Research center of Immunobiochemistry .


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Laboratory of Immunobiochemistry. Research update. Active research projects. PI Rabin MDR proteins in T cell activation Regulation of T cell responses by the Respiratory Syncytial Virus PI Slater Cockroach allergen standardization Determination of optimal surrogate test
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Lab of Immunobiochemistry Research overhaul

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Active research ventures PI Rabin MDR proteins in T cell enactment Regulation of T cell reactions by the Respiratory Syncytial Virus PI Slater Cockroach allergen institutionalization Determination of ideal surrogate test Depletion investigation of CR concentrates Endotoxin in allergen immunizations

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Publications Research Spann KM, Tran KC, Chi B, Rabin RL, Collins PL. Concealment of the acceptance of alpha, beta, and lambda interferons by the NS1 and NS2 proteins of human respiratory syncytial infection in human epithelial cells and macrophages. J Virol 2004; 78(8):4363-9. Melody K, Rabin RL, Douek D, Roederer M, Farber JM. Novel Subsets of CD4+ Memory T Cells Reveal Early Branched Pathways of T Cell Differentiation in Humans. Proc Natl Acad Sci , in press Zhang J, Alston MA, Huang H, Rabin RL. Multidrug Resistant Protein 1 (MRP1) is endless supply of human T cells, and its restraint squares T cell work, in arrangement

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Publications Review Slater JE. Recombinant allergens in the US. Techniques 2004; 32(3):209-11. Slater JE. Latex allergens. Clin Allergy Immunol 2004;18:369-86. Slater JE. Institutionalized allergen extricates in the United States. Clin Allergy Immunol 2004;18:421-32. Rabin RL. Respiratory Syncytial Virus abuses hereditary and natural hazard variables for asthma; Business Briefing, US Pediatric Care 2005, in press Rabin RL, Levinson AI, Apter AJ. Fortuitous event of immune system and unfavorably susceptible sicknesses: epidemiologic and robotic investigations, in planning

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Abstracts B Chi, M Alston, KM Spann, PL Collins, RL Rabin. A basic part for dendritic cells in immunosuppression brought about by the respiratory syncytial infection (RSV). J Allergy Clin Immunol 2005; 115:S226 NC deVore, WJJ Finlay, EN Dobrovolskaia, A Gam, JE Slater. Cloning and investigation of mono-particular single chain section variable scFv pieces that perceive German cockroach allergens Bla g 1, Bla g 2, Bla g 4, and Bla g 5. J Allergy Clin Immunol 2005; 115:S163 E Dobrovolskaia, A Gam, JE Slater. Rivalry ELISA can be a delicate strategy for the particular identification of little amounts of allergen in an intricate blend. J Allergy Clin Immunol 2005; 115:S164 J Zhang, MA Alston, H Huang, RA Houghtling, RW Pastor, RL Rabin. Human sort 1 CD4 T cell cytokine reactions are specifically subject to Multidrug Resistance Protein1. J Allergy Clin Immunol 2005; 115:S255 C Valerio, LG Arlian, JE Slater. Bacterial 16S ribosomal DNA groupings disconnected from house clean parasites. J Allergy Clin Immunol 2005; 115:S163

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Invited presentations - Rabin AAAAI Annual Meeting, March 2004: FDA Food and Cosmetics Act as it Applies to Research Studies Paul-Ehrlich-Seminar, October 2005: Recombinant and altered allergens: The US point of view

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Invited presentations - Slater American Contact Dermatitis Society, October 2004 meeting: Natural elastic latex sensitivity ACAAI Annual Meeting, November 2004, Immunotherapy Collegium: Allergen immunotherapy in the time of vulnerability AAAAI Annual Meeting, March 2005: Allergen recognizable proof

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Larry G. Arlian, PhD Director, Microbiology and Immunology, Department of Biological Sciences, Wright State University, Dayton, Ohio Patrick R. Murray, PhD Chief, Microbiology Service, NIH Clinical Center, Bethesda, Maryland Immunotherapy Committee, AAAAI Harold Nelson, MD Peter L. Collins, PhD Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, Maryland Mario Roederer, PhD Vaccine Research Center, National Institute of Allergy and Infectious Diseases, NIH, Bethesda, Maryland Outside joint efforts

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Allergy and asthma are T cell subordinate T cells are allergen responsive and emit cytokines required in the unfavorably susceptible reaction (IL-4, IL-5, and IL-13) Allergen immunotherapy works by adjusting T cell reactions Novel methodologies towards altering T cell reactions may give novel therapeutics to treatment of hypersensitive ailments and asthma The connection between respiratory viral contaminations and wheezing is interceded by T cells Understanding T cell reactions to respiratory infections will give knowledge into the components of sensitivity and asthma

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Allergy and asthma are T cell subordinate T cells are allergen responsive and discharge cytokines required in the hypersensitive reaction (IL-4, IL-5, and IL-13) Allergen immunotherapy works by changing T cell reactions Novel methodologies towards altering T cell reactions may give novel therapeutics to treatment of unfavorably susceptible illnesses and asthma The connection between respiratory viral diseases and wheezing is intervened by T cells Understanding T cell reactions to respiratory infections will give knowledge into the instruments of hypersensitivity and asthma

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MK-571, an inhibitor of Multidrug Resistant Protein 1 (MRP1) pieces T cell actuation

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Activation influences [probe] reflects quality articulation of test transporter Does test transporter tweak initiation? perception

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MDR Family Proteins that vehicle substances crosswise over cell layers, against a fixation angle, in a vitality subordinate way. ABC proteins ( A TP B inding C assette) that contain unmistakable nucleotide restricting spaces (NBD). Qualities are profoundly preserved crosswise over species. In the first place part is MDR1 (P-glycoprotein, P-gp); best substrates are huge hydrophobic cations. MDR-related Resistant Protein-1 (MRP1) depicted in 1992; substrates are natural anions, furthermore: glutathione, glucuronide, & sulfate conjugates LTC 4

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MDR1 MRP1 MRP3 MDR family quality expression in T cells 35 cycle RT-PCR particular items CD4 CD8 Cord CD4 credulous memory guileless memory B cells NK cells 3 days 0 day Monocytes ACTIN

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MK-571 pieces morphologic changes connected with PBMC initiation TSST-1 10 ng/ml Control MK-571 (100µM) 40x

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MK-571 abatements articulation of CD69 in light of superantigens

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The MRP1 inhibitor MK-571 declines IFN-g and IL-4 by superantigen invigorated CD4 T cells

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The MRP1 inhibitor MK-571 squares cytokine emission

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PHA+PGJ 2 PHA+MK PHA+CE Blank Med PHA Lane 1 2 3 4 5 6 Increased PPAR g enactment in Jurkat T cells treated with MK-571 Jurkat T cells treated with PHA for 24 hours, and afterward PPAR g agonists or MK-571 for 1 hour before gather. MK: MK-571 50uM CE: Ciglitazone 20uM PGJ 2 : Prostaglandin J 2 10uM

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Conclusions Inhibition of MRP1 with MK-571 pieces T cell initiation MK-571 does not diminish the suitability of the actuated cells (not appeared) " Washout " of MK-571 switches the hindrance of enactment (not appeared) Treatment of cells with MK-571 enacts the transcriptional repressor PPAR g Hypothesis: endogenous ligands for PPAR g are held in MRP1 blocked cells

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Allergy and asthma are T cell subordinate T cells are allergen responsive and discharge cytokines required in the unfavorably susceptible reaction (IL-4, IL-5, and IL-13) Allergen immunotherapy works by adjusting T cell reactions Novel methodologies towards changing T cell reactions may give novel therapeutics to treatment of hypersensitive illnesses and asthma The connection between respiratory viral diseases and wheezing is intervened by T cells Understanding T cell reactions to respiratory infections will give knowledge into the systems of sensitivity and asthma

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Regulation of T cell reactions by the Respiratory Syncytial Virus

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RSV restraint of T cell multiplication RSV discourages expansion of PBMC to PHA, EBV, or to RSV antigens in vitro (Roberts, 1982; Preston et al , 1992). RSV animates inhibitors of multiplication, for example, prostaglandins (Panuska et al , 1990), " IL-1 inhibitors " (Roberts et al , 1986 and interferon-a (Preston et al , 1995). Coordinate contact with RSV F (combination protein) is important and adequate to restrain multiplication of lymphocytes (Schlender et al , 2002).

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Experimental approach Published information have involved different cytokines and inhibitors, or contact reliance, most unsubstantiated or disproved Goal: Develop a rearranged model to figure out which cells are essential or potentially adequate for restraint of multiplication by RSV.

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DC + RSV L 4 hr + SEB 4 days H 3 thymidine take-up test Inhibition of SEB-incited T cell multiplication by RSV DC = monocyte-determined dendritic cells SEB = staphylococcal enterotoxin B

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RSV essentially represses CMV particular and SEB-initiated T cell expansion

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DC + DC + Virus L CD4 4 hr 4 hr + SEB 4 days + SEB 4 days H 3 thymidine take-up measure H 3 thymidine take-up examine Do CD8 T cells, NK cells, or B cells intercede immunosuppression?

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CD4 T cells and DC are adequate for concealment of expansion by RSV P>0.05 P< 0.05 P=0.003 P<0.05 SEB RSV FLU paraflu SEB RSV FLU paraflu

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Dendritic cells are beneficially tainted by GFP-RSV

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Brightfield perspective of dendritic cells

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DC + RSV ON Supernatant Add to CD4 4 days + SEB H 3 thymidine take-up measure Can suppressive action be exchanged with DC supernatants?

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Inhibition of CD4 + T cell expansion by DC supernatant P = 0.001 P>0.05 DC sup: UV RSV control RSV

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Conclusions We have disentangled the trial arrangement of RSV-interceded immunosuppression to monocyte-inferred DC and CD4 T cells Immunosuppressive action can be exchanged with supernatants from contaminated DC, and is not because of persist of infection inside the supernatants

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Laboratory of Immunobioche

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