Oral Presentations - Federation of Clinical Immunology Societies

S38 Abstracts
Detroit, MI, Kimberly Milkovich, Research Associate,
Division of Rheumatology, Case Western Reserve University,
Cleveland, OH, Benig Rodriguez, Assistant Professor, Center
for AIDS Research, Case Western Reserve University,
Cleveland, OH, Michael Lederman, Professor, Case Western
Reserve University, Center for AIDS Research, Cleveland,
OH, Marwan Abouljoud, Director of Transplant Institute,
Henry Ford Health Systems, Transplant Institute, Detroit,
MI, Donald Anthony, Assistant Professor, Case Western
Reserve University, Division of Rheumatology and Infectious
Disease, Cleveland, OH
It is difficult to detect HCV-specific T cells in the peripheral
blood of chronic HCV patients, while T cells specific
for other viruses appear intact. One explanation for this is a
compartmentalization of the immune response. Here we
performed phenotypic analysis of T cells isolated from liver,
perihepatic lymph nodes (LN), and peripheral blood of 23
HCV infected and 12 uninfected subjects undergoing liver
transplantation. Results: HCV specific IFN-γ and proliferative
responses were commonly observed in the LN of HCV infected
subjects (40% and 46% respectively), while they were
uncommonly observed in the peripheral blood and liver
(IFN-γ 16% and 25% respectively; proliferation 11% and 8%
respectively). In contrast, CMV specific IFN-γ producing
responses were not as commonly observed in the LN (30%)
compared to the periphery (58%) and liver (60%) of these
same HCV patients. Conclusions: These data are among the
first to look at HCV-specific responses in perihepatic lymph
nodes. The data indicate a selective defect in HCV, but not
CMV specific, T cell effector function in the periphery of
chronic HCV patients. The relatively common presence of
HCV-reactive T cells in the LN is consistent with T cell
activation at this site. Finally, we do not detect HCVspecific
T cells more readily in the liver compared to the
periphery or LN, indicating that hepatic lymphocytes are
defective in effector function similar to those in the
peripheral blood. The frequency of HCV-specific T cells did
not clearly correlate with the severity of recurrent HCV
after transplant.
doi:10.1016/j.clim.2007.03.284
F.72 Carbohydrate Analysis of HIV Envelope
Glycoprotein
Ingrid D. Cruzado-Park, Senior Development Scientist,
Beckman Coulter, Inc. Fullerton, CA, Munir Alam, Duke
University Medical Center, Durham, NC, Hua-Xin Liao, Duke
University Medical Center, Durham, NC, Barton Haynes,
Director, Duke University Medical Center, Human Vaccine
Institute and CHAVI, Durham, NC, Heather Desaire, Duke
University Medical Center, Durham, NC, Enrique Rabelli,
Director, Custom BioPharma Solutions, Beckman Coulter,
Inc. Miami, FL, Sybil D’Costa, Staff Advanced Research
Scientist, Beckman Coulter, Inc. Miami, FL, Michael H.
Simonian, Manager, Biomarker Discovery and Automation
Center, Beckman Coulter, Inc. Fullerton, CA, Chitra K.
Ratnayake, Team Leader, Beckman Coulter, Inc. Biomarker
Applications and Chemistry Development, Fullerton, CA
To date over 65 million people have been infected with the
human immunodeficiency virus (HIV) which continues to
claim several million lives a year. There is an urgent need to
design and develop vaccines that generate broad neutralizing
antibody enabling preventative vaccination strategies.
Although the virus has evolved to circumvent the immune
response, for example, carbohydrates on the envelope
glycoprotein of HIV serve as a strong defense against antibody
mediated responses, there is evidence that these oligomannose
components can also serve as ideal vaccine candidates
due to their ability to generate efficacious neutralizing
antibodies in HIV infected individuals (2G12 antibody) albeit
infrequently. To address the role of glycosylation in HIV
pathogenesis and immunogenicity, carbohydrate profiles
from wild type and consensus modeled, genetically engineered
envelope glycoproteins were compared. We present
data showing the N-linked oligosaccharide profiles for two
types of HIV gp140 envelope glycoproteins; JRFL is a primary
isolate and the other from a genetically engineered strain
containing a consensus sequence (ConS), which is more
effective at eliciting neutralizing antibodies. Following
treatment with PNGase F, released N-linked oligosaccharides
were labeled with fluorescent 8-aminopyrene-1,3,6-trisulfonate
and profiled using the ProteomeLab PA 800 Protein
Characterization System with laser-induced fluorescence
(LIF) detection (488 nm excitation, 520 nm emission). We
observed significant differences in the carbohydrate profiles
of these gp140 envelopes and will present and discuss those
differences. This strategy for oligosaccharide characterization
and subsequent identification may help evaluate role of
glycosylation in generating broadly neutralizing antibody
responses and enable intelligent design of carbohydratebased
epitope vaccines.
doi:10.1016/j.clim.2007.03.285
F.73 Defining the Role of BmIL-5 in Tropical
Pulmonary Eosinophilia
Sung (Steve) Kwon, University of Illinois College of Medicine
at Rockford, Rockford, IL, Gnanasekar Munirathinam,
University of Illinois College of Medicine at Rockford,
Rockford, IL, Anand Setty Balakrishnan, University of Illinois
College of Medicine at Rockford, Rockford, IL, Ramaswamy
Kalyanasundaram, University of Illinois College of Medicine
at Rockford, Rockford, IL
This study describes characterization of a novel pathogenderived
human IL-5 receptor binding molecule designated
BmIL-5. This molecule was identified in our laboratory by
screening a phage displayed cDNA expression library of the
infective larvae of the human lymphatic filarial parasite,
Brugia malayi. Soluble human IL-5 receptor (huIL-5R) was
used as the bait. This screening procedure yielded a single
clone and sequence analyses of the insert from this clone
showed a novel protein designated as “B. malayi-derived IL-
5-like molecule” (BmIL-5). Recombinant BmIL-5 was then
expressed and purified. ELISA-based binding assays confirmed
binding of rBmIL-5 to huIL-5R. Functional significance
of this binding was then evaluated using a human eosinophil
cell line, named Hs 454.T. Initial studies showed that BmIL-5