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Title : 1 (FPT)
Author(s) ,
Citation 5(3), p.115-128, 1963
Issue Date 1963-09-23
URL http://hdl.handle.net/10069/3911
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53: 115-128196311 115
1(FPT)
(:)
~~~~~----I---=~~~~~~~ ~~~~~-------~ I ~~~~ ~--~~~--I-~.--I------- .____ __
Studies on Intradermal Test for Filariasis. 1. Specificity of a peptide antigen (FPT)
prepared from Dirofilaria immitis for human filariasis. Osamu YOSHIMURA Clinical Department,
Research Institute of Endemics, Nagasaki University. (Director : Prof. Dr. D. KATAMINE)l---
.
,
,,
.
,
,
.
,(Dirofilaria
immitis,(Setaria equina,
(Setaria digitata,(Litomooides carinii
Wuchereria bancrofti,
(),(
,,
.
Taliafero. Hoffman (1930)
, Fairley (1932),(1940, -
(1942,(1943), Bozicevich (1944),
Wright (1944), Zarrow (1946),o
,1952)
,(1959)
.
I,
o(1959),(1962
.
,
,Screening test
,
.
,(1959
,
,.
2.
&).
(8
,
.
,
,
.,7O
NJIO HCl, magnetic stirrer48
,o(3,000, 1O
, pH 7.0.
,.
,3%25
magnetic stirrex-2,
.1/1o
,
.,-
43
116
,
1/100,
100IOmg (Tablel)
Table
lsolationandPurihcatioofAntigen
ms(maendfemae)Washedwthyso(cRsneseveratmesJCroonwssueJdetoandTatd(repeatedsameprocedure)JDicdavacuumdescatorJRxtRCdwiN/TiCatroomtemperaturefor48&rs.undstirCefed(3,000r.p.m.1Qmn.)ISLpernatantPrecipttedstedtop7.0Centrfegmh-_JSitpernantPrecipitateAddedsaturatedpccacidoutandstoodatoomtempeatueovernightCentrifisedL
iStipetPretateJddedandextractedwith3^HCI-AScobofo2hrsunderstTTi
Centrifuifed
JSLPeatariFreepitatConcentratedoi/1OvoumeuderreducedpressureiecnaddedlRep
ifuged/sevee-epsocedureTJJ5upnatanPrecittateJ
lTTa_
h)
,-J--
70.1
IIH
.l
J
q)
U
.
.
lV UU UV
^:ale lpgth(m)
F1 Ultraviolet absorption curve ofFPT antigen.
3.
a)
,
.
3mm
.1-2
,
.
.,
.
b)
198
0.01ml,2.or, i.or,
o.5r, o.ir4
L5.
Fig.2, i.or
of the antigen in same someples.
86m
,
,, i.or
2.Or.
o.sr, o.i
,34mm
. (Fig. 2Fig. 3)
o.1o21 o4 Mf(+]
31oMf(+)
4o oMr(+;51ooMr(+;6.o
7o-- ll
BhealthyRemarks : The inner circle represents
wheal, the outside one erythemaand black spots disappearance ofwheal respectively.
Fig, 3Responses of skin against thedifferent dilution of FPT antigen.
r
)
6
i.or5, lo
15, 20, 25, 30, 40, 50, 6O120
12,
.
,,
(), 2
118
10, 315, 120D
, 225, 330, 1
6o,
120,
,
, 25
10, 41520
1520,
o), 2
e, 0.01ml
1Or, 15 (Fig. 4)
dq)
ir/(J
o
L
q)
(
~
11
1
=
q,,
U1
LI
U
ed
>
L,1
()
-
O
1.
d
J
Ii
.
.
mm
lo
5
mm
4o
3o
2o
o
n510 15202530405060 120 12
1rS.
5 10 1520 25 30 4050 60 1212-rs.
Progress of the reactionn thecourse of time.
R)
146,
123,269
541
,15
Fig. 5
74mm5mm
21)
nJ
l!
J
1o
VA
410152025
Dianl=t-r1('ll
Fig. 5Distribution curves of increase of
wheal diameter by FPT antigen
,I5mm
,
339
-,
,
9mm
d
0 10203040506070o90
mmaKM- OfTltN11a
Fig. 6Distribution curves of the diameter
of erythema by FPT antigen
I,
.
3-5mm
,
,. 6mm
.,
2mm, 3-5mm, 6mm
,8.5#
,82.4^, 14.8#
.
3mm4mm
4mm,
94.8#,2.e,
. (Fig. 5)
,
,, 20mm
9.3#,
6.7# (Fig. 6)
dQ)~40L-dJ
Il!r,30d2oeJI1
119
b)
6oO
o
,
Omm10#, 1mm26#, 2mm44#,
3mm87#, 4mm93%,
, 7mm
.
,
,Fig- 7
.,
,99%
.2mm,4mm
,
94.0#,91.
%.,
.
(Fig. 7)
o4
Increase rl the diameter of wheal1015-20.25 mm
Fig. 7Correlation between the increase of diameter of wheal and
erythema by FPT antigen.
)
,,,,
28,2,
,,
5,35, 15
4mm.
.
-,4mm
1,
3mm
.,o)
. (Table 2)
120
Tble2
f7ijfl'ience of other helmnthsnhabited and allergic diseases upon
the i:radermal rection by FPT agen ithe non-edemiceas.
Groups
Ascarlasis
Ancylostomiasis
As. +An,
Paragommiasis
Gnathostomiasis
Contact dermatitis
Asthma bronchlale
Congelation urticaria
Penicillin allergy 1
Increase of wheal diameter
(Measurement in mm)
456-
11
11
2
d)
()
, 14
0),
(Table 3J
2,796
337(12.1-#),
JR459(16.4#)
,1780, 639%
, 319(94.8#),
417(90.8#),,
,23371363(58#)
,,
o
0),
8, 1876115(6.1
)172C9.2#)
911(48.3#)o
,l(556)L
13.8#,6.3#,
(179)T.8#, 68.2#
ao)(62)6.5
Total
4
15
3
2
4
%>54.8#,(127) 5.5#, 43.3^,[[J
(410) 2.9#, 37J(280) 0.4#,
J3.2(142)
,,
92#, 49.3^s
(120),
, 27.5^
6920, 222(24.1
), 269(29.2^), 869
94.5^
,(108)333
%,100#,(100)
28O%, 100#,(168) 23.2^, 929#,
(291) 22.4#, 93. (187)
21.9#, 89.3^,(66) 19,7%, 98 9/
,
,
2o%
a)4-6mm (+), 7-9mm(). 10mm
(W)3,0)
121
Table3
Correlation between poste rate of theradermalest and
incifff.nr.p. of mc.rnfiln.Tifi nfirrip.rx a.datfinf.x in p.nd&mic areas
,.
(2)
Ll,745,
, 3mm286
2.74.
4-6mm(+)161*, 3.3%, 13.0
7-9mm ()19.7#, 29.9#,
10mm15123,34.4#
,
. (Table 4)
1
Table 4
FrequC of the patis inclndFT3crofilaria carrers detected in
the group showi>,g differeint-s'ty of the intradermal reaction.
L"I=GIT-TTI-
ed="]17
8(1
0(184
(3)
e,,
22.5^
),
166, 10
,
, 6%10
,
1O9(90#)10mm
,3O
(Table 5)
Tabe5
Newl changed to positive for microfilara in the persons seitive
FPT antigerin the r&examiation 10 monthes later
-t~-rW~~-~I:-I:-:-t
Gus
Mn
Gus
Diaku
ko
ku
etf
ibc
.I
ib(
erig
-c
oJ
Idd
whelSCh(
>K
llol
-ge13
-ge16
es)9r
liq-i47
31
78
66
122
61*
946
7]
9;S7
Fota1166IIO6.0
*9mmofwhealdiameter
()
256
,0)s
,
(1000x),o
, 5,
161
,14891.9#,126
78.3^,
119(73.9^),
--,, 29(18.0^3
123 l[ lJ //
(4.3#).
6(3.7^) (Table 6)
,
95, 7.9%74
,. (Table 7)
24379.4#
.
,
,
(0-3mm) 1353.8%
.
Table6
Correiobetweenradermaltest(FPT}and
complementJixatiotest(CFT)inv.jil'.rial
patieits.
___=I-I-.FcFT
TI-_(_)2_18(91.9)
3
126(78.3)35161
1iTable7
Complement fixatioj,test ithe persons showing positive
for intradermal testendemic areas.
T_-_~iii==--~_==--1-1NumberofpositiveforintradermaltestiNumberofpositiveforCFtestPercent
patient
withumanifestatin58198
47::---..---_-T-I.___._.__----~~~~..~-~.I--~~~~I-II--~~-~~~
Tota124319379.4
Table8
fCamparisofintensityoftheintradermalreactioi
andcomplementjlocationtest
==1Number'Antibdytitle
ofcases;(-)!510t20i4OxCFtest(+)#
10
7
4;(9)
2)
2)17(
ll(
6(
3(2,
6,,
3.
3.Lit=o4.9)
.4)
.7)1(
3(
5(
K_I_)1
))90
73
56
53);
Total25656375915450200(78.1),5(23.0
#) 10(15.4#), 20
15.4#D
4-6mrn(44)36.7-9mm(67
) 73.%[]N
iJ, 10mm0)(132^T)
90.2^cD--.
,31oQC^o,v
40,3425,20,0)
,
-
a)o)--l.
(Table)
1.'24
5o
,i,:, I'J)i'iu>ii*!o, -. j lk'1'.,
1-j,:), l
(1961)-ju, i.:^l
T--TAirj(
(Dirofilaria immitis), ll
[il-o_)4mm*
20ran,cD>j
125
,
FPT,
.Screening test
,
6,
,
(FPT)
1)
, 1.0/0.01ml15
4mm,
94.8%
2)
3)
,
,
4)
10
10mm
5)
78.1%,
FPT
,,
,
Augustine, D, L. & Lhersson, C. : Stu-
dies on the specificity of intradermal tests in
the diagnosis of filariasis. Amer. Jour. Hyg.
43 : 3840, 19.
2)3ey,R. MB. R. Thooris,G. &Joho,
1Preliminary report on the use of the
antigen of Dirofilaria immitis as an epidemiolo-
gical tool andis a therapeutic agent in wu-
ihereria bancrojti infections in French Oceana.
Amer,, Jour. Hyg. 64 I 23-29, 1956.
3) Bzicevich, J. &Hutter, A. M. : Intra-
dermal and serological tests with Dirojilaria
immitis antigen in cases of human filariasis.
Amer. Jour. Trop. Med. 24 : 203-20, 1944,
4):
=.. 30:1136
1940
5) Dckson, .G.,unlinton, R. W. Jr.
FJchold, S. : Filariasis in defence force, Sa-
moan group. Preliminary report. U.S. N.Med.
Bull. 4 : 1240-1251, 1943.
6) Fairley, N.H : Serological and intrade-
rmal test in filariasis A preliminary report
Trans. Roy. Trop. Med. & Hyg.24:635-648,
932-
/) Huntngton,R. W- A. : Skin reaction to
Dirojilaria immitis extract. U.SN. Med. Bull
44 :07-714, 1945.
8):*==
. 2:3, 1935.
9):
. 14: 1035-1059, 1936.
1O)-,:
52:
594-602, 1961.
11),,, ]j,
,,:
. 1)
9 : 682-700, I960-
12),,,,
,:
2)0D..
12:82-87, 1963.
13),,,,
,:
. 12:77-81, 1963
14):.
oL/--. 12:14-30, 19630
15):L,-. 18
16
300-304, 1963
16') -:v ;:':/ -'-/N& i_~;],; r'[T
oo27:
9(3-922, -93.
)):0)o)
. (1)o21952.
) R,GEarly filariasis diagnosis
arid clinical findings a report of 26cases in
American troops. Amer. Jour. Trop, Med. 24
: 285--298, 19
19),,:
0). (3)0)-
0) ll :37379,962=
20) Loyc, R. G Cnara,S,N.Comple-
ment fixation in filariasis. Indian Tour. Med.
Res, 20 : 1197-i.208, 933-
21):7o)o
:10109, 1930.
22),E3,:,./
o)
Ji(I}No 6 : 188--192.
195
23),,:}j
o
]N)1Lo. 20 : 567
569, 195o
2) Roem.&Dubos, : A cent)
tion to the study of intradermal reactions in
human filariasis. Trans. Roy. SocMed. &
yg 25: 377-382,32.
25),,-:
c(IV) HW-10)
iJ"rf-?;- :;; I'I;lm
o:310-315,62
26) Sawa, T.,omo., Sato, SYmR-
moto, T, & Take, K. : I'mlnunological studies
on filariasis (I) Intradermal and precipithl
tests with Diroftlaria immitis antigen in canine
and human filariasis. Gunlna Jour Med Sci
l: 7-16, 1962
7)w T.,ono,Sato S.,Yma-
moto. T. & Tacei, K. mmunological studies
on filariasis(II)Intradermal test with purified
antigen in canne and humanfilariasis. Gunma,
Tour. Med. Sci. 1 : 16-24, 1962
28);,:Filaria
Dirofilaria immitis
, 44:147,43.
29),: Dirofilaria immitis
1. Column chromatography
)I,o.51 : 207--
209,962.
3D):,
3,29 : 36-40, 1942
31):o)
,(D--DiroHlaria immitis
, 32 : 1421-1429, 1957.
32).:
.Dirofilarift immitis
: 169-
179, 958
33):
.
o
1 :38-50, 1959.
34) Tallaro, W.I. & Hoffmen.. A.
Skin reaction to Dirojilari'i immitis in persons
infected with Wuchereria baicrofti. Jour. Prev
Med. 4 : 261-280, 1930-
35), i":
o I8(KPT) 0)
. 63 : 151-155, 1962.
36)}:
FPT
,, 64:i-4, 1962.
3),,B,
,)
,KPT0)
U), 66 : 82--86, 1963.
38)aTF-, V. G.,arren, J,, &utter,
G.W. : Studies on filariasisI. vSeological
relation-ship between antigen extracts of Wu-
chereri-bancrofi and Dirofilaria immitis, Airier.
Tour Hyg. 43 : 164-170, 1946-
39) Wght, W.& Mot-dock, J.R.Intra-
dermal reactions following the use of DirofIirii
im,miis antigen in persons infected with-
chocca vol-Is. Amer. Joi. Trop. Med. 2
' 127
: 119202, 1944-
40) Yamamura, Y,, Moawa. S.. Taka,
A. &Shoima.Studies on tuberculin
active peptide. . Purification, crystallization
and properties of tuberculin active peptide.
Proc,, Jap. Acta. 35 : 295-298.59.
41):Filaria.2
4 14: 117-124, 1959.
42),:
--
170327-31,956=
43),,:
. (1)4 :276-2l,
1955.
44).,:
()4:282-289,
1955.
45))",,,:
. cm)
Prausnitz-Kustner.6 : 449
-457, 1957.
46),:
(),
, 7:131-134, 1958
47):
cv:
,,: 333-337,
1958
48),:0)
()-
o),.8 : 44-49,
1959
49)]":,
(Rapid Flocculation Test)
,4:224-230, 1956.
50) ZaroW,. & Rifkin, H. : Observation
on the specificity and clinical use of Dirofilaria
immitis antigen in the diagnosis of human fila
riasis (Wuchereria bancrofti). Amer. Jour.
Med. Sci. 21197-102, 1946.
Summary
A peptide antigen (FPT) was prepared from the adult of Dirofilaria immitis by Yamamura-Morizawa's method which has been applied to the preparation of tuberculin peptide as shown
in Table 1. Antigen solution diluted with physiological saline solution to 1: 10,000 was
injected intradermally, just sufficient to arise a wheal of about 3.0mm diameter. Intensity
of the reaction was reflected by the increase of diameter of the wheal within 15 minutes
after injection.Intradermal test was conducted on 541 microfilaria carriers and on 269 control individuals
who were considered free from filarial infection.
Distribution curve of diameter of wheal in the microfilaria carriers showed a normal curve
with the park in the larger side, about 9.0mm. In the control group, neverthless, it ranged
from 0 to 5.0mm showing the peak in the smallest side which resembled the reaction to saline
solution tested on the population of a endemic area. There was found a close corelation
between the increase of wheal and surrounding erythema.
The normal curve presented by microfilaria carriers was corresponded to the group
sensitized by specific antigenic substances. These two different curves crossed each other
at the point of about 4.0mm. Therefore, if the reaction exceeded 4.0mm increase of diameter
of wheal was regaded as positive, 94.8% of the microfilaria carriers fell in positive reaction.
Reactivity of skin to the antigen was never influenced by other helminth infection and allergicdiseases.
Positive rate of the intradermal test assumed usually an upward curve according to theincrease of microfilaria incidence in the population. In the endemic areas, positive reaction
was often seen among those who have neither microfilaria nor clinical symptoms. These
128
positive reactions lacking filarial manifestation were also probably specific for filarial
infection, because the intradermal reaction and CF test using Dirofilaria immitis antigen
coincide with each other in 77.9% of those cases both tested.
From these results, it may be concluded that the intradermal reaction by FPT antigen is
specific for filarial infection and has a practical value for epidemiological survey in endemic
areas. (Auther)
Received for publication September 10, 1963.
