Shreffler:Notebook/Staff Meetings/2011/03/15: Difference between revisions

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* EoE
* EoE
**  Early Risk factors (recruiting, nothing for lab)
**  Early Risk factors (recruiting, nothing for lab)
** Biomarkers
** Biomarkers (recruiting, n=12)
*** EDN assays: SK will run comparison of two assays this week with AM
*** EDN assays: will follow up with FL re: his experience; SK will tentatively plan to use assay with wider dynamic range
*** Eotaxin has been difficult to measure in saliva in FL's experience; could use CBA platform for other samples
*** will not attempt to measure eotaxin from saliva at this time
*** AM will meet with MY -- ws to arrange
*** AM will order reagents for 16S cloning and plan in early April to pilot this with saliva pellet samples


* Basophil
* Basophil
** APC project
** APC project
*** SP showed data on DR kinetics by flow and localization from the Amnis run:
*** SP showed data on DR kinetics by flow comparing surface and intracellular expression at 20' and 18h from two donors -- one strongly responsive, one weakly
***: DR increases over 18 hours in an IL-3 dependent fashion
***: supports previous findings and the impression that strong responders upregulate expression of class II, where as weak responders do not
***: DR increases in activation dep manner, but decreases from 20' to 2 h
***: DR increases over 18 hours in an IL-3 dependent fashion -- saw this again
***: FcERI MFI rapidly falls and staining may become clustered -- from a few images, not quantitative analysis yet
***: DR increases in activation dep manner, but decreases from 20' to 18h
***: cell area increases (and perhaps aggregation) with activation
***: One new observation was that in the case of the weak donor, activation dependent CD63 upregulation which was not present at 20' was prominent at 18 hours; this has implications for testing -- perhaps esp for activation by weak agonists (e.g. drugs)
***: surface CD11b increases


* Allergen Adjuvant
* Allergen Adjuvant
** we hope to hear from proteonomics on new sequence information this week
** we do not expect to hear from proteonomics on new sequence information until end of month
** BR will run zymosan experiment this week
** zymosan experiment (n=1) shows lack of ALDH1A2 induction by zymosan, strong induction by fraction 3 -- lacks positive control target for zymosan (e.g. IL-10) to determine whether iDC are unresponsive to zymosan generally, or simply this pathway is not stimulated


* PM Baso
* PM Baso
** AM produced very nice anergy experiment -- key variable to prevent 'carry over' appears to be time; by waiting 90' prior to 2nd stim, controls are perfect
** AM repeated the anergy experiment, including a 90' rest between stim 1 and 2. Clear desensitization, but some 'carryover', specifically with Derp2 stim conditions (perhaps higher affinity)
**: experiment demonstrates a degree of pathway-specific, partially non-specific suppression; NP-BSA somewhat stronger anergy-inducing signal, despite weaker stim (DerpNP_06 2/25/11)
**: SP notes that we cannot distinguish in these experiments between insufficient suppression of degran in Ca free conditions versus delayed degranulation occurring upon shift into Ca
 


* JAX
* JAX
** updated scripts for JAX to run as data comes in, note nec format for pcr data
** updated scripts for JAX to run as data comes in
'''**: Note nec format for pcr data needs to be reviewed with AM'''
** AM to catch up on analysis this month
** AM to catch up on analysis this month
** requesting that K at Hopkins adds comment column for lab notes
** requesting that K at Hopkins adds comment column for lab notes (status?)
** mAb cocktail being sent; new PMA/CaI stim sent by AM
 


* PM baso
* PM baso
** AM to finish analysis this week as study nearing point of analysis
** AM to finish analysis during AAAAI


* Carboplatin
* Carboplatin
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* U19
* U19
** BR to schedule a meeting with JM
** BR to review protocol and plans to begin long term cultures, contact Dutch group
**: will have follow up meeting with JM in ~2 months




Revision as of 09:27, 15 March 2011

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15 March 2011

Agenda Items

  • EoE
    • Early Risk factors (recruiting, nothing for lab)
    • Biomarkers (recruiting, n=12)
      • EDN assays: will follow up with FL re: his experience; SK will tentatively plan to use assay with wider dynamic range
      • will not attempt to measure eotaxin from saliva at this time
      • AM will order reagents for 16S cloning and plan in early April to pilot this with saliva pellet samples
  • Basophil
    • APC project
      • SP showed data on DR kinetics by flow comparing surface and intracellular expression at 20' and 18h from two donors -- one strongly responsive, one weakly
        supports previous findings and the impression that strong responders upregulate expression of class II, where as weak responders do not
        DR increases over 18 hours in an IL-3 dependent fashion -- saw this again
        DR increases in activation dep manner, but decreases from 20' to 18h
        One new observation was that in the case of the weak donor, activation dependent CD63 upregulation which was not present at 20' was prominent at 18 hours; this has implications for testing -- perhaps esp for activation by weak agonists (e.g. drugs)
  • Allergen Adjuvant
    • we do not expect to hear from proteonomics on new sequence information until end of month
    • zymosan experiment (n=1) shows lack of ALDH1A2 induction by zymosan, strong induction by fraction 3 -- lacks positive control target for zymosan (e.g. IL-10) to determine whether iDC are unresponsive to zymosan generally, or simply this pathway is not stimulated
  • PM Baso
    • AM repeated the anergy experiment, including a 90' rest between stim 1 and 2. Clear desensitization, but some 'carryover', specifically with Derp2 stim conditions (perhaps higher affinity)
      SP notes that we cannot distinguish in these experiments between insufficient suppression of degran in Ca free conditions versus delayed degranulation occurring upon shift into Ca


  • JAX
    • updated scripts for JAX to run as data comes in

**: Note nec format for pcr data needs to be reviewed with AM

    • AM to catch up on analysis this month
    • requesting that K at Hopkins adds comment column for lab notes (status?)


  • PM baso
    • AM to finish analysis during AAAAI
  • Carboplatin
    • no changes
  • U19
    • BR to review protocol and plans to begin long term cultures, contact Dutch group
      will have follow up meeting with JM in ~2 months
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