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

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Abstract

Background: Lymphocytes proliferate considerably following appropriate stimulation in vitro. Autologous T cells are obtained from whole blood or tissue sites in relatively limited amounts. We need a method to expand these cells efficiently, study their functions and manipulate them to create appropriate cells for transferring to the patient with infection and cancer. Objectives: The aim of this study is to determine proliferation ability of two different stimulators on CD4+ lymphocytes. Methods: Lymphocytes were isolated from blood samples of healthy donors after removing adherent cells (monocytes).The efficacy of MACSiBead™ coated with anti-CD2, anti-CD3, anti-CD28 (anti-CD2/CD3/CD28) was compared with Phytohaemagglutinin A (PHA) on CD4+ lymphocytes proliferation using carboxyfluorescein diacetate succinimidyl ester (CFSE) in cell culture media. The percentage of proliferating cells was analyzed using flow cytometry. Results: Both stimulators induced extensive proliferation of CD4+ lymphocytes but proliferation ability of PHA was higher compared to stimulation by anti-CD2/CD3/CD28 MACSiBead™. The proliferation rate of cells stimulated by PHA was 93.8% ± 3.37% whereas it was 85.2% ± 4.7% in cells stimulated by anti-CD2/CD3/CD28 MACSiBead™. Conclusions: Our results show that MACSiBead™ along with PHA can be used to obtain a large number of expanded CD4+ lymphocytes.

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  • 1. Trickett A Kwan YL. T cell stimulation and expansion using anti-CD3/CD28 beads. J Immunol Methods 275 2003 251-5.

  • 2. Lamers C Van de Griend R Braakman E et al. Optimization of culture conditions for activation and large-scale expansion of human T lymphocytes for bispecific antibody-directed cellular immunotherapy. Int J Cancer 51 1992 973-9.

  • 3. Kalamasz D Long S Taniguchi R et al. Optimization of human T-cell expansion ex vivo using magnetic beads conjugated with anti-CD3 and Anti-CD28 antibodies. J Immunother 27 2004 405-18.

  • 4. Li Y Kurlander RJ. Comparison of anti-CD3 and anti-CD28- coated beads with soluble anti-CD3 for expanding human T cells: differing impact on CD8 T cell phenotype and responsiveness to restimulation. J Transl Med 8 2010 104.

  • 5. Teschner D Wenzel G Distler E et al. In vitro stimulation and expansion of human tumour-reactive CD8+ cytotoxic T lymphocytes by anti-CD3/CD28/CD137 magnetic beads. Scand J Immunol 74 2011 155-64.

  • 6. Martkamchan S Onlamoon N Wang S et al. The Effects of Anti-CD3/CD28 Coated Beads and IL-2 on Expanded T Cell for Immunotherapy. Adv Clin Exp Med 25 2016 821-8.

  • 7. Onlamoon N Boonchan M Unpol P et al. Influence of cell isolation method on the optimization of CD4+ T cell expansion using anti-CD3/CD28 coated beads Asian Pac J Allergy Immunol. 31 2013 99.

  • 8. Bernstein WB Cox JH Aronson NE et al. Immune reconstitution following autologous transfers of CD3/CD28 stimulated CD4+ T cells to HIV-infected persons. Clin Immunol 111 2004 262-74.

  • 9. Trickett AE Kwan YL Cameron B et al. Ex vivo expansion of functional T lymphocytes from HIV-infected individuals. J Immunol Methods 262 2002 71-83.

  • 10. Thomas AK Maus MV Shalaby WS et al. A cell-based artificial antigen-presenting cell coated with anti-CD3 and CD28 antibodies enables rapid expansion and long-term growth of CD4 T lymphocytes. Clin Immunol (Orlando Fla) 105 2002 259-72.

  • 11. Röth A Schneider L Himmelreich H et al. Impact of culture conditions on the proliferative lifespan of human T cells in vitro. Cytotherapy 9 2007 91-8.

  • 12. Onlamoon N Plagman N Rogers KA et al. Anti-CD3/28 mediated expansion of macaque CD4+ T cells is polyclonal and provides extended survival after adoptive transfer J Med Primatol 36 2007 206-18.

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