Vol.3, No.3, 159-161 (2011) Health
Copyright © 2011 SciRes. O penly acce ssi ble at http://www.scirp.org/journal/HEALTH/
Mycobacterium tuberculosis in the aort a of a p atient with
takayasu’s arteritis. extra pulmonary tuberculosis
María Elena Soto, Virgilia Soto, Julia Isela Martín Sandría, Ricardo Gamboa, Claudia Huesca
Immunology and Physiol og y Departa ment, Juan Badi a n o #1 Col onia Sec c i ón XVI , Tlalpan México, D. F, México.,
*Correspondi ng Author: email@example.com;
Received 15 October 2010; revised 2 December 2010; accepted 11 January 2011.
AB S T RAC T
Takayasu’s arteritis (TA), of unknown etiology,
has been related with Mycobacterium tubercu-
losis infection. The published cases are con-
troversial. A 29- year-old man died, prolapse of
the aortic valve and unspecific arteritis were
demonstrated at necropsy. Our objective was to
search in the fresh aortic tissues, the infectious
agent of tuber cul osi s through c ultur e, as well as
by performing histopathology and searching by
the molecular study, sequences of genes asso-
ciated to mycobacterium and Ziehl’s stain. Re-
sults. The bacillus of tuberculosis was observed
by the Ziehl’s stain and the sequenc es of inser-
tion IS6110 gene were identified. Our findings
support the relation of TA with possible tuber-
culosis infection.Conclusion. The possibility
that Takayasu’s arteritis is an extra pulmonary
tuberculosis must be explored.
Ke yw o r d s: Takayasu’s arteritis, extrapulmonary
tuberculosis, IS6110 and HupB sequences.
Takayasu’s arteritis (TA) is a chronic inflammatory
disease affecting the large arteries and their branches .
Patholog y is localized in the adventitia, accompanied by
mononuclear infiltration and undefined granulomas; the
intimae is usually normal but becomes altered in time due
to atherosclerosis .
TA has been related to Mycobacterium tuberculosis
infection, arterial damage depicts granulomatous lesions
similar to those of tissues with tuberculosis [3,4] and
hyperreactivity to tuberculin application has been docu-
mented [5,6,7,8].The published cases are controversial
We describe an autopsy case, in which unspecific ar-
teritis was found. We searched for tuberculous i nfect ion.
2. DESCRIPTION OF THE CASE
A 29-year-old man, without previous symptoms of tu-
berculosis or symptoms relevant to the current illness. No
relativ e wi th tu berculosis w as informed du ring questioning;
no BCG vaccination had been applied to the patient. Three
years before, the patient had referred pain in both legs
when exerting physically and he had occasional headach es.
High blood pressure had been detected in three previo us
occasions, but he received no antihypertensive treatment.
During indirect questioning, relatives informed that, an
hour before his admittance to the hospital, he had com-
plained of an intense oppressive pain in the chest and
upper abdomen, pain irradiating to both arms and neck,
with difficulty to breath, generalized pulsed headache,
and hemoptysis, he seemed disoriented.
Physical exploration revealed a hypertensive crisis,
blood pressure of 200/120 mmHg, diaphoresis, we found
it confused in time space and person; neck with yugular
ingurgita tion grade I and murmur in aortic focus the left
supraclavicular region, pulseless in the left arm. He pre-
sented cardiorespiratory failure half an hour after his
admittance, which did not revert with maneuvers of re-
suscitation. Autopsy was performed by an expert pa-
thologist, finding histopathological data of Takayasu’s
The case was commented with the rheumatologist, wh o
analyzed the post-mortem clinical file. Demographic data
recorded at his arrival at the emergency ward were: age
below 40 years, absence of pulse, aortic murmur irra-
diated to the neck, history of claudication in the lower
extremities, these data, provides four clinical criteria, for
classification of TA and the autopsy findings which are
the gold standard for Takayasu’s ar te r itis diagnosis.
Tissue samples were taken for DN A studies, searching
for sequences of the IS6110 gene associated to Myco-
bacterium tuberculosis. Samples were stains with hema-
toxylin-eosin, Schiff (periodic acid, PAS), Mason’s
trichrome, auramine-rhodamine, and Ziehl-Neelsen.
Ziehl-Neelsen stained and microscopically analyzed.
M. E. Soto et al. / Health 3 (2011) 159-161
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3. MATERIAL AND METHODS
Autopsy was performed. For the microscopic study,
3-µm histological sections of the tissue fixed in 10%
formaldehyde and embedded in paraf fin were used. Stains
used were, auramine-rhodamine, and Ziehl-Neelsen.
Tissue was embedded in paraffin, and extracted with
octane and 10% alcohol. For paraffin embedding, the Kit
Illustra Nucleon BACC Genomic was used. DNA was
extracted using a commercial kit (Illustra Nucleón Ge-
nomic, GE Healthcare, UK) following manufacturer’s
instructions. Quantification was performed and readings
were made in a spectrophotometer at 260/280 nm.
For DNA amplification of the 123 bp (IS6110) inser-
tion element, the following oligonucleotides were used:
IS6110 f (5´ CCT GCG AGC GTA GGC GTC GG-3´)
and IS6110 r (5’ CTC GT C CAG CGC CGC TT C GG-3’).
Briefly, a 25 µL reaction was set up containing 9.3 µL of
double distilled H2O, 2.5 µL of 10X buffer, 1.5 µL of 25
mM M gCl2, 25 µM (each) of the four deoxyribonucleo-
side triphosphate (Epicentre Biotechnologies Company,
USA), 2.5U Taq Polymerase (Invitrogen), forward and
reverse primer at final concentration of 10 pmol and 2.5
µL of DNA sample (400-500 ng). The following ampli-
fication cycle was used for PCR: five min 94˚C, 35
cycles each of 30 seconds at 94˚C, 30 seconds at 60˚C
and 72˚C for one min, followed by one cycle of 72˚C for
seve n mi n. We used a p osit ive co ntro l for the ana lysi s o f
mycobacterium (M. tuberculosis H37Rv) obtained from
cell lines extracted from strains of M. t uber culosis HRv37.
Macroscopic findings were: 1500 ml of blood were
found in the left pleural cavity, prolapse of the aortic
valve, hypertro ph y of the left ventricular wall. T he ao rta,
from its origin until the start of the iliac arteries, pre-
sented increased thickness and irregularity of the wall,
with some atheroma plaques, in the thoracic aorta, a
broken saccular aneurysm was found, measuring 11 × 8
cm in the major axis. Chronic granulomatous inflamma-
tion with central necrosi s was fo und in severa l segme nts
of the aorta (Figure 1). Other regions presented fibrosis
with cicatrizatio n alter ation s in the middle -vascular layer
(Figure 2(a)); acid-alcohol resistant bacilli were found
in these areas with the Ziehl-Neelsen stain ( Fi gu re 2(b)).
A blood sample was cultured in search of Mycobacte-
rium tuberculosis, which was positive.
The molecular study identified mycobacterium and the
123 bp that identifies Mycobacterium tuberculosis. Fig-
Takayasu’s arteritis is generally found in women, it is
Fi g ure 1 . Depict the autopsy specimen constituted by the heart,
the aorta, and the kidneys. Prolapse of the aortic valve, thick-
ening and irregularity of the aorta in its diverse segments, with
a noticeable constriction of the renal arteries are o bserved . The
right side depicts a close-up of the aorta and of the renal arte-
ries with evident thickening of their walls.
Figure 2. (a) It cuts to histological stained with hematoxylin
and eosin in which granulomatous chronic inflammation with
central n ecrosi s in th e thickness of the art ery is o bserved ao rta.
(b) In the black arrow can be seen an acid bacillus resistant
alcohol with the stained of Zielh-Neelsen.
M. E. Soto et al. / Health 3 (2011) 159-161
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MOLECULAR 1 2 3
Figure 3. The nitrate silver stained amplification
product of M. tuberculosis was electrophoresed on
nondenaturing 10% polyacrilamide gel. The 123
product obtained for M. tuberculosis is indicated by
the arrow. Lanes: molecu l ar si ze mar ker; 1: DN A of ba-
cillus of mycobacterium in the aorta of c as e ; 2 and 3: M.
tuberculosis DNA H37Rv taken from ce l lula r cultur e .
rare in men; the relation in Mexico is of 6:1 . Al-
though rare in men, the arterial damage progression is
similar. Mycobacterium tuberculosis induces a dominant
cellular immune response, similar to that observed in
extra- pulmonary tuberculosis. It is feasible that, in TA,
arteries are the target organs, which has been demon-
strated when the pulmonary arteries are reached by the
right blood c ir c ulation .
For more than five decades, TA has been related with
tuberculosis infection, this is still controversial because
findings differ; in isolated cases coincidence has been
reported . Recently, a prospective study pointed out
that there is no evidence for the association with Myco-
bacterium tuberculosis in tissues of injured arteries ob-
tained by biopsy .
This case presents a man without a previous history of
either arteritis o r tuberculosis; the repor ted clinical signs
and symptoms were compatible with TA, although no
imaging study was performed, the histopathological study
was made, and the latter is the gold standard for the di-
agnosis of TA.
The diagnostic suspicion and its correct classification
are necessary as a relevant initial methodology to study
the re la tion of TA with infection.
On the other side, the search for M. tuberculosis
through culture, specific stains, and modern molecular
biology techniques, which have shown diagnostic use-
fulness, could provide more reliable results for this asso-
This case shows a relation of Takayasu’s arter itis with
infection b y Mycobacterium tuberculosis, and one o f the
diverse forms of initial clinical manifestation, in the Ta-
kayasu’s arteritis. These findings must be evaluated by
means of systematic studies that would allow the repro-
ducibility o f the similar results under the same consensus
of search, with molecular techniques used in the tax-
onomic studies of complex M. tuberculosis.
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