A Human Microbiome Enhanced Campylobacter Jejuni Induced ...

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Andres Franco, MD1; 1Immunology, Hospital Universitario de Canarias,. LA LAGUNA, Spain, 2Hospital del Tórax-Ofra, Sta Cruz de Tenerife,. Spain, 3CS ...
Abstracts AB99

J ALLERGY CLIN IMMUNOL VOLUME 135, NUMBER 2

Relevance of an Antigen-Specific Liver Profile Multiplex Technique in the Diagnosis of Autoimmune Liver Diseases Yvelise Barrios, MD, PhD1, Victor Matheu, MD, PhD2, Elena Varela3, Andres Franco, MD1; 1Immunology, Hospital Universitario de Canarias, LA LAGUNA, Spain, 2Hospital del Torax-Ofra, Sta Cruz de Tenerife, Spain, 3CS Candelaria. RATIONALE: The presence of autoantibodies (autoAbs) plays a central role in the diagnosis and classification of autoimmune liver diseases (AiLD). Although the gold standard for detecting autoAb is Indirect Immunofluorescence technique (IFI) using liver, stomach and kidney tissue, there are some remarkable antigen specificities that are not easily detected with this approach. The available multiplex format of several antigens will help the clinicians to improve the correct identification of patients having different multiple autoAbs profiles. METHODS: Data was obtained from the results stored along 2013 on OpenLabTMrecords of the Immunology Section of Hospital Universitario de Canarias. Multiplex Inmunoblot was performed with serum samples using DL 1300-1601-4 G Euroline Test (Euroimmun) to identify Abs against the following antigens: AMA (M2+3E/BPO), Sp100, PML, gp210, LKM1, LC-1, and SLA/LP. RESULTS: 170 different samples from 164(132/32 female/male) patients were analysed along 2013. Positive AutoAb patients distributes as follows: 47 AMA, 28 Sp100, 16 gp210, 12 PML, 6 LC-1 and 2 LKM-1. 27 out of 77(35%) patients showed 2 or more autoAbs to different antigen specificities at the same time. CONCLUSIONS: These data suggest that an important proportion of AiLD has complex associations of autoAbs rather than single specificities; moreover some of those autoAbs are not easily detected using the IFI technique. This has importance for studies of the clinical associations and prognostic value of liver autoAbs and influences the therapeutic options of these patients.

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Cord Blood DNA Methylation of Treg Cytokine Genes Differs with Parity Orpita Nilormee1, Gabrielle A. Lockett, PhD2, Sabrina Iqbal3, John W. Holloway, PhD2, Syed H. Arshad, DM, FRCP4,5, Wilfried Karmaus, MD, DrMed, MPH6; 1University of Memphis, Memphis, 2University of Southampton, Southampton, United Kingdom, 3University Of Memphis, Memphis, TN, 4The David Hide Asthma and Allergy Research Centre, United Kingdom, 5University of Southampton, United Kingdom, 6University of Memphis, Memphis, TN. RATIONALE: Women undergo immunological changes during pregnancy including changes in the balance of TH1/TH2 cytokines. To investigate whether these changes can influence DNA-Methylation (DNA-M) in cord blood of consecutive births, we analyzed cord blood DNA-M of genes in the TH1, TH2, TH17 and Treg pathways of two consecutive deliveries. METHODS: In the Isle of Wight birth cohort, umbilical cord blood samples were collected from offspring of cohort participants to measure DNA-M using the Illumina Infinium HumanMethylation450 beadchip. Cord blood DNA-M was available for two consecutive pregnancies in seven mothers. Cytosine-phosphate-guanine (CpG) sites in TH1 (157 CpGs, 19 genes), TH2 (68 CpGs, 12 genes), TH17 (110 CpGs, 15 genes), Treg (10 CpGs, 2 genes) and in a random sample (271 CpGs) were consid_0.05), mixed linear ered. To identify CpGs with significant changes (p< models were applied to compare cord blood DNA-M of two consecutive pregnancies. RESULTS: In the last of two pregnancies, cord blood DNA-M showed significantly more CpGs of the two Treg genes (60%) with significant differences compared to CpGs of a random sample (32.47%) (p50.02). Among the six differentially methylated CpGs, three each were from FOXP3 and CTLA4, respectively. CONCLUSIONS: In cord blood, significant methylation differences in Treg genes between consecutive pregnancies suggest an adaptation in the course of two pregnancies. Future studies need to investigate whether these epigenetic changes are responsible for differential immune responses in offspring with different birth order.

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A Human Microbiome Enhanced Campylobacter Jejuni Induced Autoantibodies and Th-2 Skewing of Adaptive Immunity after Fecal Transplant Linda S. Mansfield, VMD, PhD, DACVM, Kelsey A. Brakel, BS, Ankit Malik, BS, M.S., Julia A. Bell, PhD; Michigan State University, East Lansing, MI. RATIONALE: Guillain-Barre Syndrome (GBS) is a common cause of acute generalized paralysis. Infection with the enteric pathogen C. jejuni often precedes GBS when bacterial lipo-oligosaccharide resembling host nerve gangliosides activates the immune system to produce autoantibodies. We showed that C. jejuni 11168 from an enteritis patient produced T helper-1/17 responses in C57BL/6 IL-10-/- mice, while a C. jejuni GBS patient strain (260.94) blunted Th-1/17, but enhanced Th-2 responses. Only Th-2 antibodies cross-reacted with nerve gangliosides. We hypothesized that human gut microbiota (Humicrobiota) enhances immunity to C. jejuni infection with elevated C. jejuni-specific and autoantibodies. METHODS: C57BL/6 germ-free mice were given a human fecal transplant, bred after gut microbiota stabilized, and their offspring used in a 30 day infection trial. Congenic Humicrobiota and mouse microbiota (Momicrobiota) mice were inoculated with C. jejuni enteritis strain 11168, GBS strain 260.94 or sham inoculated. Plasma was collected from all mice and levels of Th-1 and Th-2 antibody isotypes to nerve gangliosides and to C. jejuni strains were measured. IL-4 and IFN-g responses were measured in gut lamina propria cells by RT-PCR. RESULTS: Autoimmune responses were significantly elevated by the presence of Humicrobiota. Humicrobiota mice had significantly higher levels of IgG1 antibodies to C. jejuni 11168 and to GM1 and GD1a nerve gangliosides than infected Momicrobiota mice. Infected Humicrobiota mice had a 12-fold increase in IL-4 levels and decreased IFN-g levels. CONCLUSIONS: Humicrobiota enhanced C. jejuni induced autoantibodies to nerve gangliosides and Th-2 skewing of the adaptive immune response

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