• You are here
  • Forum Header
  • Topics not covered in other forums.
ratdog2112
Platinum Boarder
Posts:3612
Men never remember, but Women never forget

Re: Tell Me Something Good!!

#264010 10 months, 1 week ago
ratdog2112 wrote:
SunshineSue wrote:
I get to have lunch with gvanman tomorrow!!!

Hope everyone has some fun weekend plans!


David Nelson Band 70th Birthday last nite, awesome. Would of love to been there for that.


Lets not forget this weekend plans remember those Dad's.
"What I like best about music is when time goes away."
skidoo
Expert Boarder
Posts:898
skidoo, skidoo.....ski-doo doo doodlie doo doo doo

Re: Tell Me Something Good!!

#264015 10 months, 1 week ago
skidoo wrote:
my b cell culture looks marvelous. flow sort for resting b cells, then plate on NIH3T3tCD40L cells (200K cells per well of a 6 well plate). split every 3rd day. culture in DMEM F12 supplemented with insulin and IL-4. you end up with activated b cells expressing CD62L, CD83, CD86. they grow like gangbusters!

now to use this as antigen-presenting cells to detect antigen-specific T cells........

and it is spring here in Seattle, beautiful night last night. had a moonlight picnic down at The Cut.


check out this expansion of antigen-specific t cells. bulk t cells from whole blood were stimulated with cultured B cells (described above) pulsed with 9mer and 40mer peptides (from HIV envelope gp120....the V2 "crown"). unpulsed cultured B cells as a control. the t cells were cultured with the B cells for 8 days (supplemented with IL-2,7,15 days 3,5, and 7). at the end the t cells were stained with a tetramer containing the red peptide sequence. i used HLA matched HIV seronegative t cells as a control to test for de novo reactivity (there was none...pretty clean "background"). we are now writing a grant to get more wayne cashman so we can do more experiments. we think we will be able to detect t cells responses in vaccine trials with this assay and "map" t cell responses in natural history cohorts.


stimulatedassay.jpg
Last Edit: 10 months, 1 week ago by skidoo. Reason: trouble posting picture
The following user(s) said Thank You: Chester
Griff
Expert Boarder
Posts:803
Don't you see........

Re: Tell Me Something Good!!

#264020 10 months, 1 week ago
Football camp looked awesome Sue, congrats. What a great idea, we should have something like at the U of Wash!!

Shot an 83 last Saturday, best round for me in 4 years!! I was playing so bad that I was ready to give it up.

Have a great weekend all you other dads!!
66slim
Platinum Boarder
Posts:1419

Re: Tell Me Something Good!!

#264023 10 months, 1 week ago
skidoo wrote:
skidoo wrote:
my b cell culture looks marvelous. flow sort for resting b cells, then plate on NIH3T3tCD40L cells (200K cells per well of a 6 well plate). split every 3rd day. culture in DMEM F12 supplemented with insulin and IL-4. you end up with activated b cells expressing CD62L, CD83, CD86. they grow like gangbusters!

now to use this as antigen-presenting cells to detect antigen-specific T cells........

and it is spring here in Seattle, beautiful night last night. had a moonlight picnic down at The Cut.


check out this expansion of antigen-specific t cells. bulk t cells from whole blood were stimulated with cultured B cells (described above) pulsed with 9mer and 40mer peptides (from HIV envelope gp120....the V2 "crown"). unpulsed cultured B cells as a control. the t cells were cultured with the B cells for 8 days (supplemented with IL-2,7,15 days 3,5, and 7). at the end the t cells were stained with a tetramer containing the red peptide sequence. i used HLA matched HIV seronegative t cells as a control to test for de novo reactivity (there was none...pretty clean "background"). we are now writing a grant to get more wayne cashman so we can do more experiments. we think we will be able to detect t cells responses in vaccine trials with this assay and "map" t cell responses in natural history cohorts.


stimulatedassay.jpg



Keep up the good work Skidoo! Good luck with your grant.
And leave it on!
The following user(s) said Thank You: skidoo
Denver Man
Platinum Boarder
Posts:6948
On the day I was born daddy sat down and cried

Re: Tell Me Something Good!!

#264027 10 months, 1 week ago
cool stuff skidoo!! I really wish I was closer to the research side of things. Commercialization is generally boring yet super stressful.. not a good combination!
I went walkin' out last summer
Tryin' to find a breath of air
I went walkin' on the mountain
A friend had told me I'd find you there
The following user(s) said Thank You: skidoo
skidoo
Expert Boarder
Posts:898
skidoo, skidoo.....ski-doo doo doodlie doo doo doo

Re: Tell Me Something Good!!

#264032 10 months, 1 week ago
it's all important, just gotta keep skating, checking, throwing, kicking, punting, waiter/waitressing, slopjockeying, banging on the drum, making the donuts, splitting hairs, splitting the atom, splitting my pants....
The following user(s) said Thank You: Denver Man
Griff
Expert Boarder
Posts:803
Don't you see........

Re: Tell Me Something Good!!

#264036 10 months, 1 week ago
Cool stuff skidoo. You and DM are the smart ones here saving the world!!
The following user(s) said Thank You: Denver Man
jodiah
Expert Boarder
Posts:838

Re: Tell Me Something Good!!

#264038 10 months, 1 week ago
66slim wrote:
skidoo wrote:
skidoo wrote:
my b cell culture looks marvelous. flow sort for resting b cells, then plate on NIH3T3tCD40L cells (200K cells per well of a 6 well plate). split every 3rd day. culture in DMEM F12 supplemented with insulin and IL-4. you end up with activated b cells expressing CD62L, CD83, CD86. they grow like gangbusters!

now to use this as antigen-presenting cells to detect antigen-specific T cells........

and it is spring here in Seattle, beautiful night last night. had a moonlight picnic down at The Cut.


check out this expansion of antigen-specific t cells. bulk t cells from whole blood were stimulated with cultured B cells (described above) pulsed with 9mer and 40mer peptides (from HIV envelope gp120....the V2 "crown"). unpulsed cultured B cells as a control. the t cells were cultured with the B cells for 8 days (supplemented with IL-2,7,15 days 3,5, and 7). at the end the t cells were stained with a tetramer containing the red peptide sequence. i used HLA matched HIV seronegative t cells as a control to test for de novo reactivity (there was none...pretty clean "background"). we are now writing a grant to get more wayne cashman so we can do more experiments. we think we will be able to detect t cells responses in vaccine trials with this assay and "map" t cell responses in natural history cohorts.


stimulatedassay.jpg



Keep up the good work Skidoo! Good luck with your grant.



Skidoo that's really cool stuff and the best news I've heard all day (so far). I really hope you guys get the grant. According to the latest statistics given at this year's AIDS Walk here in Miami, HIV is currently spreading faster here in South Florida than in Nairobi. Your work could potentially save a lot of lives. As far as I'm concerned, you're the FFF "Hero of the Day".
The following user(s) said Thank You: Chester, Denver Man, skidoo
Denver Man
Platinum Boarder
Posts:6948
On the day I was born daddy sat down and cried

Re: Tell Me Something Good!!

#264059 10 months, 1 week ago
Griff wrote:
Cool stuff skidoo. You and DM are the smart ones here saving the world!!


nahhh I'm just really good at riding the coat tails of others LOL
I went walkin' out last summer
Tryin' to find a breath of air
I went walkin' on the mountain
A friend had told me I'd find you there
The following user(s) said Thank You: skidoo, Griff
skidoo
Expert Boarder
Posts:898
skidoo, skidoo.....ski-doo doo doodlie doo doo doo

Re: Tell Me Something Good!!

#264061 10 months, 1 week ago
Denver Man wrote:
Griff wrote:
Cool stuff skidoo. You and DM are the smart ones here saving the world!!


nahhh I'm just really good at riding the coat tails of others LOL


an old school bumper ride. put on your runners (better in the snow) make sure the bus driver isn't watching, grab the bumper and hang on.
Last Edit: 10 months, 1 week ago by skidoo. Reason: atrocious grammar
The following user(s) said Thank You: Denver Man, Griff
Time to create page: 0.54 seconds