Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads

Abstract

The results demonstrate that purified CD4+ T lymphocytes from immunorosette formation provided the highest CD4+ T lymphocyte expansion when stimulated with anti-CD3/CD28 coated beads. This method can be used to obtain a large number of expanded CD4+ T cells for adoptive immunotherapy.

Determination of cell expansion and surface molecule expression on anti‐CD3/28 expanded CD4+ T cells

“…Moreover, a low concentration of IL‐2 supplementation did not affect surface molecule expressions of any chemokine receptors, adhesion molecules, co‐stimulatory molecules, activation molecules, maturation markers, cytokine receptors and other molecules of the expanded cells. These findings then support the previous experiments from Onlamoon et al …”

“…The cell numbers were increased by 37‐fold after a 14‐day culture . Later, our previous study showed that CD4 + T cell stimulation with anti‐CD3/28‐coated magnetic beads to a 1:1 bead‐to‐cell ratio and culture in the absence of IL‐2 were able to prolong the cell expansion throughout a 3‐week culture with the good yield of 1000‐fold . During the cell harvest, the higher number of beads used for expansion, the higher number of beads remaining in the final products which leads to the bead accumulation in the body after the cell reinfusion tentatively causing toxicity.…”

“…With respect to the clinical uses, a large number of CD4 + T cells expanded in vitro was required for reinfusion in HIV‐infected patients; therefore, optimization of expansion protocols is warranted. There have been established in vitro culture methods for anti‐CD3/28‐stimulated CD4 + T lymphocytes providing different yields which can be related to different cell isolation methods, bead‐to‐cell ratios used for stimulation and medium supplementation. More importantly, functional‐associated phenotypic characters of the expanded cells are essential which are not only related to cell characterization but also maturation and activation stages as well as cell migration.…”

Intracellular free radical production by peripheral blood T lymphocytes from patients with systemic sclerosis: role of NADPH oxidase and ERK1/2

“…To provide a more physiological mechanism of T cell activation, we stimulated lymphocytes with magnetic beads coated with anti-CD3 and anti-CD28 antibodies, mimicking the binding of antigen-presenting cells to T cells [ 33 , 34 ]. CD3-CD28 beads stimulation induced an increase of ROS levels (Figure S3A in Additional file 4 ) and CD69 expression in SSc and normal T cells (Figure 6 C).…”

Extensive Proliferation of Cd4+ Lymphocyte by Both Phytohaemagglutinin A and Anti-Cd2/Cd3/ Cd28 Macsibeads

“…With success of T cells manipulation and stimulation methods [4] such as PHA, autologous antigen presenting cells with specifi c peptide, and anti-CD3 and anti-CD28-coated beads [7], it is possible to stimulate and expand lymphocytes to adoptive transfer [4,7]. Nowadays, adoptive transfer of T cells is known successful treatment for infections and cancer such as HIV infection and multiple myeloma [3,[8][9][10].…”