Rev. Biol. Trop.

, 45(1): 877-882,1997
Increasing of Toxoplasma gondii (Coccidia, Sarcocystidae)
infections by Trypanosoma lewisi (Kinetoplastida,
Trypanosomatidae) in white rats
OIgaM.Gucrrcro,MsacILhnchIIaandE.Abrahams
Centro de Investigacin en Enfermedades Trpicales (CIT). Departamento de Parasitologa, Facultad de Microbiologa,
Universidad de Costa Rica, San Jos Costa Rica.
(Recibido 22-IV-1996. Corregido 9-VIII-1996. Aceptado 23-IX-1996.)
Abstrat: To demonstrate Iha! T lwisi infection increases T gondii multiplicaion in white rts, groups of fve
Wistar oc Sprgue Dawley rats were inoculated with 106 T lewisi trypomastigoles and four or seven days later
infected wi!h Toxoplasm tachyzoites. Host surviva time was monitored, ad !he presence of T gondii was
confred in all dead rals by studying peritoneal exudate smears and lung tissue sections stained wih
hamatoxylin-eosin. The prsence of Toxoplasma cysts or antioies was ehecked i te brin o surviving rals.
Th nerase i observed four days afer typanosom inoculaton and is dependen! on fat strain, bul not on
inoculum size oc cat age. Humoral and cellular factors may have a role in he nerease as has been reported for
other experimental infections wit Afcan trypanosomes and T. cruzi.
Key words: Toxoplasma gondi Trpanosoma lewisi, irunosupprssion, rts.
5ncc 1- I has bcn known thaI
whIc raIs ac rcmakabIy rcssIanI Io
Toxoplasma gondii ntccIon (Kuchman
lowIcr, Lanson).
Wchavc bccn ngIocxpIan Ihc rcasons
tor such rcssIancc. n our hrsI sIudcs wc
dcmonsUaIcd, wIh conUoIIcd nocuIa, IhaI
cvcn l" IachyzoIcs do noI gvc rsc Io taIaI
ntccIon in aduII whIc raIs (ChnchIIa et al.
8). Wc showcd Ihc mporIancc ot qc
rcssmcc (LhnchIIa et al. )
macrophacsacIvIy (LhnchBa et al. 18Z),
tor IhaI naluraI rcssIancc Io gondii
sIudcsnprogcss, ncIudng ncw normaIon
abouI othcr cIors maI couId pIay an
mportanI roIcn Ihs phcnomcnom, comh
mc Iow susccpIbIIy ot whIc raIs Io
gondii.
!mmunosupprcsson ot pro|ozoa
ntccIons wIh oIhcr parasiIcs has bccn
rcportc. lor cxapIc Trpanosoma cruzi
ntcc|on appca Io bc ncnacd by somc
mmunosupprcssvc cmIs duc Io maaa
ntccuonsnhosI(KrcIII !77).
1hs cccI has bcn aIso dcmonsUakd n
Ahcan Irpanosomass (OIsson et al. .
Oar et al. Z). laIcuIaIy n
IoxopIasmoss, rcacuvaIon duc Io vrus
ntccIonwasrcportcdnnucc (lomcroy et al.
l8)andcaIs(Wiuet al. 8).Howcvcr,wc
havc noI tound oIhcr sIudcs ot gondii
mmunosupprcsson duc Io Trpanosoma
ntccIons.
Wc haVc tound |haI Trpanosoma lewisi
ncrcascs gondii ntccIon ot whIc raI,
changng Ihc susccpIbIIy ot Ihs mdcnI Io
878 RVISTA DE BIOLOGIA TROPICAL
Ihc paasIc. 5Iudcs ot d nI tacIors
cttccung Ihs phcnomcna a pnscnIcd n Ihs
papr.
MA1EK!AL^ME1HOO5
Animals: WsIar or 5praguc OawIcy r
t> wIh a IocaI chow ad wacr ad libilum
wcn uscd n Ihcsc cxpcnmcnIs. Toxoplsma
IachyzoIcs tor scroIogy tcsIs wcn obIancd
homN!HwhIcmccpcnIoncaIcxudaIc.
arasilc slrains: 1achyzoIcs ot
Toxoplasma K sUan and paaIIy
chaIcnzdJLK-Z and 1LK-J sUans (HoIsI
ad LhnchIIa , Guer et al. )
wc nocuIaIcd n d nI conccnUaIons
accordngIocachcxpcnmcnI.1hcscsUans a
manIancd by nguIa passagcs (Iwcc a wok)
nIhcacuIcphasc.
T. lewisi sIranwas IocaIIy soIaIcd hom a
Rattus noregicus, grayraI, manIancd by
wcckIypassagcnwhIcraIs.
Gcncral cxpcrimcnlal modcls: roups
ot hvc WsIar or 5praguc OawIcy raIs wm
nocuIa|cd .p. wIh " lewisi
IrypomasIgoIcs obIancd hom onc wcck
ntccIcd anmaIs ad tour or scvcn days IaIcr,
accomngIo Ihc cxprmcnIs, Ihcsc raIs wm
ntccIcd wIh dcnnI numbcr ot Toxoplasma
IachyzoIcs.
5muIIancousIy, oIhcr anmaIs wm
nouIaIcd onIy wIh gondii or lewisi,
rcprcscnIng Ihc comspondng conIroIs. Ah
ntccIonswcrcdoncnUapcrIoncaIy.
5urvvaI Imc ot raIs was monIorcd,
chcckng Ihc pnscncc ot Toxoplasma n thc
pcrIoncaIcxudaIcorIungsnah Ihc d raIs.
Lung Issuc sccIons wc sIancd by
hcmaIoxyIn-cosn Icchnquc Io dcmonsmc
Icsons. AnmaI survvngtorJ days, mc m
ponItor aII Ihc cxpcrmcnIs, wcn smdcd to
pnscncc ot branToxoplasma cysIs or tor
anIbodyprcscncc.1hcLabon mmuno-Assay
(L!A) ot known spcchcIy (LhnchIIa et al.
Z)anduscdtorusnoIhcrsIudcs(Aras e t
al. 4)waspcrtomcdtorscroIogcanaIyss.
A "I" sIudcnI IcsI tor smaII sapIcs wa
uscdtorsIaIsIcaIanaIyss.
K5UL15
Dclcrminalion o lewisi prcvious
infcclion limc: !n a conUoIIcd cxpcnmcnI,
raIs wcn ntccIcd wIh lewisi N
dcrcnI pcrods ot Imc ( Io Z dgs),
nocuIaIcd wIh gondii IachyzoIcs. O
ndcac (1abIc ) IhaI lewisi nocuIaIcd Io
raIs, tour Io sx days prcvousIy Io gondii
ntccIon produccs maor ncrcascs ot Ihc
muIIpIcaIon ot Ihs Iast parasIc (
undcrIncdnumbrs).
!noIhcr cxpcnmcnI J or b days oId raIs
w nocuIaIcd wIh IqpomasIgoIcs
tour or scvcn dgs Iakr ntcc|cd wIh "
gondi IachyzoIcs. AsIsshown n 1abIc Z,
boIh agcanmaIs nkc| wIh lewisi tour
days bctorc Io Toxoplasma ntccIon, d
carIcr(scc undcd da|a)Ihan Ihosc ntcckd
scvcn days prcvousIy wIh Uypanosomcs.
KaIsnocuIaIcd onIy wIh gondii (conUoIs)
survcd J days, cnd ot Ihc cxpcrmcnIs n
anycasc.
Ral slrain difcrcnccs (1abIc J): EIhcr
J or b day oId WsIm raIs nouIaIcd wIh
Toxoplasma, prcvousIy ntcckd wIh
lewisi, survvcd nomorcIhanZdays. On Ihc
oIhcrhand, 5pragucOawIcyraIssurvvcdmorc
IhanZJdays.!nboIhcascs, anmaIsnocuIaIcd
wIh gondii aIonc,survvcdJ0days,Ihc m
otIhc cxpcrmcnI.
oxopmmo slrain diHcrcnccs: J
appmnI mmunosupprcssvc cccI ot
lewisi wasdcmonsUaIcdtorIhrccsUansIudcd
(1abIc 4). Howcvcr Ihc survvaI Imc ot mc
KntccIcdmccwasIowcr.
Hccl o 0xopmmo inocuum: 1c
avOagc survva Imc was shorIcr n anmaIs
prcvousIyntccIcd wIhT. lewisi, ascompacd
wIh Ihc conIroIs, ndcpndcnI ot Ihc nocuIa
(J`, ", ') ot Toxoplasla Iachyzoks
(1Ic).
AIIIhc Toxoplasma nkckd raIs survvng
J days wcm posIvc, cIhcr by bran cysI
prcscnccormcL!A IcsI and many tachyzoIcs
wcn obscrvcd n pcnIonca macrophagcs
Iung Icssons (lg.) ot raIs prcvousIy
nocuIaIcdwIh lewisi.
Guerrero el aL: Incrasing of Toxoplsma 879
1ABLE.
Survival of rats* with T. gondii infection and pre-infected with T. lewisi.
Number of T lewisi Numbr of T gondi
trpomast./inoculated aimal tachyzJinoculated animal
o
10
O
10
lO
10
10
10
106
10
10
*5 rts for each goup.
Time interval between T.
lwisi ad Toxoplsma
infections (d)
O
O
O
3
4
S
6
10
14
17
21
Survival tim (d)
30
30
30
30
20.8
7
2SA
30
30
30
30
% Surviving
30 d
100
100
10
100
60
O
80
10
100
100
100
1ALEZ
Efect of strain or age of rats on T.1ewisi "immunosupression" against T. gondii, inoculted four d lter
Rat stran Age
(d)
T. lewisi infecton
Infected Non-infected
Surviva tim % surviving Surviva t % surviving
(d) (30 d) (d) (30 d)
Wistar 30
60
11.4 20 30 10
12.7 2S 30 10
Sprague
Daw1ey 30
60
23
30
AII |hc d nccs bcIwccn 'oxoplsma
muItpIca|on ot lewisi m and non-
ntccIcdamaIsshowcdal<0.001.
O!5CU55ON
AIIhough wc haVc dmonsUacd a
nmakabIc nssIacc ot whIc raIs Io
Toxoplasma ntccton (LhnchIIaet al. 8),
I s possbIc Io d I by UcaImcnI wIh
munosupprcsvc agcnIs (LhnchIIa et al.
l2).!nLcIwctoundmaIcorIsoncpccton
nduc a nvcrson ot mmunIy dunng
honIo v :rsc Io a nIapsc.
Lkcwsc, Ihc na reslstc o.
nhbIcdbyIhcsmc uamcnI (LhnchIIa et
al. I2).Howcvcr,IbsrcssIanccwascvdcnI
a scvcra dgs, whch s dcnI Io wbaI
wc rcport n Ihts smdy. 1hc cccI cxcrkd by
lewisi on Toxoplasma ntccIon o n
onIy tourIo hvcdays(1Ic l d Z) ad u
was noI dcpndcnI on paasIc sUan or agc
(1abIc J and 4). 1hs mcans maI wc ac n
pnscnccota sUong cdcc| agansI Ihc
60
10
30
30
10
10
mmunc rcsponsc. 5ncc gondii s a
nUacIIuIar paasIc n whch ccIIuIa
mmunIy s morc proIccIvc Ihan humoraI
(lrcnkcI 98), our modcI suggcsIs a
supprcssonotay ccIIuIarmantcsIaIons.
munosuprcsson n cxpnmcnIaI
IoxopIasmoss, probabIy duc to ccIIuIa
munIy mparmcnI, has bccn obscrvcd n
concomIanInccIonswIh Loupng-II V
(Kcd et al. 8) munnc lcukcma vrus,
tcIncmunodchccncyvus(WaIanabcet al.
J) a CyIomcgaIovrus (lomcroy et al.
8).
Oq et al. (2), havc shown n scvc
sIudcs a panosomc-cIcttcd munosupp-
s U Su
cncraons ot prosIagIandn-producng
macropbagcs, or a prosmIdn-ndcpcnJnI
supprcssvc mcchansm IhaI nhbI mc
cxprcsson ot J-Z rcccpIors. lurmcmorc,
Ihcy csIaIsh IhaI J a -gam
pIayanmporIanIro!cnIhcpaIhwayot1 -ccIl
(probabIy LO4) munosupprcsson. Amuo
(2) has dcmonsUak, n vcry d
cxpcnmcnIs, mc mpormcc ot LO4 ccIIs
880
Age
(d)
30
60
REVISTA DE BIOLOGIA TROPICAL
1ABLEJ
Confration that previous T.lewisi infection gives rise to increasing of T.gond multiplication in rat*
Time interal between T.
lewisi and Toxoplsma
infections (d)
o
4
7
O
4
7
T. lewisi infection
Infected
Survival time % suriving
(d) (30 d)
30 10
15 35.7
23.3 70
30 10
15.6 37.5
27.7 90
Non-infected
Surival tim % surviving
(d) (30 d)
30 10
30 10
30 10
30 10
30 10
30 10
* 5 rats foe each goup.
1ABLE4
Relation ofToxoplasma strains with the ef ectof T.lewisi parasite sinoculated four dys earlier in Wistar rats.
T gondii st T. lewisi infection (4 d before)
Non-infected Infected
Surviva time % surviving Suriva time % surviving
R
TCR-2
TCR-3
(d)
16
20.4
24
(30 d)
4
60
75
(d) (30 d)
30 10
30 10
30 10
1ABLE
Efect ofToxoplasma inoculum on "immunosupression" of Wistar rats, induced by T.lewisi, infected four dys earlier
T gondii inocula
(Tachyz.lanimal)
T. lewisi infection (4 d befor)
Non-infected Infected
Surviva tim % surviving Surviva tim
(d)
% surviving
(d) (30 d)
25 80
30 10
7 80
ad -a n Ihc mmunc nsponsc aansI
gondii. 5omc ot Ihcsc sIudcs ad oIhcrs wIh
brucei (BakhcIet al. v)as wcII as wIh
congolense (lIyn and 5Icghcn )
cxpIansomcmm0nosupprcssvcmcchansms
Iha| can b suspcK !or our Toxplasma -
lewisi modcI.5zIcnand Kcrszcnbaum (IZ)
suggcsI IhaI aIIcrauons o! Ihc cxpnsson ot
JcK, LO
4
, LO d -ZK du Io cruzi
nkcIons,couIdbcrcsponsabIcotIymphocyIc
nIcrtcrcncc, gvug nsc Io
mmunosupprcsson. 1s s anoIhcr
nIcrcsIng aspccIo O nIo accounI torour
cxpcrmcnIs.
6.4
11.4
25.3
(30 d)
O
20
10
AIbnghI and AIbrghI () havc
suggcskd IhaI n ntccIons wIh
Trpanosoma musculi, a mousc parasIc
boIogcaIIy smIar Io 1 lewisi, Ihcrc a
scvcra tacIors bIockng ot anIbody acIvIy,
nkrtcnncc maI m ccI humoraI mmunIy
supprcsson.JhscouIdbc anoIhcrcxpIanaIon
torIhcapparcnImmunosupprcssvccccI IhaI
wchavcobscrvcdnourmodcI.
Jhs phcnomcnon appcms Io bc nIakd
wIhIhcanmaI gcncIc orgcn as s shown n
1abIc J. AcIuaIIy Ihc c0I was morc cvdcnI
n WsIa Ihan n 5praguc OawIcy raIs.
Ounccs n anmaI mmunc rcsponsc Io
nUaccII0Iar paasIcs dcpcndng on animaI,
Guerrero et al: ncreasing of Toxoplasma 881
spccaIIy mcnI sIrans, havc bccn rcporu
(Handman T). 5pcctcaIIy, tor raI sUans,
n a sIudy Io dcIcmnc Pneumocystis carinii
karyoIypcs obImncd by concosIcod
UcaImcnI, Hong et al. (Z) n: onIy hvc
Io cghI wccks tor mmunosupprcsson ot
WsIa raIs. On Ihc conu a nnc Io Icn
wccks IrcaImcnI was nccccs Io obtan Ihc
saccttccItor5pragucDawIcyraIs.
!n concIuson, Ihcsc prcImnay sIudcs
dcmonsUaIc IhaI 1 lewisi ntccIons c
ncrcasc Toxoplasma muIIpIcaIonnraIs d
Ioan appacnI mmunosupprcssvc cccI. 1c
prcscncc ot many IachyzoIcs n Iung Icsons
(lg. a, b, c) as wcII as n pcrIoncaI
macrophagcs (lg. !d) oI thcsc anmaIs,
comparcd wIh Ihc conIrols, supporI Ihc
nIcrprcIaIon IhaI 1 gondii was Ihc causc ot
dcam.
lurIhcmorc survvaI ot nIs ntccIcd wIh
1 lewisi or 1 gondi aIonc IcI us Ihnk lhat
Ihc ccct s noI p by oIhcr dscasc, a
has bccn mcnIoncd tor baIoncIoss ntccuons
(lcrIa ad MaorsIon 4) whch c
modhcdIhcnaIuraIrcssIancc.
ALKOWEDGMEN5
1hswork was supportcd by granIs # 4J0-
Z-04and # 4J0-Z-0 trom thc UnvcrsIy
ot LosIa Kca. Wc Ihank !.K. lrcnkcI tor
suggcsIonsabouI Ihs manuscrpI.
KElEKNLE5
Albright, J.W. & J.E Albright. 1991. Rodent
trypanosomes: Their confict with Ihe Immune system
ofahe host Pasitol. Today 7: 137-140.
Araujo, F.G. 1992. Depletion of CD/ T cells bUI no!
inhibition of he protective activity of IN-a prevents
cure of toxoplamosis mediated by drug therapy in
mce. J.Immllnol. 149: 3003-3007.
Arias, M.L. M. Chinchilla, L. Reyes, 1. Sabah & O.M.
Guerero. 1994. Dtennation of Toxoplsma gondii
in several orgas of cattle by Caon Immlnoasay
(CIA) testing. Vet. Parasitol. 55: 133-136.
Bahiet, M. T. Olsson. P. Va der Meide, & K.
Krislensson. 1990. Depletion of CD8+ T cells
suppresses gowth of Trpanosoma brucei brucei and
interferon-gama production in nfecled rals. CIin.
Exp. Immunol. 81: 195-199.
Chinchilla, M. M. Alfaro & O.M. Guerrero 198!.
Adaptacin natural de la rata blanca a Toxoplsma
gondii. Rev. Biol. Trop. 29: 273-282.
Chinchilla, M. O.M. Guerrero & E. Solao 1982. Lack of
multiplication of Toxoplsma in macrophages of rats
in vito. J. Parasitol. 68: 952-955.
Chinchilla, M. O.M. Guerrero, L. Reyes & A. Castro.
1992. Efecto de los corticosteroides en la tasmisin
congnita de toxoplasmosis experimental. Rev. Biol.
Trop. 40: 135-137.
Chinchl a, M. L. Reyes. O.M. Guerero & F. Herndez.
1992. Specifcity of !he carbon immunoassay (CIA)
test for !he diagnosis of Toxoplasma nfeclion. Veto
Paasitol. 44: 315-320.
Darji, A. R. Lucas, S. Magez, E. Torreele, 1. Palacios, M.
Sileghem, E. Bajyana Songa, R. Haer & P. De
Baetselier. 1992. Mechanisms underlying
trpanosore-elicited iunosupprssion. Ann. Soco
Belg. Med. Trop. 72 (Supp!. 1): 27-38.
Fyn J.N. & M. Silegher. 1991. le role of the
macrophage in induction of immunosupprssion i
Trpanosoma congolense-infected caldeo Imulo
logy 74: 310-316.
Frenkel, J.K. 1985. lmmunity in toxoplasmosis. PAHO
Bulletin 19: 354-36.
Guerer, O.M. M. Chinchlla, R. Man, G. Catanella
& A. Castro. 1991. Estudio compaativo de dos cepas
de Toxoplama gondii. Paasitol. Da. 15: 97-10L
Handman, E. R. Cerig & G.F. Milchells. 1979. Murine
cutaneous leishmaniasis: Diseae parters in intact
and nude rce of various genotypes and examination
of sor differences between nona! and infected
macrophages. AJEBAK 57: 9-29.
Holst, I. & M. Chinchilla. i 990. Development and
dstrbution of cysts of an avirulent strain of
Toxoplsma gond and he humoral immune response
in rce. Rev. Biol. Trop. 38: 189-193.
Hong, S.T. lS. Ryu, lY. Chai & S.A. Lee. 1992.
Trnsmssion modes of Pneumocstis caTnii aong
rats obsered by kaotype analysis. Korean. J.
Parasit. 30: 283-287.
Kttli, A.V. 1977. Exaceration of experimental
Tranosoma cruzi infection in rce by concomitan!
maaa. J.Protozool. 24: 514-518.
Lanson, R. 1955. Toxoplasmosis in England H. Variation
factors in the pathogenesis of Toxoplsma infections:
the sudden ncrease virulence of a stin afler
passage i multimammate rls and canaries. Am.
Trop. Med. Parasitol. 49: 384-416.
Olsson, T. M. Bakhiet, C. Edlund, B. Hojeberg, P.H. Van
der Meide & K. Kristensson. 199 L Bidirectional
activating signals betwen Trpanosoma brucei and
CDg + T cells: a trypaosome released factors triggers
interferon a production Ia stlaes parasite
gowth. Eur. J. Immunol. 21: 247-2454.
Perla D. & 1. Maorton. 1941. Natural rsistance and
clnical medicine. Chapter 28. T spleen and
882 REVISTA DE BIOLOGIA TROPICA
resistance: pags 774-777. l
l
ed. Little, Brown and
Comany, Boston, 1344p.
Pomeroy, C. S.Kline, M.C. Jordan & G.A. Fllice. 1989.
Reactivation of Toxplsma gondi by
Cytomegalovirus disease in mice: Antimicrobial
actvities of macrophages. J. Infect. Dis. 160: 305-
311.
Prston, P M. K. Behbehani & D.C. Dwnonde. 1978.
Expermental cutaeous leishmaniasis: VI: Anergy
and allergy in t cellular immune response during
non-hea1ing infection in diferent strans of mce. J.
Clin. L. Immunol. 1: 207-219.
Reid, H.W. D.Buston, l. Pow & JFm1ayson. 1980.
Immunosuppression in toxoplasmosis: fther studies
on mice infected with louping-il virus. J.Med.
Microbiol 13: 313-318.
Ruchman, l. & J.C. Fowler 1951. Localizaion and
persistence of Toxplsma in tissues of experimental
infected white r. P. Soco Ept. Biol. Med. 76:
793-796.
Sztein, M.B. & F. Kierszenbaun. 1992. Suppression by
Tranosoma cr; of T -cell receptor expression by
activated hwn Iymphocytes. Immunology 77: 277-
283.
Waller, T. 1977. T India i immunoraction: a
method for t dianosis of encephalitozoonosis. Lab.
Anim. Seie. 11: 93-97.
Waaabe, H. Y. Suzuk, M. Maino & M. Fujiwara.
1993. Toxoplasma gond;i: Induction of toxoplasmic
encephalitis in mice with chronic infion by
inoculaon of a murine leukemia virs inducing
iimunodefciency. Exp. Paritol. 76: 39-45.
Witt, eJ. T.R. Moench, A.M. Gittelsohn, B.O. Bishop &
J.E.Childs. 1989. Epiderologic observations on feline
immunodefciency virus and Toxoplsma gondii
coinfection in cas in Baltimor, Md. J.A.V.M.A. 194:
229-223;