Question
Asked 18th Apr, 2018

Why T cells are stimulated by anti CD3 and CD28 in vitro? What is the principle behind this?

I need to know the principle of T-cell activation in vitro to understand a paper I am currently reading.

Most recent answer

12th Apr, 2021
Ramachandramouli Budida
Astrazeneca Gothenburg
Yamila Gazzoni I have never worked with pre-activated T cells. But I assume they need CD3/CD28 as well. Do you have a comparison? from uninfected mice? You need to figure out why do you see the dead cells... is it virus, or preactivation or culture problems etc.
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Popular answers (1)

18th Apr, 2018
Ramachandramouli Budida
Astrazeneca Gothenburg
T cells are stimulated in vitro for their expansion to have enough cells for further studies. The T cell activation needs 2 signals, 1. activation of TCR complex, 2. costimulation of CD28 by CD80 or CD86. This stimulation is normally of two kinds, antigen dependent (where you need antigen and co-stimulation from antigen presenting cells or anti-CD28 antibodies etc) and antigen independent (where you do not need an antigen but a mitogen [PHA or Con A] or anti-CD3 and anti-CD28 antibodies). Antigen dependent stimulation expands only antigen specific T cells where as antigen independent stimulation expands all T cells in the sample culture.
The main principle behind stimulation by anti-CD3 and anti-CD28 antibodies is...
1. anti-CD3 will bind to CD3 and activates TCR complex without antigenic peptide from the antigen presenting cells
2. anti-CD28 will bind to CD28 and stimulates the T cells without CD80 or CD86 from antigen presenting cells
I hope this information helps for your understanding. For more basic information follow this article.
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All Answers (12)

18th Apr, 2018
Ramachandramouli Budida
Astrazeneca Gothenburg
T cells are stimulated in vitro for their expansion to have enough cells for further studies. The T cell activation needs 2 signals, 1. activation of TCR complex, 2. costimulation of CD28 by CD80 or CD86. This stimulation is normally of two kinds, antigen dependent (where you need antigen and co-stimulation from antigen presenting cells or anti-CD28 antibodies etc) and antigen independent (where you do not need an antigen but a mitogen [PHA or Con A] or anti-CD3 and anti-CD28 antibodies). Antigen dependent stimulation expands only antigen specific T cells where as antigen independent stimulation expands all T cells in the sample culture.
The main principle behind stimulation by anti-CD3 and anti-CD28 antibodies is...
1. anti-CD3 will bind to CD3 and activates TCR complex without antigenic peptide from the antigen presenting cells
2. anti-CD28 will bind to CD28 and stimulates the T cells without CD80 or CD86 from antigen presenting cells
I hope this information helps for your understanding. For more basic information follow this article.
52 Recommendations
22nd Apr, 2018
Tsvetelina Velikova
Sofia University "St. Kliment Ohridski"
Hello, Bashar,
For the activation of T cells in vivo a minimum of two signals is required - 1. Through TCR receptor (part of which is CD3); 2. Co-stimulation (i.e., via CD28). Thus, for in vitro stimulation of T cells, this is one of the methods for stimulation by mimicking the in vivo process.
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27th Apr, 2018
David Cameron Wraith
University of Birmingham
By using either plate or bead bound anti-CD3 you are binding the CD3 component of the T cell receptor. Immobilization of the anti-CD3 is required to create a surface that mimics the immunological synapse. CD28, as you know, is the essential costimulatory molecule required to drive naive T cell proliferation. Antibodies to this can either be added to the immobilized anti-CD3, i.e. plate or bead bound, or alternatively added in solution.
7 Recommendations
13th Apr, 2019
Christos Kosmas
Metaxa Cancer Hospital
If you stimulate the T cells with anti-CD3 only, in the absence of CD28 co-stimulation or add soluble CTLA4-Ig - a CD80/86 binder and co-inhibitor of T cell stimulation, you will induce T cell anergy.
2 Recommendations
7th Dec, 2019
Saleh Alkarim
King Abdulaziz University
Hello
Yes , I agree with Tsvetelina Velikova; you need to stimulation T cells in vitro via TCR receptor and co-stimulation by CD28.
Good Luck
2 Recommendations
26th May, 2020
Leena Sapra
All India Institute of Medical Sciences
Hello,
For the activation of T cells : 2 signals are required; Signal 1 via TCR and signal 2 via co-stimulatory signals.
Thus, for stimulation under in vitro conditions we generally employ anti-CD3 and anti-CD28 antibodies.
All the Best
2 Recommendations
17th Jun, 2020
Jiang Jiahuan
Zhejiang University
Are γδ T cells also stimulated by anti CD3 and CD28 in vitro? I think γδ T didnt need CD28 stimulation. Am I right?
1 Recommendation
12th Apr, 2021
Yamila Gazzoni
National University of Cordoba, Argentina
Ramachandramouli Budida Good answer. But, What about activated T cells from infected/immunized mice for example? Do they need anti CD3/CD28,too? Because I've cultured CD8+T cells with anti CD3/CD28 from infected mice but I observed more dead cells than without stimulus.
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12th Apr, 2021
Ramachandramouli Budida
Astrazeneca Gothenburg
Yamila Gazzoni I have never worked with pre-activated T cells. But I assume they need CD3/CD28 as well. Do you have a comparison? from uninfected mice? You need to figure out why do you see the dead cells... is it virus, or preactivation or culture problems etc.
1 Recommendation

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