Expression of the Activation Antigen CD69 Predicts Functionality of in Vitro Expanded Peripheral Blood Mononuclear Cells (PBMC) from Healthy Donors and HIV-infected Patients

Overview

Journal Clin Exp Immunol

Publisher Oxford University Press

Specialty Allergy & Immunology

Date 1998 Oct 9

PMID 9764605

Citations 7

Authors

S D Nielsen

P Afzelius

A K Ersboll

J O Nielsen

J E Hansen

Affiliations

Soon will be listed here.

Abstract

Gene therapy for AIDS necessitates harvest and expansion of PBMC from HIV-infected patients. We expanded PBMC from healthy blood donors and HIV-infected patients for up to 14 days using four expansion protocols: 3 days of phytohaemagglutinin (PHA) stimulation, continuous PHA stimulation, 3 days of stimulation with anti-CD3 and anti-CD28, and continuous stimulation with anti-CD3 and anti-CD28. Functionality of PBMC was evaluated prior to and after expansion using standard proliferation assay. Phenotype and lymphocyte subset activation defined by expression of CD69 and CD25 were determined using flow cytometry. PBMC from healthy donors and HIV-infected patients were readily expanded. The best expansion was obtained using stimulation for 3 days. After expansion, functionality of PBMC measured as proliferative response was partly conserved. PBMC expanded with stimulation for 3 days exhibited more preserved functionality than PBMC stimulated continuously (P < 0.03). The mean proliferative response in each of the four different expansion protocols correlated with the mean values of CD69 expression. The proliferative responses from patients and healthy donors expanded with PHA stimulation for 3 days correlated with CD69 expression on CD4 cells (r = 0.68, P < 0.01) and on CD8 cells (r = 0.59, P < 0.03). Furthermore, expression of CD69 reliably predicted which patients and donors had highly conserved functionality after in vitro expansion. Finally, PBMC expanded with PHA stimulation for 3 days were examined for apoptosis. Only a minor fraction was primed for apoptosis, and this fraction could be significantly reduced by addition of IL-2 to the culture medium (P < 0.05). In conclusion, the feasibility of expanding PBMC from HIV patients was demonstrated. Expanded PBMC had conserved functionality. Finally, after in vitro expansion, expression of the activation antigen CD69 reliably predicted functionality of PBMC.

Citing Articles

Short-term assays for mesenchymal stromal cell immunosuppression of T-lymphocytes.

Herzig M, Christy B, Montgomery R, Cantu-Garza C, Barrera G, Lee J Front Immunol. 2023; 14:1225047.

PMID: 37822938 PMC: 10562633. DOI: 10.3389/fimmu.2023.1225047.

A method for expansion of T cells from cynomolgus monkey (Macaca fascicularis).

Gao C, Song Q, Zhang M, Li J, Yi M, Dong J In Vitro Cell Dev Biol Anim. 2018; 54(8):549-554.

PMID: 30112696 DOI: 10.1007/s11626-018-0278-y.

CD32 expression is associated to T-cell activation and is not a marker of the HIV-1 reservoir.

Badia R, Ballana E, Castellvi M, Garcia-Vidal E, Pujantell M, Clotet B Nat Commun. 2018; 9(1):2739.

PMID: 30013105 PMC: 6048139. DOI: 10.1038/s41467-018-05157-w.

Exploitation of stable nanostructures based on the mouse polyomavirus for development of a recombinant vaccine against porcine circovirus 2.

Fraiberk M, Hajkova M, Krulova M, Kojzarova M, Drda Moravkova A, Psikal I PLoS One. 2017; 12(9):e0184870.

PMID: 28922413 PMC: 5602543. DOI: 10.1371/journal.pone.0184870.

Factors associated with the elevated percentage of CD4CD69 T cells in maintained hemodialysis patients.

Chen R, Xiang F, Hu J, Cao X, Tan X, Jia P Ren Fail. 2017; 39(1):547-554.

PMID: 28726529 PMC: 6014306. DOI: 10.1080/0886022X.2017.1349672.

References

Lopez-Cabrera M, Santis A, Fernandez-Ruiz E, Blacher R, Esch F, Sanchez-Mateos P . Molecular cloning, expression, and chromosomal localization of the human earliest lymphocyte activation antigen AIM/CD69, a new member of the C-type animal lectin superfamily of signal-transmitting receptors. J Exp Med. 1993; 178(2):537-47. PMC: 2191117. DOI: 10.1084/jem.178.2.537. View

Wesselborg S, Janssen O, Kabelitz D . Induction of activation-driven death (apoptosis) in activated but not resting peripheral blood T cells. J Immunol. 1993; 150(10):4338-45. View

Boise L, Minn A, June C, LINDSTEN T, Thompson C . Growth factors can enhance lymphocyte survival without committing the cell to undergo cell division. Proc Natl Acad Sci U S A. 1995; 92(12):5491-5. PMC: 41721. DOI: 10.1073/pnas.92.12.5491. View

Wilson C, Wong J, Girard D, Merrill D, Dynan M, An D . Ex vivo expansion of CD4 lymphocytes from human immunodeficiency virus type 1-infected persons in the presence of combination antiretroviral agents. J Infect Dis. 1995; 172(1):88-96. DOI: 10.1093/infdis/172.1.88. View

Maino V, Suni M, Ruitenberg J . Rapid flow cytometric method for measuring lymphocyte subset activation. Cytometry. 1995; 20(2):127-33. DOI: 10.1002/cyto.990200205. View

van Lunzen J, Schmitz J, Dengler K, Kuhlmann C, Schmitz H, Dietrich M . Investigations on autologous T-cells for adoptive immunotherapy of AIDS. Adv Exp Med Biol. 1995; 374:57-70. DOI: 10.1007/978-1-4615-1995-9_6. View

Jagers E, De Brabander M, Baisier A, de Cree J, Verhaegen H, Verbiest W . A simple and rapid flow cytometric method to measure lymphocyte activation in HIV+ subjects. Diminished response to pokeweed mitogen in early disease. Anal Cell Pathol. 1995; 8(4):345-55. View

Krowka J, Cuevas B, Maron D, Steimer K, Ascher M, Sheppard H . Expression of CD69 after in vitro stimulation: a rapid method for quantitating impaired lymphocyte responses in HIV-infected individuals. J Acquir Immune Defic Syndr Hum Retrovirol. 1996; 11(1):95-104. DOI: 10.1097/00042560-199601010-00013. View

Riddell S, Elliott M, Lewinsohn D, Gilbert M, Wilson L, Manley S . T-cell mediated rejection of gene-modified HIV-specific cytotoxic T lymphocytes in HIV-infected patients. Nat Med. 1996; 2(2):216-23. DOI: 10.1038/nm0296-216. View

10.

Woffendin C, Ranga U, Yang Z, Xu L, Nabel G . Expression of a protective gene-prolongs survival of T cells in human immunodeficiency virus-infected patients. Proc Natl Acad Sci U S A. 1996; 93(7):2889-94. PMC: 39729. DOI: 10.1073/pnas.93.7.2889. View

11.

Levine B, Mosca J, Riley J, Carroll R, Vahey M, Jagodzinski L . Antiviral effect and ex vivo CD4+ T cell proliferation in HIV-positive patients as a result of CD28 costimulation. Science. 1996; 272(5270):1939-43. DOI: 10.1126/science.272.5270.1939. View

12.

Walker R . A phase I/II pilot study of the safety of the adoptive transfer of syngeneic gene-modified cytotoxic T lymphocytes in HIV-infected identical twins. Hum Gene Ther. 1996; 7(3):367-400. DOI: 10.1089/hum.1996.7.3-367. View

13.

Adachi Y, Oyaizu N, Than S, McCloskey T, Pahwa S . IL-2 rescues in vitro lymphocyte apoptosis in patients with HIV infection: correlation with its ability to block culture-induced down-modulation of Bcl-2. J Immunol. 1996; 157(9):4184-93. View

14.

Mardiney 3rd M, Brown M, Fleisher T . Measurement of T-cell CD69 expression: a rapid and efficient means to assess mitogen- or antigen-induced proliferative capacity in normals. Cytometry. 1996; 26(4):305-10. DOI: 10.1002/(SICI)1097-0320(19961215)26:4<305::AID-CYTO11>3.0.CO;2-V. View

15.

Caruso A, Licenziati S, Corulli M, Canaris A, De Francesco M, Fiorentini S . Flow cytometric analysis of activation markers on stimulated T cells and their correlation with cell proliferation. Cytometry. 1997; 27(1):71-6. DOI: 10.1002/(sici)1097-0320(19970101)27:1<71::aid-cyto9>3.0.co;2-o. View

16.

Nielsen S, Nielsen J, Hansen J . In vitro separation and expansion of CD4 lymphocytes from HIV-infected individuals without activation of HIV infection. J Immunol Methods. 1997; 200(1-2):107-12. DOI: 10.1016/s0022-1759(96)00193-7. View

17.

Bass H, Fahey J, Nishanian P, Detels R, Cumberland W, Kemeny M . Relation of impaired lymphocyte proliferative function to other major human immunodeficiency virus type 1-induced immunological changes. Clin Diagn Lab Immunol. 1997; 4(1):64-9. PMC: 170477. DOI: 10.1128/cdli.4.1.64-69.1997. View

18.

PRINCE H . CD69 expression reliably predicts the anti-CD3-induced proliferative response of lymphocytes from human immunodeficiency virus type 1-infected patients. Clin Diagn Lab Immunol. 1997; 4(2):217-22. PMC: 170505. DOI: 10.1128/cdli.4.2.217-222.1997. View

19.

Afzelius P, Nielsen S, Hofmann B, Nielsen J . The serotonin analogue buspirone increases the function of PBMC from HIV-infected individuals in vitro. Scand J Infect Dis. 1997; 29(2):117-20. DOI: 10.3109/00365549709035870. View

20.

Lorenz H, Hieronymus T, Grunke M, Manger B, Kalden J . Differential role for IL-2 and IL-15 in the inhibition of apoptosis in short-term activated human lymphocytes. Scand J Immunol. 1997; 45(6):660-9. DOI: 10.1046/j.1365-3083.1997.d01-443.x. View