U of rTth polymer-
ase, and 3 µl (5–50 ng) of total RNA. The thermal cycling con-
ditions on the 7700 were as follows: 50°C for two minutes,
58°C for 30 minutes, 95°C for five minutes, followed by 40
cycles of 94°C for 20 seconds and 60°C for one minute.
Controls for TaqMan RT-PCR included the following: no
template control (water added as template), no amplification
control (omission of rTth polymerase), irrelevant target prim-
ers and specific TaqMan probe (human papillomavirus 16,
human herpes virus 8 primers), probe only control (omit PCR
primers), human RNA control, spiked RNA control, and
asymmetric TaqMan PCR (TaqMan PCR with one or other
primer and specific TaqMan probe).
A gene dosage correction was carried out using glyceralde-
hyde phosphate dehydrogenase as a housekeeping gene.Mea-
sles virus quantitative TaqMan RT-PCR was performed by gen-
erating standard curves for the F and H genes.Taqman RT-PCR
standards were generated by cloning the F and H gene specific
PCR products into a vector using the TOPO TA cloning®system
(Invitrogen, Groningen, the Netherlands), according to the
manufacturer’s instructions. Plasmids containing the PCR
chain reaction (RT-PCR) amplicons generated using RNA extracted from fresh frozen terminal ileum biopsies from affected children and
subacute sclerosing panencephalitis (SSPE) brain tissue and measles infected Vero cells. M denotes 100 base pair molecular weight marker;
lanes 1–6 show F gene PCR amplicons from the following RNA samples: measles infected Vero cells, SSPE brain, affected children 1–4,
respectively; lane 7, no template control. Lanes 8–14 show H gene PCR amplicons generated from RNA samples in the same order as above.
For one sample, RT-PCR for the F gene failed (lane 5). (B) Southern blot of the agarose gel described in (A) probed with a digoxigenin (DIG)
labelled F gene specific probe as described in materials and methods. (C) Southern blot of the agarose gel described in (A) stripped and
reprobed with a DIG labelled H gene specific probe.
(A) Agarose gel electrophoresis of measles virus Fusion (F) gene and haemagglutinin (H) gene reverse transcription polymerase
86 Uhlmann, Martin, Sheils, et al
inserts were then in vitro transcribed into cRNA using the
Riboprobe®in vitro transcription system (Promega, Madison,
Wisconsin, USA). Serial dilutions of this cRNA were used to
generate standard curves.
RT in situ PCR
RT in situ PCR facilitates low copy gene detection and permits
cellular localisation within tissues, with a reported sensitivity
of one viral genome copy/cell.8–10Sections were dewaxed in
xylene and taken through a series of graded alcohols. Endog-
enous avidin and biotin activity was blocked using the Dako
biotin blocking system (Dako, Glostrup, Denmark). Sections
were digested with proteinase K (300 µg/ml) for 17 minutes at
After pretreatment, MV RNA was amplified using the
following protocol: 58°C for 45 minutes and 94°C for five min-
utes, followed by 25 cycles of 94°C for 45 seconds, 60°C for 45
seconds,and 72°C for 45 seconds.After amplification,sections
were fixed in 100% ethanol and air dried.
Hybridisation was carried out with a 5′ biotinylated
oligonucleotide probe using previously published protocols.8
Hybrid detection was achieved using a three step immunocy-
amplification.12Alkaline phosphatase was detected with
nitrobluetetrazolium (NBT) and bromochloroindoylphos-
phate (BCIP) as chromogen. Endogenous alkaline phos-
phatase was blocked using levamisole, an endogenous AP
inhibitor (Dako), during chromogenic detection.
Reaction optimisation experiments were initially carried
out using formalin fixed, paraffin wax embedded, measles
infected Vero cells, and a variety of probe concentrations
(1 µg/ml, 1.5 µg/ml, 2 µg/ml). A concentration of 1 µg/ml
yielded optimal signals and was used in subsequent experi-
ments. Biopsies from 73 affected children and five normal
controls were examined.
Controls for RT in situ PCR included the following: MV
infected and uninfected Vero cells, a mixed population of
infected and uninfected Vero cells, MV N gene primers and an
irrelevant probe (nonsense pyruvate dehyrogenase probe),
and irrelevant primers and N gene specific probe. Hybridisa-
tion control experiments were performed using a histone
mRNA probe. Other control experiments included RNase
digestion of MV infected Vero cells before RT in situ PCR.
Combined RT in situ PCR and immunohistochemistry
To examine MV signal localisation, RT in situ PCR for the MV
N gene was performed as described above on tissue sections
following immunohistochemistry using the follicular den-
dritic cell CNA 42 monoclonal antibody13(Dako). For in situ
hybridisation a 5′ biotin labelled oligonucleotide probe was
used as above.
The MV hybridisation signal was developed with horserad-
ish peroxidase and aminoethyl carbazole (AEC;Vector Labora-
tories, Burlingame, California, USA), and the dendritic cell
AP (Dako) and NBT and BCIP (Roche Molecular Biochemi-
cals) as substrate.
Approval for these studies was obtained from the ethical prac-
tices committee of the Royal Free, Hampstead, NHS Trust.
Fully informed, written parental consent was obtained from
all trust patients including controls.
Overall, 75 of 91 affected children had MV RNA in their ileal
lymphoid tissue compared with five of 70 in the control
patient cohort (Fisher exact test, p < 0.0001; Analyse-it Soft-
ware, General 1.62).
A total of six different PCR primer sets were optimised by
solution phase RT-PCR to amplify the MV F, H, and N genes
from RNA extracted from MV infected Vero cells (fig 1).
The specificity of the primer/probe sets to detect MV F and
H genes was established using RNA extracted from: (1) cryo-
preserved ileal biopsy material from four affected children,(2)
SSPE brain, and (3) MV infected Vero cells (fig 2A). All four
patient samples were positive for MV F and H genes by
TaqMan RT-PCR. Amplicon specificity was confirmed by
Southern blot analysis using F and H gene specific probes (fig
2B,C). No template controls run in parallel were negative.
Seventy of 91 affected children were positive for MV
compared with four of 70 controls as analysed by TaqMan
RT-PCR (table 2). MV copy number in positive biopsies was
generally low, but ranged from 1 to 3 × 105copies of MV/ng of
total RNA. Of the paediatric control group, MV was not
detected in normal children or children with isolated ileal
LNH. However, four of 26 appendicectomy samples harboured
the MV genome (table 2).
Summary of TaqMan RT-PCR and RT in situ
In situ PCR
Mild non-specific changes
70 (91) 42 (57)
Total number of patients tested in parentheses.
NT not tested; RT-PCR, reverse transcription polymerase chain
probe for the detection of the measles virus Nucleocapsid (N) gene in infected Vero cells. Optimal cytoplasmic staining is achieved in (A)
(1 µg/ml), higher probe concentrations in panels (B) (1.5 µg/ml) and (C) (2 µg/ml) showed increasing non-specific nuclear staining.
Reverse transcription (RT) in situ polymerase chain reaction (PCR) experiment using a single stranded biotinylated oligonucleotide
Pathogenesis of new variant inflammatory bowel disease87
RT in situ PCR optimisation experiments were performed as
described (fig 3). A probe concentration of 1 µg/ml yielded
optimal signals and was used in subsequent experiments.
In MV infected Vero cells, MV amplicons were identified as
a cytoplasmic signal (fig 3). In SSPE brain material, discrete,
intense foci of MV amplicons were detected in grey matter
using RT in situ PCR (fig 4A).Signal was not detected in simi-
larly processed normal brain, or when irrelevant PCR primers
were used on sections of SSPE brain (fig 4B).
Of the 57 tissue biopsies from affected children, 42 were
positive for the MV N gene by RT in situ PCR and amplicons
could be detected in serial sections of ileal biopsies. Four sam-
ples were inconclusive for the presence of MV because of high
background staining,and 11 biopsies were negative for MV.Of
the control group, one of five children with histologically nor-
mal small and large bowel mucosa had detectable MV N gene
RNA, present in a distribution that was identical to that seen
in biopsies from affected children (table 2). MV amplicon
localised to reactive follicle centres, and was associated with
cells possessing dendritic processes (fig 4C,E) and some lym-
phocytes (fig 4F).No MV signal was seen in serial control sec-
tions where irrelevant PCR primers were used (fig 4D). Addi-
tional RT in situ PCR control experiments were performed on
measles infected Vero cells. No signal was obtained on MV
infected Vero cells after RNase digestion (fig 5).
Overall, 91 biopsies have been examined, 56 of which were
analysed by a combination of in cell RT-PCR (MV N gene) and
TaqMan RT-PCR. Thirty seven of these biopsies were positive
for MV and five were negative for MV using both techniques,
six were positive by TaqMan RT-PCR but negative by in situ
PCR, and five were positive by in situ PCR only (table 2).
We describe an association between persistent MV infection
and ileocolonic lymphonodular hyperplasia and ileocolitis in
children with developmental disorder.1The molecular data
indicate the presence of MV genomes in 75 of 91 affected chil-
dren with the disorder compared with five of 70 control chil-
dren. In addition, there appears to be a strong segregation of
Nucleocapsid (N) gene detection in subacute sclerosing panencephalitis brain tissue showing intense, discrete black signals within the brain
parenchyma. (B) Negative control section of (A) treated identically but using irrelevant PCR primers; no MV signal was seen (original
magnification, ×400). (C) MV N gene RT in situ PCR combined with in situ hybridisation in a reactive ileal lymphoid follicle centre from an
affected child. A positive signal radiates from an intense central core in an apparently beaded, fibrillary pattern (original magnification,
×1000). (D) RT in situ PCR on a serial section of (C) constituting a negative control (irrelevant PCR primers) (original magnification, ×1000). (E)
Combined RT in situ PCR for the MV N gene and immunohistochemistry with monoclonal antibody CNA 42, a follicular dendritic cell marker. A
single cell is shown, present in a reactive follicle centre from an affected child. Once again, the MV genomic signal (red) appears to exhibit a
central pattern that would be consistent with the cell body, whereas CNA 42 (blue/black) delineates dendritic processes radiating from the cell
body (enlarged from original magnification of ×1000). (F) RT in situ PCR shows a single mature lymphocyte infected with MV RNA in a
(A) Reverse transcription (RT) in situ polymerase chain reaction (PCR) combined with in situ hybridisation for measles virus (MV)
on MV infected Vero cells (A) with and (B) without an RNase digestion step. (A) No signal was seen when RNase digestion was performed
before in situ RT-PCR. (B) Shows a cytoplasmic signal in the absence of an RNase digestion step.
Reverse transcription (RT) in situ polymerase chain reaction (PCR) control experiment for measles virus (MV) Nucleocapsid (N) gene
88 Uhlmann, Martin, Sheils, et al
the disease phenotype with male children,in keeping with the
reported male predominance of the developmental disorder.
MV localisation was demonstrated in tissue biopsies by RT
in situ PCR and combined immunohistochemistry. The signal
had a fibrillary character, and appeared to associate with the
dendritic cell matrix confirmed by immunohistochemical
staining using a specific CNA 42 antibody (fig 4E).
MV was predominantly detected in dendritic cells in
reactive follicular hyperplastic centres in ileal biopsies from
affected children (fig 4C,E), but was also identified in mature
lymphocytes in these hyperplastic areas (fig 4F). This result
indicates a possible interaction between MV and the immune
response in the pathogenesis of ileocolitis in these children.
MV may be a potential “immunological trigger” in the patho-
genesis of lymphoid hyperplasia and ileocolitis. Potential
initiators for this type of immunological response are
dendritic cells. Dendritic cells capture and process viral
antigens in the periphery, express costimulatory molecules,
and serve as vehicles for viral antigens to the mediators of
immunity (B and T cells) in lymphoid tissue.As a result,cyto-
kines are released and initiate an immune response.14
MV localisation in follicular dendritic cells mirrors the
human immunodeficiency virus type 1 (HIV-1) infection pat-
terns seen in HIV-1 enteropathy. The presence of MV antigen
in follicular dendritic cells may reflect a transient stage in the
progression from latent to persistent MV infection.15–17The
hypothetical parallel with HIV infection is interesting: HIV,
like measles virus, potentially disrupts cellular immunity, and
induces follicular hyperplasia and lymphadenopathy in the
early stage of infection. This is associated with expansion of
the follicular dendritic cell network and trapping of HIV
within germinal centres.15–17During the early latent phase of
infection, HIV antigens are detectable upon the surface of fol-
licular dendritic cells15in a pattern similar to that reported
here for MV. Such a location may favour the induction of
immunological tolerance and failure of viral clearance. The
mechanisms by which MV, immunological abnormalities, and
chronic intestinal pathology may be linked are currently not
known. Precedents for delayed intestinal and immunological
sequelae to MV exposure include chronic immunodeficiency,
diarrhoeal disease, and death following early natural measles
exposure.18Natural measles infection induces an initial T
helper cell type 1 (Th1) response and protective cytotoxic
immunity (characterised by the classic measles rash and
gastrointestinal upset), followed by a prolonged Th2 response
with antibody production.19In most individuals, this leads to
lifelong immunity without delayed pathological sequelae.
Nonetheless, immune activation during the measles attack is
set against a profound reduction in non-specific cellular
immune responsiveness, reflecting the potent immunosup-
pressive properties of MV.Recent studies suggest that this may
be achieved through impaired interleukin 12 (IL-12) produc-
tion by infected dendritic cells,20and blocking of IL-2 receptor
α expression by activated T cells.21In such circumstances,
impaired Th1 immunity, with a shift towards a dominant Th2
response, might occasionally contribute not only to establish-
ing persistent infection,but also to delayed immunopathology.
“Measles virus may be a potential immunological trigger
in the pathogenesis of lymphoid hyperplasia and
MV was not detected in three children with isolated LNH or
in biopsies from normal children. Of the 26 children who
underwent apendicectomy,only four harboured MV RNA.The
presence of measles virus in the intestine of a small minority
of apparently healthy children is not surprising in light of the
finding of Warthin-Finkeldy giant cells in inflamed appendi-
ces in children with measles infection. However, the preva-
lence of persistent MV infection in the general population is
not known and warrants further investigation.
These preliminary studies have focused principally on MV.
We have not excluded the presence of alternative infections.
Viruses may persist elsewhere, or exert a transient effect not
requiring subsequent persistence.One such transient risk may
be that concurrent exposure to measles and another infection
may increase the risk of MV establishing persistent infection:
this atypical pattern of exposure has been identified as a risk
factor for both SSPE (chicken pox and encephalitogenic
Our study raises many questions—most importantly, does
MV play an aetiological role in intestinal inflammation in
association between MV infection and ileocolonic lymphon-
odular hyperplasia and ileocolitis in children with develop-
This work was supported by the John Ellerman Foundation, Welton
Foundation, Medical Interventions for Autism, and Visceral.
V Uhlmann, L Pilkington, I Silva, A Killalea, J J O’Leary, Department
of Pathology, Coombe Women’s Hospital, Dublin 8, Ireland
C M Martin, O Sheils, Department of Histopathology, Trinity College,
S B Murch, Department of Paediatric Gastroenterology, the Royal Free
Hospital, London, UK
J Walker-Smith, M Thomson, A J Wakefield, Department of
Medicine, The Royal Free Hospital and University College Medical
School, London, UK
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Take home messages
• Of 91 patients with a histologically confirmed diagnosis of
ileal lymphonodular hyperplasia and enterocolitis, 75 were
positive for measles virus in their intestinal tissue compared
with five of 70 controls
• Measles virus was found within the follicular dendritic cells
and some lymphocytes in foci of reactive follicular
• These data confirm an association between the presence of
measlesvirus and gut
Pathogenesis of new variant inflammatory bowel disease89
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23 Detels R, Brody JA, McNew J, et al. Further epidemiological studies
of subacute sclerosing panencephalitis. Lancet 1973;2:11–14.
What turns neutrophils on
more responsive.Understanding how immune complexes and neutrophils interact,leading to production
of tissue damaging enzymes and oxidants, and the molecular control of the interactions are potential
keys to new therapeutic treatments.
Fossati et al used in vitro chemiluminescence to detect production of oxidants by washed blood
neutrophils in their unprimed state or primed with granulocyte-macrophage colony stimulating
factor—a cytokine in synovial fluid in rheumatoid arthritis—when incubated with synthetic insoluble or
soluble immune complexes. By comparing the reaction kinetics of chemiluminescence with substrates
available to the neutrophils either extracellularly or intracellularly and with different specific scavengers
of oxidants, they were able to unravel the processes.
Insoluble complexes activated unprimed or primed neutrophils to produce oxidants that remained
intracellular whereas soluble complexes reacted only with primed neutrophils but did so rapidly and
transiently,the oxidants being secreted.Insoluble complexes also required FcyRIIIb but not FcyRII func-
tion whereas soluble complexes required both. In tests to detect granule enzymes insoluble complexes
activated secretion with unprimed or primed neutrophils, although the reaction kinetics differed from
those for the oxidants. Soluble complexes activated secretion only with primed neutrophils, leading the
researchers to conclude that neutrophil activation occurs by different pathways for insoluble or soluble
immune complexes and can be significantly affected by cytokines.
m Annals of the Rheumatic Diseases 2002;61:13–19.
nsoluble and soluble immune complexes in synovial fluid in rheumatoid arthritis activate neutrophils
differently,researchers in Liverpool,UK,have found.Immune complexes and neutrophils are abundant
in synovial fluid in rheumatoid arthritis, as are cytokines, which can prime neutrophils making them
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