Journal of Clinical Virology - Volume 82 - Supplement 1 - September 2016

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Abstracts / Journal of Clinical Virology 82S (2016) S1–S142 S29
increased levels of CXCL13, the main finding of low sensitivity for
diagnosing LNB using previously proposed cut-offs, is caused by the
relatively low increase of CXCL13 in the LNB patients in this mate-
rial. However, if only including patients where sampling was done
in the first week after onset of facial palsy, the sensitivity increases
to 92.9% at 61 pg/mL and 64.3% at 1224 pg/mL respectively. If CSF
CXCL13 is to be used in the clinical setting, further studies on CNS
infections with similar clinical presentations are needed, as the dif-
ficulties presented here requires a thorough understanding of the
limitations of the analysis.
Abstract no: 149
Presentation at ESCV 2016: Poster 16
Evaluation of HPV16-specific central memory T
cell response in healthy subjects and patients
with head–neck cancer
I. Cassaniti ∗, S.A. Calarota ,
K.M.G. Adzasehoun , C. Fornara , G. Comolli ,
L. Barzon , P. Pedrazzoli , F. Baldanti
1 Molecular Virology Unit, Microbiology and Virology
Department, Fondazione IRCCS Policlinico San
Matteo, Via Taramelli 5, 27100 Pavia, Italy
2 Experimental Research Laboratories, Biotechnology
Area, Fondazione IRCCS Policlinico San Matteo, Viale
Golgi 19, 2, Italy
3 Department of Molecular Medicine, University of
Padova, Padova, Italy
4 Division of Medical Oncology, Department of
Hematology and Oncology, Fondazione IRCCS
Policlinico San Matteo, Pavia, Italy
5 Department of Clinical, Surgical, Diagnostic and
Pediatric Sciences, University of Pavia, 27100 Pavia,
Italy
Background: Growing evidences support the etiologic role of
HPV16 in the onset of Oropharyngeal Head andNeck Cancer (OSCC).
However HPV-related OSCC show better prognosis compared to
HPV-unrelated OSCC. Recently, prophylactic vaccine has been rec-
ommended for preventing high-risk anogenital HPV infections.
HPV L1 antibodies are generally weak or absent after natural infec-
tion, while high concentrations of anti-L1 antibodies are detectable
about onemonth after the three-dose of prophylactic vaccine. It has
been established that cell-mediated immune response is crucial in
the control of HPV infection and correlated diseases. The princi-
pal aim of our study was to evaluate and characterize the memory
HPV16-specific T-cell response in patients with OSCC.
Material and methods: PBMC from twenty-five patients with
OSCC (8 females and 17 males, median age 60 years, range 40–85)
were tested. Ten of 25 (40%) had biopsy-proven HPV16-related
OSCC, while the remaining 15 (60%) had HPV-unrelated head–neck
cancer. Additionally, long-termHPV16-specific T-cell memory was
evaluated in healthy controls. Among 33 healthy subjects, six
(18.2%) were males and 27 (81.8%) females (median age 24, range
21–26). Seven out of 33 (21.2%) subjects were vaccinated for high
risk HPV types about 5–10 years earlier. HPV16-specific antibod-
ies were quantified by neutralization assay and a cultured ELISPOT
assay was developed to evaluate HPV16-specific central memory
T-cell response. In detail, peripheral blood mononuclear cells were
opportunely stimulated for 10 days in culture and restimulated for
24 h in ELISPOT plates. Pools of peptides (15 aa length with an 11
amino acid overlap) including full-length L1, E6 and E7 proteins
were used as stimuli. S. aureus enterotoxin B (SEB) was used as
positive control.
Results: A statistic difference was observed between E6-
specific T-cell response in HPV16-related OSCC (median 462.0 IQR
86.0–1733.0 net spots/million PBMC* PI) and patients without
HPV16-related OSCC (median 4.0 IQR 0.0–29.0 net spots/million
PBMC* PI) p = 0.0067. A trend of significativitywas observedwith L1
(median 280.0 IQR 12–985 vs 1428 IQR 202–3533 net spots/million
PBMC* PI; p = 0.0631) and E7-specific peptide pool (median 5.0
IQR 0.0–87.0 vs 60.0 IQR 28.0–607.0 net spots/million PBMC* PI;
p = 0.0951) between the two groups of subjects. Among healthy
subjects, the median antigen-specific T-cell response was signifi-
cantly higher in HPV16 seropositive subjects for all antigen tested
(respectively p = 0.0124; p = 0.0039; p = 0.0179) suggesting the per-
sistence of an expandable long-term T-cell memory to HPV16 in
healthy subjects.
Conclusion:In conclusion, we observed an increased IFN- pro-
ducing T cells after long period stimulation with HPV16-specific
antigens in patients with HPV16 related head neck cancer. The role
of memory cellular response in progression and severity of cancer
has to be clarify.
Abstract no: 155
Presentation at ESCV 2016: Poster 17
Evaluation of a rapid test for the detection of
Tick Borne Encephalitis (TBE) IgM in serum and
CSF
Lisa Vennberg ∗, Katrin Forsström , Lena Rova ,
Oskar Ekelund , Martin Sundqvist
1 Faculty of Medicine and Health, Department of
Laboratory Medicine, Örebro University, Örebro,
Sweden
2 Department of Clinical Microbiology Kronoberg
County, Karlskrona, Sweden
Introduction: The incidence of Tick-borne encephalitis (TBE)
is increasing with 150–300 cases reported in Sweden annually.
The clinical picture can be hard to differentiate from other causes
of encephalitis and a rapid reliable diagnosis is therefore impor-
tant. The laboratory diagnosis of TBE relies on ELISA-based testing
to determine specific anti-TBE IgM and IgG in serum and in
CSF. The aim of this study was to evaluate the ReaScan TBE
IgM rapid test (Reagena, Toivala, Finland), a qualitative immune-
chromatographic lateral flow assay for the rapid detection of TBE
IgM in serum and CSF.
Materials and methods: The material consisted of two blinded
panels of serum and CSF. (1) 16 serum samples previously analyzed
for TBE IgM and IgG using ELISA (Euroimmun) at the department of
Clinical Microbiology Kronoberg County. Six of these were positive
for TBE IgM, three of which had a matching CSF sample. (2) Seven
(7) serum samples (6 with matching CSF samples) from patients
diagnosed with TBE in Örebro County during 2015 based on IgM
positivity (Immunozym) performed at the Dept of Clinical Virology,
Karolinska University Hospital. All samples were analyzed using
the ReaScan TBE IgM rapid test according to the manufacturer’s
instructions.
Results: The results obtained using ReaScan were in full con-
cordance with the Euroimmun IgM assay for all 16 serum-samples
from Kronoberg County. Of the 7 serum samples from Örebro 5
were positive for IgM with both ReaScan and Euroimmun with the
remaining 2 samples being classified as Equivocal with Reascan
and negative with Euroimmun. Of the 9 CSF samples tested, 2 were