PO-13: Control of HIV-1 replication in the humanized BLT mouse model after vaccination with
TBK1-engineered DCs is associated with polyfunctional T cell responses
TBK-1
Marta Calvet-MirabentEnrique Martin-Gayo Ph.D.
Hospital Universitario de la PrincesaUniversidad Autónoma de Madrid
GeSIDA 2019
Polyfunctional CD8 T cells
TBK-1
Martin-Gayo, et al, Genome Biol 2018
Objective:Can we enhance DC function
in vivo by artificially activating TBK-1?
TLR3STING
RIG-ITBK-1
IFNb
MDA-5
cGAS
DAI
RNAsensors
DNAsensors
2´3´-cAMPagonist
Poly I:C
GeSIDA 2019
Methods:Experimental Design of in vivo DC vaccine test in
the BLT-mouse modelFetal Liver
CD34+ HSC
NSG Rag2KOIL2RgcKO
Fetal Liver
Fetal Thymus
Media
STING
Poly I:C+
Gag pool
Gag pool 8x h-BLT
8x h-BLT
8x h-BLT
7-10 days
pre-vaccination
In vitro DC geneation
f h-BLT
16 Weeks
3/6Wk
HIV-110,000 TCID50 I.V.
h-BLT: VL
HIV-CTL, CD4Tcell
3/6Wk
HIV-1
3/6Wk
HIV-1
h-BLT: VLHIV-CTL, CD4Tcell
h-BLT: VLHIV-CTL, CD4Tcell
TBK-1
Martin-Gayo, et al, Genome Biol 2018
GeSIDA 2019
MED GAG GAG+ADJ104
105
106
107
Vita
l Load (
copie
s/m
l)
Copy of VL Mice EG1 at 3Weeks Post infection
0.2345
0.2345 0.0207 *
6x105
Plasma Viral Load at 3 weeks p.i.
Vir
al L
oad
(c
op
ies/
ml)
107
106
105
104
MED GAG ADJ
ns
ns *
RESULTS: Mice treated with DC activated with TBK-1 adjuvants display partial control of Viral loads and less severe CD4 T cell depletion
MED d
0
MED w
5
MED w
6 #
GAG d0
GAG w
5
GAG w
6#
GAG
+ADJ d
0
GAG
+ADJ w
5
GAG
+ADJ w
60.0
0.5
1.0
1.5
2.05
10
Fo
ld c
han
ge in
C
D4+
T c
ell
co
un
ts (
cells
/ul) 0.0078
0.0781
0.0078
0.0156
0.0547
0.4609
****
*
D0 W5 W6 D0 W5 W6 D0 W5 W6
DC-Med DC-Gag DC-Gag+ADJ
ns
ns nsFold change in CD4+ T cell counts
GeSIDA 2019
Red: 4 cytokines
Green: 3 cytokines
Blue: 2 cytokines
Grey: 1 cytokines
MED GAG GAG+ADJ
CD8
Chi-square **
Proportions TNFa CD107a
RESULTS: BLT mice vaccinated with DC treated with TBK1 adjuvants display
superior multicytokine polyfunctional T cell profiles
GeSIDA 2019
CD8 T cell aggregation and exclussion of p24+ cells in BLT lymph nodes
DAPI CD8 HIV P24 40x
p24+cells(more excluded in ADJ?)
CD8 clusters(higher in ADJ?)
GeSIDA 2019
Conclusions
1-Vaccination of BLT mice with DC treated with TBK-1 adjuvants is associated with partial control of plasma viral loads at early time points of infection and less severe depletion of CD4 T cells.
2- Mice vaccinated with DCs stimulated with TBK-1 adjuvants exhibit higher levels of polyfunctional CTL responses in the blood at early time points after infection with HIV-1.
3-Clusters of CD8 T cells appear to be enriched in the Lymph nodes of vaccinated animals and this phenomena seems to correlated with a higher level of exclusion of P24+ cells within the tissue.
GeSIDA 2019
Acknowledgements
Martin-Gayo’s LabEnrique Martín Gayo
Cristina Delgado Arévalo
Marta Calvet Mirabent
Ildefonso Sánchez Cerrillo
Lorena Bruzon Hernández
Servicio de Inmunología
Francisco Sánchez Madrid
Servicio de Enfermedades
Infecciosas
Ignacio de los Santos
Jesús Sanz
Lucio Garcia Fraile
Hospital Gregorio
Marañón
María Ángeles Muñoz
Instituto de Salud Carlos III
José Alcami Pertejo
Hospital Vall d´Hebron,
Barcelona
María José Buzón
FundingAtracción de Talento
Program- Government
of Madrid-CAM
NIH-R21 (R21AI140930)
Programa Retos Sociedad Ministerio Ciencia, InnovaciónUniversidades
Becas Gilead GLD19/00168
GeSIDA 2019
