probiotic and il-21 treatment promotes th17 cell recovery in arv-treatment of pigtail macaques...
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Probiotic and IL-21 Treatment Promotes Th17 Cell Recovery in ARV-Treatment of
Pigtail Macaques
Alexandra M. OrtizLaboratory of Jason Brenchley
July 23, 2014
Abstract #: WEAA0104
Th17 Cells and Probiotics in HIV-associated Intestinal Immune Dysfunction
• HIV is characterized by persistent GI immune dysfunction.• Preferential Th17 loss
• Th17 are antimicrobial CD4+ T-cells essential for intestinal immunity.• Generated in the context of commensal signaling
• Immune dysfunction correlates with microbial translocation and dysbiosis
• Probiotic supplementation of chronically-infected, ARV-treated macaques:• Increased colonic CD4+ T-cell function and frequency• Increased function but not frequency of colonic Th17 cells
• IL21 therapy in acutely SIV-infected macaques promotes Th17 retention during treatment (Pallikkuth PLoSPathog 2013)
(Klatt JCI 2013)
Experimental Design
• Animals: 11 Pigtail Macaques (Maccaca nemestrina)
• 6 animals treated with ARV + Probiotics (Pb) + IL-21, 5 ARV-only controls
• Infection: 3000 TCID50, SIVmac239 i.v.
• ARVs: 20-30 mg/kg PMPA i.v., 30mg/kg FTC i.v., 240 mg L’812 – daily
• Probiotics: 1011 live bacteria (VSL#3) oral - daily
• Bifidobacterium: breve, longum and infantis
• Lactobacillus: acidophilus, plantarum, paracasei, bulgaricus
• Streptococcus thermophilus
• IL-21: 50 ug/kg IL-21, s.c. – 5 weekly doses, early and late chronic
• RM IL-21 fused to RM IgG2 Fc, produced in S-2 cells
IL-21 and Probiotic Therapy do not Interfere with Viral Suppression
Vir
al Lo
ad (
log c
opie
s/m
L)
Day (relative to infection)
PTA2P031
Treatment initiation
(late) IL21(early) IL21
IL-21and Probiotic Therapy Improve Intestinal CD4+ T-Cell Frequency
Jejunum Rectal Biopsy
Day (relative to infection)
Perc
en
t C
D4
+ o
fC
D3
+ T
-cells
ARV onlyARV+Pb+IL21
PTA2P031
PTA2P031
IL-21and Probiotic Therapy do not Induce CD4+ T-cell Activation
PBMC LN
BAL Jej RB
Day (relative to infection)
Perc
en
t K
i67
+ o
f C
D4
+
TM
ARV onlyARV+Pb+IL21
IL-21and Probiotic Therapy Improve Intestinal Th17 Frequency
Perc
en
t IL
17
+ o
fm
em
ory
CD
4+
T-C
ells
Day (relative to infection)
Jejunum Rectal Biopsy
ARV onlyARV+Pb+IL21
IL-21and Probiotic Therapy Improve Polyfunctional Th17 Recovery
Perc
en
t Po
lyfu
nct
ion
al of
CD
3+
T-C
ells
Day (relative to infection)
Jejunum Rectal Biopsy
ARV onlyARV+Pb+IL21
IL-21 and Probiotic Therapy Moderately Reduced Colonic IDO Expression
ARV onlyARV+Pb+IL21
Perc
en
t ID
O+
Tis
sue
Colon Mesenteric LN
IL-21and Probiotic Therapy Reduce Comorbidities
Perc
ent
Inci
den
ce-F
ree A
nim
als
Day (relative to infection)
Conclusions and Future Directions
• Therapeutic treatment with probiotics and IL21 improved disease progression in ARV-treated macaques as measured by:– Intestinal CD4+ T-cell recovery– Improved intestinal Th17 frequency and functionality– Decreased incidence of health complications
• By what mechanism are Th17 cells expanded?• Would treatment promote enhanced protection against
microbial challenge?• Does probiotic and IL21 treatment reduce microbial
translocation and dysbiosis?
Acknowledgements
Brenchley Lab• Jason Brenchley• Nina Calantone• Kirby Carmack• Sarah DiNapoli• Zachary Klase• Molly Perkins• Carol Vinton
Veterinary StaffHeather CroniseRichard HerbertJoanna Swerczek
LMM Core
NCI-NIH• Jake Estes
Emory University – IL21• Mirko Paiardini• Francois Villinger
Gilead – PMPA/FTC• Michael Miller
Merck – L’812• Daria Hazuda
Sigma-Tau – VSL#3• Claudio De Simone