J ALLERGY CLIN IMMUNOL

VOLUME 125, NUMBER 2

Abstracts AB77

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302 Congenital NEMO Alteration at Position 223 causesEctodermal Dysplasia and Immunodeficiency withNormogammaglobulinemia

G. Karamchandani-Patel1, R. U. Sorensen1, E. Hanson2, R. Saltzman2,

J. Orange2; 1Louisiana State University Health Sciences Center, New Or-

leans, LA, 2University of Pennsylvania School of Medicine, Philadelphia,

PA.

RATIONALE: Hypomorphic mutations in the NEMO gene result in

a variable syndrome of somatic and immunological abnormalities. It is

becoming increasingly apparent that there are clinically relevant geno-

type-phenotype associations in this disease. We studied two unrelated

boys with novel NEMO mutations resulting in alterations at position 223

to determine if they had similar phenotypes.

METHODS: Clinical and immunologic characteristics as well as NF-kB

activation were defined in two patients with novel NEMO mutations pre-

dicted to affect residue 223. Since this specific region of the NEMO protein

has not been previously affected, consideration was given to a new geno-

type-phenotype correlation.

RESULTS: Both patients were diagnosed at age two with hypohydrotic

ectodermal dysplasia. Immunoglobulin levels, lymphocyte subpopulations

and lymphocyte proliferation studies were normal. Patient 1 had recurrent

sinopulmonary infections and pneumococcal antibody titers protective to

only 1/14 serotypes. Patient 2 had a necrotizing soft tissue MRSA infection

at 16 months and streptococcal angiosus bacteremia with subdural empy-

ema at 26 months.

DNA sequencing of the NEMO gene in both patients revealed mutations in

exon 5 corresponding to position 223. Patient 1 had a 3 nucleotide deletion,

c.667_669delGAG predicting the deletion of glutamine 223. Patient 2 had

a missense mutation resulting in a nucleotide 667G>A which predicts

Glu223>Lys substitution. Infectious susceptibility in both patients im-

proved after immunoglobulin replacement therapy.

CONCLUSIONS: Two different novel mutations affecting NEMO posi-

tion 223 resulted in clinically relevant and similar phenotypes. This under-

scores the consideration of genotype-phenotype correlations in this disease

and defines residue 223 as dispensable for quantitative, but not qualitative

immunoglobulin production.

303 A Comparison Of The Effects Of TNF, IL-12/IL-23p40, AndTGFb1 Peptide-based Vaccines In The Amelioration Of TNBS-induced Chronic Colitis

q. guan1,2, Y. Ma2, C. Hillman2, A. G. Ma1,2, G. Zhou2, G. Qin3, C. R.

Weiss1,2, Z. Peng1,2; 1Department of Immunology, University of Mani-

toba, Winnipeg, MB, CANADA, 2Department of Pediatrics and Child

Health, University of Manitoba, Winnipeg, MB, CANADA, 3Department

of Pathology, University of Manitoba, Winnipeg, MB, CANADA.

RATIONALE: TNF, IL-12, IL-23 and TGFb1 are key cytokines in the

pathogenesis of Crohn’s disease. We previously developed three mouse cy-

tokine peptide-based virus-like particle vaccines against TNF, the p40 sub-

unit shared by IL-12 and IL-23, and TGFb1, respectively. These vaccines

elicited specific antibodies to the target cytokine and their antisera in-

hibited the bioactivity of the target. Here, we evaluated the effects of the

vaccines separately and paralelly in murine chronic colitis, induced by in-

trarectal administrations of trinitrobenzene sulfonic acid (TNBS).

METHODS: Mice were subcutaneously immunized with vaccine, vaccine

carrier or saline three times and then intra-rectally administered TNBS

weekly for 7 weeks. Serum cytokine-specific IgG, body weight, histology,

collagen, and cytokines in colon tissue, and Th1 and Th17 cells in mesen-

teric lymph nodes (MLN) were measured.

RESULTS: Vaccines induced high and long-lasting specific IgG anti-

bodies to target cytokine. Vaccinated mice had less body weight loss and

a decrease of inflammation, collagen deposition, and expression of p40,

IL-23 and IL-17 proteins in colon tissues, compared with carrier and saline

groups. Among them, p40 vaccine showed relatively stronger effects in

body weight gain and colon inflammation than the other 2 vaccines. In

p40 vaccinated mice, more parameters were analyzed. p40 vaccine de-

creased mRNA expression of p40, IL-10 and Bcl-2 and down-regulated

the percentages of Th1 and Th17 cells in MLN. CONCLUSIONS:

Targeting TNF or IL-12/IL-23 or TGFb1, especially IL-12/IL-23, by

employing a vaccine is effective in the amelioration of intestinal inflammation

and fibrosis, providing a novel approach in the long-term treatment of Crohn’s

disease.

304 Human Papillomavirus (HPV)-Specific T-cells RecognizingDominant E2/E6 Epitopes Elicit Reduced IFN-g in Patientswith Recurrent Respiratory Papillomatosis (RRP)

E. James1, J. A. DeVoti2,3, D. W. Rosenthal3,4, L. J. Hatam3, B. M. Stein-

berg2,5, A. Abramson5, W. W. Kwok1, V. R. Bonagura3,4; 1Benaroya

Research Institute, Seattle, WA, 2Feinstein Institute for Medical Research,

North Shore-LIJ Health System, Manhasset, NY, 3Division of Allergy/

Immunology, North Shore LIJ Health System, Manhasset, NY, 4Elmezzi

Graduate School of Molecular Medicine, North Shore LIJ Health System,

Manhasset, NY, 5Department of Otolaryngology, Long Island Jewish

Medical Center, New Hyde Park, NY.

RATIONALE: RRP is an HPV-related disease, in which HLA-

DRB1*0102 (DR0102) and DRB1*0301 (DR0301) haplotypes are associ-

ated with increased severity. TH1-like responses to E6/E2 proteins have

been shown to protect against papillomavirus-induced disease in animals.

Therefore, dysfunctional TH1 responses may be related to RPP disease se-

verity in DR0102 and DR0301 subjects.

METHODS: Using tetramer guided epitope mapping, we identified im-

munogenic peptides within HPV-11 early proteins (E6/E2) restricted by

DR0102 and/or DR0301. Peptide binding, tetramer staining, and prolifer-

ation assays identified minimal epitopes within these peptides. The cyto-

kine profile of sorted tetramer positive T-cell lines from RPP patients

(n56) and controls (n510) was measured using a cytokine capture assay,

to determine E2/E6-responsive T-cell polarization.

RESULTS: Two distinct E6/E2 peptides (E6113-132 and E21-20) con-

tained DR0102 and DR0301 restricted epitopes respectively. An additional

peptide (E2281-300) contained an epitope presented by both DR0102 and

DR0301. Minimal epitopes within these peptides bound to recombinant

DR protein, gave positive tetramer staining for sorted T-cell lines, and eli-

cited T-cell proliferation in both RPP patients and HLA-matched healthy

controls. While the magnitude of responses to these epitopes was similar

in both groups, IFN-g secretion was substantially lower in T-cell lines

isolated from RPP patients.

CONCLUSIONS: CD4+ T-cells specific for E2/E6 epitopes are easily

detected in RPP patients and healthy controls with DR0102 and DR0301

haplotypes. However, RPP patients exhibit HPV-specific, immune dysre-

gulation, indicated by a lack of IFN-g secretion. Therefore, therapeutic

vaccination or other interventions that repolarize T-cell responses and

restore TH1-like cytokine responses to HPV proteins could improve

outcomes for RPP patients.