Medical Journal of Babylon - 2004 Volume - 1 - no.3 and 4
‫ – ا ﻠﺪ اﻷول – اﻟﻌﺪد اﻟﺜﺎﻟﺚ واﻟﺮاﺑﻊ‬٢٠٠٤ ‫ﳎﻠﺔ ﺑﺎﺑﻞ اﻟﻄﺒﻴﺔ‬
Immunology of E. coli Persistant pyuria
Ibrahim M.S. Shnawa* And **Baha H.H. Al Amidi
*Babylon University , College of Science Biology Department Hilla P.O. box. 4 Iraq.
And **Public Health Laboratory/Hilla
MJB
Abstract
Fourty Persistent Pyuria Patients were diagnosed as E. coli persistant pyuria . Of which 33 were
Immune (In a sense of mounting normal adapted immune response) and seven were
immunocompromised (In a sense of mounting subnormal adapted immune response .The female / Male
ratio was 13/27 . The patients age range was 30-70 years .The median total serum protein was 7.274
g/dl . Likewise the median serum total globulin was 4.302 g/dl . while the median of systemic to the
median of mucosal globulin ratio was 0.69 . These scores were generally higher than those of normal
control subjects . The median immunoglobulin class concentrations were 2.339, 0.195 and 0.309 g/dl to
classes IgG , IgM and IgA respectively .
E. coli immunodominant epitopes were intiating specific circulating and mucosal agglutinin as well as
significant
leucocytes
inhibition
factors
in
immune
patients
and
nonsignificant
in
immunocompromized .
Thus, E. coli epitopes could be of T dependent type and activate either or both Th1 and Th2
lymphocyte, or it might be of T independent epitope type .
‫اﻟﺧﻼﺻﺔ‬
. ‫ ﻣـﻧﻬم ﻣﻧﯾﻌـون وﺳـﺑﻊ ﻣﻧﺧﻔﺿـﻲ اﻟﻣﻧﺎﻋـﺔ‬٣٣ ‫ ﻛـﺎن‬. ‫ﺟرى ﺗﺷﺧﯾص ارﺑﻌون ﻣرﯾﺿـﺎ ﺑﺑﯾﻠـﺔ ﻗﯾﺣﯾـﺔ ﻣﺳـﺗدﯾﻣﺔ ﻣﺷـﺗرﻛﺔ ﻣـﻊ اﺷـرﻛﯾﺎ اﻟﻘوﻟـون‬
،٢٧٤‫ ﻛ ــﺎن وﺳ ــﯾط ﺗرﻛﯾ ــز اﻟﺑ ــروﺗﯾن اﻟﻛﻠ ــﻲ اﻟﻣﺻ ــﻠﻲ‬. ‫ ﺳ ــﻧﺔ‬٧٠-٣٠ ‫ واﻟﻔﺋ ــﺔ اﻟﻌﻣرﯾ ــﺔ ﺑﻣ ــدى‬٢٧ ‫ اﻟ ــﻰ‬١٣ ‫ﻛﺎﻧ ــت ﻧﺳ ــﺑﺔ اﻻﻧ ــﺎث ﻟﻠ ــذﻛور‬
‫ وﺗﻌـد ﻫـذﻩ‬٠,٧٢ ‫ ووﺳـﯾط ﺗرﻛﯾـز اﻟﻛﻠوﺑﯾـوﻟﯾن ﻓـﻲ اﻟﻣﺧـﺎط ﻫـﻲ‬٤،٣٠٢ ‫ ووﺳﯾط ﺗرﻛﯾز اﻟﻛﻠوﺑﯾوﻟﯾن اﻟﻣﻧﺎﻋﻲ اﻟﻛﻠﻲ اﻟﻣﺻﻠﻲ‬. ‫دﻟﺗر‬/‫ﻏم‬٧
. ‫اﻟﻘﯾم ﺑﺷﻛل ﻋﺎم اﻋﻠﻰ ﻣﻣﺎ ﻫو ﻋﻠﯾﻪ ﺑﯾن اﻷﺷﺧﺎص اﻷﺳوﯾﺎء ﻓﻲ ﻣﺟﻣوﻋﺔ اﻟﺳﯾطرة‬
. ‫دﻟﺗر‬/‫ ﻏم‬٠,٣٠٩ ‫ و‬٠،١٩٥;٢،٣٣٩‫ ﻋﻠﻰ اﻟﺗواﻟﻲ‬IgA,IgM,IgG ‫وظﻬر ﺑﺎن وﺳﯾط ﺗرﻛﯾز أﺻﻧﺎف اﻟﻛﻠوﺑﯾوﻟﯾن اﻟﻣﻧﺎﻋﻲ‬
٣٠٠
‫ – ا ﻠﺪ اﻷول – اﻟﻌﺪد اﻟﺜﺎﻟﺚ واﻟﺮاﺑﻊ‬٢٠٠٤ ‫ﳎﻠﺔ ﺑﺎﺑﻞ اﻟﻄﺒﻴﺔ‬
Medical Journal of Babylon - 2004 Volume - 1 - no.3 and 4
‫وﯾﺑدو ﺑﺎن اﻟذرى اﻟﻣﺳﺗﺿدﯾﺔ ذات اﻟﺳـﯾﺎدة اﻟﻣﻧﺎﻋﯾـﺔ ﻗـد ﺣﻔـزت ﺣﺻـول اﺿـداد ﺗﻼزﯾﻧﯾـﺔ ﻣﺗﺧﺻﺻـﺔ دوارة وﻣﺧﺎطﯾـﺔ ﻣﺗﺧﺻﺻـﻪ ﺑﺎﺷـرﻛﯾﺎ‬
‫ وﻗـد أﺣـدث ﻣﻌـﺎﻣﻼت ﺗﺛﺑـﯾط‬. ‫اﻟﻘوﻟون وﺑﻌﯾﺎرات ﻣﻌﻧوﯾﺔ ﻣن اﻟﻣﺟﻣوﻋﺔ اﻷوﻟـﻰ ﻣـن اﻟﻣـرض وﺑﺷـﻛل اﻗـل ﺑﺎﻟﻣﺟﻣوﻋـﺔ اﻟﺛﺎﻧﯾـﺔ ﻣـن اﻟﻣرﺿـﻰ‬
‫اﻟﻬﺟرة اﻟﺧﻼﯾﺎ اﻟﺑﯾض ﻣﻌﻧوﯾﺔ وﻏﯾر ﻣﻌﻧوﯾﺔ ﻋﻠﻰ اﻟﺗواﻟﻲ ﻟﻣﺟﺎﻣﯾﻊ اﻟﻣرﺿﻰ ﻋﻠﻰ اﻟﺗواﻟﻲ وﺑﻬذا ﻓﻣن اﻟﻣﻣﻛن إن ﺗﻛون اﻟـذرى اﻟﻣﺳﺗﺿـدﯾﺔ‬
. ‫ أو ﻣﻌﺗﻣدة وﻏﯾر ﻣﻌﺗﻣدة ﻋﻠﻰ اﻟﺧﻼﯾﺎ اﻟﺗﺎﺋﯾﺔ‬. Th1 ,Th2 ‫اﻟﺳﺎﺋدة ﻣﻧﺎﻋﯾﺎ ﺗﻧﺷط ﺧﻼﯾﺎ ﺗﺎﺋﯾﺔ ﻣﺳﺎﻋدة‬
Introduction
The
and
The
immunology of uropathogenic E. coli
processed
allover the world have been tackled by
quadrate streaking technique [23] as
several workers [ 1,2,3,4 ] . In this
well as by indirect broth enrichment
area
,
pathogensis
however
urine
for
samples
culture
were
by
direct
,
among
the
and quadrate streaking technique [24] .
coli
has
been
Growth on blood and MacConkey agar
documented [5,6] Mahdi [7] has been
plates were checked for metallic sheen
studied
muucosal
on Eosin methylene blue agar . Pure
immune responses in persistant pyuria
isolates were identified by classical
patients .
and miniaturized biochemical tests
uropathogens
E
E.
coli
specific
The objective of the present
work
was
immunology
to
investigate
of
E.
coli
[25] . Whole cell somatic E. coli
the
antigens were prepared by heat killing
human
at 60 C ْ ◌ for one hour at waterbath
persistant pyuria patients .
[26] . Cell free culture filtrate was
Materials And Methods
made from 24 broth culture after
Fourty persistant pyuria patient were
centerfugation for 10 min. at 5000
investigated . From each of which
RPM . Supernates were Millipore
clean catch midstream urine & blood
(0.22
with and without anticoagulant were
unite
collected [8, 9] . The normal human
preparation was used as sensitizer in
subjects were sampled as for test
leukocyte
group.
leukocyte inhibition test was done by
The
bacteriology
and
) filtered in microfiltration
of
injection
type
inhibition
The
capillary
performed
standardized
blood leukocyte and mucosal leukocyte
and were
[9 , 29] . The immunoglobulin classes
methodology [23 – 34]
briefed in the followings :
were
determined
immunoglobulin
٣٠١
using
.
This
immunology of these samples were
by
method
test
.
peripheral
by specific
partigen
anti
plates
‫ – ا ﻠﺪ اﻷول – اﻟﻌﺪد اﻟﺜﺎﻟﺚ واﻟﺮاﺑﻊ‬٢٠٠٤ ‫ﳎﻠﺔ ﺑﺎﺑﻞ اﻟﻄﺒﻴﺔ‬
Medical Journal of Babylon - 2004 Volume - 1 - no.3 and 4
according to Mancini and Laurel [30] .
to 1.197 , 0.295 and 0.294 g/dl for
The
IgG , IgM and IgA respectively in
separation
of
mucosal
immunoglobulin (MIG) was done by
controls
.The
6% PEG 6000 [31] . MIG was partially
circulating
and
characterized as ; dialysis for three
titres , were 360 and 40 respectively .
days using saline for two shifts and
The ratio of systemic to mucosal
distilled water for one shift [32] ,
antibody titre was 9:1 for the immune
protein
patient
determination
by
biurate
group
E.
coli
mucosal
and
specific
agglntinin
6:1
for
the
method [33] , slide agglutination ,
immunocompromised group . There
standard tube agglutination with and
was a linear correlation between MIG
without 2ME treatment using heat
concentrations and the E. coli specific
killed somatic antigen of the causal E.
antibody titers, as simple regression
coli bacteria [34] statistical analysis
analysis showed :
was made as in Dawed and Al-
Serum
Y=-1596.7 + 45.012 X
Yas[10] .
Y=0.6263
Results
Fc=24.533 at p=0.01
From these fourty patients .Thirty three
Ft= 7.35 at p=0.01
were
Mucosa Y=-8.828 + 65.715 X
immune
and
seven
were
immunocompromized . E. coli were
r = 0.61497
identified from the urine cultures of
Fc = 23.11274 at p=0.01
these patients (table .1 ) .The female /
Ft=7.35 at p=0.01
male ratio 13/27 . The age range of
The leucocyte inhibition factors in
these patients were from 30 to 70
peripheral
years .
leucocytes were 0.57 , 0.5 for the
The
median
total
serum
blood
and
mucosal
protein
immune and 0.85 as well as 0.77 for
concentrations were 7.274 g/dl while
immunocompromy patient respective-
that for total globulin concentrations
ely . In other word it was significant in
was 4.302 g/dl . The median mucosal
immune patient and non signification
globulin concentrations were 0.072
in the other group .
g/dl . The albumin to globulin ratio
Discussion
was 0.69 . The median concentrations
Uropathogenic E. coli was documented
for
( table1,2,Fig 1 ) the pathogenesis in
immunoglobulin
concentrations
were
class
2.339,0.195
,
these
0.309 g/dl in patient which corresponds
cases
exogenous
٣٠٢
can
and
be
ascending
descending
‫ – ا ﻠﺪ اﻷول – اﻟﻌﺪد اﻟﺜﺎﻟﺚ واﻟﺮاﺑﻊ‬٢٠٠٤ ‫ﳎﻠﺔ ﺑﺎﺑﻞ اﻟﻄﺒﻴﺔ‬
Medical Journal of Babylon - 2004 Volume - 1 - no.3 and 4
haematogenouse
and
/
or
lymphogenous [11-13] . The
net
2-
Mulvey
,
M.A.;Schilling
,
J.D.;Martinez , J.J. and Hultgren , S.J.
outcome of host – parasite interaction
P.N.A.S , 2000, 97(16):8747.
depends on balance versus imbalance
3-
between the pathogen and the
Hultgren
host.
Shilling , J.D.;Mulve , M.and
,
S.J.
Urology
Thus the persistant E. coli infection
2001( supplement 6A) .57:56.
may be due to antibody
4-
coated
.
Garica,M.I.;Joure,M.;Nataro
,
bacteria , cryptic organisms and / or
J.P.;Gouna,P.Bouguenec,C.L.;
cell well defective state [6,2,3,13]
letters .2000. 111.
The virulence associated antigens of
5-
uropathogenic E cloli are Exo and
1996.37(1):29.
Endo-toxin , adhesion factors , K
6-
antigen [1,15,16]. In these, uropathic
Babylon University .
E. coli the immunodominant epitopes
7-
could be of a T – dependent and T
abylon university . Babylon .
independent types or they may be of
8-
activation potential to Th1 and Th2
Diagnosis of Infectious Diseases .
cells [17] In uropathy , howover , E.
2Dnded
coli epitopes may be K,O and / or F
Publishing Co,I.N.C.
components . To date knowledge has
9-
indicated that one or more of these
1980 .Selecte
antigens may be protective and helpful
immunology
in diagnosis , Thus E. coli in these
company . san franscico .
persistant pyuria cases , stimulated
10- Dawed
mucosal and systemic response at both
1990
humeral and cellular levels , since they
agricultural
induced specific antibody response and
University – Iraq .
significant LIF both at mucosal and
11- Hooton
systemic levels
Chemo2000.45 (supplement):1.
[13,18-22] .
12- Uelling,D.T.;Johnson,D.B.;Hopki
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‫ – ا ﻠﺪ اﻷول – اﻟﻌﺪد اﻟﺜﺎﻟﺚ واﻟﺮاﺑﻊ‬٢٠٠٤ ‫ﳎﻠﺔ ﺑﺎﺑﻞ اﻟﻄﺒﻴﺔ‬
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‫ – ا ﻠﺪ اﻷول – اﻟﻌﺪد اﻟﺜﺎﻟﺚ واﻟﺮاﺑﻊ‬٢٠٠٤ ‫ﳎﻠﺔ ﺑﺎﺑﻞ اﻟﻄﺒﻴﺔ‬
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Table 1: Biochemical charcteriztion of uropathic E. coli
Gram reaction
_
Shape
Rod
Rod
M
M
Motility
Facult.O2
Growth
agar
_
on
MacConkey
Lactose fermenting colonies
Capsule
-
-
Catalase
-
-
Oxidase
_
-
Urease
(+)
Indole
+
+
MR
+
+
VP
_
-
Citrate
-
-
Lactase
+
+
Glucose
+
+
٣٠٥
‫ – ا ﻠﺪ اﻷول – اﻟﻌﺪد اﻟﺜﺎﻟﺚ واﻟﺮاﺑﻊ‬٢٠٠٤ ‫ﳎﻠﺔ ﺑﺎﺑﻞ اﻟﻄﺒﻴﺔ‬
Medical Journal of Babylon - 2004 Volume - 1 - no.3 and 4
Table 2: Immunology of E. coli human persistant pyuria in an immune and immuno
compromised patients
Character
E. coli
control
F/M
13/27
3/5
___________________________________________________
Age range
1
1-20
1
2
21-29
17
3
30-39
16
3
40-49
1
3
50-59
4
60-69
___________________________________________________
median of
total serum protein g/dl
7.274
7.027
total serum globulin g/dl
4.302
3.492
mucosal globulin
g/dl
0.72
0.02
S/M globulins
5.945
17.46
serum alb /serum globulin
0.69
0.101
median IgG g/dl
IgM g/dl
IgA g/dl
median of serum specific
agglutinin. Titre
median of mucosal specific.
Agglutinin .
Median of serum specific.
Agglutinin .Titre/comp.
Median of mucosal specific
Agglutinin .Titre/comp
systemic LIF
systemic LIF Com
mucosal LIF
mucosal LIF Comp*
* Comp = immunocompromised patient
2.339
0.145
0.305
1.197
0.245
0.294
360
-
40
-
240
-
40
-
0.57
0.85
0.5
0.77
٣٠٦
-
‫ – ا ﻠﺪ اﻷول – اﻟﻌﺪد اﻟﺜﺎﻟﺚ واﻟﺮاﺑﻊ‬٢٠٠٤ ‫ﳎﻠﺔ ﺑﺎﺑﻞ اﻟﻄﺒﻴﺔ‬
Medical Journal of Babylon - 2004 Volume - 1 - no.3 and 4
Mucosal antibody
titer
80
60
y = -8.828+65.715x
r = 0.6149
40
20
0
0.00
0.20
0.40
0.60
0.80
1.00
Table (1) Th
A
800
Mucosal anti body concentration g/dL
600
Systemic Anti body titer
400
y = -159607+45.012x
r = 0.6263
200
B
0
0.00
10.00
20.00
30.00
٣٠٧
40.00
50.00