hiv/t. gondii co-infected patients produce lower levels of ifn-γ in response to t. gondii antigens,...
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HIVHIV//T. gondiiT. gondii co-infected patients co-infected patients
produce lower levels of IFN-γ produce lower levels of IFN-γ in response to in response to T. gondiiT. gondii antigens, antigens,
even in the early stage of viral infectioneven in the early stage of viral infection
E. Escobar, M. Alfonzo, M. Fernández, J. Camacho, E. Escobar, M. Alfonzo, M. Fernández, J. Camacho, Y. Roldán, B. Alarcón de Noya, M.E. de QuesadaY. Roldán, B. Alarcón de Noya, M.E. de Quesada
UCV UCV // IVIC IVIC
VenezuelaVenezuela
eescobar@ivic.ve WELBA104
HIV InfectionHIV Infection
Central Nervous SystemCentral Nervous System
• HIV: Neurotropic and NeuropathicHIV: Neurotropic and Neuropathic– Mild DisturbanceMild Disturbance• FatigueFatigue• Sensitive / Cognitive ImpairmentSensitive / Cognitive Impairment• Motor DysfunctionMotor Dysfunction• Behavioral ChangeBehavioral Change
– Severe AffectionSevere Affection• HIV-Associated DementiaHIV-Associated Dementia
T. gondii T. gondii InfectionInfection
• Asymptomatic in Immunocompetent Hosts Asymptomatic in Immunocompetent Hosts – Tissue CystsTissue Cysts• Central Nervous SystemCentral Nervous System• Cardiac MuscleCardiac Muscle• EyeEye
• Reactivates in Immunodeficient HostsReactivates in Immunodeficient Hosts– Toxoplasmic EncephalitisToxoplasmic Encephalitis– CardiopathyCardiopathy– ChorioretinitisChorioretinitis
CoinfectionCoinfectionHIV HIV // Toxoplasma gondiiToxoplasma gondii
T. gondiiT. gondii Stage Interconversion Stage Interconversion
TachyzoiteTachyzoite BradyzoiteBradyzoite
Cellular StressCellular Stress•Heat ShockHeat Shock•Presence of Nitric OxidePresence of Nitric Oxide•Mitochondrial InhibitionMitochondrial Inhibition•Extreme pH Extreme pH (Low/High)(Low/High)
ImmunodeficiencyImmunodeficiency•Lack of T CellsLack of T Cells•Lack of IL-12, IFN-Lack of IL-12, IFN-γγ, , TNF-TNF-αα•Lack of Nitric OxideLack of Nitric Oxide
ControlControl
ReactivationReactivation
Trends in Parasitology 2002; 16(5):198-201.
ObjetiveObjetive
• Evaluate anti-Evaluate anti-T. gondiiT. gondii specific immune specific immune response in HIVresponse in HIV//T. gondiiT. gondii coinfected coinfected patientspatients**
* Several stages of viral infection* Several stages of viral infection
* No HAART nor anti-* No HAART nor anti-T. gondiiT. gondii treatment treatment
– In vitroIn vitro production of IFN-γ and TNF-α under production of IFN-γ and TNF-α under parasitic stimulation was assessed.parasitic stimulation was assessed.
Patients and Control GroupsPatients and Control Groups
ControlsControlsControlsControls PatientsPatientsPatientsPatients
HIV (-)HIV (-)HIV (-)HIV (-) HIV (+)HIV (+)HIV (+)HIV (+)
C1C1T. gondiiT. gondii (-) (-)
C1C1T. gondiiT. gondii (-) (-)
C2C2T. gondiiT. gondii (+) (+)
C2C2T. gondiiT. gondii (+) (+)
P1P1T. gondiiT. gondii (-) (-)
P1P1T. gondiiT. gondii (-) (-)
P2P2T. gondiiT. gondii (+) (+)
P2P2T. gondiiT. gondii (+) (+)
AACD4 > 350/ulCD4 > 350/ul
AACD4 > 350/ulCD4 > 350/ul
BBCD4 200-350/ulCD4 200-350/ul
BBCD4 200-350/ulCD4 200-350/ul
CCCD4 <200/ulCD4 <200/ul
CCCD4 <200/ulCD4 <200/ul
Cell CultureCell Culture
PBMCPBMC1 x 101 x 1055 cells cells
200 ul200 ul
PBMCPBMC1 x 101 x 1055 cells cells
200 ul200 ul
Medium AloneMedium AloneRPMI, 10% FCSRPMI, 10% FCS
Medium AloneMedium AloneRPMI, 10% FCSRPMI, 10% FCS
PHAPHA5 ug/ml5 ug/ml
PHAPHA5 ug/ml5 ug/ml
SATg*SATg*1 ug/ml1 ug/ml
**Soluble Antigens of Soluble Antigens of T. gondiiT. gondii RH strain RH strain
SATg*SATg*1 ug/ml1 ug/ml
**Soluble Antigens of Soluble Antigens of T. gondiiT. gondii RH strain RH strain
CultureCulture72 hours72 hours
37°C, 5% CO37°C, 5% CO22
CultureCulture72 hours72 hours
37°C, 5% CO37°C, 5% CO22
SupernatantsSupernatantsCitokine DeterminationsCitokine Determinations
(Flow cytometry-(Flow cytometry-CBA, Becton-DickinsonCBA, Becton-Dickinson))
SupernatantsSupernatantsCitokine DeterminationsCitokine Determinations
(Flow cytometry-(Flow cytometry-CBA, Becton-DickinsonCBA, Becton-Dickinson))
RESULTSRESULTS
IFN-IFN-γγ Production Production
0
250
500
10000
20000
30000
40000
50000
60000
C1 (VIH-/Tx-; n=4)C2 (VIH-/Tx+; n=5)P1A (VIH+/Tx-, CD4>350/ul; n=5)P1B (VIH+/Tx-; CD4 200-350/ul; n=4)P1C (VIH+/Tx-; CD4<200/ul; n=3) P2A (VIH+/Tx+; CD4 >350/ul; n=5)P2B (VIH+/Tx+; CD4 200-350/ul; n=6) P2C (VIH+/Tx+; CD4 <200/ul; n=4)
Co
nce
ntr
atio
n (
pg
/ml)
IFN-gamma
Medium PHA SATg
IFN-IFN-γγ Production under SATg Stimulation Production under SATg Stimulation
0
250
500
750
10000
20000
30000
40000
C1 (VIH-/Tx-; n=4)C2 (VIH-/Tx+; n=5)P1A (VIH+/Tx-, CD4>350/ul; n=5)P1B (VIH+/Tx-; CD4 200-350/ul; n=4)P1C (VIH+/Tx-; CD4<200/ul; n=3) P2A (VIH+/Tx+; CD4 >350/ul; n=5)P2B (VIH+/Tx+; CD4 200-350/ul; n=6) P2C (VIH+/Tx+; CD4 <200/ul; n=4)
Co
nce
ntr
atio
n (
pg
/ml)
IFN-gamma
SATg
* p = 0.0152
* p = 0.0173
* p = 0.0131
* p = 0.00952
TNF-α ProductionTNF-α Production
0
50
100
150
200
750
1500
2250
3000
C1 (VIH-/Tx-; n=4)C2 (VIH-/Tx+; n=6)P1A (VIH+/Tx-, CD4>350/ul; n=5)P1B (VIH+/Tx-; CD4 200-350/ul; n=4)P1C (VIH+/Tx-; CD4<200/ul; n=3) P2A (VIH+/Tx+; CD4 >350/ul; n=4)P2B (VIH+/Tx+; CD4 200-350/ul; n=5) P2C (VIH+/Tx+; CD4 <200/ul; n=4)
Co
nce
ntr
atio
n (
pg
/ml)
TNF-alpha
Medium PHA SATg
TNF-α Production under SATg StimulationTNF-α Production under SATg Stimulation
0
25
50
75
100250
500
750
1000
1250
1500
C1 (VIH-/Tx-; n=4)C2 (VIH-/Tx+; n=6)P1A (VIH+/Tx-, CD4>350/ul; n=5)P1B (VIH+/Tx-; CD4 200-350/ul; n=4)P1C (VIH+/Tx-; CD4<200/ul; n=3) P2A (VIH+/Tx+; CD4 >350/ul; n=4)P2B (VIH+/Tx+; CD4 200-350/ul; n=5) P2C (VIH+/Tx+; CD4 <200/ul; n=4)
Co
nce
ntr
atio
n (
pg
/ml)
TNF-alpha
SATg
* p = 0.0381
* p = 0.0303
* p = 0.0095
* p = 0.0159
IFN-γ and TNF-αIFN-γ and TNF-α Production Production under SATg stimulationunder SATg stimulation
• Coinfected Patients (P2) higher than P1Coinfected Patients (P2) higher than P1
• Coinfected Patients Lower than ControlsCoinfected Patients Lower than Controls
• Coinfected Patients Affected from early Coinfected Patients Affected from early stagesstages
ConclusionsConclusions
• Defects in IFN-γ (and TNF-α) production Defects in IFN-γ (and TNF-α) production in response to in response to Toxoplasma gondiiToxoplasma gondii may may impair anti-parasitic response in impair anti-parasitic response in coinfected patients since early stages of coinfected patients since early stages of viral infection viral infection
• Eventual reactivation Eventual reactivation (even partial / limited) (even partial / limited)
of brain parasitic latent infection may of brain parasitic latent infection may contribute to develop neurological signs contribute to develop neurological signs and symptoms even before toxoplasmic and symptoms even before toxoplasmic encephalitis appears.encephalitis appears.
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