849
Pyroptosis, production of reactive oxygen species and synthesis of
nitric oxide, and inflammatory response were activated in survival
patients at D0 and D7. Furthermore, bacterial infection inhibition
was found in survivals. Interestingly, similar functions were also
identified in non-survivals but differences were the activation
or inhibition z scores. These results highlight the activation of
inflammasome during sepsis, with a different degree of modulation
in survivors and non-survivors. Further validation of some genes at
protein level is under process.
AS01-adjuvanted vaccine induces a transient innate immune
response in humans
Smolen, K.
1
, Herve, C.
2
, Brouwer,M.
1
, Taton,M.
1
, Auquier, P.
2
, Fissette, L.
2
,
Marchant, A.
1
, Van Belle, P.
2
, Burny, W.
2
, Didierlaurent, A.
2
, Willems, F.
2
1
ULB,InstituteforMedicalImmunology,Gosselies,Belgium,
2
GSK,
Rixensart,Belgium
AS01 is an Adjuvant System containing 3-
O
-desacyl-4’-
monophosphoryl lipid A (MPL),
Quillaja saponaria
Molina, fraction
21(QS-21) and liposome. AS01 has been selected for the clinical
development of several candidate vaccines including the malaria
and varicella zoster vaccine. AS01 promotes both humoral and
cellular responses. Yet, little is known about the immunological
mechanisms in humans by which AS01 induces effective immune
responses. Given the fundamental role of the innate immune
system in controlling the adaptive immune responses, we have
conducted a phase II randomized, single centre, single-blind study
(NCT01777295) to characterize the kinetics of the early immune
response to immunizationwithanAS01-adjuvantedvaccine.
60 HBV-naive adults (18-45 years old) received 2 doses of Hepatitis
B virus surface antigen (HbsAg) adjuvanted with AS01 B (50µg
MPL, 50µg QS-21) or aluminium hydroxide at days 0 and 30. All
subjects received a saline placebo (baseline) 30 days prior vaccine
administration.Whole blood samples were collected at defined time
points (0h to 7 days) before and after placebo, dose 1 and dose2.
Immunization with AS01 B -adjuvanted vaccine leads to an early
and transient inflammatory response in blood, characterized by
changes in some cytokines, innate cell number and activation.
Cellular variations were specifically reflected in monocyte subsets-
particularly intermediate monocytes- and NK cells, indicative of
the stimulation of the innate immune system. Although restricted
to HBsAg and a naive population, identification of these early
signatures may provide new insights into the mechanisms of AS01-
adjuvantedvaccineinducedprotectiveimmuneresponse.
Analysis of
CTLA-4
gene polymorphisms (-319 C/T, +49 A/G,
CT60 G/A) in primary Sjögren´s syndrome
Carrillo-Ballesteros,F.J.
1
,Muñoz-Valle,J.F.
1
,Palafox-Sánchez,C.A.
1
,
Valle,Y.
1
,López-Villalobos,E.F.
1
,Badial-Hernández, M.F.
1
,Hernández-
Martínez,M.A.
1
,Rodríguez-Machuca,V.U.
1
,Vázquez-Villamar,M.
1
,
Orozco-Barocio,G.
2
,Oregon-Romero,E.
1 1
UniversidaddeGuadalajara,
InstitutodeInvestigaciónenCienciasBiomédicas(IICB),Guadalajara,
Mexico,
2
HospitalGeneral deOccidente,SSJ.,Zapopan,Mexico
Introduction:
Primary Sjögren´s syndrome (pSS) is an autoimmune
disease affecting salivary and lacrimal glands. It is characterized
by a progressive lymphocytic infiltration, mainly T cells. Cytotoxic
T-Lymphocyte Antigen 4 (CTLA-4) plays a critical role in the
prevention of autoimmune pathology by a critical inhibitory role in
T cell responses resulting in diminished proliferation and cytokine
production. Polymorphic sites located at -319C/T, +49A/G and
CT60G/A in the
CTLA-4
gene have been associated with elevated
CTLA-4 serumlevels (sCTLA-4) andautoimmunity.
Objective:
Determine the frequency of CTLA-4 polymorphism in pSS.
Methodology:
The study included 111 Mexican mestizo patients
with pSS, classified according to the American-European Consensus
criteria. As control group, 122 clinically healthy subjects were
included. The polymorphisms were genotyped by PCR/RFLP
technique. Statistical analysis was carried out with
Pasw statistics
18
andEmHapFre softwares.
Results:
Polymorphisms were in Hardy-Weinberg equilibrium
and showed moderate linkage disequilibrium
(50%, p=0.009)
. The
most frequent haplotypes were: CAG(34%), CAA(26%), CGG(18%),
and CGA(17%). Patients that carried CAA haplotype showed high
sCTLA-4levels,lymphocyticinfiltratedandautoantibodyproduction
without statistic significance. Inaddition, high levelsof sCTLA-4were
observed inpSSpatients carriers of -319CT genotype (p=0.018
vs
CC)
and+49AAgenotype
(p=0.013
vs
GG).
Conclusion:
Our results suggest that
CTLA-4
polymorphisms -319
and +49 could have a role for the development of pSS. However, the
lowassociations of sCTLA-4with clinical parameters suggest that the
potentialeffectof
CTLA-4
polymorphisms needs tobe investigated in
subsequent studies inMexican patients in order to elucidate it.
Simple and effective tumour immunotherapy using
intratumoural complete Freund´s adjuvant
Fahrer,A.
1
,Carroll,C.
1
,Andrew,E.
1
,Orange,M.
2
,Allavena,R.
3
1
TheAustralianNationalUniversity,ResearchSchoolofBiology,Acton,
Australia,
2
ArlesheimKlinik, Onkologie, Arlesheim, Switzerland,
3
University
of Queensland, School of Veterinary Science, Gatton, Australia
We demonstrate that intra-tumoural injection of Complete Freund´s
Adjuvant can result in a potent anti-tumour immune response. In
order to test the efficacy of this treatment, we have initiated pre-
clinicaltrialsinthreespecies:mice,dogsandhorses.Efficacy hasbeen
demonstrated in a range of solid tumours, including mastocytoma,
mammary tumours and melanoma. Complete tumour regressions
have been observed in all three species. Evidence of systemic
immune responses (regression of non-injected metastases) have
also been observed. We characterise the immune cells infiltrating
mouse tumours after treatment. Finally we provide case reports on
the treatment of four human cancer patients; suffering from lung
cancer, metastatic osteosarcoma, or breast cancer, with one patient
havingapartial response to treatmentwith intra-tumoural Complete
Freund´s Adjuvant.
Taken together, our data demonstrate that our treatment has major
anti-tumour effects in a proportion of treated animals, and is safe for
use inhumancancer patients. Further trials inhumancancer patients
are therefore strongly warranted. Our novel treatment provides a
simple and inexpensive cancer immunotherapy, applicable toawide
range of solid tumours, and of potential benefit to cancer patients
around theworld, includingpatients fromdeveloping countries.